Scleral Collagen Cross-linking Research Articles

Permeation characteristics and cross-linking efficacy of iontophoresis-assisted riboflavin delivery for accelerated riboflavin-ultraviolet A scleral collagen cross-linking in porcine eyes

The purpose of this study is to investigate whether the iontophoresis-assisted riboflavin delivery to posterior sclera with less delivery time, can achieve the same riboflavin permeation efficiency as the passive soaking way, and its effect on the mechanical properties of posterior sclera for accelerated scleral collagen cross-linking (A-SXL). In this study, 0.1% riboflavin solution was applied into the posterior sclera of porcine eyes either by the iontophoresis-assisted or passive soaking method, with delivery time of 5, 7.5, 10, 12.5, 15, 17.5, and 20 min, respectively. The fluorescence intensity and the distribution of riboflavin concentration in the 10 μm frozen sections of the sclera were evaluated by fluorescence inverted microscope. The posterior sclera with riboflavin treatment through either the iontophoresis-assisted or the passive soaking method for different durations ranging from 5 to 20 min was treated with ultraviolet A (UVA) irradiation at an intensity of 10 mW/cm2 for 9 min. The elastic modulus was determined at the physiological strain level using the uniaxial tensile test after ASXL. The results showed that the fluorescence intensity of riboflavin increased by prolonging the delivery time in both the iontophoresis and passive soaking groups, and the permeation depth of riboflavin remained constant over 15 min. The fluorescence intensity in the iontophoresis group was significantly higher than in the passive soaking group at 12.5 min and 15 min, respectively. The elastic modulus at 12.5 min in the iontophoresis group was significantly higher than in the passive soaking group at the same delivery time and showed no significant difference compared to the passive soaking group at 20 min. In conclusion, it indicated that iontophoresis-assisted delivery could not only shorten the surgery time but also achieve similar mechanical performance to the passive soaking method in ASXL.

Scleral collagen crosslinking as a promising direction of sclera-strengthening treatment of progressive myopia

The review presents recent research works on new technologies of scleral collagen crosslinking, a promising approach to sclerastrengthening treatment of progressive myopia. We assess the advantages and limitations of a number of experimental techniques of photochemical and medicinal crosslinking of the sclera, as well as donor tissue crosslinking aimed at optimizing the plastic material for sclera-strengthening surgery. Successful experiments and effective first clinical implementation results of the approach can be viewed as the basis for its further improvement and wider introduction of the technology into ophthalmological practice.

Effects of riboflavin/ultraviolet-A(UVA) scleral crosslinking on the mechanical behavior of the scleral fibroblasts of lens-induced myopia Guinea pigs

Myopia is becoming increasingly severe, and studies have shown that the cellular mechanics of scleral fibroblasts are altered following myopia. Scleral UVA-Riboflavin Collagen Crosslinking（sCXL） is a promising treatment for myopia prevention and control of axial growth. Understanding the mechanical properties of scleral fibroblasts is crucial, as it influences the cellular response and limits the extent of molecular deformation triggered. Thus, our study aimed to investigate the effect of mechanical properties of scleral fibroblasts in a lens-induced myopic guinea pig model following sCXL. For this purpose, we performed the 0.1% riboflavin/UVA scleral crosslinking (365 nm,3 mW/cm2,30 min) in the right eyes of guinea pigs in Group CXL. In Group LIM, the right eyes were only administrated negative lens for 6 weeks. No treatment was performed in both eyes of the guinea pigs in group Control. The scleral fibroblasts were isolated and cultured from the scleral tissue at the cross-linking area in Group CXL and the corresponding area in Group LIM and control. The curve of the length of microtubules inhaled by cells under negative pressure was measured by a microaspiration-based isolation technique, and the equilibrium Young's modulus and apparent viscosity of scleral fibroblasts were calculated by formula fitting. The equilibrium Young's modulus of scleral fibroblasts in group CXL was significantly lower than that in the LIM group (P < 0.01, two-sample t-test between pairs）, and there was no significant difference between groups CXL and control. The results show that sCXL can effectively moderate the phenomenon that scleral fibroblasts are not easy to deform after myopia. The apparent viscosity modulus in the CXL group was higher than the groups' control and LIM. Taken together, our data demonstrate the biomechanics of the scleral fibroblasts altered after Riboflavin/UVA scleral collagen cross-linking in a lens-induced myopia model.

