Journal of Functional Biomaterials

13 pages, 2535 KiB

Open AccessArticle

Anti-Inflammatory, Antipyretic, and Analgesic Potential of Chitin and Chitosan Derived from Cockroaches (Periplaneta americana) and Termites

by Khushbakht Asad, Sumaira Shams, Eliana Ibáñez-Arancibia, Patricio R. De los Ríos-Escalante, Farhad Badshah, Farooq Ahmad, Muhammad Salman Khan and Asar Khan

J. Funct. Biomater. 2024, 15(3), 80; https://doi.org/10.3390/jfb15030080 - 21 Mar 2024

Cited by 5 | Viewed by 2301 | Correction

Abstract

The chitin and chitosan biopolymers are extremely valuable because of their numerous industrial and pharmacological uses. Chitin and chitosan were extracted from the exoskeleton of Periplaneta americana (cockroaches) and termites using various acid and alkali techniques. The extraction process involves an initial demineralization [...] Read more.

The chitin and chitosan biopolymers are extremely valuable because of their numerous industrial and pharmacological uses. Chitin and chitosan were extracted from the exoskeleton of Periplaneta americana (cockroaches) and termites using various acid and alkali techniques. The extraction process involves an initial demineralization step, during which integument dry powder was subjected to 500 mL (2.07 mol/L) of concentrated HCl at 100 degrees Celsius for 30 min, followed by meticulous rinsing with distilled water to restore the pH to its baseline. Deproteinization was conducted at 80 degrees Celsius using 500 mL (1 mol/L) of NaOH solution, which was repeated for 24 h. A total of 250 mL (0.06 mol/L) of NaOH was added at 100 degrees Celsius for 4 h to obtain chitosan, followed by extensive washing and subsequent drying. FTIR analysis was used to identify the functional groups in Periplaneta americana and termites. The crystallinity of these biopolymers, which have a face-centered cubic structure, was determined by X-ray diffraction analysis. This study assessed the analgesic properties of chitin and chitosan via an acetic-acid-induced writhing test in mice, revealing a significant reduction in writhing behavior following the chitin and chitosan extract. Notably, chitin exhibits the highest degree of analgesic activity compared to chitosan. Both chitin and chitosan show anti-inflammatory effects, with chitosan absorbing proton ions at sites of inflammation, while chitin effectively inhibits ear edema and elicits an analgesic response in mice. Furthermore, the present study revealed antipyretic activity, with termite chitin demonstrating the most significant effect at a concentration of 500 µL/mL, followed by chitosan and chitin at 100 µL/mL. These findings indicate the potential of using chitin and chitosan derived from termites and Periplaneta americana as natural anti-inflammatory compounds, implying prospective uses in anti-inflammatory, antipyretic, and analgesic capabilities. Full article

(This article belongs to the Section Biomaterials and Devices for Healthcare Applications)

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(A) Chitin solution (B) chitosan solution (C) termite’s chitin solution derived from Periplaneta americana and termites. Full article ">Figure 2
FT–IR spectra of (A) chitin and chitosan (B) termite chitin derived from Periplaneta americana and termites. Full article ">Figure 3
XRD analysis of (A) chitin, (B) chitosan, and (C) termite chitin derived from Periplaneta americana and termites. Full article ">Figure 4
Analgesic activity graph of chitin and chitosan and termite’s chitin derived from Periplaneta americana and termites. Full article ">Figure 5
Anti-inflammatory graph of chitin, chitosan, and termite’s chitin extracted from Periplaneta americana and termites. Full article ">Figure 6
(A) Histological section of the ear at 50 µL/mL extracts (magnification power = 50×). (B) Extract inhibition at 100 µL/mL (magnification power = 50×). (C) Extract inhibition at 500 µL/mL (magnification power = 100×). (D) Termite’s chitin at 500 µL/mL (magnification power = 100×). (E) Shows a positive control (magnification power = 100×), which is an enlarged section of the tissue shown in (B) and demonstrates a parallel reduction in swelling in a different part of the same tissue. The red arrows indicate the reduction of swelling in specific areas. Full article ">

26 pages, 3537 KiB

Open AccessEditor’s ChoiceReview

Advancements in Biomedical Applications of Calcium Phosphate Glass and Glass-Based Devices—A Review

by Jawad T. Pandayil, Nadia G. Boetti and Davide Janner

J. Funct. Biomater. 2024, 15(3), 79; https://doi.org/10.3390/jfb15030079 - 21 Mar 2024

Cited by 8 | Viewed by 2753

Abstract

Calcium phosphate (CaP) glass has recently gained popularity as a promising material for a wide range of biomedical applications. Recent developments have seen CaP glasses moving from a passive implant material to an active degradable material, particularly as a major constituent of bioresorbable [...] Read more.

Calcium phosphate (CaP) glass has recently gained popularity as a promising material for a wide range of biomedical applications. Recent developments have seen CaP glasses moving from a passive implant material to an active degradable material, particularly as a major constituent of bioresorbable photonic devices. This holds great promise in advanced biomedical applications, since the main constituents of CaP glasses are present in the human body. In this review, the progressive advancements in the biomedical applications of calcium phosphate glass-based devices over the past 50 years are discussed. An overview of their role as reinforcing agents and the studies on doping their matrices for ion releasing and drug and gene delivery are reviewed. Recent applications of CaP glass and fibers in soft-tissue engineering and their potential for optical quality bioresorbable devices are then discussed along with the current challenges and potential future directions, emphasizing the promising role of CaP glass in the next generation of biomaterials. Considering their progress and potential in performing several biomedical functionalities over time, CaP glass-based devices hold promise for becoming enabling tools as an implantable, bioresorbable, multifunctional class of devices in future biomedicine. Full article

(This article belongs to the Special Issue Bioactive Glasses in Medical Applications)

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5 pages, 203 KiB

Open AccessEditorial

Bone Regeneration and Repair Materials

by Marcio Mateus Beloti and Adalberto Luiz Rosa

J. Funct. Biomater. 2024, 15(3), 78; https://doi.org/10.3390/jfb15030078 - 21 Mar 2024

Cited by 1 | Viewed by 2348

Abstract

Bone tissue has a remarkable ability to regenerate following injury and trauma [...] Full article

(This article belongs to the Special Issue Bone Regeneration and Repair Materials)

19 pages, 5486 KiB

Open AccessArticle

Precision Engineering of Chondrocyte Microenvironments: Investigating the Optimal Reaction Conditions for Type B Gelatin Methacrylate Hydrogel Matrix for TC28a2 Cells

by Qichan Hu, Marc A. Torres, Hongjun Pan, Steven L. Williams and Melanie Ecker

J. Funct. Biomater. 2024, 15(3), 77; https://doi.org/10.3390/jfb15030077 - 20 Mar 2024

Cited by 3 | Viewed by 2533

Abstract

Gelatin methacrylate (GelMA) is a photocrosslinkable biomaterial that has gained widespread use in tissue engineering due to its favorable biological attributes and customizable physical and mechanical traits. While GelMA is compatible with various cell types, distinct cellular responses are observed within GelMA hydrogels. [...] Read more.

Gelatin methacrylate (GelMA) is a photocrosslinkable biomaterial that has gained widespread use in tissue engineering due to its favorable biological attributes and customizable physical and mechanical traits. While GelMA is compatible with various cell types, distinct cellular responses are observed within GelMA hydrogels. As such, tailoring hydrogels for specific applications has become imperative. Thus, our objective was to develop GelMA hydrogels tailored to enhance cell viability specifically for TC28a2 chondrocytes in a three-dimensional (3D) cell culture setting. We investigated GelMA synthesis using PBS and 0.25M CB buffer, analyzed the mechanical and physical traits of GelMA hydrogels, and evaluated how varying GelMA crosslinking conditions (GelMA concentration, photoinitiator concentration, and UV exposure time) affected the viability of TC28a2 chondrocytes. The results revealed that GelMA synthesis using 0.25M CB buffer led to a greater degree of methacrylation compared to PBS buffer, and the LAP photoinitiator demonstrated superior efficacy for GelMA gelation compared to Irgacure 2959. Additionally, the stiffness, porosity, and swelling degree of GelMA hydrogels were predominantly affected by GelMA concentration, while cell viability was impacted by all crosslinking conditions, decreasing notably with increasing GelMA concentration, photoinitiator concentration, and UV exposure time. This study facilitated the optimization of crosslinking conditions to enhance cell viability within GelMA hydrogels, a critical aspect for diverse biomedical applications. Full article

(This article belongs to the Section Biomaterials for Tissue Engineering and Regenerative Medicine)

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Schematic illustration of GelMA hydrogel synthesis. Full article ">Figure 2
The appearance of reaction solutions during GelMA synthesis. (a) Appearance after a 3 h reaction of gelatin and methacrylic anhydride at 500 rpm at 50 °C. (b) Appearance after centrifugation of the reacted mixture at 3500 rpm for 3 min. Full article ">Figure 3
Verification of GelMA synthesis after a 3 h reaction. (a) 1H NMR spectra of GelMA and gelatin. (b) Degree of substitution assessed through TNBS assay at different pH conditions. (c) Degree of substitution assessed through TNBS assay at different ratios of MAA/gelatin. Data in (b,c) is presented as mean ± standard error, and statistical significance (* p < 0.05, ** p < 0.01) was determined using one-way ANOVA. Full article ">Figure 4
Compressive moduli of GelMA hydrogels when crosslinked at the following conditions: (a) 10% GelMA, UV exposure for 2 min; (b) 10% GelMA, LAP concentration of 0.025%; (c) UV exposure for 2 min, LAP concentration of 0.025%. All the data is presented as mean ± standard error, and statistical significance (* p < 0.05, ** p < 0.01) was determined using one-way ANOVA. Full article ">Figure 5
Physical properties of GelMA hydrogels. (a) SEM images of 10% GelMA subjected to 2 min of UV exposure with varying concentrations of LAP. (b) SEM images of 10% GelMA with 0.025% LAP under varying UV exposure times. (c) SEM images of GelMA hydrogels with different gel concentrations using 0.025% LAP and a UV exposure time of 2 min. (d) Swelling ratio of GelMA hydrogels with different gel concentrations using 0.025% LAP and a UV exposure time of 2 min. Data in (d) is presented as mean ± standard error, and statistical significance (* p < 0.05, ** p < 0.01) was determined using one-way ANOVA. Full article ">Figure 6
Cell viability measured by MTT. (a) Effect of photoinitiator LAP on cell viability assessed after 24 h and 48 h incubation, respectively. (b) Effect of UV exposure time on cell viability assessed after 24 h and 48 h incubation, respectively. (c) The combined effect of UV and LAP on cell viability assessed after 24 h of incubation. The dotted line represents 70% cell viability. Full article ">Figure 7
Cell behavior in GelMA hydrogels. (a) Cell viability by live/dead fluorescence staining. Live cells are green (top row), and dead cells are red (center row). The bottom row displays an overlay of the first and second rows. The scale bar is 200 μM. (b) Cell viability by alamarBlue assay. Data is presented as mean ± standard error, and statistical significance (* p < 0.05, ** p < 0.01) was determined using one-way ANOVA. (c) H&E staining of cell-laden GelMA hydrogels. Green circles indicate mitosis, and red circles indicate cellular fragmentation. The scale bar is 100 μM. Full article ">

14 pages, 5480 KiB

Open AccessArticle

Shear Stress Quantification in Tissue Engineering Bioreactor Heart Valves: A Computational Approach

by Raj Dave, Giulia Luraghi, Leslie Sierad, Francesco Migliavacca and Ethan Kung

J. Funct. Biomater. 2024, 15(3), 76; https://doi.org/10.3390/jfb15030076 - 20 Mar 2024

Viewed by 2067

Abstract

Tissue-engineered heart valves can grow, repair, and remodel after implantation, presenting a more favorable long-term solution compared to mechanical and porcine valves. Achieving functional engineered valve tissue requires the maturation of human cells seeded onto valve scaffolds under favorable growth conditions in bioreactors. [...] Read more.

