Small-diameter acellular porcine corneal stroma for peripheral corneal ulceration treatment

AIM:To evaluate the clinical efficacy of small-diameter acellular porcine corneal stroma(SAPS)for the treatment of peripheral corneal ulceration(PCU).METHODS:This retrospective clinical study included 18 patients(18 eyes)with PCU between April 2018 and December 2020.All patients had PCU and underwent lamellar keratoplasty with SAPS.Observation indicators included preoperative and postoperative best-corrected visual acuity(BCVA)and transparency of SAPS.The infection control rate in the surgical eye-lesion area was also calculated.RESULTS:Eighteen patients underwent lamellar keratoplasty with SAPS to treat PCU.None of the patients experienced rejection after 6mo(18/18)and 12mo(16/16)of follow-up.The BCVA(0.47±0.30)at the 6mo followup after operation was significantly improved compared with the baseline(0.99±0.80),and the difference was statistically significant(Z=-3.415,P<0.05).The BCVA at the 12mo follow-up after operation was not statistically significant compared to the 6mo(Z=0,P=1).With time,the SAPS graft gradually became transparent.At the 6mo(18/18)and 12mo(16/16)follow-up,none of the patients had recurrent corneal infection.CONCLUSION:SAPS is clinically effective in the treatment of PCU,improving the patient’s BCVA and reducing the incidence of rejection after keratoplasty.

Stromal lenticule addition keratoplasty with corneal crosslinking for corneal ectasia secondary to FS-LASIK:a case series

●AIM:To explore the clinical efficacy and safety of stromal lenticule addition keratoplasty(SLAK)with corneal crosslinking(CXL)on patients with corneal ectasia secondary to femtosecond laser-assisted in situ keratomileusis(FS-LASIK).●METHODS:A series of 5 patients undertaking SLAK with CXL for the treatment of corneal ectasia secondary to FS-LASIK were followed for 4-9mo.The lenticules were collected from patients undertaking small incision lenticule extraction(SMILE)for the correction of myopia.Adding a stromal lenticule was aimed at improving the corneal thickness for the safe application of crosslinking and compensating for the thin cornea to improve its mechanical strength.●RESULTS:All surgeries were conducted successfully with no significant complications.Their best corrected visual acuity(BCVA)ranged from 0.05 to 0.8-2 before surgery.The pre-operational total corneal thickness ranged from 345-404μm and maximum keratometry(Kmax)ranged from 50.8 to 86.3.After the combination surgery,both the corneal keratometry(range 55.9 to 92.8)and total corneal thickness(range 413-482μm)significantly increased.Four out of 5 patients had improvement of corneal biomechanical parameters(reflected by stiffness parameter A1 in Corvis ST).However,3 patients showed decreased BCVA after surgery due to the development of irregular astigmatism and transient haze.Despite the onset of corneal edema right after SLAK,the corneal topography and thickness generally stabilized after 3mo.●CONCLUSION:SLAK with CXL is a potentially beneficial and safe therapy for advanced corneal ectasia.Future work needs to address the poor predictability of corneal refractometry and compare the outcomes of different surgical modes.

Covering corneal stromal lenticule for macular hole in pathological myopia

AIM:To evaluate the clinical effect of a new surgery technique(covering corneal stromal lenticule,CSL)for macular hole(MH)in pathological myopia.METHODS:This was a prospective non-randomized series case study.Fourteen eyes of 14 patients whose axial length were more than 29 mm and suffered from MH and macular hole retinal detachment(MHRD)were included in this study.All cases were treated with 25-gauge pars plana vitrectomy(PPV)with internal limiting membrane(ILM)peeling,covering CSL and C_(3)F_(8) gas tamponade.These cases were followed for 6mo,and the best-corrected visual acuity(BCVA),healing status of MH,the reattached rate of retinal detachment(RD),and reoperation rate were analyzed.RESULTS:All cases were successfully performed the surgery and the postoperative follow-up was completed.After surgery,MHs were healed in all 14 eyes(100%,14/14)after assessed by optical coherence tomography.The reattachment of retina was achieved in all 6 eyes(100%,6/6)with MHRD.BCVA was improved in 12 eyes(85.71%,12/14),and had no significant change in 2 eyes(14.29%,2/14).The overall mean BCVA was improved from 1.80±0.77 to 0.82±0.46 logMAR(F=10.46,P<0.01).No serious complications occurred in all cases.CONCLUSION:The new surgery technique(covering CSL)has high reattached rate of RD and high healing rate of MH in pathological myopia in the preliminary study.And it can effectively improve the visual function of patients.This new technique offers meaningful new ideas for treating refractory MH in pathological myopia.