Effects of riboflavin/ultraviolet-A scleral collagen cross-linking on regional scleral thickness and expression of MMP-2 and MT1-MMP in myopic guinea pigs.

To investigate the effects of scleral collagen cross-linking (SXL) using riboflavin and ultraviolet A (UVA) light on the scleral thickness of different regions and expression of matrix metalloproteinase 2 (MMP-2) and membrane-type MMP-1 (MT1-MMP) in guinea pigs with lens-induced myopia. Forty-eight 4-week-old guinea pigs were assigned to three groups (n = 16 per group): SXL group, lens-induced myopia (LIM) group, and control group. The sclera of the right eye of the guinea pig in the SXL group was surgically exposed, riboflavin was dropped on the treatment area for 10 minutes before the 30-minute UVA irradiation. The same surgical procedure was performed in the LIM group without UVA irradiation. The -10.00 D lenses were then placed on the right eyes of guinea pigs in the SXL and LIM groups for six weeks. The control group received no treatment. The left eyes were untreated in all groups. The ocular axial length (AXL) and refraction were measured at 4 weeks and 10 weeks of age. 10-week-old guinea pigs were sacrificed, and the right eyes were enucleated and evenly divided for preparation of hematoxylin and eosin (HE) stained sections, quantitative real-time polymerase chain reaction (qPCR) and western blotting. The scleral thickness of different regions was measured on HE stained sections. The temporal half of the sclera was harvested to measure the expression of MMP-2 and MT1-MMP by qPCR and western blotting. The AXL was significantly shorter, and the degree of myopic refraction was significantly lower in the SXL group than those in the LIM group at 10 weeks of age. The scleral thickness of the cross-linked area was significantly greater in the SXL group than that of the corresponding area in the LIM group, while the scleral thickness of the untreated nasal side was not significantly different between the SXL group and the LIM group. The expression of MMP-2 and MT1-MMP of the cross-linked sclera was significantly downregulated compared with that of the corresponding area in the LIM group. Riboflavin/UVA SXL could slow myopia progression and thicken the cross-linked sclera in guinea pigs, which might be related to the downregulation of MMP-2 and MT1-MMP expression during the scleral remodeling process.

Efficacy and safety of microbial transglutaminase-induced scleral stiffening invivo

The purpose of this study was to investigate the efficacy and safety of microbial transglutaminases (mTGases) during scleral collagen cross-linking (CXL) in vivo. Sixteen New Zealand white albino rabbits were treated with sub-Tenon's injections of 2 ml of 1 U/ml mTGases in the right eye and 2 ml of phosphate buffer saline (PBS) in the left eye. The rabbits were killed 2 weeks after the injection, and all eyeballs, including some scleral strips, were processed. The elastic modulus was measured with a biomaterials tester. Histopathological analysis and transmission electron microscopy (TEM) were used for the morphological observations. The elastic modulus of the mTGase-treated sclera was 15.79 ± 2.93 MPa, and that of the control was 6.91 ± 2.23 MPa, indicating an increase of 129% after the mTGases treatment (P < 0.05). The density of the scleral collagen bundles and diameter of the collagen fibrils increased compared with those in the control group. No apoptosis was detected in the retina or posterior sclera by TUNEL staining, and no histological damage was observed on the TEM scan. This study is based on a short-term study on animal models. These results indicate that mTGase-mediated scleral CXL is a promising approach to effectively stiffen the sclera and safe enough for retina, and may be a useful treatment modality for strengthening scleral tissue.

Clinical Feasibility and Safety of Scleral Collagen Cross-Linking by Riboflavin and UltravioletA in Pathological Myopia Blindness: A Pilot Study.