Tissue-engineered heart valves can grow, repair, and remodel after implantation, presenting a more favorable long-term solution compared to mechanical and porcine valves. Achieving functional engineered valve tissue requires the maturation of human cells seeded onto valve scaffolds under favorable growth conditions in bioreactors. The mechanical stress and strain on developing valve tissue caused by different pressure and flow conditions in bioreactors are currently unknown. The aim of this study is to quantify the wall shear stress (WSS) magnitude in heart valve prostheses under different valve geometries and bioreactor flow rates. To achieve this, this study used fluid–structure interaction simulations to obtain the valve’s opening geometries during the systolic phase. These geometries were then used in computational fluid dynamics simulations with refined near-wall mesh elements and ranges of prescribed inlet flow rates. The data obtained included histograms and regression curves that characterized the distribution, peak, and median WSS for various flow rates and valve opening configurations. This study also found that the upper region of the valve near the commissures experienced higher WSS magnitudes than the rest of the valve. Full article

(This article belongs to the Special Issue Applications of Biomaterials on Vascular Tissue Engineering)

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Valves representing pediatric, adolescent, and adult sizes: 12.3, 18.45, and 24.6 mm, respectively. Full article ">Figure 2
Isometric and planform (top) views of GOA. Full article ">Figure 3
Summary of the geometries included in the CFD simulation to quantify WSS. Numbers represent the GOA in mm2. Full article ">Figure 4
WSS histogram for the 24.6 mm valve. Flow rate is arranged in the vertical direction and increases down the columns. % GOA is arranged in the horizontal direction. The area of each histogram is individually normalized to 1. Full article ">Figure 5
WSS histogram for the 18.45 mm valve. Flow rate is arranged in the vertical direction and increases down the columns. % GOA is arranged in the horizontal direction. The area of each histogram is individually normalized to 1. Full article ">Figure 6
WSS histogram for the 12.3 mm valve. Flow rate is arranged in the vertical direction and increases down the columns. % GOA is arranged in the horizontal direction. The area of each histogram is individually normalized to 1. Full article ">Figure 7
(a) Typically observed WSS pattern in the valve simulations. (b) Altered WSS pattern due to separation of flow from the valve surface for smaller valves with high flow rates. WSS legend: orange indicates regions of higher WSS and deep blue regions of lower WSS. The image is presented for illustrative purposes to depict the typical distribution of WSS observed during this study; therefore, specific values are not shown. Full article ">Figure 8
Regression plot: median (50th) percentile WSS. The normalized root mean squared error (NRMSE) value is normalized by range. Full article ">Figure 9
Regression plot: 99th percentile WSS. Full article ">Figure 10
Comparing histograms of selected data points close to each other in <a href="#jfb-15-00076-f008" class="html-fig">Figure 8</a>. (a) Point 1 [10.45, 1.227], Point 2 [10.47, 1.231], Percentage Difference [0.179, 0.302]; (b) Point 1 [10.69, 1.423], Point 2 [10.71, 1.424], Percentage Difference [0.163, 0.022]; (c) Point 1 [11.06, 2.163], Point 2 [11.08, 2.165], Percentage Difference [0.137, 0.063]; (d) Point 1 [11.54, 2.338], Point 2 [11.57, 2.333], Percentage Difference [0.261, 0.237]. Full article ">

12 pages, 5026 KiB

Open AccessArticle

Toxicological Evaluation toward Refined Montmorillonite with Human Colon Associated Cells and Human Skin Associated Cells

by Zhou Wang, Yibei Jiang, Guangjian Tian, Chuyu Zhu and Yi Zhang

J. Funct. Biomater. 2024, 15(3), 75; https://doi.org/10.3390/jfb15030075 - 20 Mar 2024

Cited by 4 | Viewed by 1664

Abstract

Montmorillonite has been refined to overcome uncertainties originating from different sources, which offers opportunities for addressing various health issues, e.g., cosmetics, wound dressings, and antidiarrheal medicines. Herein, three commercial montmorillonite samples were obtained from different sources and labeled M1, M2, and M3 for [...] Read more.

Montmorillonite has been refined to overcome uncertainties originating from different sources, which offers opportunities for addressing various health issues, e.g., cosmetics, wound dressings, and antidiarrheal medicines. Herein, three commercial montmorillonite samples were obtained from different sources and labeled M1, M2, and M3 for Ca-montmorillonite, magnesium-enriched Ca-montmorillonite, and silicon-enriched Na-montmorillonite, respectively. Commercial montmorillonite was refined via ultrasonic scission-differential centrifugation and labeled S, M, or L according to the particle sizes (small, medium, or large, respectively). The size distribution decreased from 2000 nm to 250 nm with increasing centrifugation rates from 3000 rpm to 12,000 rpm. Toxicological evaluations with human colon-associated cells and human skin-associated cells indicated that side effects were correlated with excess dosages and silica sand. These side effects were more obvious with human colon-associated cells. The microscopic interactions between micro/nanosized montmorillonite and human colon-associated cells or human skin-associated cells indicated that those interactions were correlated with the size distributions. The interactions of the M1 series with the human cells were attributed to size effects because montmorillonite with a broad size distribution was stored in the M1 series. The M2 series interactions with human cells did not seem to be correlated with size effects because large montmorillonite particles were retained after refining. The M3 series interactions with human cells were attributed to size effects because small montmorillonite particles were retained after refining. This illustrates that toxicological evaluations with refined montmorillonite must be performed in accordance with clinical medical practices. Full article

(This article belongs to the Special Issue Molecular Mechanisms and Biological Procedures of Biomaterials in Medical Applications)

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Characterization and toxicological evaluation of MMT via (A) XRD, (B) FTIR, and (C) CCK-8. M1-RAW: unrefined Ca-MMT; M2-RAW: unrefined magnesium-enriched Ca-MMT; M3-RAW: unrefined silicon-enriched Na-MMT. The P value was determined by ANOVA and Student’s t tests. *** p < 0.0005. Full article ">Figure 2
Characterization of refined MMT via (A) SEM, (B) DSL, and (C) XRD. The refined MMTs with small, medium, and large particle sizes were labeled S, M, and L, respectively. Full article ">Figure 3
Toxicological effects of refined MMT on human colon-associated cells (HIEC 6 and NCM460) and human skin-associated cells (HaCaT and HUVECs). The p value was determined by ANOVA and Student’s t tests. ** p < 0.005, *** p < 0.0005. Full article ">Figure 4
SEM images of MMT–human cell interactions. Unrefined Ca-MMT was labeled M1-RAW, and refined M1-RAW with small particle sizes was labeled M1-S. Unrefined magnesium-enriched Ca-MMT was labeled M2-RAW, and refined M2-RAW with small particle sizes was labeled M2-S. Unrefined silicon-enriched Na-MMT was labeled M3-RAW, and refined M3-RAW with small particle sizes was labeled M3-S. The thumbnail showed an enlarged image featuring a white arrow indicating the presence of MMT. Full article ">

27 pages, 4488 KiB

Open AccessArticle

A Comprehensive Study on Folate-Targeted Mesoporous Silica Nanoparticles Loaded with 5-Fluorouracil for the Enhanced Treatment of Gynecological Cancers

by Aliyah Almomen and Adel Alhowyan

J. Funct. Biomater. 2024, 15(3), 74; https://doi.org/10.3390/jfb15030074 - 20 Mar 2024

Cited by 4 | Viewed by 2412

Abstract

Background: Gynecological cancers are a significant public health concern, accounting for 40% of all cancer incidence and 30% of deaths in women. 5-Fluorouracil (5-FU) can be used with chemotherapy to improve treatment in advanced-stage gynecological cancer. Mesoporous silica nanoparticles (MSNs) can improve drug [...] Read more.

Background: Gynecological cancers are a significant public health concern, accounting for 40% of all cancer incidence and 30% of deaths in women. 5-Fluorouracil (5-FU) can be used with chemotherapy to improve treatment in advanced-stage gynecological cancer. Mesoporous silica nanoparticles (MSNs) can improve drug effectiveness and reduce toxicity. Folic acid can target folate receptors in epithelial malignancies like ovarian and cervical cancer. Methods: The mixture of MSN-NH2 was synthesized by dissolving N-lauroylsarcosine sodium in a water–ethanol mixture, adding APTES and TEOS, and heating at 80 °C for 18 h, before being fully characterized. The drug is loaded into a 5-FU solution and functionalized with folate. The drug release mechanism, as well as ex vivo intestinal permeation from MSN-NH2 formulations, was tested. The cell viability study of the nanoparticles was evaluated in various cancer cell lines, and the cellular uptake was measured indirectly using HPLC. Results: The study analyzed the amine content, propylamine loading, and drug loading capacity of MSN-NH2 nanoparticles. It found that the loading of propylamine was around 0.733 mmol/g, and the surface density was 0.81 molecules/nm. The study also showed that the surface decoration of MSN-NH2 with folic acid was successfully achieved. The release rate of 5-FU from MSN-NH2 was slow and controlled, with a slower rate at pH 5.5. The study found that the amin surface functionalization of MSN-NH2 nanoparticles can reduce potential toxicity in ovarian and cervical cancer cells. Conclusions: Based on the results, the encapsulation of 5-FU and functionalization of MSN-NH2 with folic acid can serve as potential carriers for 5-FU in treating gynecological cancer. Full article

(This article belongs to the Special Issue Nanoparticles and Nanocompounds for Cancer Therapy)

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18 pages, 5077 KiB

Open AccessEditor’s ChoiceArticle

Antibacterial and Antibiofouling Activities of Carbon Polymerized Dots/Polyurethane and C₆₀/Polyurethane Composite Films

by Zoran M. Marković, Milica D. Budimir Filimonović, Dušan D. Milivojević, Janez Kovač and Biljana M. Todorović Marković

J. Funct. Biomater. 2024, 15(3), 73; https://doi.org/10.3390/jfb15030073 - 17 Mar 2024

Cited by 11 | Viewed by 2242

Abstract

The cost of treatment of antibiotic-resistant pathogens is on the level of tens of billions of dollars at the moment. It is of special interest to reduce or solve this problem using antimicrobial coatings, especially in hospitals or other healthcare facilities. The bacteria [...] Read more.

The cost of treatment of antibiotic-resistant pathogens is on the level of tens of billions of dollars at the moment. It is of special interest to reduce or solve this problem using antimicrobial coatings, especially in hospitals or other healthcare facilities. The bacteria can transfer from medical staff or contaminated surfaces to patients. In this paper, we focused our attention on the antibacterial and antibiofouling activities of two types of photodynamic polyurethane composite films doped with carbon polymerized dots (CPDs) and fullerene C₆₀. Detailed atomic force, electrostatic force and viscoelastic microscopy revealed topology, nanoelectrical and nanomechanical properties of used fillers and composites. A relationship between the electronic structure of the nanocarbon fillers and the antibacterial and antibiofouling activities of the composites was established. Thorough spectroscopic analysis of reactive oxygen species (ROS) generation was conducted for both composite films, and it was found that both of them were potent antibacterial agents against nosocomial bacteria (Klebsiela pneumoniae, Proteus mirabilis, Salmonela enterica, Enterococcus faecalis, Enterococcus epidermis and Pseudomonas aeruginosa). Antibiofouling testing of composite films indicated that the CPDs/PU composite films eradicated almost completely the biofilms of Pseudomonas aeruginosa and Staphylococcus aureus and about 50% of Escherichia coli biofilms. Full article

(This article belongs to the Special Issue Photodynamic Therapy of Cancer, Microbes and Viruses)

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Top view AFM images of CPD nanoparticles: (a) height retrace mode, (b) phase retrace mode, (c) viscoelastic measurement of Young’s modulus and (d) profile of Young’s modulus. Full article ">Figure 2
Top view AFM images of neat PU (a), CPDs/PU (b) and C60/PU (c) composite films. Scale bar is 400 nm. Full article ">Figure 3
Top view AFM images of surface morphology (left column) and corresponding EFM phase retrace mode images of surface of CPDs (a,b), CPDs/PU (d,e) and C60/PU composite films (g,h). Dark voids represent CPDs themselves (b), and CPDs in the interior of polymer matrix (d) and C60 into the polymer matrix (f). (c,f,i) show profiles of CPDs, CPDs inside PU and C60 in the interior of PU. Tip voltage was 3 V; room humidity was 35%. Full article ">Figure 4
(a) Absorption spectra of SOSG used as fluorescence probe at 530 nm in the presence of CPDs/PU samples, (b) absorption spectra of SOSG at 533 nm in the presence of C60/PU samples, (c) production rate of singlet oxygen of CPD/PU and C60/PU samples vs. time, (d) photobleaching of ABDA in the presence of CPD/PU samples, (e) photobleaching of ABDA in the presence of C60/PU samples. All absorbance spectra of ABDA were recorded at 398 nm, normalized at the start of the irradiation and averaged over several repeat experiments. Full article ">Figure 5
(a) PL intensity spectra of h-TA at time intervals of 0–90 min and (b) PL intensity spectra of h-TA at time intervals of 0–10 min under excitation of 330 nm. Both samples were irradiated with blue light at 470 nm, 3 W. Full article ">Figure 6
Antibiofouling effect of CPD/PU and C60/PU composite films on P. aureginosa (a), S. aureus (b) and E. coli (c) bacterial biofilms with and without blue light irradiation for 1 h. Absorbance is an indicator of the quantity of biofilm formed. Full article ">Figure 7
Cytotoxicity of PU (control), CPD/PU and C60/PU composite film samples without blue light irradiation determined as a percentage of viable HaCaT cells. The dashed lines highlight the limits of viability according to EN ISO 10993-5: viability >80 corresponds to no cytotoxicity, >60–80 mild cytotoxicity, >40–60 moderate toxicity and <40 severe toxicity. Full article ">

15 pages, 2586 KiB

Open AccessEditor’s ChoiceArticle

Reduction in Pathogenic Biofilms by the Photoactive Composite of Bacterial Cellulose and Nanochitosan Dots under Blue and Green Light

by Danica Z. Zmejkoski, Nemanja M. Zdravković, Milica D. Budimir Filimonović, Vladimir B. Pavlović, Svetlana V. Butulija, Dušan D. Milivojević, Zoran M. Marković and Biljana M. Todorović Marković

J. Funct. Biomater. 2024, 15(3), 72; https://doi.org/10.3390/jfb15030072 - 14 Mar 2024

Cited by 4 | Viewed by 2074

Abstract

In this study, nanochitosan dots (ChiDs) were synthesized using gamma rays and encapsulated in bacterial cellulose (BC) polymer matrix for antibiofilm potential in photodynamic therapy. The composites were analyzed for structural changes using SEM, AFM, FTIR, XRD, EPR, and porosity measurements. Additionally, ChiD [...] Read more.

