Search Results (14)

Repeatable and reliable assessment of corneal biomechanics with spatial resolution remains a challenge. Vibrational Optical Computerized Tomography (V-OCT), based on sound-wave elastography, has made it possible to investigate the natural resonant modes of the cornea and obtain the elastic moduli non-invasively. This pilot study presents a characterization of four corneal vibrational modes from aberrometric, geometrical, and biomechanical approaches in the living human cornea of five healthy volunteers by combining a corneal sound-wave generator, dual Placido–Scheimpflug corneal imaging, and the Ocular Response Analyzer (ORA) devices. Sound-induced corneal wavefront aberration maps were reconstructed as a function of sound frequency and isolated from the natural state. While maps of low-order aberrations (LOA) revealed symmetric geometrical patterns, those corresponding to high-order aberrations (HOA) showed complex non-symmetric patterns. Corneal geometry was evaluated by reconstructing corneal elevation maps through biconical fitting, and the elastic and viscous components were calculated by applying the standard linear solid model to the ORA measurements. The results showed that sound-wave modulation can increase high-order corneal aberrations significantly. Two frequencies rendered the corneal shape more prolate (50 Hz) and oblate (150 Hz) with respect to the baseline, respectively. Finally, both the elastic and viscous properties are sensitive to sound-induced vibrational modes, which can also modulate the corneal stress-strain response. The cornea exhibits natural resonant modes influenced by its optical, structural, and biomechanical properties. Full article

Terahertz (THz) waves reside in the electromagnetic spectrum between the microwave and infrared bands. In recent decades, THz technology has demonstrated its potential for biomedical applications. With the highly unique characteristics of THz waves, such as the high sensitivity to water and optimal spatial resolution coupled with the characteristics of the human cornea, such as its high water content, THz technology has been explored as a potential modality to assess corneas and corneal diseases. This systematic review provides an overview of the characteristics of THz waves, the safety profile of THz technology in the field of ophthalmology, and its clinical applications, including the objective evaluation of the corneal hydration, tear film, dry eye disease, corneal endothelium, corneal elasticity, and scarring. The paper also presents our viewpoint on the present challenges and future directions of THz technology prior to its broader integration into clinical practice. Full article

We have compared the biomechanical properties of human and porcine corneas using vibrational optical coherence tomography (VOCT). The elastic modulus of the cornea has been previously reported in the literature to vary from about several kPa to more than several GPa based on the results of different techniques. In addition, the formation of corneal cones near the central cornea in keratoconus has been observed in the clinic. Measurements of the resonant frequency and morphology of human and porcine corneas were used to evaluate the role of the limbus in corneal stabilization, the effect of Bowman’s layer, and the effect of collagen content on the low-strain corneal biomechanics. The results of these studies indicate that limbus stability plays an important anatomic role in preventing folding, corneal slippage, and cone formation. Machine learning studies of both human and porcine corneas indicate that Bowman’s membrane, like that of the collagen fibrils found in the anterior corneal stroma, contributes to the 110–120 Hz resonant frequency peak. Finite element and SOLIDWORKS models of normal and keratoconus corneas suggest that the deformation of the cornea is the highest at the central zone and is higher in keratoconus corneas compared to normal controls. VOCT results suggest that although collagen fibril slippage occurs first at the limbus, cone formation in keratoconus occurs centrally/paracentrally, where stress concentration and deformation due to intraocular forces are the highest. Cone formation occurs at the points of maximum curvature. Results of these studies indicate the elastic modulus of cornea fibrillar collagen dictates the corneal elastic modulus at low strains. These results suggest that tension in the cornea at the limbus results in deformation into the low modulus region of the J-shaped stress–strain curve, resulting in an in vivo strain of less than about 10%. We propose that tension in the cornea provides a baseline force that regulates corneal epithelial regeneration as well as corneal lamellae composition and matrix turnover. Full article

The cornea is the optical window to the brain. Its optical and structural properties are responsible for optical transparency and vision. The shape, elasticity, rigidity, or stiffness are due to its biomechanical properties, whose stability results in ocular integrity and intraocular pressure dynamics. Here, we report in vivo observations of shape changes and biomechanical alterations in the human cornea induced by acoustic wave pressure within the frequency range of 50–350 Hz and the sound pressure level of 90 dB. The central corneal thickness (CCT) and eccentricity (e²) were measured using Scheimpflug imaging and biomechanical properties [corneal hysteresis (CH) and intraocular pressure (IOP)] were assessed with air-puff tonometry in six young, healthy volunteers. At the specific 150 Hz acoustic frequency, the variations in e² and CCT were 0.058 and 7.33 µm, respectively. Biomechanical alterations were also observed in both the IOP (a decrease of 3.60 mmHg) and CH (an increase of 0.40 mmHg). Full article