Treatment of superficial corneal opacities with corneal stromal lenticule obtained through SMILE surgery

AIM:To evaluate the clinical efficacy and feasibility of superficial corneal opacities treated by excimer laser phototherapeutic keratectomy(PTK)combined with small incision lenticule extraction(SMILE)-derived corneal stromal lenticule transplantation.METHODS:A retrospective interventional case series of nine patients aged 12-59y with superficial corneal opacity caused by different pathologies who underwent standardized PTK combined with SMILE-derived corneal stromal lenticule transplantation was examined.Lenticule patches were fixed with fibrin glue.All patients underwent pre-and post-operative clinical assessments at different times for up to 12mo.Slit lamp microscopy,corneal density,uncorrected distance visual acuity(UDVA),corrected distance visual acuity(CDVA),and anterior segment optical coherence tomography(AS-OCT)were examined.RESULTS:The patients’mean age was 36.00±5.80(12-59)y.Seven eyes(77.8%)gained UDVA and CDVA at the last measurement compared to their preoperative levels.The densities of the total cornea,the total anterior corneal layer,and the anterior corneal layers of 0-2 and 2-6 mm decreased significantly by 12.4%,27.5%,46.7%,and 32.8%,respectively.After human allogeneic transplantation,the implanted lenticules of all eyes were clearly visible by AS-OCT and remained transparent without displacement or graft rejection.The thickness of the central cornea and corneal lenticule transplants were stable throughout the entire postoperative period.One case experienced the postoperative complication of delayed corneal epithelial healing.CONCLUSION:PTK combined with SMILE-derived corneal lenticule transplantation improves long-term visual acuity.Therefore,it is a new,safe,and effective method for treating superficial corneal opacity.

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.

Establishment of an untransfected human corneal stromal cell line and its biocompatibility to acellular porcine corneal stroma

AIM: To establish an untransfected human corneal stromal (HCS) cell line and characterize its biocompatibility to acellular porcine corneal stoma (aPCS). METHODS: Primary culture was initiated with a pure population of HCS cells in DMEM/F12 media (pH 7.2) containing 20% fetal bovine serum and various necessary growth factors. The established cell line was characterized by growth property, chromosome analysis, tumorigenicity assay, expression of marker proteins and functional proteins. Furthermore, the biocompatibility of HCS cells with aPCS was examined through histological and immunocytochemistry analyses and with light, electron microscopies. RESULTS: HCS cells proliferated to confluence 2 weeks later in primary culture and have been subcultured to passage 140 so far. A continuous untransfected HCS cell line with a population doubling time of 41.44 hours at passage 80 has been determined. Results of chromosome analysis, morphology, combined with the results of expression of marker protein and functional proteins suggested that the cells retained HCS cell properties. Furthermore, HCS cells have no tumorigenicity, and with excellent biocompatibility to aPCS. CONCLUSION: An untransfected and non-tumorigenic HCS cell line has been established, and the cells maintained positive expression of marker proteins and functional proteins. The cell line, with excellent biocompatibility to aPCS, might be used for in vitroreconstruction of tissue-engineered HCS.

Acellular ostrich corneal stroma used as scaffold for construction of tissue-engineered cornea

