EXPERIMENTAL STUDY ON THE CORNEAL ENDOTHELIUM OF TRAUMATIC CATARACT

The cell morphology of corneal endothelium in 84 mice with experimental traumatic cataract was investigated with stained corneal buttons. In the experimental group, the boundaries between adjacent corneal endothelial cells were significantly distorted, some cell boundaries manifested degenerative changes that led to coalescence of the cells. The mean density and mean area of endothelial cells of the controls showed significant difference from those of the experimental group during the 12 weeks of observ...

Evaluation of corneal endothelium after UVA/riboflavin cross-linking in thin keratoconic corneas

Dear Sir,UVA/riboflavin cross-linking （CXL） has been used clinically applied for the treatment of keratoconus and corneal edema via enhancement of corneal stiffness The safety of the corneal endothelium is of prime importance during CXL treatment. In clinical practice, a corneal thickness （CT） of 400um has traditionally been regarded as the minimum treatable thickness, thereby avoiding damage to the corneal endothelium Although CXL has been applied to thinner corneas, using a hypoosmotic solution onto cornea and inducing edema . CXL safety still needs further evaluation because of lower relative concentration of collagen in the hydrated stroma . This study aims to evaluate the changes of corneal endothelial density （ECD） in cases where the CT is 〈400 um before iatrogenic corneal swelling and CXL treatment.

Short term effects of small incision lenticule extraction surgery on corneal endothelium

AIM： To assess the effects of small incision lenticule extraction（SMILE） surgery on the corneal endothelium at1 d to 1mo postoperatively.·METHODS： A retrospective, observational study was conducted on 47 patients（47 eyes） who received SMILE surgery. Patients were grouped according to contact lens wear condition. The corneal endothelium was examined preoperatively and at 1d, 1wk and 1mo postoperatively.The corneal endothelium was analyzed for endothelial cell density（ECD）, percentage of hexagonal cells, and coefficient of variation（CV） of cell size.·RESULTS： There were no significant decrease in the ECD, percentage of hexagonal cells or increase in CV at1 d, 1wk and 1mo postoperatively（P 〉0.05）. However,there was a small increase of ECD by 2.88% in contact lens wearers（78.26±113.62 cell/mm2, P 〈0.05）.· CONCLUSION： SMILE has no significant adverse effects on the corneal ECD and morphology during 1mo follow-up time.

Differences in energy and corneal endothelium between femtosecond laser-assisted and conventional cataract surgeries：prospective,intraindividual,randomized controlled trial

AIM： To compare intraoperative phacoemulsification parameters and its effect on the corneal endothelium of eyes undergoing femtosecond laser-assisted cataract surgery（FLACS） versus conventional phacoemulsification（CP） cataract surgery.METHODS： Two hundred eyes from one hundred patients were included in a prospective, non-blinded, randomized, controlled, intraindividual clinical study. One hundred eyes underwent FLACS while their one hundred fellow eyes underwent CP. All surgeries were performed using the Victus？ femtosecond laser platform and Infinity？ Vision System phacoemulsification machine. Primary outcome measure was endothelial cell density 6 mo after surgery. Secondary outcome measures included central corneal thickness（CCT）, average cell area, standard deviation, coefficient of variation and hexagonality before surgery and 6 mo after surgery and endothelial cell density loss during this period were also evaluated. Intraoperative efficiency parameters [cumulative dissipated energy（CDE）, total intraocular surgery time, total ultrasound time, total phacoemulsification time, total torsional energy time, total aspiration time, ultrasound energy, torsional amplitude and fluid required during surgery] were also collated. RESULTS： Data from these patients was not considered for analysis. Data from 92 patients were analysed. Postoperative endothelial cell density（cells/mm2） between groups（2211.88±392.49 CP; 2246.31±403.48 FLACS） was not statistically significant（P=0.869）. Total ultrasound time, torsional energy time, CDE and fluid requirements were significantly lower the FLACS group（P〈0.05）. Other parameters did not show statistically significant difference between FLACS and CP.CONCLUSION： FLACS displays significant improvements in phacoemulsification parameters in comparison to CP. There are no significant differences in corneal endothelium measures between FLACS and CP.