In this study, nanochitosan dots (ChiDs) were synthesized using gamma rays and encapsulated in bacterial cellulose (BC) polymer matrix for antibiofilm potential in photodynamic therapy. The composites were analyzed for structural changes using SEM, AFM, FTIR, XRD, EPR, and porosity measurements. Additionally, ChiD release was assessed. The results showed that the chemical composition remained unaltered, but ChiD agglomerates embedded in BC changed shape (1.5–2.5 µm). Bacterial cellulose fibers became deformed and interconnected, with increased surface roughness and porosity and decreased crystallinity. No singlet oxygen formation was observed, and the total amount of released ChiD was up to 16.10%. Antibiofilm activity was higher under green light, with reductions ranging from 48 to 57% under blue light and 78 to 85% under green light. Methicillin-resistant Staphylococcus aureus was the most sensitive strain. The new photoactive composite hydrogels show promising potential for combating biofilm-related infections. Full article

(This article belongs to the Section Biomaterials for Cancer Therapies)

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Graphical abstract
Full article ">Figure 1
Scanning electron microscopy (SEM) and atomic force microscopy (AFM) images of BC-ChiD_control (a,d); BC-ChiD_blue (b,e); and BC-ChiD_green (c,f) samples. Full article ">Figure 2
FTIR spectra (a) of BC-ChiD_control (black curve), BC-ChiD_blue (blue curve), and BC-ChiD_green (green curve). All spectra are displaced for clarity. XRD patterns (b) of BC-ChiD_blue and BC-ChiD_green samples. Full article ">Figure 3
Porosity (a) of BC-ChiD_control, BC-ChiD_blue, and BC-ChiD_green composite hydrogels. (* p < 0.05; *** p < 0.001). EPR signal intensity (b) of control samples, as well as BC-ChiD_blue and BC-ChiD_green samples. Full article ">Figure 4
In vitro release profile of ChiD from BC-ChiD composites exposed to green and blue light (a) and ChiD release data fitted to Korsmeyer–Peppas mathematical kinetic model (b). Full article ">Figure 5
AFM images of E. coli and MRSA biofilms before the application of BC-ChiD composites ((a,d), respectively) and after the application of BC-ChiD under blue ((b,e), respectively) and green ((c,f), respectively) light. Full article ">

14 pages, 1969 KiB

Open AccessArticle

Bonding Pretreatment of Aesthetic Dental CAD-CAM Materials through Surface Etching with a Mixed Aqueous Solution of Ammonium Fluoride and Ammonium Hydrogen Sulfate

by Yusaku Nishizawa, Tatsuo Kawamoto and Hiroshi Ikeda

J. Funct. Biomater. 2024, 15(3), 71; https://doi.org/10.3390/jfb15030071 - 14 Mar 2024

Viewed by 1927

Abstract

Hydrofluoric acid (HF) is commonly used as an etchant for the pretreatment of dental computer-aided design/computer-aided manufacturing (CAD-CAM) materials, such as glass-ceramics and resin composites. Despite its effectiveness, the harmful and hazardous nature of HF has raised significant safety concerns. In contrast, ammonium [...] Read more.

Hydrofluoric acid (HF) is commonly used as an etchant for the pretreatment of dental computer-aided design/computer-aided manufacturing (CAD-CAM) materials, such as glass-ceramics and resin composites. Despite its effectiveness, the harmful and hazardous nature of HF has raised significant safety concerns. In contrast, ammonium fluoride (AF) is known for its relatively low toxicity but has limited etching capability. This study explored the potential of ammonium hydrogen sulfate (AHS), a low-toxicity and weak acid, to enhance the etching ability of aqueous AF solutions for the bonding pretreatment of CAD-CAM materials. This study investigated five types of aesthetic CAD-CAM materials: lithium disilicate glass, feldspathic porcelain, polymer-infiltrated ceramic networks, resin composites, and zirconia. Seven experimental etchants were prepared by varying the amount of AHS added to aqueous AF solutions, with each etchant used to etch the surfaces of the respective CAD-CAM materials. The treated surfaces were analyzed using scanning electron microscopy and confocal laser scanning microscopy. Additionally, the shear bond strength (SBS) of the CAD-CAM materials treated with a luting agent (resin cement) was evaluated. The results indicated that the AF1/AHS3 (weight ratio AF:AHS = 1:3) etchant had the most substantial etching effect on the surfaces of silica-containing materials (lithium disilicate glass, feldspathic porcelain, polymer-infiltrated ceramic networks, and resin composites) but not on zirconia. The SBS of the materials treated with the AF1/AHS3 etchant was comparable to that of the commercial HF etchant. Hence, an AF/AHS mixed solution could effectively etch silica-containing CAD-CAM materials, thereby enhancing their bonding capabilities. Full article

(This article belongs to the Special Issue State of the Art in Dental Materials)

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Typical failure mode of the cement-debonded sample surfaces after the shear bond strength test; (a) adhesive failure and (b) cohesive failure. The surfaces were observed using the optical microscope with 30× magnification. Full article ">Figure 2
Relationship between pH values and weight ratio of ammonium fluoride (AF) and ammonium hydrogen sulfate (AHS) in experimental etchant. Full article ">Figure 3
SEM images for CAD-CAM material surfaces after treatment with experimental etchants and commercial HF etchant. Full article ">Figure 4
Ra values for CAD-CAM material surfaces after treatment with experimental etchants or commercial HF etchant: (a) lithium disilicate glass, (b) feldspathic porcelain, (c) polymer-infiltrated ceramic network, (d) resin composite, and (e) zirconia. The different alphabetic letters in the figure indicate significant differences between the groups (p < 0.05, Tukey’s test). Full article ">Figure 5
SBS values for CAD-CAM material surfaces treated with the experimental AF1/AHS3 etchant or commercial HF etchant: (a) lithium disilicate glass, (b) feldspathic porcelain, (c) polymer-infiltrated ceramic network, (d) resin composite, and (e) zirconia. The different alphabetic letters in the figure indicate significant differences between the groups (p < 0.05, Tukey’s test). Full article ">

12 pages, 1244 KiB

Open AccessArticle

Evaluation of Antimicrobial Properties, Cell Viability, and Metalloproteinase Activity of Bioceramic Endodontic Materials Used in Vital Pulp Therapy

by Felipe Immich, Durvalino de Oliveira, Juliana Silva Ribeiro de Andrade, Andressa da Silva Barboza, Carlos Enrique Cuevas-Suárez, Adriana Fernandes da Silva, Wellington Luiz de Oliveira da Rosa, Álvaro Henrique Borges, Neftali Lenin Villarreal Carreno, Evandro Piva and Rafael Guerra Lund

J. Funct. Biomater. 2024, 15(3), 70; https://doi.org/10.3390/jfb15030070 - 14 Mar 2024

Cited by 3 | Viewed by 2501 | Correction

Abstract

This study aimed to evaluate the antimicrobial properties, cell viability, and matrix metalloproteinase (MMP) inhibition capacity of several endodontic materials aimed at vital pulp therapy: Pro Root MTA^®, EndoSequence^®, Biodentine^®, MTA Angelus^®, TheraCal LC^® [...] Read more.

This study aimed to evaluate the antimicrobial properties, cell viability, and matrix metalloproteinase (MMP) inhibition capacity of several endodontic materials aimed at vital pulp therapy: Pro Root MTA^®, EndoSequence^®, Biodentine^®, MTA Angelus^®, TheraCal LC^®, and BioC Repair^®. The materials were prepared according to the manufacturer’s instructions. Antimicrobial tests were conducted using a microcosm biofilm model, cell viability was assessed using murine fibroblasts (L929), and MMP activity was analyzed through electrophoresis. The results showed that BioC Repair^®, Biodentine^®, and EndoSequence^® exhibited similar antimicrobial properties, while MTA Angelus^® and ProRoot MTA^® had inferior results but were comparable to each other. In terms of cell viability, no significant differences were observed among the materials. EndoSequence^® demonstrated the highest MMP inhibition capacity. In conclusion, BioC Repair^®, Biodentine^®, EndoSequence^®, and TheraCal^® showed better antimicrobial properties among the tested materials. The materials did not exhibit significant differences in terms of cytotoxicity. However, EndoSequence^® displayed superior MMP inhibition capacity. Full article

(This article belongs to the Section Dental Biomaterials)

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18 pages, 3793 KiB

Open AccessEditor’s ChoiceArticle

Formulative Study and Characterization of Novel Biomaterials Based on Chitosan/Hydrolyzed Collagen Films

by Tomás Martínez Rodríguez, Caterina Valentino, Francisco Ramón Rodríguez Pozo, Pablo Hernández Benavides, Francisco Arrebola Vargas, José Manuel Paredes, Claro Ignacio Sainz-Díaz, Guillermo R. Iglesias, Silvia Rossi, Giuseppina Sandri, María del Mar Medina Pérez and Carola Aguzzi

J. Funct. Biomater. 2024, 15(3), 69; https://doi.org/10.3390/jfb15030069 - 11 Mar 2024

Cited by 2 | Viewed by 2247

Abstract

To date, the need for biomaterials capable of improving the treatment of chronic skin wounds remains a clinical challenge. The aim of the present work is to formulate and characterize chitosan (Cs)/hydrolyzed collagen (HC) films as potential biomaterials with improved mechanical and hydration [...] Read more.