AIM： To assess acellular ostrich corneal matrix used as a scaffold to reconstruct a damaged cornea. METHODS： A hypertonic saline solution combined with a digestion method was used to decellularize the ostrich cornea. The microstructure of the acellular corneal matrix was observed by transmission electron microscopy （TEM） and hematoxylin and eosin （H＆E） staining. The mechanical properties were detected by a rheometer and a tension machine. The acellular corneal matrix was also transplanted into a rabbit cornea and cytokeratin 3 was used to check the immune phenotype, RESULTS： The microstructure and mechanical properties of the ostrich cornea were well preserved after the decellularization process, in vitro, the methyl thiazolyl tetrazoUum results revealed that extracts of the acellular ostrich corneas （AOCs） had no inhibitory effects on the proliferation of the corneal epithelial or endothelial cells or on the keratocytes, The rabbit lamellar keratoplasty showed that the transplanted AOCs were transparent and completely incorporated into the host cornea while corneal turbidity and graft dissolution occurred in the acellular porcine cornea （APC） transplantation, The phenotype of the reconstructed cornea was similar to a normal rabbit cornea with a high expression of cytokeratin 3 in the superficial epithelial cell layer, CONCLUSION： We first used AOCs as scaffolds to reconstruct damaged corneas. Compared with porcine corneas, the anatomical structures of ostrich corneas are closer to those of human corneas. In accordance with the principle that structure determines function, a xenograft lamellar keratoplasty also confirmed that the AOC transplantation generated a superior outcome compared to that of the APC graft.

Clinical outcomes after implantation of a new intrastromal corneal ring with 140-degree of arc in patients with corneal ectasia

AIM：To evaluate the clinical and tomographic outcomes after implantation of a new intrastromal corneal ring segment（ICRS） with 140-degrees of arc in eyes with corneal ectasia.METHODS：We evaluated patients with corneal ectasia implanted with Ferrara 140° ICRS from April 2010 to February 2015.Outcome measures included preoperative and postoperative corrected distance visual acuity（CDVA）,keratometry simulated（K） reading,tomographic astigmatism and asphericity.All patients were evaluated using the Pentacam Scheimpflug system.RESULTS：The study evaluated 58 eyes.The mean followup was 16.81±10.8 mo.The CDVA（logM AR） improved from 0.5±0.20（20/60） to 0.3±0.21（20/40）（P〈0.01）.The average K reduced from 49.87±7.01 to 47.34±4.90 D（P〈0.01）.The asphericity changed from-0.60±0.86 to-0.23±0.67 D（P〈0.01）.The mean preoperative tomographic astigmatism decreased from-8.0±3.45 to-4.53±2.52 D（P〈0.01）.CONCLUSION：The new ICRS model with 140-degrees of arc effectively improve the visual acuity and reduce the high astigmatism usually found in patients with corneal ectasia.

Local anesthetic lidocaine induces apoptosis in human corneal stromal cells in vitro

AIM:To demonstrate the apoptosis-inducing effect of iidocalne on human corneal stromal(HCS)cells fn vitm,and provide experimental basis for safety anesthetic usage In clinic of ophthalmology.METHODS:In vitro cultured HCS cells were treated with lidocaine at different doses and times,and their morphology was monitored successively with inverted phase contrast microscopy.The membrane permeability of them was detected by acridine orange/ethidium bromide(AO/EB)double staining.The DNA fragmentation of them was examined by agarose gel electrophoresis,and their ultrastructure was observed by transmission electron microscopy(TEM),respectively.RESULTS:Exposure to lidocaine at doses from0.3125g/L to 20g/L induced morphological changes of HCS cells such as cytoplasmic vacuolation,cellular shrinkage,and turning round,and elevated membrane permeability of these cells in AO/EB staining.The change of morphology and membrane permeability was doseand time-dependent,while lidocaine at dose below0.15625g/L could not induce these changes.Furthermore,lidocaine induced DNA fragmentation and ultrastructural changes such as cytoplasmic vacuolation,structural disorganization,chromatin condensation,and apoptotic body appearance of the cells.CONCLUSION:Lidocaine has significant cytotoxicity on human corneal stromal cells in vitro in a dose-and time-dependent manner by inducing apoptosis of these cells.The established experimental model and findingsbased on this model here help provide new insight into the apoptosis-inducing effect of local anesthetics in eye clinic.