Effect of topical 0.05% cyclosporine A on corneal endothelium in patients with dry eye disease

· AIM: To determine the effect of topical 0.05% cyclosporine A (CsA) on corneal endothelium in patients with dry eye disease. · METHODS: Observational, prospective, case series study. Fifty-five eyes of 29 consecutive patients (9 males and 20 females; median age: 66.8 years, interquartile range: 61 -73.2 years) with moderate -severe dry eye disease were evaluated. All patients were treated with topical 0.05% CsA ophthalmic emulsion twice a day in addition to lubricant eyedrops 5 times a day. The follow- up period was 12 months. Before treatment and at 3 and 12 months post -treatment central corneal specular microscopy was performed. The endothelial cell density (ECD), coefficient of variation of cell size (CoV), and percentage of hexagonal cells (Hex %) were analyzed. ·RESULTS: The median ECDs pre-treatment and at 3 and 12 months post-treatment were 2 352.5/mm 2 (inter- quartile range, 2 178 -2548.5), 2 364/mm 2 (interquartile range, 2 174.25 -2 657.5), and 2 366 cells/mm 2 (inter - quartile range, 2 174.75-2 539.75), respectively (P=0.927, one way ANOVA). The median CoVs pre-treatment and at 3 and 12 months post -treatment were 34.5 (interquartile range, 30 -37), 35 (interquartile range, 30 -38), and 34 (interquartile range, 30.75-38.25), respectively (P=0.7193, one way ANOVA). The median Hex % values pre - treatment and at 3 and 12 months post -treatment were 53 (interquartile range, 47 -58), 54 (interquartile range, 45.75 -59), and 50.5 (interquartile range, 45.75 -58), respectively (P=0.824, one way ANOVA). · CONCLUSION: Treatment of patients with dry eye disease for 12 months with topical 0.05% CsA does not seem to cause substantial changes on corneal endothelium.

Transplanting embryonic stem cells onto damaged human corneal endothelium

AIM To investigate whether human embryonic stem cells(hESCs) could be made to attach, grow and differentiate on a human Descemet's membrane(DM).METHODS Spontaneously differentiated hESCs were transferred onto a human corneal button with the endothelial layer removed using ocular sticks. The cells were cultured on a DM for up to 15 d. The genetically engineered hESC line expressed green fluorescent protein, which facilitated identification during the culture experiments, tissue preparation, and analysis. To detect any differentiation into human corneal endothelial-like cells, we analysed the transplanted cells by immunohistochemistry using specific antibodies.RESULTS We found transplanted cells form a single layer of cells with a hexagonal shape in the periphery of the DM. The majority of the cells were negative for octamer-binding transcription factor 4 but positive for paired box 6 protein, sodium potassium adenosine triphosphatase(NaKATPase), and Zona Occludens protein 1. In four of the 18 trials, the transplanted cells were found to express CK3, which indicates that the stem cells differentiated into corneal epithelial cells in these cases. CONCLUSION It is possible to get cells originating from hESCs to become established on a human DM, where they grow and differentiate into corneal endothelial-like cells in vitro.

STUDY IN CYTOTOXICITY OF GENTAMYCIN TO CORNEAL EPITHELIUM AND ENDOTHELIUM IN TISSUE CULTURE

A study in cytotoxicity of gentamyein to tissue-cultured bovine corneal endothelial cells and rabbit corneal epithelial cells is reported. When the cultured cells reached confluence, they were exposed to tissue culture media containing gentamycin for 6 hours. We founl that 0.5% gentamycin caused significant damage to corneal epithelial cells---diffuse plasmolysis, with scattered cell necrosis and 5% loss.While corneal endothelial cells were exposed to 1.6 mg/ml gentamycin, extensive cell loss (approximate- ly 15%) was observed. The damaged cells recovered their normal morphology after 24 hours. When the concentration of gentamycin increased twice, serious damage to cells occured. The area of cell loss reached 40%, and the recovery of cellular morphology Was much slower. This study demonstrates that gentamycin potential cytotoxicity to corneal epithelium and endothelium, suggesting that gentamycin should be rationally used in the treatment of ocular diseases.