A strain gradient was created by punching a hole in the center of a stretched elastic polydimethylsiloxane membrane to determine the effect of different strains on cultured human keratocytes (HK). In this study, two stretching methods were used: continuous stretching and cyclic stretching. Continuous stretching is relatively static, while acyclic stretching is relatively dynamic. These methods, respectively, represented the effects of high intraocular pressure and rubbing of the eyes on corneal cells. Image processing codes were developed to observe the effects of stress concentration, shear stress, continuous stretching, and cyclic stretching on HKs. The results demonstrate that stretching and shear stress are not conducive to the proliferation of corneal cells and instead cause cell death. A 10% strain had greater inhibitory effects than a 3% strain on cell proliferation. Cell survival rates for continuous stretching (static) were higher than those for cyclic stretching (dynamic). The stretching experiment revealed that cyclic stretching has a greater inhibitory effect on the growth and proliferation of corneal cells than continuous stretching. Accordingly, it shows that cyclic loading is more harmful than high intraocular pressure (static loading) to corneal cells. Full article

Optical properties of the cornea are responsible for correct vision; the ultrastructure allows optical transparency, and the biomechanical properties govern the shape, elasticity, or stiffness of the cornea, affecting ocular integrity and intraocular pressure. Therefore, the optical aberrations, corneal transparency, structure, and biomechanics play a fundamental role in the optical quality of human vision, ocular health, and refractive surgery outcomes. However, the inter-relationships of those properties are not yet reported at a macroscopic scale within the hierarchical structure of the cornea. This work explores the relationships between the biomechanics, structure, and optical properties (corneal aberrations and optical density) at a macro-structural level of the cornea through dual Placido–Scheimpflug imaging and air-puff tonometry systems in a healthy young adult population. Results showed correlation between optical transparency, corneal macrostructure, and biomechanics, whereas corneal aberrations and in particular spherical terms remained independent. A compensation mechanism for the spherical aberration is proposed through corneal shape and biomechanics. Full article

Pterygium is a benign fibrovascular lesion of the bulbar conjunctiva with frequent involvement of the corneal limbus. Its pathogenesis has been mainly attributed to sun exposure to ultraviolet-B radiation. Obtained evidence has shown that it is a complex and multifactorial process which involves multiple mechanisms such as oxidative stress, dysregulation of cell cycle checkpoints, induction of inflammatory mediators and growth factors, angiogenic stimulation, extracellular matrix (ECM) disorders, and, most likely, viruses and hereditary changes. In this review, we aim to collect all authors’ experiences and our own, with respect to the study of fibroelastic ECM of pterygium. Collagen and elastin are intrinsic indicators of physiological and pathological states. Here, we focus on an in-depth analysis of collagen (types I and III), as well as the main constituents of elastic fibers (tropoelastin (TE), fibrillins (FBNs), and fibulins (FBLNs)) and the enzymes (lysyl oxidases (LOXs)) that carry out their assembly or crosslinking. All the studies established that changes in the fibroelastic ECM occur in pterygium, based on the following facts: An increase in the synthesis and deposition of an immature form of collagen type III, which showed the process of tissue remodeling. An increase in protein levels in most of the constituents necessary for the development of elastic fibers, except FBLN4, whose biological roles are critical in the binding of the enzyme LOX, as well as FBN1 for the development of stable elastin. There was gene overexpression of TE, FBN1, FBLN5, and LOXL1, while the expression of LOX and FBLN2 and -4 remained stable. In conclusion, collagen and elastin, as well as several constituents involved in elastic fiber assembly are overexpressed in human pterygium, thus, supporting the hypothesis that there is dysregulation in the synthesis and crosslinking of the fibroelastic component, constituting an important pathogenetic mechanism for the development of the disease. Full article

The development of commercial collagen inks for extrusion-based bioprinting has increased the amount of research on pure collagen bioprinting, i.e., collagen inks not mixed with gelatin, alginate, or other more common biomaterial inks. New printing techniques have also improved the resolution achievable with pure collagen bioprinting. However, the resultant collagen constructs still appear too weak to replicate dense collagenous tissues, such as the cornea. This work aims to demonstrate the first reported case of bioprinted recombinant collagen films with suitable optical and mechanical properties for corneal tissue engineering. The printing technology used, aerosol jet^® printing (AJP), is a high-resolution printing method normally used to deposit conductive inks for electronic printing. In this work, AJP was employed to deposit recombinant human collagen type III (RHCIII) in overlapping continuous lines of 60 µm to form thin layers. Layers were repeated up to 764 times to result in a construct that was considered a few hundred microns thick when swollen. Samples were subsequently neutralised and crosslinked using EDC:NHS crosslinking. Nanoindentation and absorbance measurements were conducted, and the results show that the AJP-deposited RHCIII samples possess suitable mechanical and optical properties for corneal tissue engineering: an average effective elastic modulus of 506 ± 173 kPa and transparency ?87% at all visible wavelengths. Circular dichroism showed that there was some loss of helicity of the collagen due to aerosolisation. SDS-PAGE and pepsin digestion were used to show that while some collagen is degraded due to aerosolisation, it remains an inaccessible substrate for pepsin cleavage. Full article