To investigate the feasibility and safety of scleral ultravioletA (UVA) cross-linking (scleral CXL) on pathologically blindness. This was a prospective, observational clinical study. Five patients with monocular blindness due to pathological myopic maculopathy were enrolled. Eyes with best corrected visual acuity (BCVA) under 0.05 were defined as experimental eyes. The fellow eyes were defined as control eyes. Patients first underwent posterior scleral reinforcement (PSR) surgery in the control eye. Thereafter, scleral CXL surgery was performed in the experimental eye on the same day. Visual acuity, BCVA, slit lamp biomicroscopic examination, intraocular pressure measurement, corneal specula microscopies, axis length measurement, funduscopy with pupil dilation, color fundus photography, full-field flash electroretinography, optical coherence tomography, and color Doppler flow imaging were performed at baseline, 1week, 1month, 3months, 6months, and 12months after surgery. No signs of inflammation were observed after operation and throughout the follow-up period. Retinoschisis was improved, while choroidal neovascularization fibrosis and retinal and choroidal atrophy were unchanged after scleral CXL. There were no statistically significant differences in the ophthalmic artery, central retinal artery, and posterior ciliary artery parameters of color Doppler flow imaging or in retinal thickness, within experimental and control eyes, at baseline, 1week, 1month, 3months, or 12months (P > 0.05). This pilot study verified the feasibility and safety of scleral CXL on human blindness. The UVA-CXL on the sclera of human eyes seems to have the same effect as PSR in preventing progressive pathological myopia in the future. Chinese Clinical Trial Registry (ChiCTR2100042422).

Scleral Remolding-Related Gene Expression After Scleral Collagen Cross-Linking Using Ultraviolet A and Riboflavin in Myopic Guinea Pig Model

Purpose This study was conducted to evaluate scleral remolding-related gene expression after scleral collagen cross-linking (SCXL) using ultraviolet A (UVA) and riboflavin in lens-induced myopia (LIM) guinea pigs. Methods A total of 100 4-week-old pigmented guinea pigs were randomly divided into five groups (n = 20): SCXL + LIM, LIM, SCXL, Sham, and Control. Refraction, anterior chamber depth (ACD), lens thickness (LT), vitreous chamber depth (VCD), and axial length (AL) were measured using streak retinoscope and A-scan ultrasonography. SCXL was performed using 0.1% riboflavin solution and 365 nm UVA irradiation. Lens-induced myopia was achieved by wearing −10 D concave lenses. Quantitative real-time PCR (qPCR) and western blot were used to measure mRNA and protein levels, respectively. Results Myopia was successfully induced in the LIM group, while myopic refraction was higher and ACD and AL were shorter in SCXL + LIM compared with LIM, suppressing myopia progression. The scleral COL1A1 mRNA levels were significantly decreased and MMP2 and ACTA2 mRNA levels were significantly increased in LIM compared with other groups, while COL1A1 mRNA levels were increased and MMP2 and ACTA2 mRNA levels were decreased in SCXL + LIM compared with LIM. The scleral COL1A1 protein levels were significantly increased at 1 week and 4 weeks and MMP2 protein levels were significantly decreased at 1 week in SCXL compared with SCXL + LIM, LIM and Control. MMP2 protein levels were significantly decreased in SCXL + LIM and SCXL compared with LIM at 4 weeks. The differences in TGFB1, BMP2, CCN2, ITGA2, and ITGB1 mRNA levels and ACTA2 protein levels between the five groups were not significantly different. Conclusion SCXL using UVA and riboflavin could influence the expression of scleral remolding-related genes, including COL1A1, MMP2, TIMP2, and ACTA2, and thus contribute to improving collagen synthesis and reducing collagen degradation and might have an effect on slowing myopia progression.

Advances in preventing myopia by scleral collagen crosslinking

<p indent="0mm">The prevention of myopia is crucial due to its increasing prevalence globally. Myopia development is associated with scleral remodeling and reduced scleral biomechanical properties. Scleral collagen crosslinking is a new, effective approach to prevent myopia clinically, including photo-crosslinking by light irradiation (ultraviolet A and riboflavin scleral collagen crosslinking, and blue light and riboflavin scleral collagen crosslinking) and crosslinking by biological crosslinkers (genipin, carbohydrate, FARs, and Tgases). This article summarizes the relevant research on the application scheme, validity, and safety of scleral collagen crosslinking to prevent myopia.