Clinical outcomes after intrastromal corneal ring segments reoperation in keratoconus patients

AIM:To evaluate the clinical outcomes after Ferrara intrastromal corneal ring segments(ICRS)reoperation in patients with keratoconus.METHODS:A total of 37 keratoconus eyes implanted with intrastromal corneal ring segments,which had an ICRS exchange,addition,reposition or removal were evaluated.Uncorrected visual acuity(UCVA),best corrected visual acuity(BCVA),keratometry(K),asphericity(Q)and pachymetry at the thinnest point(PTP)of the cornea were evaluated using a corneal tomography(Oculus Pentacam,USA)RESULTS:The mean follow-up time after the reoperation was 30.5±9.7 months.The mean UCVA improved from 20/300 to 20/80(P=0.005);the mean BCVA improved from 20/160 to 20/50(P=0.0002),the mean keratometry reduced from 49.33±4.19D to 46.16±3.90D(P=0.0001),the mean pachymetry at the thinnest point increased from 450±42.9μm to 469±40.8μm(P=0.0001).The asphericity increased from-0.84±0.74 to-0.35±0.81(P=0.15)and the spherical equivalent reduced from-4.64±4.87D to-3.04±3.45D(P=0.137).The changes in the asphericity and spherical equivalent were not statistically significant.CONCLUSION:Ferrara ICRS implantation showed to be a reversible and readjustable surgical procedure for keratoconus treatment.Good outcomes can be obtained even after removal,addition,reposition or exchange of ICRS.

Corneal stromal mesenchymal stem cells: reconstructing a bioactive cornea and repairing the corneal limbus and stromal microenvironment

Corneal stroma-derived mesenchymal stem cells(CS-MSCs) are mainly distributed in the anterior part of the corneal stroma near the corneal limbal stem cells(LSCs). CS-MSCs are stem cells with self-renewal and multidirectional differentiation potential. A large amount of data confirmed that CS-MSCs can be induced to differentiate into functional keratocytes in vitro, which is the motive force for maintaining corneal transparency and producing a normal corneal stroma. CS-MSCs are also an important component of the limbal microenvironment. Furthermore, they are of great significance in the reconstruction of ocular surface tissue and tissue engineering for active biocornea construction. In this paper, the localization and biological characteristics of CS-MSCs, the use of CS-MSCs to reconstruct a tissue-engineered active biocornea, and the repair of the limbal and matrix microenvironment by CS-MSCs are reviewed, and their application prospects are discussed.

Preliminary results of a new intrastromal corneal ring segment as a tissue saving procedure in photorefractive keratectomy to correct moderate to high myopia

AIM:To evaluate the clinical results after implantation of a new intrastromal corneal ring segment(ICRS)associated with photorefractive keratectomy(PRK)to correct high myopia(HM)patients with thin corneas.METHODS:We evaluated 42 eyes of 23 HM patients that had ICRS implantation followed by PRK.The mean age of patients was 29.1±7.12 y(range 18 to 40 years old).Uncorrected visual acuity(UCVA),best corrected visual acuity(BCVA),keratometry,spherical equivalent,pachymetry,and aberrometry were compared using ANOVA with repeated measurements evaluated preoperatively and at last follow-up visit after the procedures.The refractive predictability and simulated/real corneal ablation were also assessed.RESULTS:The mean follow-up time after PRK was 6.8±1.6 mo.The mean preoperative UCVA improved from 20/800 preoperative to 20/100 after ICRS and 20/35 after PRK.The mean preoperative BCVA was 20/25(range from 20/30 to 20/20)and remained unchanged after ICRS implantation.Following the PRK the mean BCVA was 20/25(range from 20/30 to 20/20).The mean spherical equivalent decreased from-7.25±1.12(range-5.00 to-9.00)preoperatively to-3.32±1.0(range-2.00 to-5.00)postoperatively(P<0.001)after ICRS implantation and decreased from-2.44±1.51 preoperatively to 0.32±0.45(range-0.625 to 0.875)postoperatively(P<0.001)after PRK.The change in BCVA and topographic astigmatism was statistically significant(P<0.0001).CONCLUSION:ICRS in HM associated with PRK can be a tissue saving procedure and an alternative surgical option for correction of moderate to high myopia.