There is a growing need for novel in vitro corneal models to replace animal-based ex vivo tests in drug permeability studies. In this study, we demonstrated a corneal mimetic that models the stromal and epithelial compartments of the human cornea. Human corneal epithelial cells (HCE-T) were grown on top of a self-supporting porcine collagen-based hydrogel. Cross-sections of the multi-layers were characterized by histological staining and immunocytochemistry of zonula oc-cludens-1 protein (ZO-1) and occludin. Furthermore, water content and bssic elastic properties of the synthetized collagen type I-based hydrogels were measured. The apparent permeability coefficient (P_app) values of a representative set of ophthalmic drugs were measured and correlated to rabbit cornea P_app values found in the literature. A multilayered structure of HCE-T cells and the expression of ZO-1 and occludin in the full thickness of the multilayer were observed. The hydrogel-based corneal model exhibited an excellent correlation to rabbit corneal permeability (r = 0.96), whereas the insert-grown HCE-T multilayer was more permeable and the correlation to the rabbit corneal permeability was lower (r = 0.89). The hydrogel-based human corneal model predicts the rabbit corneal permeability more reliably in comparison to HCE-T cells grown in inserts. This in vitro human corneal model can be successfully employed for drug permeability tests whilst avoiding ethical issues and reducing costs. Full article

An important component of tissue engineering (TE) is the supporting matrix upon which cells and tissues grow, also known as the scaffold. Scaffolds must easily integrate with host tissue and provide an excellent environment for cell growth and differentiation. Human amniotic membrane (hAM) is considered as a surgical waste without ethical issue, so it is a highly abundant, cost-effective, and readily available biomaterial. It has biocompatibility, low immunogenicity, adequate mechanical properties (permeability, stability, elasticity, flexibility, resorbability), and good cell adhesion. It exerts anti-inflammatory, antifibrotic, and antimutagenic properties and pain-relieving effects. It is also a source of growth factors, cytokines, and hAM cells with stem cell properties. This important source for scaffolding material has been widely studied and used in various areas of tissue repair: corneal repair, chronic wound treatment, genital reconstruction, tendon repair, microvascular reconstruction, nerve repair, and intraoral reconstruction. Depending on the targeted application, hAM has been used as a simple scaffold or seeded with various types of cells that are able to grow and differentiate. Thus, this natural biomaterial offers a wide range of applications in TE applications. Here, we review hAM properties as a biocompatible and degradable scaffold. Its use strategies (i.e., alone or combined with cells, cell seeding) and its degradation rate are also presented. Full article

Whether hyperoxia affects the refraction in neonatal and adult mice is unknown. The mice exposed to 85% oxygen at postnatal 8 days (P8d) for 3 days and the mice exposed to normal air were assigned to the neonatal hyperoxia and normoxia groups, respectively. The refraction, the corneal curvature radius (CR) and the axial length (AL) were measured at P30d and P47d. Postnatal 6 weeks (P6w) adult mice were divided into the adult hyperoxia and normoxia groups. These parameters were measured before oxygen exposure, after 1 and 6 weeks, and every 7 weeks. The lens elasticity was measured at P7w and P26w by enucleation. The neonatal hyperoxia group showed a significantly larger myopic change than the neonatal normoxia group (P47d −6.56 ± 5.89 D, +4.11 ± 2.02 D, p < 0.001), whereas the changes in AL were not significantly different (P47d, 3.31 ± 0.04 mm, 3.31 ± 0.05 mm, p = 0.852). The adult hyperoxia group also showed a significantly larger myopic change (P12w, −7.20 ± 4.09 D, +7.52 ± 2.54 D, p < 0.001). The AL did not show significant difference (P12w, 3.44 ± 0.03 mm, 3.43 ± 0.01 mm, p = 0.545); however, the CR in the adult hyperoxia group was significantly smaller than the adult normoxia group (P12w, 1.44 ± 0.03 mm, 1.50 ± 0.03 mm, p = 0.003). In conclusion, hyperoxia was demonstrated to induce myopic shift both in neonatal and adult mice, which was attributed to the change in the CR rather than the AL. Elucidation of the mechanisms of hyperoxia and the application of this result to humans should be carried out in future studies. Full article