Changes in intraocular pressure and ocular pulse amplitude of rhesus macaques after blue light scleral cross-linking

BackgroundScleral cross-linking can enhance the biomechanical strength of the sclera and is expected to be a new operative method for the prevention of myopia. However, studies investigating the changes in intraocular pressure (IOP) and ocular pulse amplitude (OPA) after blue light-riboflavin induced scleral collagen cross-linking (SXL) in rhesus monkeys are limited. This study aimed to investigate the changes in IOP and OPA in three-year-old rhesus macaques 1 week, 1 month, and 3 months after blue light-riboflavin SXL.MethodsSeven three-year-old rhesus macaques (14 eyes) were randomly divided into two groups, with 4 monkeys in group A (8 eyes) and 3 monkeys in group B (6 eyes). The right eye of each rhesus macaque was used as the experimental eye, whereas the left eye was used as the control. In group A, one quadrant of each right eye was irradiated. In group B, two quadrants of each right eye and one quadrant of each left eye were irradiated. The IOP and OPA of both eyes were measured in all seven rhesus macaques before SXL and 1 week, 1 month, and 3 months postoperatively, and differences in the IOP and OPA between the experimental and control eyes were evaluated via the paired t test.ResultsIn groups A and B, there were no significant differences between the experimental and control eyes in the IOP or OPA before SXL or 1 week, 1 month, or 3 months postoperatively (P > 0.05).ConclusionsThe IOP and OPA are not significantly affected in 1 vs 0 or in 1 vs 2 quadrants of blue light SXL.

Finite element analysis of the effect of local posterior sclera collagen cross-linking on eyeball shape

China is the country with high incidence of high myopia in the world. High myopia can cause severe vision impairment. So far, there is no effective treatment for high myopia in clinic. Scleral collagen cross-linking surgery has been proven to be effective in preventing animal eye axial elongation in vitro and in vivo. However, the influence of posterior scleral collagen cross-linking on the deformation of the whole eyeball is still unclear. In this study, finite element simulation were used to analyze the changes of eyeball shape and the position of light casting on the retina after posterior sclera cross-linking, and the mathematical algorithm was written to verify their similarity. The results showed that the shape of the whole eyeball was still very similar before and after cross-linking, and the diopter of the eyeball after cross-linking had little change, which had almost no effect on the position of light projection on the retina. Our results indicate that posterior sclera cross-linking wouldn't lead to distortion to the optometry, that is, the increase of elastic modulus in local scleral tissue after cross-linking wouldn't cause new problem of optometry and vision.

Shaping Eyeballs by Scleral Collagen Cross-Linking: A Hypothesis for Myopia Treatment.

The global prevalence of myopia has brought to the attention of the different eye and vision specialists, who make way to control its progression. Evidence have shown that a proactive reshaping of the eyeball is the core point of myopia developing process, which particularly includes the weakening, thinning, and expanding of the sclera. Thus, the sclera is considered to be a prime target for therapeutic manipulation in halting progressive myopia. In the past decades, corneal collagen cross-linking has been applied in clinical practice for treating aberrant corneal remodeling diseases. In this article, we hypothesize that scleral collagen cross-linking (SXL) has a huge potential in stabilizing myopic process by shaping the eyeball and preventing the aberrant scleral remodeling. In contrast with the current methods of optometry correction, such as physiotherapy, pharmacotherapy, spectacles, contact lenses, refractive surgeries, etc., eyeball-shaping method using SXL is a fundamental intervention which aims at the pathogenesis of progressive visual loss of myopia. Compared with the current posterior scleral reinforcement, the most advantage of SXL is that there is no allotransplant into the myopic eye, which means less expenditure, lower risk, and easier to handle in operating.

Экспериментальное обоснование кросслинкинга склеры

Purpose. To substantiate the efficacy and safety of scleral collagen cross-linking with riboflavin and ultraviolet radiation. Material and methods. The study was conducted on cadaver porcine eyes and rabbit eyes. Results. It has been experimentally proven that crosslinking with riboflavin / ultraviolet A leads to an increase in the biomechanical strength of the scleral tissue, changing its structure and leading to an increase in the packing density of collagen fibers and an increase in the diameter of collagen fibrils, which confirms the formation of additional cross-links between scleral macromolecules. The absence of any pathological changes in the fibers of the fibrous membrane of the eye was established. The permeability of scleral tissue for ultraviolet radiation of the A range was determined. Based on the data obtained, a formula was proposed for calculating the threshold value of the intensity of ultraviolet radiation during the SCXL procedure. Using the methods of light microscopy, electroretinography and optical coherence tomography, it was revealed that SCXL with riboflavin / ultraviolet A under the studied parameters is safe for the structures of the eye (cornea, retina and optic nerve). A gentle technology for photopolymerization of the sclera with the use of the “UFalink S” device has been developed, the advantage of which is the ability to carry out ultraviolet irradiation in hard-to-reach areas of the eyeball. At the same time, an effective increase in the biomechanical strength characteristics of the scleral tissue was proved and the absence of a damaging effect on the structures of the eye was demonstrated. Conclusion. A clinical study of the scleral crosslinking procedure with riboflavin / ultraviolet A with the proposed parameters: irradiance of 3 mW / cm², irradiation time 30 minutes and preliminary saturation of the sclera with 0.1% aqueous solution of riboflavin for 20 minutes, which in experiments have shown their effectiveness and safety, is recommended. During the procedure, it is recommended to use a water-based photosensitizer riboflavin, without the addition of dextran, which leads to dehydration of the sclera and an increase in its permeability to ultraviolet light, which increases the risk of damage to the inner layers of the eye. Key words: scleral crosslinking, ultraviolet A, riboflavin, experiment, myopia.