To date, the need for biomaterials capable of improving the treatment of chronic skin wounds remains a clinical challenge. The aim of the present work is to formulate and characterize chitosan (Cs)/hydrolyzed collagen (HC) films as potential biomaterials with improved mechanical and hydration performances compared to single component formulations. Films were made by the solvent casting method, with or without glycerin and/or PEG1500 as plasticizers, resulting in a total of eight formulations. All films were characterized by their physico-chemical characteristics and their mechanical and hydration features. A full factorial design was also used to statistically assess the effect of HC concentration, type and concentration of plasticizers and their possible interactions on mechanical and swelling behaviors. Solid state characterization confirmed the hybrid nature of the films, with suggested electrostatic interactions between Cs and HC. Mechanical and swelling properties, along with the analysis of the experimental design, allowed the identification of formulations containing high HC concentration (2% w/v) and glycerin or glycerin/PEG1500 as more suitable candidates for skin wound treatment. Finally, viability assay of immortalized human keratinocytes (HaCaT) showed no statistical differences in cell survival compared to the complete culture medium, suggesting their potential as a promising tool for biomedical applications. Full article

(This article belongs to the Special Issue Natural Product-Based Biomaterials for Advanced Wound Dressings)

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FTIR spectra of the film formulations and their pristine components. Full article ">Figure 2
TGA curves of the film formulations and their pristine components: complete curves (a) and detail from 35 °C to 135 °C (b). Full article ">Figure 3
Physical appearance of the films. Full article ">Figure 4
Film swelling capacity profiles (mean values ± s.d.; n = 3). Full article ">Figure 5
Interaction plots between GLY and PEG1500 for EAB % (a) and swelling capacity (b). Full article ">Figure 6
Response surface plots for EAB % (a) and swelling capacity (b), obtained from the analysis of the full factorial design (response values from all the studied film formulations are given in the plots). Full article ">Figure 7
SEM microphotographs of F4 (a) and F6 (b) film samples. Full article ">Figure 8
Viability of HaCaT cells after 24 h and 72 h of incubation with F4, F6 and pristine Cs films; CM was considered as control (mean values ± e.s.; n = 9). Full article ">

68 pages, 4001 KiB

Open AccessSystematic Review

Surface Treatment of Dental Mini-Sized Implants and Screws: A Systematic Review with Meta-Analysis

by Ana Luísa Figueiredo, Raquel Travassos, Catarina Nunes, Madalena Prata Ribeiro, Mariana Santos, Flavia Iaculli, Anabela Baptista Paula, Carlos Miguel Marto, Francisco Caramelo, Inês Francisco and Francisco Vale

J. Funct. Biomater. 2024, 15(3), 68; https://doi.org/10.3390/jfb15030068 - 10 Mar 2024

Viewed by 2961

Abstract

Miniscrews are devices that allow for absolute skeletal anchorage. However, their use has a higher failure rate (10–30%) than dental implants (10%). To overcome these flaws, chemical and/or mechanical treatment of the surface of miniscrews has been suggested. There is no consensus in [...] Read more.

Miniscrews are devices that allow for absolute skeletal anchorage. However, their use has a higher failure rate (10–30%) than dental implants (10%). To overcome these flaws, chemical and/or mechanical treatment of the surface of miniscrews has been suggested. There is no consensus in the current literature about which of these methods is the gold standard; thus, our objective was to carry out a systematic review and meta-analysis of the literature on surface treatments of miniscrews. The review protocol was registered (PROSPERO CRD42023408011) and is in accordance with the PRISMA guidelines. A bibliographic search was carried out on PubMed via MEDLINE, Cochrane Library, Embase and Web of Science. The initial search of the databases yielded 1684 results, with 98 studies included in the review, with one article originating from the search in the bibliographic references of the included studies. The results of this systematic review show that the protocols of miniscrew surface treatments, such as acid-etching; sandblasting, large-grit and acid-etching; photofunctionalization with ultraviolet light; and photobiomodulation, can increase stability and the success of orthodontic treatment. The meta-analysis revealed that the treatment with the highest removal torque is SLA, followed by acid-etching. On the other hand, techniques such as oxidative anodization, anodization with pre-calcification and heat treatment, as well as deposition of chemical compounds, require further investigation to confirm their effectiveness. Full article

(This article belongs to the Special Issue Orthodontics Materials and Technologies)

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14 pages, 2086 KiB

Open AccessArticle

Effect of DMSO on Structural Properties of DMPC and DPPC Liposome Suspensions

by Luísa M. P. F. Amaral, Maria Rangel and Margarida Bastos

J. Funct. Biomater. 2024, 15(3), 67; https://doi.org/10.3390/jfb15030067 - 10 Mar 2024

Cited by 3 | Viewed by 2502

Abstract

The study and characterization of the biophysical properties of membranes and drug–membrane interactions represent a critical step in drug development, as biological membranes act as a barrier that the drug must overcome to reach its active site. Liposomes are widely used in drug [...] Read more.

The study and characterization of the biophysical properties of membranes and drug–membrane interactions represent a critical step in drug development, as biological membranes act as a barrier that the drug must overcome to reach its active site. Liposomes are widely used in drug delivery to circumvent the poor aqueous solubility of most drugs, improving systemic bioavailability and pharmacokinetics. Further, they can be targeted to deliver to specific disease sites, thus decreasing drug load, and reducing side effects and poor adherence to treatment. To improve drug solubility during liposome preparation, DMSO is the most widely used solvent. This raises concern about the potential effect of DMSO on membranes and leads us to investigate, using DSC and EPR, the influence of DMSO on the behavior of lipid model membranes of DMPC and DPPC. In addition, we tested the influence of DMSO on drug–membrane interaction, using compounds with different hydrophobicity and varying DMSO content, using the same experimental techniques. Overall, it was found that with up to 10% DMSO, changes in the bilayer fluidity or the thermotropic properties of the studied liposomes were not significant, within the experimental uncertainty. For higher concentrations of DMSO, there is a stabilization of both the gel and the rippled gel phases, and increased bilayer fluidity of DMPC and DPPC liposomes leading to an increase in membrane permeability. Full article

(This article belongs to the Special Issue Liposomal Nanomedicine: Applications for Drug Delivery and Cancer Therapy)

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Full article ">Figure 1
Typical EPR spectra of 5-DSA in: (A) in solution, (B) DMPC liposome at T = 310 K, where 2Amax is the maximum hyperfine splitting. The structure of 5-DSA is shown below. Full article ">Figure 2
The DSC heating thermograms for DMPC (A,B) and DPPC (C,D) unilamellar vesicles (LUVs), with increasing amounts of DMSO, added at room temperature (~22 °C) after liposome preparation, immediately prior to the DSC run (“DMSO added after preparation”). The DMSO content is expressed in the legends, in (v/v) %. The curves are already corrected for blank (buffer/buffer run), and baseline subtracted. Full article ">Figure 3
DSC heating thermograms of DMPC multilamellar vesicles, MLVs, “with DMSO added to already formed liposomes” plotted as a function of DMSO content. The DMSO content is expressed in the legends, in (v/v)%. The curves are already corrected for blank (buffer/buffer run) and baseline subtracted. Insert: enlargement of the pre-transition for the different DMSO contents. Full article ">Figure 4
DSC heating thermograms of the DMPC multilamellar vesicles, MLVs, with increasing amounts of DMSO, using “DMSO added on liposome preparation, incorporation method”. Insert: enlargement of the pre-transition for the various DMSO contents. The DMSO content is expressed in the legends, in (v/v)%. The curves are already corrected for blank (buffer/buffer run) and baseline subtracted. Full article ">Figure 5
Effect of increasing amounts of DMSO on 5-DSA degree of anisotropy (2Amax) in DMPC liposomes at T = 310 K (fluid phase, Lα). ** p ≤ 0.01; **** p ≤ 0.0001. Full article ">Figure 6
EPR spectrum of spin labelled DPPC liposomes at T = 323 K, with DMSO contents: (A)—0%; (B)—40%; (C)—5-DSA in HEPES/DMSO solution. Full article ">Scheme 1
Methodology adopted for liposome samples preparation. (a) LUVs with “DMSO added after preparation”, (b) MLVS with DMSO added to already formed liposomes, and (c) MLVs with “DMSO added on liposome preparation, incorporation method”. In sample (a,b) DMSO is added to the liposome suspensions just prior to the DSC experiment. Full article ">

24 pages, 20720 KiB

Open AccessArticle

Bone Marrow-Derived Mesenchymal Stem Cell-Laden Nanocomposite Scaffolds Enhance Bone Regeneration in Rabbit Critical-Size Segmental Bone Defect Model

by Elangovan Kalaiselvan, Swapan Kumar Maiti, Shivaraju Shivaramu, Shajahan Amitha Banu, Khan Sharun, Divya Mohan, Sangeetha Palakkara, Sadhan Bag, Monalisa Sahoo, Suresh Ramalingam and Jürgen Hescheler

J. Funct. Biomater. 2024, 15(3), 66; https://doi.org/10.3390/jfb15030066 - 10 Mar 2024

Cited by 4 | Viewed by 2496

Abstract

Bone regeneration poses a significant challenge in the field of tissue engineering, prompting ongoing research to explore innovative strategies for effective bone healing. The integration of stem cells and nanomaterial scaffolds has emerged as a promising approach, offering the potential to enhance regenerative [...] Read more.

Bone regeneration poses a significant challenge in the field of tissue engineering, prompting ongoing research to explore innovative strategies for effective bone healing. The integration of stem cells and nanomaterial scaffolds has emerged as a promising approach, offering the potential to enhance regenerative outcomes. This study focuses on the application of a stem cell-laden nanomaterial scaffold designed for bone regeneration in rabbits. The in vivo study was conducted on thirty-six healthy skeletally mature New Zealand white rabbits that were randomly allocated into six groups. Group A was considered the control, wherein a 15 mm critical-sized defect was created and left as such without any treatment. In group B, this defect was filled with a polycaprolactone–hydroxyapatite (PCL + HAP) scaffold, whereas in group C, a PCL + HAP-carboxylated multiwalled carbon nanotube (PCL + HAP + MWCNT-COOH) scaffold was used. In group D, a PCL + HAP + MWCNT-COOH scaffold was used with local injection of bone morphogenetic protein-2 (BMP-2) on postoperative days 30, 45, and 60. The rabbit bone marrow-derived mesenchymal stem cells (rBMSCs) were seeded onto the PCL + HAP + MWCNT-COOH scaffold by the centrifugal method. In group E, an rBMSC-seeded PCL + HAP + MWCNT-COOH scaffold was used along with the local injection of rBMSC on postoperative days 7, 14, and 21. For group F, in addition to the treatment given to group E, BMP-2 was administered locally on postoperative days 30, 45, and 60. Gross observations, radiological observation, scanning electron microscopic assessment, and histological evaluation study showed that group F displayed the best healing properties, followed by group E, group D, group C, and B. Group A showed no healing with ends blunting minimal fibrous tissue. Incorporating growth factor BMP-2 in tissue-engineered rBMSC-loaded nanocomposite PCL + HAP + MWCNT-COOH construct can augment the osteoinductive and osteoconductive properties, thereby enhancing the healing in a critical-sized bone defect. This novel stem cell composite could prove worthy in the treatment of non-union and delayed union fractures in the near future. Full article

(This article belongs to the Special Issue Application of Biomaterials in Tissue Engineering and Regenerative Medicine)

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(A1) Nanocomposite polycaprolactone (PCL) + hydroxyapatite (HAP) scaffold and (B1) nanocomposite PCL + HAP + multiwalled carboxylated carbon nanotube scaffold surgically placed in the critical-sized defect in the radius bone. Scanning electron microscope (SEM) image of PCL + HAP scaffold (A2: 1000× and A3: 2000×). SEM image of PCL + HAP + MWCNT-COOH scaffold (B2: 1000× and B3: 2000×). Full article ">Figure 2
Histogram (mean ± SD) showing (A) pain score and (B) swelling score of various treatment groups at different time intervals. Line graph (mean ± SD) showing (C) exudation score and (D) lameness score of various treatment groups at different time intervals. Full article ">Figure 3
Histogram (mean ± SD) showing (A) serum alkaline phosphatase (IU/L), (B) serum phosphorus concentration (mg/dL), and (C) serum ionized calcium (mg/dL) of various treatment groups at different time intervals. Full article ">Figure 4
(A) Medio-lateral radiographs showing healing status of different groups at various time intervals (pre, day 0, day 30, day 60, and day 90). Histogram (mean ± SD) showing radiographic (B) periosteal reaction score, (C) osteotomy line, and (D) critical-size defect (CSD) filling % score of various treatment groups at different time intervals. Full article ">Figure 5
Scanning electron microscope (SEM) images depicting healed bone tissue samples from different groups on day 90. Full article ">Figure 6
(A) Histopathological images depicting healed bone tissue samples from different groups on day 90, stained with hematoxylin and eosin (H&E) for microscopic analysis (scale bar: 400 µm). (B) Histopathological images depicting healed bone tissue samples from different groups on day 90, stained with Masson’s trichrome for microscopic analysis (scale bar: 400 µm). Histogram (mean ± SD) showing the (C) histopathological score and (D) total histopathological score of various treatment groups on day 90. Full article ">

16 pages, 6751 KiB

Open AccessArticle

In Vitro Effects of Weissella cibaria CMU and CMS1 on Receptor Activator of NF-κB Ligand (RANKL)-Induced Osteoclast Differentiation

by Geun-Yeong Park, Jeong-Ae Park and Mi-Sun Kang

J. Funct. Biomater. 2024, 15(3), 65; https://doi.org/10.3390/jfb15030065 - 8 Mar 2024

Viewed by 1770

Abstract

Excessive osteoclast activity can promote periodontitis-associated bone destruction. The inhibitory mechanisms of Weissella cibaria strains CMU and CMS1 against periodontitis have not yet been fully elucidated. In this study, we aimed to investigate whether heat-killed (HK) W. cibaria CMU and CMS1 or their [...] Read more.