Hammered silver appearance of the corneal endothelium in Fuchs uveitis syndrome:A case report

BACKGROUND Hammered silver appearance of the corneal endothelium is considered a characteristic change in iridocorneal-endothelial syndrome.Herein we report an interesting case of hammered silver appearance of the corneal endothelium in Fuchs uveitis syndrome(FUS).CASE SUMMARY A 49-year-old man with progressive vision loss in the right eye for one year was admitted to our hospital.The clinical manifestations of the patient’s right eye were mild conjunctival hyperemia,scattered stellate keratic precipitates on the corneal endothelium,normal depth anterior chamber,2+cellular reaction in the aqueous humor,diffuse iris depigmentation,absence of synechia,Koeppe nodules,opalescent lens,and vitreous opacity.FUS and a complicated cataract were diagnosed based on the typical clinical manifestations.The corneal endothelial changes were recorded in detail by slit-lamp examination,specular microscopy,and in vivo confocal microscopy before cataract extraction,revealing a hammered silver appearance of the corneal endothelium in the affected eye,a wide-band dark area,as well as irregular corneal endothelial protuberances and dark bodies of various sizes.Subsequently,the patient underwent phacoemulsification combined with intraocular lens implantation,and his postoperative visual acuity recovered to 1.0.CONCLUSION Hammered silver appearance of the corneal endothelium in FUS,which is considered a more serious manifestation of endothelial damage,is rare and may be caused by many irregular protrusions in the corneal endothelium.

AB086.Influence of the intraocular pressure on the expression of tight junctions in the corneal endothelium

Background:Cells are influenced by their environment.In vivo,the corneal endothelium is subjected to intraocular pressure(IOP).The purpose of this project was to evaluate in vitro,the effect of the IOP on the formation of tight junctions in the corneal endothelium.Methods:Cultivated corneal endothelial cells(P2-P3;n=6 populations)were seeded on devitalized on corneas(n=10 pairs).Native corneas and devitalized corneas were respectively used as positive(n=2 pairs)and negative controls(n=3 pairs).Corneas were placed in artificial anterior chambers and subjected to a hydrostatic pressure between 0.3 and 0.4 psi during 4-5 days.Unpressured control corneas were maintained in cell culture dishes.Pictures of the corneas were taken following the experiment to assess stromal transparency.Morphology,corneal thickness and distribution of ZO-1,n-cadherin,b-catenin,NaK ATPase pump and HCO3-cotransporter were evaluated by electron microscopy,histological staining and immunofluorescences.Results:Pressure treated corneas were more transparent than the controls.Thickness was accordingly reduced by 38.4%±4.9%for cultivated endothelium and 32.2%±2.7%for native endothelium.Negative controls change in transparency and thickness were marginal.Pressure treated cells showed none or at most marginal difference in morphology and expression of ZO-1,n-cadherin,b-catenin,NaK ATPase pump and HCO3-cotransporters and failed to recreate a phenotype similar to native corneas.Pressure however increased cortical localisation of the protein ZO-1 in both cultivated and native endothelium.Conclusions:These results suggest that anterior chamber hydrostatic pressure may enhance endothelial functionality by modulating the distribution of tight junction’s proteins.