Effect of enzyme-induced collagen crosslinking on porcine sclera

This study aims to evaluate the effect of a new type of collagen crosslinking (CXL) mediated by microbial transglutaminases (Tgases) on sclera. Porcine eyes were divided into two groups according to the different crosslinking procedures used: the double-sided CXL group (D-CXL group) and the single-sided CXL group (S-CXL group). In the D-CXL group, 4.0 × 14.0 mm scleral strips harvested from 40 porcine eyeballs were incubated with 1 U/ml Tgases for 30 min at 37 °C. Parallel scleral strips from the same eyeball were incubated with PBS under the same conditions as the controls. In the S-CXL group, 80 whole globes were directly incubated with 1 U/ml Tgases and PBS as the controls for 30 min at 37 °C. After incubation, 4.0 × 14.0 mm scleral strips were cut from each eyeball. Biomechanical testing and light microscopy were used. In the D-CXL group, the general elastic modulus of the Tgases-treated scleral strips was 14.89 ± 6.05 MPa, and the controls was 6.72 ± 2.58 MPa, indicating an increase of 121% with Tgases treatment. In the S-CXL group, the general elastic modulus of the Tgases-treated scleral strips was 12.88 ± 4.29 MPa, and the controls was 7.00 ± 2.45 MPa, indicating an increase of 84% with Tgases treatment. In both the D-CXL and S-CXL groups, significant increases in scleral rigidity were observed compared to that of the respective controls (P < 0.05). The histology indicated increased collagen bundle density, decreased interfibrillar spaces and increased interlamellar spaces after CXL. In conclusion, scleral collagen crosslinking mediated by Tgases produced a significant increase in biomechanical strength.

Differential Regional Stiffening of Sclera by Collagen Cross-linking

ABSTRACTPurpose: Corneal collagen cross-linking by ultraviolet light activation of riboflavin has been used clinically to enhance corneal stiffness. We sought to determine if cross-linking differentially affects scleral regions.Methods: Adjacent, parallel strips of sclera were cut from superolateral, superomedial, inferolateral, and inferomedial quadrants of posterior and equatorial sclera of 12 human cadaver eyes. One of each pair served as control while the other was cross-linked by immersion in 0.1% riboflavin and 365 nm exposure at 6 mW/cm2 irradiance for 30 min. Behavior of strips was characterized using a microtensile load cell. Preloaded strips were imaged using orthogonally mounted cameras and optical coherence tomography to determine specimen dimensions including cross-sectional area. Tension was measured during 0.1 mm/s constant rate elongation.Results: Young’s modulus (YM), the slope of the relationship relating tensile stress to strain, was calculated at 8% strain, and increased significantly after cross-linking (P < .001). In posterior sclera, mean (± standard error of mean, SEM) YM is increased in the superolateral, superomedial, inferolateral, and inferomedial quadrants by 46 ± 15%, 32 ± 11%, 67 ± 20%, and 53 ± 11%, respectively. In equatorial sclera, YM is increased by 139 ± 43%, 68 ± 27%, 143 ± 92%, and 68 ± 14%, respectively. The YM of pooled equatorial quadrants increased significantly more than that of the pooled posterior quadrants.Conclusions: Scleral collagen cross-linking by ultraviolet activation of riboflavin differentially increases scleral YM more in the equatorial than posterior sclera, and most in the lateral, equatorial sclera. Cross-linking might be used to arrest progressive myopia or to prevent staphyloma formation.

Effects of scleral collagen crosslinking with different carbohydrate on chemical bond and ultrastructure of rabbit sclera: Future treatment for myopia progression.