Excessive osteoclast activity can promote periodontitis-associated bone destruction. The inhibitory mechanisms of Weissella cibaria strains CMU and CMS1 against periodontitis have not yet been fully elucidated. In this study, we aimed to investigate whether heat-killed (HK) W. cibaria CMU and CMS1 or their respective cell-free supernatants (CFSs) inhibit osteoclast differentiation and bone resorption in response to receptor activator of nuclear factor kappa-B ligand (RANKL)-treated RAW 264.7 cells. TRAP (tartrate-resistant acid phosphatase) staining and bone resorption assays revealed that both HK bacteria and CFSs significantly suppressed the number of TRAP-positive cells, TRAP activity, and bone pit formation compared to the RANKL-treated control (p < 0.05). HK bacteria dose-dependently inhibited osteoclastogenesis while selectively regulating certain genes in CFSs (p < 0.05). We found that disrupting the direct interaction between HK bacteria and RAW 264.7 cells abolished the inhibitory effect of HK bacteria on the expression of osteoclastogenesis-associated proteins (c-Fos, nuclear factor of activated T cells c1 (NFATc1), and cathepsin K). These results suggest that dead bacteria suppress osteoclast differentiation more effectively than the metabolites and may serve as beneficial agents in preventing periodontitis by inhibiting osteoclast differentiation via direct interaction with cells. Full article

(This article belongs to the Special Issue Functional Biomaterials for Regenerative Dentistry)

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19 pages, 6053 KiB

Open AccessArticle

The Potential Teeth Bleaching and Halitosis Prevention Effects of Pediococcus inopinatus THK-30, a Kimchi-Derived Lactic Acid Bacterium: In Vitro Study

by Trang Thi Minh Nguyen, Qiwen Zheng, Eun-Ji Yi, Arce Defeo Bellere, Xiangji Jin, Hong-Yong Kim and Tae-Hoo Yi

J. Funct. Biomater. 2024, 15(3), 64; https://doi.org/10.3390/jfb15030064 - 7 Mar 2024

Viewed by 2582

Abstract

Background: Recent developments in addressing dental aesthetic concerns, encompassing issues like teeth discoloration and halitosis, underscore the demand for safer alternative solutions. Purpose: This study aims to confirm the effects of lactic acid bacteria (LAB) from kimchi on artificial teeth bleaching and their [...] Read more.

Background: Recent developments in addressing dental aesthetic concerns, encompassing issues like teeth discoloration and halitosis, underscore the demand for safer alternative solutions. Purpose: This study aims to confirm the effects of lactic acid bacteria (LAB) from kimchi on artificial teeth bleaching and their potential impact in terms of preventing halitosis-related bacteria. Materials and Methods: To evaluate the antimicrobial effects against oral pathogens, disc diffusion tests and broth microdilution methods were used. Additionally, crystal violet analysis was performed to confirm the biofilm inhibition effect. The bleaching effects on stained artificial teeth were analyzed using the CIEDE2000 colorimetric method. Statistical analyses were performed using GraphPad Prism 9 with one-way and two-way ANOVA, with the significance level set at α < 0.05. Results: The strain THK-30, isolated from kimchi, exhibited antibacterial activity against Streptococcus mutans, Porphyromonas gingivalis, and Fusobacterium nucleatum, and was identified as Pediococcus inopinatus. Moreover, THK-30 showed a synergistic antibacterial effect against Gram-negative oral pathogens with 8% sodium hexametaphosphate (SHMP). In the stained artificial teeth bleaching test and artificial teeth biofilm inhibition test, the cell-free supernatant of THK-30 displayed significant teeth bleaching effects and caused the inhibition of biofilm formation, both independently and in combination with SHMP 8%. Conclusions: This study has demonstrated the potential applicability of LAB in teeth discoloration and halitosis. These findings are poised to provide a foundation for the development of research pertaining to the control of oral bacteria. Full article

(This article belongs to the Special Issue Biomaterials in Conservative Dentistry and Prosthodontics)

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14 pages, 4002 KiB

Open AccessEditor’s ChoiceArticle

Water-Soluble Quaternary and Protonable Basic Chitotriazolans: Synthesis by Click Chemistry Conversion of Chitosan Azides and Investigation of Antibacterial Activity

by Sankar Rathinam, Romano Magdadaro, Martha Á. Hjálmarsdóttir and Már Másson

J. Funct. Biomater. 2024, 15(3), 63; https://doi.org/10.3390/jfb15030063 - 5 Mar 2024

Cited by 4 | Viewed by 1936

Abstract

The azide transfer reaction and copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC) can be used to convert the amino groups in chitosan to triazole 1,2,3-moieties. The resulting polymer has been named chitotriazolan. This synthesis was performed with six different quaternary ammonium alkynes and three amine alkynes [...] Read more.

The azide transfer reaction and copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC) can be used to convert the amino groups in chitosan to triazole 1,2,3-moieties. The resulting polymer has been named chitotriazolan. This synthesis was performed with six different quaternary ammonium alkynes and three amine alkynes to obtain a series of nine water-soluble chitotriazolan derivatives. The structure and complete conversion of the azide were confirmed by FT-IR and proton NMR spectroscopy. The derivatives were investigated for antibacterial activity against S. aureus, E. faecalis, E. coli, and P. aeruginosa. The activity of the quaternized chitotriazolan derivatives varied depending on the structure of the quaternary moiety and the species of bacteria. The basic protonable derivatives were less active or inactive against the bacteria. Full article

(This article belongs to the Special Issue Biomedical Applications of Chitin and Chitosan-II)

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18 pages, 4730 KiB

Open AccessArticle

Hybrid Coatings Based on Polyvinylpyrrolidone/Polyethylene Glycol Enriched with Collagen and Hydroxyapatite: Incubation Studies and Evaluation of Mechanical and Physiochemical Properties

by Dagmara Słota, Josef Jampilek and Agnieszka Sobczak-Kupiec

J. Funct. Biomater. 2024, 15(3), 62; https://doi.org/10.3390/jfb15030062 - 1 Mar 2024

Viewed by 2006

Abstract

Coating materials offers an intriguing solution for imparting inert implants with additional bioactive characteristics without changing underlying parameters such as mechanical strength. Metallic implants like endoprostheses or polymeric implants can be coated with a thin layer of bioactive film capable of stimulating bone-forming [...] Read more.

Coating materials offers an intriguing solution for imparting inert implants with additional bioactive characteristics without changing underlying parameters such as mechanical strength. Metallic implants like endoprostheses or polymeric implants can be coated with a thin layer of bioactive film capable of stimulating bone-forming cells to proliferate or release a drug. However, irrespective of the final implantation site of such a coating biomaterial, it is necessary to conduct detailed mechanical and physicochemical in vitro analyses to determine its likely behavior under biological conditions. In this study, polymeric and composite coatings with hydroxyapatite obtained under UV light underwent incubation tests in four different artificial biological fluids: simulated body fluid (SBF), artificial saliva, Ringer’s fluid, and water (as the reference fluid). The potentiometric and conductometric properties, sorption capacity, and degradation rate of the coatings were examined. Furthermore, their hardness, modulus of elasticity, and deformation were determined. It was demonstrated that the coatings remained stable in SBF liquid at a pH value of around 7.4. In artificial saliva, the greatest degradation of the polymer matrix (ranging between 36.19% and 39.79%) and chipping of hydroxyapatite in the composite coatings were observed. Additionally, the effect of ceramics on sorption capacity was determined, with lower capacity noted with higher HA additions. Moreover, the evaluation of surface morphology supported by elemental microanalysis confirmed the appearance of new apatite layers on the surface as a result of incubation in SBF. Ceramics also influenced mechanical aspects, increasing hardness and modulus of elasticity. For the polymer coatings, the value was 11.48 ± 0.61, while for the composite coating with 15% ceramics, it increased more than eightfold to a value of 93.31 ± 11.18 N/mm². Based on the conducted studies, the effect of ceramics on the physicochemical as well as mechanical properties of the materials was determined, and their behavior in various biological fluids was evaluated. However, further studies, especially cytotoxicity analyses, are required to determine the potential use of the coatings as biomaterials. Full article

(This article belongs to the Section Bone Biomaterials)

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15 pages, 13306 KiB

Open AccessArticle

A Comparison of the Structure and Selected Mechanical Properties of Cr/Co Alloys Obtained by Casting and Selective Laser Melting

by Leszek Klimek, Barbara Bułhak and Beata Śmielak

J. Funct. Biomater. 2024, 15(3), 61; https://doi.org/10.3390/jfb15030061 - 1 Mar 2024

Cited by 3 | Viewed by 2269 | Correction

Abstract

Selective laser melting (SLM) technologies are becoming increasingly popular. The aim of the work is to compare the metallographic structure, hardness, and selected strength properties of alloys obtained by casting and by SLM, with a particular emphasis on fatigue strength. Twenty Cr/Co alloy [...] Read more.

Selective laser melting (SLM) technologies are becoming increasingly popular. The aim of the work is to compare the metallographic structure, hardness, and selected strength properties of alloys obtained by casting and by SLM, with a particular emphasis on fatigue strength. Twenty Cr/Co alloy bars were made by casting or SLM, and samples of appropriate dimensions were prepared for individual tests. The microstructures of the samples were tested by metallography, and then tested for hardness, impact strength, tensile strength, bending strength, and fatigue strength; they were also subjected to fracture after bending, tensile, fatigue, and impact tests, with the resulting fractures examined by scanning electron microscopy (SEM). Primary dendrites and small amounts of gas bubbles were present in the cast samples ground lengthwise. The SEM samples were more finer grained and uniform. Compared to the casting samples, the SLM samples demonstrated higher hardness, lower mean impact strength and higher tensile strength. The casting samples also displayed lower mean elongation values. The casting samples demonstrated slightly higher fatigue strength. The fractures of the casting samples showed an interdendritic character with clearly visible dendrites at the fracture, while those of the SLM samples were also intergranular, but finer grained. SLM generally results in better strength properties, while casting obtains slightly greater fatigue strength. Full article

(This article belongs to the Special Issue Additive Manufacturing of Dental Materials)

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28 pages, 2015 KiB

Open AccessReview

Three-Dimensional Printing Methods for Bioceramic-Based Scaffold Fabrication for Craniomaxillofacial Bone Tissue Engineering

by Zeeshan Sheikh, Vasudev Vivekanand Nayak, Umer Daood, Anupreet Kaur, Hanan Moussa, Abbas Canteenwala, Pierre-Luc Michaud, Ísis de Fátima Balderrama, Edisa de Oliveira Sousa, Nick Tovar, Andrea Torroni, Michael Glogauer, Huzefa Talib, Paulo G. Coelho and Lukasz Witek

J. Funct. Biomater. 2024, 15(3), 60; https://doi.org/10.3390/jfb15030060 - 1 Mar 2024

Cited by 2 | Viewed by 4472

Abstract

Three-dimensional printing (3DP) technology has revolutionized the field of the use of bioceramics for maxillofacial and periodontal applications, offering unprecedented control over the shape, size, and structure of bioceramic implants. In addition, bioceramics have become attractive materials for these applications due to their [...] Read more.

Three-dimensional printing (3DP) technology has revolutionized the field of the use of bioceramics for maxillofacial and periodontal applications, offering unprecedented control over the shape, size, and structure of bioceramic implants. In addition, bioceramics have become attractive materials for these applications due to their biocompatibility, biostability, and favorable mechanical properties. However, despite their advantages, bioceramic implants are still associated with inferior biological performance issues after implantation, such as slow osseointegration, inadequate tissue response, and an increased risk of implant failure. To address these challenges, researchers have been developing strategies to improve the biological performance of 3D-printed bioceramic implants. The purpose of this review is to provide an overview of 3DP techniques and strategies for bioceramic materials designed for bone regeneration. The review also addresses the use and incorporation of active biomolecules in 3D-printed bioceramic constructs to stimulate bone regeneration. By controlling the surface roughness and chemical composition of the implant, the construct can be tailored to promote osseointegration and reduce the risk of adverse tissue reactions. Additionally, growth factors, such as bone morphogenic proteins (rhBMP-2) and pharmacologic agent (dipyridamole), can be incorporated to promote the growth of new bone tissue. Incorporating porosity into bioceramic constructs can improve bone tissue formation and the overall biological response of the implant. As such, employing surface modification, combining with other materials, and incorporating the 3DP workflow can lead to better patient healing outcomes. Full article

(This article belongs to the Special Issue Biologics and Devices for Periodontal and Peri-Implant Reconstruction)

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11 pages, 2697 KiB

Open AccessEditor’s ChoiceArticle

Injectable Micro-Hydrogel for DNA Delivery: A Promising Therapeutic Platform

by Sunghyun Moon and Jong Bum Lee

J. Funct. Biomater. 2024, 15(3), 59; https://doi.org/10.3390/jfb15030059 - 1 Mar 2024

Viewed by 2141

Abstract

Utilizing the immune system as a strategy for disease prevention and treatment is promising, especially with dendritic cells (DCs) playing a central role in adaptive immune responses. The unique properties of DCs drive interest in developing materials for cell-based therapy and immune modulation. [...] Read more.