The role of corneal endothelium in macular corneal dystrophy development and recurrence

Macular corneal dystrophy(MCD)is a progressive,bilateral stromal dystrophic disease that arises from mutations in carbohydrate sulfotransferase 6(CHST6).Corneal transplantation is the ultimate therapeutic solution for MCD patients.Unfortunately,postoperative recurrence remains a significant challenge.We conducted a retrospective review of a clinical cohort comprising 102 MCD patients with 124 eyes that underwent either penetrating keratoplasty(PKP)or deep anterior lamellar keratoplasty(DALK).Our results revealed that the recurrence rate was nearly three times higher in the DALK group(39.13%,9/23 eyes)compared with the PKP group(10.89%,11/101 eyes),suggesting that surgical replacement of the corneal endothelium for treating MCD is advisable to prevent postoperative recurrence.Our experimental data confirmed the robust m RNA and protein expression of CHST6 in human corneal endothelium and the rodent homolog CHST5 in mouse endothelium.Selective knockdown of wild-type Chst5 in mouse corneal endothelium(AC^(siChst5)),but not in the corneal stroma,induced experimental MCD with similar extracellular matrix synthesis impairments and corneal thinning as observed in MCD patients.Mice carrying Chst5 point mutation also recapitulated clinical phenotypes of MCD,along with corneal endothelial abnormalities.Intracameral injection of wild-type Chst5 rescued the corneal impairments in AC^(siChst5)mice and retarded the disease progression in Chst5 mutant mice.Overall,our study provides new mechanistic insights and therapeutic approaches for MCD treatment by highlighting the role of corneal endothelium in MCD development.

Effect of mitomycin C on the corneal endothelial cells of Saudi patients with myopia after transepithelial photorefractive keratectomy:a two-armed cohort study

AIM:To study the effect of mitomycin C(MMC)applied during transepithelial photorefractive keratectomy(TPRK)on the corneal endothelium one week(W1)and three months(M3)after surgery and its determinants.METHODS:In this two-armed cohort study conducted in 2022,eyes treated with MMC during TPRK(group 1)were compared with eyes not treated with MMC(group 2).The corneal endothelial cell(EC)count,EC density(ECD;cells/mm2),average(μm2),standard deviation(μm2),coefficient of variation(CV%),ECmax,ECmin,and EC percentage of hexagonality were estimated at W1 and M3.The postoperative changes in the EC count in the two groups were compared and correlated with the other independent variables.RESULTS:Group 1 had 26 eyes,and group 2 had 78 eyes.All TPRK indices were significantly higher for the eyes in group 1 than for those in group 2.The MMC usage was not a significant predictor of the change in ECD(P=0.644),change in CV(P=0.374),and change in the percentage of hexagonality of EC(P=0.164)at W1.However,the use of MMC was a significant predictor of changes in CV(P=0.014)and the change in the percentage of hexagonality of EC(P=0.039)at M3.The duration of laser exposure and the size of the optical zone influenced the correlation of MMC use with the changes in EC indices,postoperatively.CONCLUSION:The use of MMC doesn’t affect ECD,CV,and percentage of hexagonality at W1 if other surgical indices are considered.At M3 after operating myopic eyes by TPRK,MMC significantly influence the CV and percentage of hexagonality.The duration of the laser application and the size of the optical zone should be considered when determining the effect of MMC on the EC indices.

Surgical approaches to correct corneal astigmatism at time of cataract surgery: a mini-review

Among refractive errors,astigmatism is the most common optical aberration,where refraction changes in different meridians of the eye.It causes blurred vision at any distance and includes corneal,lenticular,and retinal astigmatism.Cataract surgery used to cause a progressive increase in the pre-exisiting corneal astigmatism because of creating a surgically induced astigmatism,for example,a large size surgery incision.The development of surgical techniques during last decades has made cataract surgery interchange to treat preoperative corneal astigmatism at time of surgery.Nowadays,three surgical approaches can be used.By placing a sutureless clear corneal incision on the steep meridian of the cornea,a preoperative corneal astigmatism less than 1.0 D can be corrected.Single or paired peripheral corneal relaxing incisions(PCRIs)provide 1.0-3.0 D corneal astigmatism correction.PCRIs are typically used for treating 1.0-1.5 D of regular corneal astigmatism,if more than 2.0 D,the risk of overcorrection and irregular astigmatism is increased.When toric intraocular lenses(IOLs)are unavailable in markets,PCRIs are still a reasonable option for patients with up to 3.0 D of pre-existing corneal astigmatism.Toric IOLs implantation can correct 1.0-4.5 D of corneal astigmatism.Several IOLs are approved to correct a high degree of corneal astigmatism with cylinder power up to 12.0 D.These approaches can be used alone or in combination.

