Understanding the Role of Surface Modification of Randomized Trabecular Titanium Structures in Bone Tissue Regeneration: An Experimental Study
<p>The macrophoto shows a 3D scaffold with a native surface. This prototype, 6.2 mm in diameter and 11 mm in length, was produced for the in vivo study experiment.</p> "> Figure 2
<p>The microphoto shows the surface topography of the native surface that gained roughness through the sand-blasting process. SEM analysis, total magnification 600×.</p> "> Figure 3
<p>Small three-dimensional pores are observed on the surface of all trabeculae (range: 845 nm–11.2 µm) in the treated surface. SEM analysis, total magnification 2000×.</p> "> Figure 4
<p>Three-dimensional reconstruction by a confocal microscope (objective 100×) of the native surface profile (<b>left</b>) and the treated surface (<b>right</b>). The color scale indicates the height of each point.</p> "> Figure 5
<p>Compression test results for cubic specimens with native surfaces. The figure shows the increasing trend of the compression load (N) that the specimens faced before their failure: after a linear (elastic) trend, the load reaches its maximum value (peak) before the failure of the specimen. The displacement represents the deformation value of the specimen before it is crushed. All specimens failed within 0.4 mm of displacement. Each colored line represents the increasing trend of compression load of a specimen.</p> "> Figure 6
<p>Compression test results for cubic specimens with treated surfaces. The explanation of the graphic is provided in <a href="#medicina-58-00315-f005" class="html-fig">Figure 5</a>. Each colored line represents the increasing trend of compression load of a specimen.</p> "> Figure 7
<p>AlamarBlue<sup>®</sup> test results. Axis-X shows time and axis-Y shows the percentage of incremented viability compared to CNTR data.</p> "> Figure 8
<p>SAOS2 cells seeded on (<b>A</b>) the Ti smooth surface and (<b>B</b>) the Ti trabecular structure with a native surface. SEM analysis, total magnification of 1300×.</p> "> Figure 9
<p>The panel shows overviews of two representative sections of (<b>A</b>) a trabecular structure with treated surfaces and (<b>B</b>) a trabecular structure with native surfaces per cortical bone. New compact bone surrounds the implant, englobing the specimens and filling the macropores of the implant. In both cases, the process of bone deposition does not seem wholly terminated and new lamellar bone seems in a phase of organization around the titanium trabeculae of the scaffold (<b>C</b>,<b>D</b>). Furthermore, newly regenerated bone appears well in contact and osseointegrated with the implant. Toluidine blue/pyronin yellow: (<b>A</b>,<b>B</b>) total magnification 13× and (<b>C</b>,<b>D</b>) total magnification 100×.</p> "> Figure 10
<p>The panel shows the overviews of two representative sections of (<b>A</b>) a trabecular structure with treated surfaces and (<b>B</b>) a trabecular structure native surfaces per spongious bone. New bony trabeculae surround the titanium trabeculae, running into and among the macropores of the implant toward the implant surface (<b>C</b>,<b>D</b>). As reported for cortical bone, in both groups, the process of bone deposition is ongoing and new trabeculae are in a phase of maturation around the implant (<b>C</b>,<b>D</b>). Furthermore, newly formed bone comes into contact with the implant surface, osseointegrating with it. Toluidine blue/pyronin yellow: (<b>A</b>,<b>B</b>) total magnification 13× and (<b>C</b>,<b>D</b>) total magnification 100×.</p> "> Figure 11
<p>Both images show spaces occupied by the medulla. In treated surface samples, in cortical bone (<b>A</b>), neurovascular Haversian canals are characterized by blood vessels (red arrows), observable in longitudinal sections. In contrast, many vessels are detected in the medullary spaces of spongious bone (<b>B</b>) (red arrows). In both groups, no inflammatory infiltrate is observed. Toluidine blue/pyronin yellow: (<b>A</b>) total magnification 150× and (<b>B</b>) total magnification 100×.</p> "> Figure 12
<p>The panel highlights cell activity during the modeling and remodeling process. In (<b>A</b>) (chemically treated group) and (<b>B</b>) (untreated group), bony islands in the phase of formation and mineralization surrounded by osteoblast cells (light-blue arrows) are visible. In (<b>C</b>), a cutting cone is characterized by multinucleated cells (red arrows) housed in Howship lacunae (yellow arrows) and osteoblasts (light-blue arrows) that all together maintain the bone structure. (<b>D</b>) Osteocyte cells in different grades of maturation: the red arrow indicates less mature osteocytes housed in large lacunae, identified by the irregular shape, immersed in a violet matrix rich in collagen fibers. Close to the immature lacunae, mature tapered lacunae are visible, with osteocytes housed in calcified bone matrix (light brown). Toluidine blue/pyronin yellow, (<b>A</b>) total magnification 320×, (<b>B</b>) total magnification 320×, (<b>C</b>) total magnification 400×, and (<b>D</b>) total magnification 500×.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Design of the Study
- -
- Evaluation of a randomized trabecular titanium structure with the native surface versus a randomized trabecular titanium structure with the surface chemically treated, describing mechanical and morphological characteristics and cytocompatibility by means of an in vitro test and finally by a preclinical in vivo experiment in a sheep model.