Cytotoxicity of pilocarpine to human corneal stromal cells and its underlying cytotoxic mechanisms

AIM： To examine the cytotoxic effect of pilocarpine, an anti-glaucoma drug, on human corneal stromal（HCS）cells and its underlying cytotoxic mechanisms using an in vitro model of non-transfected HCS cells.· METHODS： After HCS cells were treated with pilocarpine at a concentration from 0.15625 g/L to 20.0 g/L,their morphology and viability were detected by light microscopy and MTT assay. The membrane permeability,DNA fragmentation and ultrastructure were examined by acridine orange（AO）/ethidium bromide（EB） double-staining. DNA electrophoresis and transmission electron microscopy（TEM）, cell cycle, phosphatidylserine（PS）orientation and mitochondrial transmembrane potential（MTP） were assayed by flow cytometry（FCM）. And the activation of caspases was checked by ELISA.· RESULTS： Morphology observations and viability assay showed that pilocarpine at concentrations above0.625 g/L induced dose- and time-dependent morphological abnormality and viability decline of HCS cells. AO/EB double-staining, DNA electrophoresis and TEM noted that pilocarpine at concentrations above 0.625 g/L induced dose- and/or time-dependent membrane permeability elevation, DNA fragmentation, and apoptotic body formation of the cells. Moreover, FCM and ELISA assays revealed that 2.5 g/L pilocarpine also induced S phase arrest, PS externalization, MTP disruption, and caspase-8,-9 and-3 activation of the cells.· CONCLUSION： Pilocarpine at concentrations above0.625 g/L（1/32 of its clinical therapeutic dosage） has a dose- and time-dependent cytotoxicity to HCS cells by inducing apoptosis in these cells, which is most probably regulated by a death receptor-mediated mitochondrion-dependent signaling pathway.

Cell viability and extracellular matrix synthesis in a co-culture system of corneal stromal cells and adipose-derived mesenchymal stem cells

AIM：To investigate the impact of adipose-derived mesenchymal stem cells（ADSCs） on cell viability and extracellular matrix（ECM） synthesis of corneal stromal cells（CSCs）. METHODS：ADSCs and CSCs were obtained from the corneas of New Zealand white rabbits and indirectly cocultured in vitro. The proliferative capacity of CSCs in the different groups was assessed by CCK-8 assays. Annexin V-fluorescein isothiocyanate（FITC）/proliferation indices（PI） assays were used to detect the apoptosis of CSCs. The expression levels of matrix metalloproteinase（MMP）, such as MMP1, MMP2, MMP9, and collagens were also evaluated by Western blot. RESULTS：ADSCs significantly promoted proliferation and invasion of CSCs in the indirect co-culture assays. The co-cultural group displayed much higher ability of proliferation, especially under the co-culture conditions of ADSCs for 3d, compared with that CSCs cultured alone. The PI of CSCs in the co-culture system were increased approximately 3-8-fold compared with the control group. A significant change was observed in the proportions of cells at apoptosis（early and late） between the negative control group（6.34% and 2.06%） and the ADCSs-treated group（4.69% and 1.59%）. The expression levels of MMPs were down regulated in the co-culture models. Compared with the control group, the decrease intensities of MMP-1, MMP-2 and MMP-9 in CSCs/ADSCs group were observed, 3.90-fold, 1.09-fold and 3.03-fold, respectively. However, the increase intensities of collagen type（I, II, III, IV, and V） in CSCs were observed in CSCs/ADSCs group, 3.47-fold,4.30-fold, 2.35-fold, 2.55-fold and 2.43-fold, respectively, compared to that in the control group. The expressions of aldehyde dehydrogenase and fibronectin in CSCs were upregulated in the co-culture models.CONCLUSION：ADSCs play a promotive role in CSCs＇ growth and invasion, which may be partially associated with MMPs decrease and collagens increase, resulting in a positive participation in the plasticity and ECM synthesis of CSCs. This provided a new insight into the extensive role of ADSCs in CSCs and a potential molecular target for corneal therapy.

Effects of corneal stromal cell-and bone marrow-derived endothelial progenitor cell-conditioned media on the proliferation of corneal endothelial cells