Utilizing the immune system as a strategy for disease prevention and treatment is promising, especially with dendritic cells (DCs) playing a central role in adaptive immune responses. The unique properties of DCs drive interest in developing materials for cell-based therapy and immune modulation. Injectable systems require syringe-compatible scaffolds, while hydrogels, like alginate, known for their programmability and biocompatibility, offer a versatile platform for immune medicine enhancement through easy preparation and room-temperature cross-linking. In this study, we synthesized alginate balls loaded with DCs or cytosine–phosphorothioate–guanine deoxyribonucleotide (CpG DNA) microparticles, aiming for long-term immune cell culture with potential immune stimulation effects. Encapsulated DCs exhibited proliferation within the alginate balls for up to 7 days, and CpG MPs were uniformly dispersed, which can facilitate uptake by DCs. This was supported by the result that DCs effectively phagocytosed CpG microparticles in a 2D environment. After the uptake of CpG MPs, the alginate balls with CpG-MP-uptaken DCs were synthesized successfully. The injectable properties of the alginate balls were easily modulated by adjusting the syringe needle gauges. This innovative strategy holds substantial promise for advancing medical treatments, offering effective and comfortable solutions for controlled immune modulation. Full article

(This article belongs to the Special Issue Injectable and Biodegradable Hydrogels for Biomedical Applications)

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(A) Schematic illustration of the synthesis process of DC-loaded alginate balls, illustrating the key steps involved in encapsulating DCs within the alginate matrix (created with BioRender.com). (B) Digital images of alginate balls loaded with DCs. (C) This set of images demonstrates the viability and stability of DCs over an extended period within the alginate matrix (scale bar: 40 μm). Full article ">Figure 2
(A) Schematic illustration of the fabrication of CpG DNA microparticles by Rolling Circle Amplification. (B) Fluorescence images of the CpG MPs. The particles were stained with SYBR Green I (scale bar: 10 μm). (C) SEM images of the CpG MPs (scale bar: 1 μm). (D,E) The sizes and zeta-potential of the CpG MPs. Data were obtained by dynamic light scattering. (F) Schematic illustration of the fabrication process for small CpG-microparticle (MP)-loaded alginate balls, illustrating the key steps involved in the synthesis (created with BioRender.com). (G) Digital images of the injectable features of CpG-MP-loaded alginate balls. (H) Visualization of small CpG-MP-loaded alginate balls through digital images of Cy5-labeled CpG MPs loaded into the alginate balls and corresponding fluorescence images revealing the distribution of CpG MPs within the small alginate ball (scale bar: 20 μm). Full article ">Figure 3
(A) Schematic illustration depicting the process of DCs phagocytosing CpG MPs (created with BioRender.com). (B) Microscopic analysis combining DIC and fluorescence images to show the uptake of CpG MPs by DCs (scale bar: 20 μm). (C) Confocal microscopy images providing detailed visualization of the internalization of CpG MPs by DCs (scale bar: 20 μm). (D) Schematic illustration of CpG-MP-uptaken DC-loaded alginate ball. (E) Microscopic analysis combining DIC and fluorescence images to show the loading of CpG-MP-uptaken DCs into the alginate ball (scale bar: 10 μm). (F) Merged images of DIC and fluorescence images of DCs inside the different layers of the alginate ball in the same spot (Scale bar: 20 μm). Full article ">

15 pages, 4174 KiB

Open AccessArticle

Gold Nanoparticle-Based Colorimetric and Fluorescent Dual-Mode Lateral Flow Immunoassay for SARS-CoV-2 Detection

by Ying Gan, Hefan Zhang, Jing Liu, Fuqin He, Fengheng Li, Ao Li, Man Xing, Dongming Zhou, Shan-Yu Fung and Hong Yang

J. Funct. Biomater. 2024, 15(3), 58; https://doi.org/10.3390/jfb15030058 - 27 Feb 2024

Cited by 2 | Viewed by 2523

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection caused the COVID-19 pandemic, impacting the global economy and medical system due to its fast spread and extremely high infectivity. Efficient control of the spread of the disease relies on a fast, accurate, and convenient [...] Read more.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection caused the COVID-19 pandemic, impacting the global economy and medical system due to its fast spread and extremely high infectivity. Efficient control of the spread of the disease relies on a fast, accurate, and convenient detection system for the early screening of the infected population. Although reverse transcription–quantitative polymerase chain reaction (RT-qPCR) is the gold-standard method for SARS-CoV-2 RNA analysis, it has complex experimental procedures and relies on expensive instruments and professional operators. In this work, we proposed a simple, direct, amplification-free lateral flow immunoassay (LFIA) with dual-mode detection of SARS-CoV-2 RNA via direct visualization as well as fluorescence detection. The viral RNA was detected by the designed DNA probes to specifically hybridize with the conserved open reading frame 1ab (ORF1ab), envelope protein (E), and nucleocapsid (N) regions of the SARS-CoV-2 genome to form DNA–RNA hybrids. These hybrids were then recognized by the dual-mode gold nanoparticles (DMNPs) to produce two different readout signals. The fluorescence characteristics of different sizes of GNPs were explored. Under the optimized conditions, the LFIA presented a linear detection range of 10⁴–10⁶ TU/mL with a limit of detection (LOD) of 0.76, 1.83, and 2.58 × 10⁴ TU/mL for lentiviral particles carrying SARS-CoV-2 ORF1ab, E, and N motifs, respectively, in the fluorescent mode, which was up to 10 times more sensitive than the colorimetric mode. Furthermore, the LFIA exhibited excellent specificity to SARS-CoV-2 in comparison with other respiratory viruses. It could be used to detect SARS-CoV-2 in saliva samples. The developed LFIA represents a promising and convenient point-of-care method for dual-mode, rapid detection of SARS-CoV-2, especially in the periods with high infectivity. Full article

(This article belongs to the Special Issue Biomaterials for Diagnostic, Therapeutic, and Theranostic Applications)

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The physicochemical characterization of DMNPs and the evaluation of their performance in RNA detection. (a) A schematic diagram of DMNPs’ synthesis. The TEM images of GNP5 (b), GNP13 (c), and GNP20 (d). (e) The fluorescence intensity scan of three sizes of GNPs with Cy5. Excitation scan wavelength: 550–700 nm (emission wavelength: 750 nm). Emission scan wavelength: 650–800 nm (excitation wavelength: 600 nm). (f) Fluorescence images of dried droplets of GNP5–Cy5, GNP13–Cy5, and GNP20–Cy5 on the nitrocellulose membrane. (g) The hydrodynamic diameter of GNPs and DMNPs by DLS measurement. Scale bar: 50 nm. (h) Bright-field (left) and fluorescent (right) images of DMNP-based LFIA for direct RNA detection. (i) Colorimetric and (j) fluorescence analysis of (h). ΔB = |Bbackground − Btest line|. N ≥ 3 per group; * p < 0.05; ** p < 0.01. Full article ">Figure 2
LFIA-based detection of SARS-CoV-2 lentiviral particles. (a) A schematic of lentiviral particle packaging workflow created with BioRender.com. (b) Representative fluorescent images showing the HEK293T cells transfected with GFP-expressing lentiviral particles at a 1:40 dilution. Scale bar: 50 μm. (c) Representative images of the flow cytometric analysis of the HEK293T cells transfected with the GFP-expressing lentiviral particles at a 1:40 dilution. (d) Bright-field (left) and fluorescent (right) images of LFIA for ORF1ab lentiviral particle detection. (e) Colorimetric and (f) fluorescent analysis of (d). ΔB = |Bbackground − Btest line|. N ≥ 3 per group; ** p < 0.01. Full article ">Figure 3
Optimization of the LFIA fabrication and operation conditions for sensing lentiviral RNA. The optimization curves and representative photographs (inset) of (a) the S9.6 antibody concentrations (0.5, 1, and 2 mg/mL) on test line, (b) DMNP concentrations (35, 70, and 140 nM), (c) incubation times (15, 30, 60, and 90 min) of DNA probes with RNA released from SARS-CoV-2 lentiviral particles, (d) NaCl concentrations (0.3, 0.6, and 1 M) of hybridization, and the addition of (e) S9.6 antibody (2, 4, and 8 μL) and (f) Cy5 (0.9, 1.8, and 3.6 μL) onto the GNPs. The ORF1ab lentiviral particles were used in the optimization experiments. N ≥ 2 per group; ns: not significant; * p < 0.05. Full article ">Figure 4
The sensitivity of LFIA in detecting three different types of SARS-CoV-2 RNA-carrying lentiviral particles. The colorimetric analysis and representative photographs (inset) of LFIA for the detection of (a) ORF1ab, (c) N, and (e) E regions of the SARS-CoV-2 RNA-carrying lentiviral particles. Fluorescence analysis and representative photographs (inset) of (b) ORF1ab, (d) N, (f) E regions of the SARS-CoV-2 RNA-carrying lentiviral particles. ΔB = |Bbackground − Btest line|. N ≥ 3 per group. Full article ">Figure 5
Specificity of LFIA in detecting SARS-CoV-2 RNA-carrying lentiviral particles in comparison with two respiratory DNA/RNA viruses (negative control viruses). (a) Colorimetric and (b) fluorescence analysis. Inset: Representative photographs with the test line labelled with red arrow. The concentration of H7N9 and HAdV7 viruses was 105 TU/mL. The SARS-CoV-2 group contained 105 TU/mL of ORF1ab-, E-, and N-carrying lentiviral particles to simulate the complete viral genome. The control group refers to the detection buffer. ΔB = |Bbackground − Btest line|. N ≥ 3 per group; *** p < 0.001. Full article ">Figure 6
The performance of LFIA in SARS-CoV-2 detection in saliva samples. (a) The schematic diagram of the workflow for SARS-CoV-2 lentiviral particles (105 TU/mL of mixture of ORF1ab, N, and E lentiviral particles) spiked in the saliva. (b) Colorimetric and (c) fluorescence analysis. Inset: Representative photographs with the test line labelled with a red arrow. ΔB = |Bbackground − Btest line|. N ≥ 3 per group; * p < 0.05. Full article ">Scheme 1
The schematic workflow of the proposed dual-mode LFIA for the detection of SARS-CoV-2. (a) Genome map showing the ORF1ab, E, and N DNA probes that hybridize with the three gene regions (ORF1ab, E, and N) in the SARS-CoV-2 RNA genome. (b) The workflow of LFIA. SARS-CoV-2 RNA is released and bound with DNA probes forming DNA–RNA hybrids, which can be captured by S9.6 antibodies on the test line of the strip. Upon the DMNPs binding, the colorimetric and fluorescent signals can be obtained. Full article ">

19 pages, 4533 KiB

Open AccessArticle

Characterization and In Vitro Evaluation of Porous Polymer-Blended Scaffolds Functionalized with Tricalcium Phosphate

by Iwona Pudełko-Prażuch, Mareeswari Balasubramanian, Sundara Moorthi Ganesan, Stanisław Marecik, Kamila Walczak, Kinga Pielichowska, Suvro Chatterjee, Ravichandran Kandaswamy and Elżbieta Pamuła

J. Funct. Biomater. 2024, 15(3), 57; https://doi.org/10.3390/jfb15030057 - 26 Feb 2024

Cited by 5 | Viewed by 2291

Abstract

Bone tissue is one of the most transplanted tissues. The ageing population and bone diseases are the main causes of the growing need for novel treatments offered by bone tissue engineering. Three-dimensional (3D) scaffolds, as artificial structures that fulfil certain characteristics, can be [...] Read more.