Estimating the corneal elasticity can provide valuable information for corneal pathologies and treatments. Ophthalmologic pathologies will invariably cause changes to the elasticity of the cornea. For example, keratoconus and the phototoxic effects of ultraviolet radiation usually increase the corneal elasticity. This makes a quantitative estimation of the elasticity of the human cornea important for ophthalmic diagnoses. The present study investigated the use of a proposed high-resolution shear wave imaging (HR-SWI) method based on a dual-element transducer (comprising an 8-MHz element for pushing and a 32-MHz element for imaging) for measuring the group shear wave velocity (GSWV) of the human cornea. An empirical Young’s modulus formula was used to accurately convert the GSWV to Young’s modulus. Four quantitative parameters, bias, resolution, contrast, and contrast-to-noise ratio (CNR), were measured in gelatin phantoms with two different concentrations (3% and 7%) to evaluate the performance of HR-SWI. The biases of gelatin phantoms (3% and 7%) were 5.88% and 0.78%, respectively. The contrast and CNR were 0.76, 1.31 and 3.22, 2.43 for the two-side and two-layer phantoms, respectively. The measured image resolutions of HR-SWI in the lateral and axial directions were 72 and 140 μm, respectively. The calculated phase SWV (PSWV) and their corresponding Young’s modulus from six human donors were 2.45 ± 0.48 m/s (1600 Hz) and 11.52 ± 7.81 kPa, respectively. All the experimental results validated the concept of HR-SWI and its ability for measuring the human corneal elasticity. Full article

A silk protein, fibroin, was isolated from the cocoons of the domesticated silkworm (Bombyx mori) and cast into membranes to serve as freestanding templates for tissue-engineered corneal cell constructs to be used in ocular surface reconstruction. In this study, we sought to enhance the attachment and proliferation of corneal epithelial cells by increasing the permeability of the fibroin membranes and the topographic roughness of their surface. By mixing the fibroin solution with poly(ethylene glycol) (PEG) of molecular weight 300 Da, membranes were produced with increased permeability and with topographic patterns generated on their surface. In order to enhance their mechanical stability, some PEG-treated membranes were also crosslinked with genipin. The resulting membranes were thoroughly characterized and compared to the non-treated membranes. The PEG-treated membranes were similar in tensile strength to the non-treated ones, but their elastic modulus was higher and elongation lower, indicating enhanced rigidity. The crosslinking with genipin did not induce a significant improvement in mechanical properties. In cultures of a human-derived corneal epithelial cell line (HCE-T), the PEG treatment of the substratum did not improve the attachment of cells and it enhanced only slightly the cell proliferation in the longer term. Likewise, primary cultures of human limbal epithelial cells grew equally well on both non-treated and PEG-treated membranes, and the stratification of cultures was consistently improved in the presence of an underlying culture of irradiated 3T3 feeder cells, irrespectively of PEG-treatment. Nevertheless, the cultures grown on the PEG-treated membranes in the presence of feeder cells did display a higher nuclear-to-cytoplasmic ratio suggesting a more proliferative phenotype. We concluded that while the treatment with PEG had a significant effect on some structural properties of the B. mori silk fibroin (BMSF) membranes, there were minimal gains in the performance of these materials as a substratum for corneal epithelial cell growth. The reduced mechanical stability of freestanding PEG-treated membranes makes them a less viable choice than the non-treated membranes. Full article

A bi-functional epoxy-based cross-linker, 1,4-Butanediol diglycidyl ether (BDDGE), was investigated in the fabrication of collagen based corneal substitutes. Two synthetic strategies were explored in the preparation of the cross-linked collagen scaffolds. The lysine residues of Type 1 porcine collagen were directly cross-linked using l,4-Butanediol diglycidyl ether (BDDGE) under basic conditions at pH 11. Alternatively, under conventional methodology, using both BDDGE and 1-Ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) as cross-linkers, hydrogels were fabricated under acidic conditions. In this latter strategy, Cu(BF₄)₂·XH₂O was used to catalyze the formation of secondary amine bonds. To date, we have demonstrated that both methods of chemical cross-linking improved the elasticity and tensile strength of the collagen implants. Differential scanning calorimetry and biocompatibility studies indicate comparable, and in some cases, enhanced properties compared to that of the EDC/NHS controls. In vitro studies showed that human corneal epithelial cells and neuronal progenitor cell lines proliferated on these hydrogels. In addition, improvement of cell proliferation on the surfaces of the materials was observed when neurite promoting laminin epitope, IKVAV, and adhesion peptide, YIGSR, were incorporated. However, the elasticity decreased with peptide incorporation and will require further optimization. Nevertheless, we have shown that epoxy cross-linkers should be further explored in the fabrication of collagen-based hydrogels, as alternatives to or in conjunction with carbodiimide cross-linkers. Full article