The Limbal Niche and Its Role in Maintaining Corneal Regeneration

In recent years, stem cells have been a focal point in research designed to evaluate the efficacy of ophthalmologic therapies, specifically those for corneal conditions. The corneal epithelium is one of the few regions of the body that maintains itself using a residual stem cell population within the adjacent limbus. Stem cell movement has additionally captivated the minds of researchers due to its potential application in different body regions. The cornea is a viable model for varying methods to track stem cell migratory patterns, such as lineage tracing and live imaging from the limbus. These developments have the potential to pave the way for future therapies designed to ensure the continuous regeneration of the corneal epithelium following injury via the limbal stem cell niche. This literature review aims to analyze the various methods of imaging used to understand the limbal stem cell niche and possible future directions that might be useful to consider for the better treatment and prevention of disorders of the cornea and corneal epithelium. .

Clinical associations of corneal neuromas with ocular surface diseases

Corneal neuromas,also termed microneuromas,refer to microscopic,irregula rly-shaped enlargements of terminal subbasal nerve endings at sites of nerve damage or injury.The formation of corneal neuromas results from damage to corneal nerves,such as following corneal pathology or corneal or intraocular surge ries.Initially,denervated areas of sensory nerve fibers become invaded by sprouts of intact sensory nerve fibers,and later injured axons regenerate and new sprouts called neuromas develop.In recent years,analysis of corneal nerve abnormalities including corneal neuromas which can be identified using in vivo confocal microscopy,a non-invasive imaging technique with microscopic resolution,has been used to evaluate corneal neuropathy and ocular surface dysfunction.Corneal neuromas have been shown to be associated with clinical symptoms of discomfort and dryness of eyes,and are a promising surrogate biomarker for ocular surface diseases,such as neuropathic corneal pain,dry eye disease,diabetic corneal neuropathy,neurotrophic keratopathy,Sjogren's syndrome,bullous keratopathy,post-refra ctive surgery,and others.In this review,we have summarized the current literature on the association between these ocular surface diseases and the presentation of corneal microneuromas,as well as elaborated on their pathogenesis,visualization via in vivo confocal microscopy,and utility in monitoring treatment efficacy.As current quantitative analysis on neuromas mainly relies on manual annotation and quantification,which is user-dependent and labor-intensive,future direction includes the development of artificial intelligence software to identify and quantify these potential imaging biomarkers in a more automated and sensitive manner,allowing it to be applied in clinical settings more efficiently.Combining imaging and molecular biomarkers may also help elucidate the associations between corneal neuromas and ocular surface diseases.

The Mean Central Cornea Thickness and Associated Factors among Adult Patients Attending Mulago Hospital Eye Clinic Kampala, Uganda