AIM： To explore the effects of conditioned media on the proliferation of corneal endothelial cells （CECa） and to compare the efficiency of different conditioned media （CM）. METHODS： Rat CECs, corneal stromal cells （CSCs）, bone marrow -derived endothelial progenitor cells （BEPCs）, and bone marrow-derived mesenchymal stem cells （BMSCs） were isolated and cultured in vitra CM was collected from CSCs, BEPCs, and BMSCSo CECs were cultivated in different culture media. Cell morphology was recorded, and gene and protein expression were analyzed.~ RESULTS： After grown in CM for 5d, CECs in each experimental group remained polygonal, in a cobblestone- like monolayer arrangement. Immunocytofluorescence revealed positive expression of Na＋/K＋-ATP, aquaporin 1 （AQP1）, and zonula occludens 1 （ZO-1）. Based on quantitative polymerase chain reaction （qPCR） analysis, Na ＋/K ＋-ATP expression in CSC-CM was notably upregulated by 1.3-fold （＋0.036） （P〈0.05, n=3）. The expression levels of ZO-1, neuron specific enolase （NSE）, Vimentin, paired homebox 6 （PAX6）, and procollagen type VII （COL8A1） were notably upregulated in each experimental group. Each CM had a positive effect on CEC proliferation, and CSC-CM had the strongest effect on proliferation.~ CONCLUSION： CSC-CM, BEPC-CM, and BMSC-CM not only stimulated the proliferation of CECs, but also maintained the characteristic differentiated phenotypes necessary for endothelial functions. CSC-CM had the most notable effect on CEC proliferation. KEYWORDS： conditioned medium; corneal endothelial cell; corneal stromal cell; bone marrow-derived endothelial progenitor cell; proliferation

Aligned Fibrous Scaffold Induced Aligned Growth of Corneal Stroma Cells in vitro Culture

To investigate the contribution of fibre arrangement to guiding the aligned growth of corneal stroma cells,aligned and randomly oriented fibrous scaffolds of gelatin and poly-L-lactic acid（PLLA） were fabricated by electrospinning.A comparative study of two different systems with corneal stroma cells on randomly organized and aligned fibres were conducted.The efficiency of the scaffolds for inducing the aligned growth of cells was assessed by morphological observation and 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl-tetrazolium bromide（MTT） assay.Results show that the cells cultured on both randomly oriented and aligned scaffolds maintained normal morphology and well spreading as well as long term proliferation.Importantly,corneal stroma cells grew high orderly on the aligned scaffold,while the cells grew disordered on the randomly oriented scaffold.Moreover,the cells exhibited higher viability in aligned scaffold than that in randomly oriented scaffold.These results indcate that electrospinng to prepare aligned fibrous scaffolds has provided an effective approach to the aligned growth of corneal stroma cells in vitro.Our findings that fiber arrangement plays a crucial role in guiding the aligned growth of cells may be helpful to the development of better biomaterials for tissue engineered cornea.

Anti-inflammatory potential of human corneal stroma-derived stem cells determined by a novel in vitro corneal epithelial injury model

BACKGROUND An in vitro injury model mimicking a corneal surface injury was optimised using human corneal epithelial cells(hCEC).AIM To investigate whether corneal-stroma derived stem cells(CSSC) seeded on an amniotic membrane(AM) construct manifests an anti-inflammatory, healing response.METHODS Treatment of hCEC with ethanol and pro-inflammatory cytokines were compared in terms of viability loss, cytotoxicity, and pro-inflammatory cytokine release, in order to generate the in vitro injury. This resulted in an optimal injury of 20%(v/v) ethanol for 30 s with 1 ng/mL interleukin-1(IL-1) beta. Co-culture experiments were performed with CSSC alone and with CSSC-AM constructs.The effect of injury and co-culture on viability, cytotoxicity, IL-6 and IL-8 production, and IL1 B, TNF, IL6, and CXCL8 mRNA expression were assessed.RESULTS Co-culture with CSSC inhibited loss of hCEC viability caused by injury. Enzyme linked immunosorbent assay and polymerase chain reaction showed a significant reduction in the production of IL-6 and IL-8 pro-inflammatory cytokines, and reduction in pro-inflammatory cytokine mRNA expression during co-culture with CSSC alone and with the AM construct. These results confirmed the therapeutic potential of the CSSC and the possible use of AM as a cell carrier for application to the ocular surface.CONCLUSION CSSC were shown to have a potentially therapeutic anti-inflammatory effectwhen treating injured hCEC, demonstrating an important role in corneal regeneration and wound healing, leading to an improved knowledge of their potential use for research and therapeutic purposes.