Bone tissue is one of the most transplanted tissues. The ageing population and bone diseases are the main causes of the growing need for novel treatments offered by bone tissue engineering. Three-dimensional (3D) scaffolds, as artificial structures that fulfil certain characteristics, can be used as a temporary matrix for bone regeneration. In this study, we aimed to fabricate 3D porous polymer scaffolds functionalized with tricalcium phosphate (TCP) particles for applications in bone tissue regeneration. Different combinations of poly(lactic acid) (PLA), poly(ethylene glycol) (PEG with molecular weight of 600 or 2000 Da) and poly(ε-caprolactone) (PCL) with TCP were blended by a gel-casting method combined with rapid heating. Porous composite scaffolds with pore sizes from 100 to 1500 µm were obtained. ATR-FTIR, DSC, and wettability tests were performed to study scaffold composition, thermal properties, and hydrophilicity, respectively. The samples were observed with the use of optical and scanning electron microscopes. The addition of PCL to PLA increased the hydrophobicity of the composite scaffolds and reduced their susceptibility to degradation, whereas the addition of PEG increased the hydrophilicity and degradation rates but concomitantly resulted in enhanced creation of rounded mineral deposits. The scaffolds were not cytotoxic according to an indirect test in L929 fibroblasts, and they supported adhesion and growth of MG-63 cells when cultured in direct contact. Full article

(This article belongs to the Special Issue Functional Scaffolds for Bone and Joint Surgery)

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13 pages, 4856 KiB

Open AccessArticle

Analysis of Bone Mineral Density and Bone Quality of Cortical Bone in the Human Hyoid Body and Histological Observation of the Entheses

by Masaaki Kasahara, Tomoko Someya, Kei Kitamura, Genji Watanabe, Satoru Matsunaga, Shinichi Abe and Masayuki Hattori

J. Funct. Biomater. 2024, 15(3), 56; https://doi.org/10.3390/jfb15030056 - 22 Feb 2024

Cited by 2 | Viewed by 1887

Abstract

The hyoid is the only bone in the human body that is completely independent, not forming a joint with any other bone; its position is maintained by the suprahyoid and infrahyoid muscles, as well as several ligaments. The purpose of this study was [...] Read more.

The hyoid is the only bone in the human body that is completely independent, not forming a joint with any other bone; its position is maintained by the suprahyoid and infrahyoid muscles, as well as several ligaments. The purpose of this study was to ascertain the effect of the functional pressure arising from these muscles and ligaments on the hyoid body structure from its bone mineral density, bone quality, and histological observations. The area between the mesial-most part of each lesser horn and the center of the hyoid body was divided equally into four measurement regions. We conducted histological investigations at each measurement region and observed the entheses. To analyze bone mass and bone quality, we also measured bone mineral density (BMD) and analyzed biological apatite (BAp) crystallite orientation in the same regions. Histological observations identified periosteal insertions and fibrocartilaginous entheses. There was no significant difference in BMD between any of the measurement regions, but the preferential orientation of BAp crystallites was stronger in the infrahyoid muscles and ligaments, where fibrocartilaginous entheses are found, than in other places. This suggests that the functional pressure at these sites might exert a major effect not only on the morphological characteristics of the entheses but also on bone quality. Full article

(This article belongs to the Special Issue Feature Papers in Bone Biomaterials)

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Setting of the coordinate axes. Full article ">Figure 2
Designation of the region of interest in the human hyoid bone. (A) The region of interest was the body of the hyoid between the mesial-most parts of the lesser horns bilaterally. The measurement regions comprised the area of the hyoid body (HB) between the mesial-most part of the lesser horn and the center of the HB divided into four equal parts (α, β, γ, and δ). (B) Four points around the circumference of the cortical bone of the HB were used for analysis. Point a; uppermost part of the center of the cortical bone of the HB (superior HB), Point b; anterior-most part of the center of the cortical bone of the HB (anterior HB), Point c; bottommost part of the center of the cortical bone of the HB (inferior HB), Point d; posterior -most part of the center of the cortical bone of the HB (posterior HB) The equals sign (=) indicates lines of equal length. Full article ">Figure 3
Evaluation of scanning electron microscope (SEM) images. Full article ">Figure 4
Hematoxylin-eosin-stained sagittal sections from each measurement region (α, β, γ, δ). Full article ">Figure 5
Hematoxylin-eosin-stained and toluidine blue-stained sections showing the morphology of the entheses around the hyoid body (HB). Full article ">Figure 6
Evaluation of BMD at each measurement point in each measurement region of the hyoid. Full article ">Figure 7
Diffraction intensity ratios along the X-, Y-, and Z-axis, indicating the preferential orientation of BAp at each measurement point in each measurement region of the hyoid. Full article ">

20 pages, 6235 KiB

Open AccessArticle

Generation of Pearl/Calcium Phosphate Composite Particles and Their Integration into Porous Chitosan Scaffolds for Bone Regeneration

by Zhiyi Li, Ihtesham Ur Rehman, Rebecca Shepherd and Timothy E. L. Douglas

J. Funct. Biomater. 2024, 15(3), 55; https://doi.org/10.3390/jfb15030055 - 21 Feb 2024

Cited by 2 | Viewed by 2284

Abstract

Bone tissue engineering using osteoconductive scaffolds holds promise for regeneration, with pearl powder gaining interest for its bioactive qualities. This study used freeze drying to create chitosan (CS) scaffolds with pearl/calcium phosphate (p/CaP) powders, mimicking bone tissue structurally and compositionally. Characterization included scanning [...] Read more.

Bone tissue engineering using osteoconductive scaffolds holds promise for regeneration, with pearl powder gaining interest for its bioactive qualities. This study used freeze drying to create chitosan (CS) scaffolds with pearl/calcium phosphate (p/CaP) powders, mimicking bone tissue structurally and compositionally. Characterization included scanning electron microscopy (SEM) and mechanical testing. X-ray diffraction (XRD) Fourier-transform infrared–photoacoustic photo-acoustic sampling (FTIR−PAS), and FTIR- attenuated total reflectance (FTIR-ATR) were used to characterize p/CaP. In vitro tests covered degradation, cell activity, and SEM analysis. The scaffolds showed notable compressive strength and modulus enhancements with increasing p/CaP content. Porosity, ranging from 60% to 90%, decreased significantly at higher pearl/CaP ratios. Optimal cell proliferation and differentiation were observed with scaffolds containing up to 30 wt.% p/CaP, with 30 wt.% pearl powder and 30 wt.% p/CaP yielding the best results. In conclusion, pearl/calcium phosphate chitosan (p/CaP_CS) composite scaffolds emerged as promising biomaterials for bone tissue engineering, combining structural mimicry and favourable biological responses. Full article

(This article belongs to the Special Issue Hydroxyapatite Composites for Biomedical Application)

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12 pages, 4380 KiB

Open AccessArticle

Active and Passive Mineralization of Bio-Gide^® Membranes in Rat Calvaria Defects

by Karol Ali Apaza Alccayhuaman, Patrick Heimel, Stefan Tangl, Stefan Lettner, Carina Kampleitner, Layla Panahipour, Ulrike Kuchler and Reinhard Gruber

J. Funct. Biomater. 2024, 15(3), 54; https://doi.org/10.3390/jfb15030054 - 21 Feb 2024

Cited by 3 | Viewed by 1953

Abstract

Bio-Gide^® is a collagen membrane routinely used in guided bone regeneration. Recent studies have shown that this collagen membrane has osteoconductive properties, meaning that it can support the growth of new bone. However, it has also been observed that the collagen membrane [...] Read more.

Bio-Gide^® is a collagen membrane routinely used in guided bone regeneration. Recent studies have shown that this collagen membrane has osteoconductive properties, meaning that it can support the growth of new bone. However, it has also been observed that the collagen membrane has areas of mineralized fibers which can occur spontaneously and independently of osteoblasts. To better understand how this works, we established a model using minced collagen membranes to reduce the active mineralization of intact collagen membranes in favor of passive mineralization. We thus compared the original intact membrane with a minced collagen membrane in a 5 mm calvarial defect model in Sprague Dawley rats. After three weeks of healing, histology and microcomputed tomography (μCT) were performed. Histological analysis confirmed the osteoconductive properties, with new bone growing inside the intact collagen membrane. However, in minced collagen membranes, the osteoconductive properties were restricted to the defect margins. Interestingly, histology revealed large mineralized areas indicating passive mineralization with no signs of bone formation. In the μCT analysis, the intact collagen membranes caused a higher median mineralized volume (1.5 mm³) compared with the minced group (0.4 mm³), but this lacked significance (p = 0.09). The μCT analysis needs to be interpreted carefully, particularly in defects filled with minced membranes, considering that the mineralized tissue may not necessarily be bone but also the result of passive mineralization. Taken together, the findings suggest that Bio-Gide^® collagen membranes support bone formation while also exhibiting potential for passive mineralization. Full article

(This article belongs to the Special Issue Biomaterials for Oral Reconstructive Treatment)

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Histology planning in a 3D reconstruction for precise plane placement (the plane in red). The 3D reconstruction depicts a sagittal plane parallel to the midline suture precisely centered within the defect. The image demonstrates the seamless alignment between the µCT slide and the histological slide, indicated by the highlighted red area denoting the position of the plane. Full article ">Figure 2
The µCT parameters were assessed in conjunction with histological findings (A,B). The mineralized volume (MV) within the entire defect exhibited similar measurements, with notable variability among samples when comparing intact collagen membranes with the minced collagen membrane group (C). The MV/TV fraction displayed a similar trend (D). However, the Tb.Th showed a slightly superior outcome in the intact group compared with the minced group (E). Full article ">Figure 3
Three-dimensional visualization depicting floating mineralization and defect coverage. Rendered 3D images showcase the distribution of floating mineralization, in which mineralized areas are not connected to the pristine bone, and the extent of defect coverage. In the intact collagen membranes (A), a cohesive surface with minimal floating mineralization is observed. Conversely, the minced collagen membranes (B) display numerous disconnected “floating” islands. The statistical analysis reveals a significant difference in floating mineralization (p = 0.04) (C). Additionally, defect coverage appears more robust in the intact group compared with the minced group but did not reach statistical significance (D). Full article ">Figure 4
Representative microphotograph depicts the histological characteristics of the intact collagen membrane (A). Periosteal new bone growth is evident toward the collagen membrane (B) and toward the center (C), resembling an attempt to bridge new bone formation from opposing sites (D). This hybrid bone exhibits embedded collagen fibers from the membrane within the newly formed bone, showcasing distinctive features (D). The intense purple stain is characteristic for less mineralization (C,D); hence, more dye is taken up by newly mineralized tissue compared with the more mineralized original pristine bone. Full article ">Figure 5
Representative microphotographs depict new bone formation both beneath and above the intact collagen membrane (A). Notably, a heavily stained woven bone indicates signs of resorption (B). The newly formed bone beneath the membrane displays characteristics of lamellar bone, while above the membrane, a distinctive hybrid bone is observed, intermixed with collagen fibers (C). The growth of bone from the edges appears to exert pressure on the collagen membrane, particularly when it is more mature (D). Details with arrows marking the locations of the different tissues are in <a href="#app1-jfb-15-00054" class="html-app">Supplementary Figure S1</a>. Full article ">Figure 6
The microphotograph illustrates a minced collagen membrane (A), revealing areas of bone formation adjacent to the periosteal bone, exhibiting aspects reminiscent of partially mineralized cartilage (B). The collagen membrane pieces resemble floes within the defect (C). The newly formed bone exhibits characteristics akin to hybrid bone, as the collagen fibers appear embedded within the newly formed bone structure (D). Details with arrows marking the locations of the different tissues are in <a href="#app1-jfb-15-00054" class="html-app">Supplementary Figure S2</a>. Full article ">Figure 7
The representative microphotograph (A) reveals a minced collagen membrane displaying evident passive mineralization. Clusters of mineralized fibers (B) are distributed throughout, resembling bone tissue. However, there is an absence of bone cells around these fibers or an osteoid matrix (C). The mineralization of collagen fibrils interconnects the pieces of the collagen membrane (D). Details with arrows marking the locations of the different tissues are in <a href="#app1-jfb-15-00054" class="html-app">Supplementary Figure S3</a>. Full article ">

12 pages, 2742 KiB

Open AccessEditor’s ChoiceArticle

Polycarbonate-Based Copolymer Micelles as Biodegradable Carriers of Anticancer Podophyllotoxin or Juniper Extracts

by Radostina G. Kalinova, Ivaylo V. Dimitrov, Diana I. Ivanova, Yana E. Ilieva, Alexander N. Tashev, Maya M. Zaharieva, George Angelov and Hristo M. Najdenski

J. Funct. Biomater. 2024, 15(3), 53; https://doi.org/10.3390/jfb15030053 - 21 Feb 2024

Cited by 3 | Viewed by 2356

Abstract

Podophyllotoxin (PPT) is used in the industrial production of efficient anticancer, antiviral and other drugs. Sinopodophyllum hexandrum or Podophyllum peltatum are natural sources of PPT, but at present they are considered as endangered species. Their PPT content is variable, depending on the growing [...] Read more.