Background: The mean central corneal thickness (CCT) values are known to have an influence on intraocular pressure measurement and vary widely with age, sex and ethnicity. Little is known about the pattern of CCT in central Uganda. Aim: To determine the mean central corneal thickness and its associated factors among adult patients attending Mulago Hospital eye clinic, Kampala Uganda. Design and Method: A cross sectional study was done from August to October 2016 on 270 adult patients selected by systemic random sampling. The study included all adult patients who attended the eye clinic during the study period and consented to the study excluding those who had conditions causing abnormal thinning, thickening or irregularity of the cornea. The participants were evaluated by history taking and ocular examination including visual acuity assessment, slit lamp examination, central cornea thickness measurement, intraocular pressure measurement and funduscopy. Data were collected using a pretested questionnaire, entered using Epidata and analyzed by STATA11.0. Results: Of the 270 patients examined, 61% were female, 39% male, the mean age being 42 years ranging from 18 to 96 years. The mean central corneal thickness for both eyes was 539.3 (SD ± 41.65 μm) ranging from 332 μm to 701 μm and mean intraocular pressure was 15 (SD ± 5.34) mmHg. Gender, ethnicity, IOP, glaucoma, hypertension, diabetes, refractive error, were not statistically significantly associated with central cornea thickness. However central cornea thickness was statistically significantly associated with systolic blood pressure (r = -0.24, p = 0.025) and age (p = 0.002). A one-year increase in age reduced the mean central cornea thickness by 0.54 μm. Conclusion: The average CCT of Ugandan adult population was 539.3 ± 41.65 μm which falls in the category of normal central cornea thickness. Central corneal thickness reduced with increasing age and increasing systolic blood pressure.

Effects of moxibustion at 46°C on blood lipids and related indicators of thoracic aortic endothelium in a hyperlipidemia rat model

Objective:To observe the effects of moxibustion at different temperatures(38℃,46℃)on blood lipids,endothelial morphology of the thoracic aorta,serum endothelin-1(ET-1),calcitonin gene-related peptide(CGRP),nitric oxide(NO),and endothelial NO synthase(eNOS)in hyperlipidemic rats.Methods:Using the random number table method,60 Sprague Dawley rats were randomly and evenly divided into blank,model,38℃-moxibustion,and 46℃-moxibustion groups.Rats in the 3 experimental groups were fed a high-fat feed to model hyperlipidemia in rats.Rats in the 38℃-moxibustion and 46℃-moxibustion groups were moxibustion on the Shenque and bilateral Zusanli acupoints for 10 minutes each,once every other day for 4 weeks,at temperatures of 38±1℃ and 46±1℃.After that,rat blood samples were collected to detect blood lipids and ET-1,CGRP,eNOS and NO.Take the endotheal tissue of the thoracic aorta to do HE staining.Results:(1)The serum total cholesterol,triglycerides,and low-density lipoprotein cholesterol of rats in the 46℃-moxibustion group were significantly lower than those in the model and 38℃-moxibustion groups.(2)Revealed by hematoxylin and eosin staining,showed necrosis in the local vascular endothelial cells and mild inflammatory cell infiltration in the tunica adventitia of the hyperlipidemic rats.These endothelial morphologies did not improve significantly after moxibustion at 38℃ but did improve at 46℃.(3)Compared with the blank group,serum ET-1 was significantly higher and serum CGRP,NO,eNOS were significantly lower in the model group.Compared with the model and the 38℃-moxibustion groups,serum ET-1 was significantly lower and serum CGRP,NO,eNOS were significantly higher in 46℃-moxibustion groups.Conclusion:Moxibustion at 46℃ effectively regulated blood lipids,improved the morphology of the vascular endothelium,and protected vascular endothelial function.

Research progress on animal models of corneal epithelial-stromal injury

A corneal epithelial-stromal defect is recognized as a major contributor to corneal scarring.Given the rising prevalence of blindness caused by corneal scarring,increasing attention has been focused on corneal epithelialstromal defects.Currently,the etiology and pathogenesis of these defects remain inadequately understood,necessitating further investigation through experimental research.Various modeling methods exist both domestically and internationally,each with distinct adaptive conditions,advantages,and disadvantages.This review primarily aims to summarize the techniques used to establish optimal animal models of corneal epithelial-stromal injury,including mechanical modeling,chemical alkali burns,post-refractive surgery infections,and genetic engineering.The intention is to provide valuable insights for studying the mechanisms underlying corneal epithelial-stromal injury and the development of corresponding therapeutic interventions.