Postoperative changes in corneal epithelial and stromal thickness profiles after SMILE in high myopic eyes

Background:To observe changes in the epithelial and stromal thickness after small incision refractive lenticule extraction(SMILE)and investigate their relationship with the different refractive error.Methods:One hundred and eighty eyes of 90 patients with a manifest refraction spherical equivalent(MRSE)of-6.36±1.53 diopters(D)were included.The eyes were assigned to the moderate myopic group(MRSE-3.00 to-6.00 D),high myopic group(MRSE-6.00 to-8.00 D)and super-high myopic group(MRSE above-8.00 D).The spectral-domain optical coherence tomography(SD-OCT)measured corneal and epithelial thickness in 17 zones preoperatively and 1,3,and 6 months postoperatively.Stromal thickness was calculated by subtracting the epithelial thickness from the total corneal thickness.The observed changes were correlated with the degree of myopia corrected.Results:MRSE showed significant differences between 3 and 6 months in the super-high myopic group(P=0.024).At 6 months,a statistically significant epithelial thickness increase was observed in the central zone(7.18%for moderate,10.23%for high,and 13.76%for super-high myopia,P<0.05 for all groups).The peripheral thickness decreased between 3 and 6 months in the high myopia and super-high myopia groups(P<0.05,respectively).A positive correlation between MRSE corrected and the postoperative epithelial thickening was observed in the central(r2=0.551,P<0.05).Compared to 1 month values,the central stromal thickness showed a decrease(3.2±4.5μm)at 3 months and an increase(4.4±4.9μm)at 6 months.The stroma thickened in moderate and high myopic groups but the thickness reduction were in super-high myopia group at 6 months paracentrally and peripherally.Conclusions:Significant thickness changes in the epithelium and stroma were detected during the 6 months after SMILE.Preliminary results suggest that epithelial and stromal thickness profile changes after SMILE may have an impact on the refractive outcome in the long-term postoperative period,especially in super-higher degrees of myopia.

The lived experiences of patients undergoing acellular porcine corneal stroma transplantation

To explore the lived experiences of patients undergoing acellular porcine corneal stroma（APCS）transplantation,a descriptive,qualitative design was performed.A purposive sample of 13 patients who underwent APCS transplantation to treat progressive infectious keratitis were enrolled in the semi-structured,open-ended interviews.The taped and transcribed interviews were analyzed using a thematic analysis approach.Alterations in the transparency of APCS grafts were accompanied by a gradual improved visual acuity（before surgery：1.38±0.91 logMAR;3mo postoperatively ：0.40±0.24 logMAR, respectively）.Accordingly,in terms of lived experiences,the patients generally reported＂negative＂experiences before the operation and during the early postoperative period,but this was greatly improved 3mo after surgery.Four main themes were derived：anxiety and fear,stigma,lifestyle change,and gratitude and insights. Conclusively,health care professionals should provide holistic care for patients,proactively promoting patients’physical and mental health.

TGFBI and CHST6 gene analysis in Chinese stromal corneal dystrophies

AIM:To investigate whether mutations in TGFBI gene or CHST6 gene correlated with stromal corneal dystrophies(CD) in 8 Chinese probands.· METHODS:Eight unrelated patients with stromal corneal dystrophies were recruited in this study;all affected members were assessed by completely ophthalmologic examinations.Genomic DNA was extracted from peripheral leukocytes,17 exons of TGFBI gene and the exon of CHST6 gene were amplified by polymerase chain reaction(PCR),sequenced directly and compared with the reference database.· RESULTS:Three heterozygous mutations in TGFBI gene were identified in six patients:c.370C>T(p.Arg124Cys) was found in exon 4 of TGFBI gene in three members,c.371G>A(p.Arg124His) was found in one patient;c.1663C>T(p.Arg555Trp) was found in exon 12 in other two members.In addition,four polymorphisms with the nucleotide changes rs1442,rs1054124,rs4669,and rs35151677 were found in TGFBI gene.Mutations were not identified in the rest of 2 affected individuals in TGFBI gene or CHST6 gene.· CONCLUSION:Within these patients,R124C,R124H and R555W mutations were co-segregated with the disease phenotypes and were specific mutations for lattice corneal dystrophy type I(LCD I),Avellino corneal dystrophy(ACD,GCDⅡ),granular corneal dystrophy type I(GCD I),respectively.Our study highlights the prevalence of codon 124 and codon 555 mutations in the TGFBI gene among the Chinese stromal corneal dystrophies patients.·