Podophyllotoxin (PPT) is used in the industrial production of efficient anticancer, antiviral and other drugs. Sinopodophyllum hexandrum or Podophyllum peltatum are natural sources of PPT, but at present they are considered as endangered species. Their PPT content is variable, depending on the growing conditions. Searching for new sources of PPT, some representatives of the genus Juniperus were found to exhibit efficient PPT biosynthesis. However, PPT is highly toxic and poorly soluble in water compound, which limits its clinical applications. In this connection, amphiphilic polymer micelles are considered to be suitable PPT carriers, aimed at increase in water solubility and decrease in toxicity. The present research deals with the evaluation of MPEG–polycarbonate block copolymer micelles loaded with PPT or juniper extracts. The active component-loaded polymer nanocarriers were characterized by dynamic and electrophoretic light scattering, as well as by transmission electron microscopy. The active component loading efficiency and loading capacity were also determined. Highly efficient antiproliferative activity of the loaded micelles was determined in a panel of cancer cell lines. The obtained amphiphilic nanocarriers, loaded with PPT-containing bioactive components, have application in future in vivo preclinical trials of their pharmacokinetics and pharmacodynamics as potential therapeutical agents in the prospective nanomedicine. Full article

(This article belongs to the Special Issue Synthetic Polymers for the Delivery of Vaccines and Therapeutics)

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Specimens of Juniperus virginiana (A) and Juniperus sabina var. balkanensis (B), whose leaf extracts, containing podophyllotoxin (C), were used for loading into biodegradable amphiphilic MPEG-b-PC diblock copolymer nanocarriers. Full article ">Figure 2
Structure of a biodegradable amphiphilic MPEG-b-PC diblock copolymer, used in this study to obtain podophyllotoxin- or juniper extract-loaded nanocarriers. Full article ">Figure 3
Presentation of the size distributions (a) and zeta potentials (b) obtained from dynamic and electrophoretic light scattering analyses of the corresponding micelles: empty (MPEG-b-PC: d = 44 nm, PdI 0.245, ζ = −0.44 mV), JV-extract loaded (MPEG-b-PC/JV: d = 46 nm, PdI 0.246, ζ = 0.96 mV), JS-extract loaded (MPEG-b-PC/JS: d = 48 nm, PdI 0.272, ζ = 0.27 mV) and PPT-loaded (MPEG-b-PC/PPT: d = 70 nm, PdI 0.226, ζ = −4.39 mV) block copolymer micelles. Full article ">Figure 4
Stability of the copolymer micelles loaded with JS-leaf extract, JV- leaf extract or PPT, assessed by DLS measurements in aqueous media after various time intervals of incubation at 4 °C. Full article ">Figure 5
Transmission electron microscopy images of the corresponding micelles: (a) empty (d = 45.87 ± 4.68 nm); (b) PPT-loaded (d = 71.35 ± 5.14 nm); (c) JS-extract loaded (d = 53.18 ± 2.58 nm); and (d) JV-extract loaded (d = 46.96 ± 6.89 nm) block copolymer (MPEG-b-PC) micelles. Full article ">Figure 5 Cont.
Transmission electron microscopy images of the corresponding micelles: (a) empty (d = 45.87 ± 4.68 nm); (b) PPT-loaded (d = 71.35 ± 5.14 nm); (c) JS-extract loaded (d = 53.18 ± 2.58 nm); and (d) JV-extract loaded (d = 46.96 ± 6.89 nm) block copolymer (MPEG-b-PC) micelles. Full article ">

23 pages, 9147 KiB

Open AccessArticle

Influence of Hybrid Surface Modification on Biocompatibility and Physicochemical Properties of Ti-6Al-4V ELI Titanium

by Anna Woźniak, Weronika Smok, Janusz Szewczenko, Marcin Staszuk and Grzegorz Chladek

J. Funct. Biomater. 2024, 15(3), 52; https://doi.org/10.3390/jfb15030052 - 20 Feb 2024

Cited by 4 | Viewed by 2371

Abstract

Titanium-based materials are the most widely used materials in biomedical applications. However, according to literature findings, the degradation products of titanium have been associated with potential allergic reactions, inflammation, and bone resorption. The corrosion process of Ti-6Al-4V in the human body environment may [...] Read more.

Titanium-based materials are the most widely used materials in biomedical applications. However, according to literature findings, the degradation products of titanium have been associated with potential allergic reactions, inflammation, and bone resorption. The corrosion process of Ti-6Al-4V in the human body environment may be exacerbated by factors such as reduced pH levels and elevated concentrations of chloride compounds. Coatings made of biopolymers are gaining attention as they offer numerous advantages for enhancing implant functionality, including improved biocompatibility, bioactivity, wettability, drug release, and antibacterial activity. This study analyzes the physicochemical and electrochemical behavior of the Ti-6Al-4V ELI alloy subjected to PCL and PCL/TiO₂ deposition by the electrospinning method. To characterize the polymer-based layer, tests of chemical and phase composition, as well as surface morphology investigations, were performed. Wetting angle tests were conducted as part of assessing the physicochemical properties. The samples were subjected to corrosion behavior analysis, which included open circuit potential measurements, potentiodynamic tests, and the electrochemical impedance spectroscopy method. Additionally, the quantification of released ions post the potentiodynamic test was carried out using the inductively coupled plasma atomic emission spectrometry (ICP–AES) method. Cytotoxicity tests were also performed. It was found that surface modification by depositing a polymer-based layer on the titanium substrate material using the electrospinning method provides improved corrosion behavior, and the samples exhibit non-toxic properties. Full article

(This article belongs to the Special Issue Advanced Biomaterials and Biotechnology: Applications in Dental Medicine)

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14 pages, 6794 KiB

Open AccessArticle

Toxicity Assessment of New Ag-ZnO/AgO Nanocomposites: An In Vitro and In Vivo Approach

by José Rodrigues do Carmo Neto, Pablo Igor Ribeiro Franco, Yarlla Loyane Lira Braga, Jordana Fernandes de Oliveira, Hugo Felix Perini, Luís Fernando Duarte Albuquerque, Danieli Brolo Martins, Fernanda Rodrigues Helmo, Anderson Assunção Andrade, Marina Pacheco Miguel, Mara Rúbia Nunes Celes, Thiago Lopes Rocha, Anielle Christine Almeida Silva, Juliana Reis Machado and Marcos Vinícius da Silva

J. Funct. Biomater. 2024, 15(3), 51; https://doi.org/10.3390/jfb15030051 - 20 Feb 2024

Cited by 2 | Viewed by 2347

Abstract

Zinc oxide nanoparticles (ZnO NPs) are metal oxide nanomaterials, which are important for several applications: antibacterial, anthelmintic, antiprotozoal and antitumoral, among others. These applications are mainly related to the ability to spontaneously produce and induce the production of reactive oxygen species that are [...] Read more.

Zinc oxide nanoparticles (ZnO NPs) are metal oxide nanomaterials, which are important for several applications: antibacterial, anthelmintic, antiprotozoal and antitumoral, among others. These applications are mainly related to the ability to spontaneously produce and induce the production of reactive oxygen species that are important components for the destruction of pathogens and tumor cells. While trying to potentiate ZnO NPs, studies have associated these NPs with silver oxide (AgO) or silver (Ag) NPs. It has already been reported that this combination (Ag-ZnO/AgO NPs) is able to enhance the microbicidal potential. Although possessing much potential for several purposes, it is important to evaluate whether this association also poses the risk of toxicity to cells and experimental models. Therefore, this work aimed to evaluate the toxicity of various Ag-ZnO/AgO NP nanocomposites, in vitro and in vivo. Accordingly, ZnO nanocrystals and nanocomposites with various concentrations of AgO (ZnO:5Ag, ZnO:9Ag or ZnO:11Ag) were used in different cytotoxicity models: Galleria mellonella (G. mellonella), cell lines (VERO and RAW 264.7) and C57BL/6 mice. In the G. mellonella model, four concentrations were used in a single dose, with subsequent evaluation of mortality. In the case of cells, serial concentrations starting at 125 µg/mL were used, with subsequent cytotoxicity assessment. Based on the safe doses obtained in G. mellonella and cell models, the best doses were used in mice, with subsequent evaluations of weight, biochemistry as also renal and liver histopathology. It was observed that the toxicity, although low, of the nanocomposites was dependent upon the concentration of AgO used in association with ZnO NPs, both in vitro and in vivo. Full article

(This article belongs to the Special Issue Nanostructured Materials/Biomaterials for Healthcare Applications)

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Graphical abstract

Graphical abstract
Full article ">Figure 1
Impact of treatment with ZnO nanocrystals (A), or nanocomposites of ZnO:5Ag (B), ZnO:9Ag (C) or ZnO:11Ag (D) on the survival of Galleria mellonella (nm = 30/concentration). Treatments were administered via the left last proleg, in a single dose, at concentrations of 5 mg/kg, 10 mg/kg, 50 mg/kg or 100 mg/kg, and survival was followed for 192 h/8 days. Survival curve construction was performed using Kaplan-Meier analysis, and significant differences were determined using the log-rank test. Differences were considered statistically significant at p < 0.05: (a) difference between treatments and vehicle of PBS; (b) difference between treatment and concentration of 5 mg/kg; (c) difference between treatment and concentration of 10 mg/kg vehicle; and (d) difference between treatment and concentration of 50 mg/kg. Full article ">Figure 2
Impact of treatment with ZnO nanocrystals (A), or nanocomposites of ZnO:5Ag (B), ZnO:9Ag (C) or ZnO:11Ag (D) on weight gain of male C57Bl/6 mice (n = 4). Treatments were administered via gavage for seven consecutive days at doses of 5 mg/kg/d or 10 mg/kg/d, and weight was collected on the first day before treatment, after 8 days (A,B) or after 20 days (E). A two-way ANOVA test (Sidak’s multiple comparisons test) was performed. Differences were considered statistically significant when p < 0.05. * indicates a difference between treatments and vehicles; (a) the difference between 20 days and 0 days; (b) the difference between 8 days and 20 days. Full article ">Figure 3
HAI of liver (A) and kidney (B) histopathological changes in male C57Bl/6 mice (n = 4) after treatment with zinc oxide nanocrystals and ZnO:9Ag or ZnO:11Ag nanocomposites. Treatments were administered via gavage for seven consecutive days at doses of 5 mg/kg/d or 10 mg/kg/d, followed by animal euthanasia (8 days) and liver and kidney collection for subsequent histological analyses. A one-way ANOVA test was performed, and differences were considered statistically significant at p < 0.05. * indicates a difference between treatments and vehicles. Full article ">Figure 4
Photomicrographs of the livers of male C57Bl/6 mice after treatment with ZnO nanocrystals or ZnO:9Ag or ZnO:11Ag nanocomposites: group treated with PBS (A); groups treated with ZnO9Ag, 5 mg/kg/d (B) and 10 mg/kg/d (C); groups treated with ZnO11:Ag, 5 mg/kg/d (D) and 10 mg/kg/d (E). Treatments were administered via gavage for seven consecutive days at doses of 5 mg/kg/d or 10 mg/kg/d, followed by animal euthanasia (8 days) and liver and kidney collection for subsequent histological analyses. Full article ">Figure 5
Photomicrographs of the kidneys of male C57Bl/6 mice after treatment with ZnO nanocrystals or ZnO:9Ag or ZnO:11Ag nanocomposites: group treated with PBS (A); groups treated with ZnO9Ag, 5 mg/kg/d (B) and 10 mg/kg/d (C); groups treated with ZnO11:Ag, 5 mg/kg/d (D) and 10 mg/kg/d (E). Treatments were administered via gavage for seven consecutive days at doses of 5 mg/kg/d or 10 mg/kg/d, followed by animal euthanasia (8 days) and liver and kidney collection for subsequent histological analyses. Full article ">

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J. Funct. Biomater., Volume 15, Issue 3 (March 2024) – 31 articles

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