BackgroundMyopia is the most common ocular disorder and is mainly caused by axial elongation of the sclera. If the stiffness of sclera increased, it can inhibit myopia progression. The aim of this study is to compare the effect of the collagen crosslinking with different types and concentrations of carbohydrates on chemical bond and ultrastructural change of rabbit sclera.MethodsNine New Zealand white rabbits were treated with five, sequential sub-Tenon injections of 0.15 mL solutions of ribose, sucrose, and glycogen of 0.1, 0.2 and 0.4 M concentration at the right eye over 14 days. Ten weeks after the last injection, the rabbits were sacrificed and chemical bond and ultrastructural changes were compared with those of the untreated left sclera using Raman spectroscopy, atomic force microscopy (AFM), and histology.ResultsRaman spectroscopy of the control and cross-linked rabbit sclera tissue revealed different types of collagen interactions. Raman shift of 919 cm-1 (C-C stretching and vibration of the proline ring in collagen) was the highest in ribose, followed by sucrose and glycogen. Total energy intensity was also highest in ribose, followed by sucrose and glycogen, and showed a tendency to increase at higher concentrations. AFM revealed interlocking arrangements of collagen fibrils. The collagen fibril diameter was 105.6 ± 21.2 nm, 109.4 ± 28.8 nm, 113.1 ± 30.8 nm and 137.6 ± 25.3 nm for control group, 0.4 M glycogen, sucrose, and ribose, respectively. Histology indicated increased density of the collagen bundle and no increase in inflammatory cell recruitment compared to control at high concentrations of ribose.ConclusionsScleral crosslinking using glycation increased the scleral biomechanical rigidity and these results were particularly pronounced in ribose. Scleral crosslinking using glycation may be a promising method for inhibiting high myopia progression.

Research advances in corneal and scleral collagen cross-linking with ultraviolet A and riboflavin

In recent years, the use of collagen cross-linking with ultraviolet A and riboflavin is being developed universally due to its significance in the improvement of the mechanical strength of collagen fibers in clinical and basic research. It has been used in treating some corneal diseases including keratoconus, keratectasia and keratohelcosis. Thus, it might become a new treatment to prevent progressive myopia by strengthening scleral tissue. This article focuses on the effectiveness and safety of ultraviolet A-riboflavin cross-linking, and discusses the application prospect, focal points and problems related to cross-linking therapy. (Chin J Ophthalmol, 2018, 54:948-953).

Posterior staphyloma in pathologic myopia.

A posterior staphyloma is an outpouching of a circumscribed region of the posterior fundus and has been considered a hallmark of pathologic myopia. Occurring in highly myopic eyes, it is histologically characterized by a relatively abrupt scleral thinning starting at the staphyloma edge, a pronounced de-arrangement of scleral collagen fibrils and a marked choroidal thinning, which is the most marked at the staphyloma edge and which occurs in addition to the axial elongation-associated choroidal thinning. Besides in highly myopic eyes, a posterior staphyloma can be found in non-highly myopic eyes in association with retinitis pigmentosa or localized defects of Bruch's membrane in the cases of which it is not associated with a marked choroidal thinning. The diagnosis of posterior staphylomas is considered best made by wide-field optical coherence tomography, because wide-field optical coherence tomography encompasses the entire extent of the most predominant type of staphylomas (i.e., the wide macular type) and since it also has a sufficiently high resolution of images (in contrast to ultrasonography, computed tomography and three-dimensional magnetic resonance imaging). While the etiology of posterior staphylomas has remained unclear, local choroidal factors and a locally decreased biomechanical resistance of the sclera against a posteriorly expanding Bruch's membrane have been one of the assumed pathogenic parameters. For the therapy of staphylomas, scleral reinforcement strategies such as by posterior encircling bands, posterior scleral collagen cross-linking or scleral regeneration have been discussed or performed, however, with the pathogenesis being elusive, the therapy of staphylomas has remained undetermined.