Single-cell RNA sequencing in cornea research:Insights into limbal stem cells and their niche regulation

The corneal epithelium is composed of stratified squamous epithelial cells on the outer surface of the eye,which acts as a protective barrier and is critical for clear and stable vision.Its continuous renewal or wound healing depends on the proliferation and differentiation of limbal stem cells(LSCs),a cell population that resides at the limbus in a highly regulated niche.Dysfunction of LSCs or their niche can cause limbal stem cell deficiency,a disease that is manifested by failed epithelial wound healing or even blindness.Nevertheless,compared to stem cells in other tissues,little is known about the LSCs and their niche.With the advent of single-cell RNA sequencing,our understanding of LSC characteristics and their microenvironment has grown considerably.In this review,we summarized the current findings from single-cell studies in the field of cornea research and focused on important advancements driven by this technology,including the heterogeneity of the LSC population,novel LSC markers and regulation of the LSC niche,which will provide a reference for clinical issues such as corneal epithelial wound healing,ocular surface reconstruction and interventions for related diseases.

Central Corneal Thickness of Diabetic Patients in Yaoundé: Case Control Study

Background: To compare central corneal thickness (CCT) values measured in diabetic eyes and compare the CCT values in normal patients without diabetes. Methods: A total of 83 diabetic patients and 83 non-diabetic patients were prospectively enrolled in this comparative case series. CCT was measured using a Topcon CT-1P non-contact automated pachymeter, and values were compared. Results: Mean age was 57 ± 11 years, with a range of 23 and 80 years. The mean CCT was 508.87 ± 35.83 μm for diabetic patients and 513.41 ± 37.22 μm for non-diabetic patients. There was no statistically significant difference between the CCT of diabetic patients compared to non-diabetic patients (p = 0.260). We did not find a significant correlation between the CCT of diabetic patients and clinical characteristics of diabetes. However, there was a positive and statistically significant linear correlation between the CCT of diabetic patients and intra ocular pressure (IOP). Conclusion: The mean CCT in diabetic patients was lower than in non-diabetic control patients, although there was no statistically significant difference.

Preliminary studies of constructing a tissue-engineered lamellar corneal graft by culturing mesenchymal stem cells onto decellularized corneal matrix

AIM:To construct a competent corneal lamellar substitute in order to alleviate the shortage of human corneal donor.METHODS:Rabbit mesenchymal stem cells(MSCs)were isolated from bone marrow and identified by flow cytometric,osteogenic and adipogenic induction.Xenogenic decellularized corneal matrix(XDCM)was generated from dog corneas.MSCs were seeded and cultured on XDCM to construct the tissueengineered cornea.Post-transplantation biocompatibility of engineered corneal graft were tested by animal experiment.Rabbits were divided into two groups then underwent lamellar keratoplasty(LK)with different corneal grafts:1)XDCM group(n=5):XDCM;2)XDCM-MSCs groups(n=4):tissue-engineered cornea made up with XDCM and MSCs.The ocular surface recovery procedure was observed while corneal transparency,neovascularization and epithelium defection were measured and compared.In vivo on focal exam was performed 3 mo postoperatively.RESULTS:Rabbit MSCs were isolated and identified.Flow cytometry demonstrated isolated cells were CD90 positive and CD34,CD45 negative.Osteogenic and adipogenic induction verified their multipotent abilities.MSC-XDCM grafts were constructed and observed.In vivo transplantation showed the neovascularization in XDCMMSC group was much less than that in XDCM group postoperatively.Post-transplant 3-month confocal test showed less nerve regeneration and bigger cell-absent area in XDCM-MSC group.CONCLUSION:This study present a novel corneal tissue-engineered graft that could reduce post-operatively neovascularization and remain transparency,meanwhile shows that co-transplantation of MSCs may help increase corneal transplantation successful rate and enlarge the source range of corneal substitute to overcome cornea donor shortage.