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- Sample preparation;
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- Mechanical and morphological evaluation;
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- In vitro biological assay;
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- In vivo preclinical experiment.
2.2. Sample Preparation
2.2.1. Randomized Trabecular Titanium Structure with the Native Surface
2.2.2. Randomized Trabecular Titanium Structure with an Increased Micro-Roughness Surface
2.3. Mechanical and Morphological Evaluation
2.3.1. Scanning Electron Microscopy
2.3.2. Surface Roughness Characterization
2.3.3. Compression Test
2.4. In Vitro Biological Assay
2.4.1. Cytotoxicity Test: In Vitro Evaluation
2.4.2. Cells Adhesion on the Implant Surface
2.5. In Vivo Preclinical Experiment
2.5.1. Surgical Procedure
2.5.2. Histological Process
2.5.3. Histological Assessment
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- Qualitative evaluation of cellular and tissue reaction
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- Histomorphometrical evaluation of osseointegration around the implant
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- Qualitative and semi-quantitative analysis of the regenerated area
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- Semi-quantitative analysis of the regenerated area
2.6. Statistical Analysis
3. Results
3.1. Mechanical and Morphological Evaluation
3.1.1. Implant Surface SEM
3.1.2. Profilometry
3.1.3. Compression Test
3.2. In Vitro Biological Assay
3.2.1. Cytotoxicity Test: In Vitro Evaluation
3.2.2. Cells Adhesion on the Implant Surface
3.3. In Vivo Preclinical Experiment
3.3.1. Histological Assessment
3.3.2. Qualitative Evaluation of Cellular and Tissue Reaction
3.3.3. Histomorphometric Evaluation of Osseointegration around the Implants
3.3.4. Qualitative Analysis of the Regenerated Area
3.3.5. Semi-Quantitative Analysis of the Regenerated Area
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
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6 Weeks | ||||
---|---|---|---|---|
Bone | BIC | BIn | ||
Mean | SD | Mean | SD | |
Cortical Bone | ||||
Treated | 74.79% | 16.92% | 73.72% | 11.47% |
Native | 77.10% | 16.21% | 81.58% | 9.87% |
Spongious Bone | ||||
Treated | 27.20% | 3.84% | 29.25% | 3.88% |
Native | 30.43% | 9.80% | 33.53% | 20.05% |
Lamellar Bone | Woven Bone | Osteoid | Soft Tissue | |
---|---|---|---|---|
Cortical Bone | ||||
Treated | 3.63 | 1.5 | 1.13 | 1.38 |
Native | 3.83 | 1.5 | 1,00 | 1.13 |
Spongious Bone | ||||
Treated | 1.13 | 1.25 | 2.38 | 3.00 |
Native | 1.63 | 1.5 | 1.63 | 3.38 |
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Canciani, E.; Ragone, V.; Biffi, C.A.; Valenza, F.; D’Ambrosi, R.; Olimpo, M.; Cristofalo, A.; Galliera, E.; Dellavia, C. Understanding the Role of Surface Modification of Randomized Trabecular Titanium Structures in Bone Tissue Regeneration: An Experimental Study. Medicina 2022, 58, 315. https://doi.org/10.3390/medicina58020315
Canciani E, Ragone V, Biffi CA, Valenza F, D’Ambrosi R, Olimpo M, Cristofalo A, Galliera E, Dellavia C. Understanding the Role of Surface Modification of Randomized Trabecular Titanium Structures in Bone Tissue Regeneration: An Experimental Study. Medicina. 2022; 58(2):315. https://doi.org/10.3390/medicina58020315
Chicago/Turabian StyleCanciani, Elena, Vincenza Ragone, Carlo Alberto Biffi, Fabrizio Valenza, Riccardo D’Ambrosi, Matteo Olimpo, Aurora Cristofalo, Emanuela Galliera, and Claudia Dellavia. 2022. "Understanding the Role of Surface Modification of Randomized Trabecular Titanium Structures in Bone Tissue Regeneration: An Experimental Study" Medicina 58, no. 2: 315. https://doi.org/10.3390/medicina58020315
APA StyleCanciani, E., Ragone, V., Biffi, C. A., Valenza, F., D’Ambrosi, R., Olimpo, M., Cristofalo, A., Galliera, E., & Dellavia, C. (2022). Understanding the Role of Surface Modification of Randomized Trabecular Titanium Structures in Bone Tissue Regeneration: An Experimental Study. Medicina, 58(2), 315. https://doi.org/10.3390/medicina58020315