Scleral Cross-Linking Using Glyceraldehyde for the Prevention of Axial Elongation in the Rabbit: Blocked Axial Elongation and Altered Scleral Microstructure

ABSTRACTBackground: This study aims to assess the efficacy of the scleral collagen cross-linking method using glyceraldehyde solution for prevention of lens-induced axial elongation in New Zealand rabbits and investigate the biochemical and microstructural changes that occur.Methods: The right eyes of New Zealand rabbits aged seven weeks were randomly divided into three groups: the cross-linking group (n = 6), non-crosslinking group (n = 5), and untreated control group (n = 5). Eyes in cross-linking and non-crosslinking groups were treated with a −8.00 Diopter spherical lens over the course of two weeks. The cross-linking effects were achieved by a sub-Tenon’s injection of 0.15 ml 0.5 M glyceraldehyde to eyes in the CL group. Ocular parameters were measured on the 1st, 7th, and 14th days. Biomechanical testing, light and electronic microscopy were used.Results: Following the cross-linking treatment, eyes in the cross-linking group had a shorter axial length compared to those in the non-crosslinking group (p = 0.006). Collagen fibrils larger than 240 nm were observed in the scleral stroma of cross-linking group, which were absent in the scleral stroma of the non-crosslinking and untreated control group. The mean ultimate stress and Young’s modulus was significantly greater in the cross-linking group compared to those in the non-crosslinking and untreated control group (p < 0.05). No histological damage observed in the retina or choroid.Conclusions: This study demonstrates that lens-induced axial elongation in rabbits can be effectively blocked by cross-linking using glyceraldehyde, with anatomical and mechanical modification and no deleterious effects.

Study of retina and choroid biological parameters of rhesus monkeys eyes on scleral collagen cross-linking by riboflavin and ultraviolet A.

To evaluate ocular fundus biological changes after scleral collagen cross-linking (CXL) with riboflavin/ ultraviolet A (UVA) on rhesus monkeys in vivo by analyzing retina and choroid biological parameters. Six 3-year-old male rhesus monkeys (12 eyes) were observed in this study, with scleral CXL procedures applied on superior temporal equatorial sclera on random eyes of all rhesus. Optical coherence tomography (OCT) and optical coherence tomography angiography (OCTA) examination were performed before and 1 week, 1 month, 3 months and 6 months after CXL. The thickness of retina and choroid and the flow density of retinal superficial vascular networks were analyzed respectively in different regions after CXL. As for retina thickness and flow density of retinal superficial vascular networks, no statistical difference was noted between CXL eyes and control eyes at 1 day, 1week, 1 month, 3 months and 6 months (P>0.05). Among choroid parameters, the choroidal thickness in 1500μm temporal to the fovea center of CXL eyes revealed a significant reduction in 1 week postoperatively (P<0.05), but it subsequently increased from 1 month postoperatively, and no statistical difference was found between two groups in the following periods (P>0.05). The choroidal thickness nearby crosslinked region may change temporarily following scleral CXL, and it might recover gradually after 1 month postoperatively. The vascular flow density and thickness of retina were not affected by scleral CXL. Further study should be performed to evaluate the potential adverse effects at the direct vicinity of the application site and the long-term effect of scleral CXL in clinical application.

Iontophoresis-assisted accelerated riboflavin/ultraviolet A scleral cross-linking: A potential treatment for pathologic myopia

Scleral collagen cross-linking is one of the most promising treatments to control the pathologic process of myopia. However, the exact procedure and its impact on animal models of myopia are still to be explored. We modified the scleral riboflavin/ultraviolet A (UVA) cross-linking procedure with an iontophoresis-assisted drug delivery system and an accelerated UVA irradiation (10 mW/cm2, 9 min) and applied this treatment to an animal model of myopia. Ninety-six New Zealand White rabbits developed relatively stable myopia by visual deprivation and then underwent the modified scleral cross-linking surgery. All the statistics and sample collection were obtained from 4 postoperative time points (1-day, 10-day, 1-month and 3-month groups). We found that the ultimate stress, Young's modulus and physiological Young's modulus of treated myopia sclera were significantly increased and maintained in 4 groups. The abnormal elongation of the myopic eye was effectively controlled 1 month after the treatment and even almost halted 3 months after the treatment. The histochemical assay revealed no notable post-surgery damage or apoptosis in the retina and choroid. Vigorous collagen synthesis was observed in scleral fibroblasts of the treated samples but were rarely observed in the untreated ones under electron microscopy. Furthermore, the remarkable difference in collagen gene expression and protein content between treated and untreated samples also indicated that an alteration in collagen metabolism may be triggered by the treatment. The effectiveness and safety exploration suggested that the modified scleral cross-linking procedure may be a potential method to control the pathologic process of myopia.