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In vitro transdifferentiation of corneal epithelial-like cells from human skin-derived precursor cells 被引量:4
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作者 Sarawut Saichanma Ahnond Bunyaratvej Monnipha Sila-asna 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2012年第2期158-163,共6页
The damage of human corneal cells encounter with the problem of availability of corneal cells for replacement. Limitation of the source of corneal cells has been realized. An attempt of development of corneal epitheli... The damage of human corneal cells encounter with the problem of availability of corneal cells for replacement. Limitation of the source of corneal cells has been realized. An attempt of development of corneal epithelial-like cells from the human skin-derived precursor (hSKPs) has been made in this study. Combination of three essential growth factors: epidermal growth factor (EGF), keratinocyte growth factor (KGF) and hepatocyte growth factor (HGF) could demonstrate successfully induction of hSKPs to differentiation into corneal cells.The induced cells expressed the appearance of markers of corneal epithelial cells as shown by the presence of keratin 3 (K3) by antibody label and Western blot assay. The K3 gene expression of induced hSKPs cells as shown by reverse transcription-polymerase chain reaction (RT-PCR) technology was also demonstrated. The presence of these markers at both gene and protein levels could lead to our conclusion that the directional transdifferentiation of hSKPs cells into corneal epithelial cells was successfully done under this cell induction protocol. The finding shows a newly available stem cell source can be obtained from easily available skin. Cells from autologous human skin might be used for corneal disorder treatment in future clinical application. 展开更多
关键词 corneal epithelial-like cell human skin-derived precursor cell TRANSDIFFERENTIATION
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Differentiation of human amniotic epithelial cells into corneal epithelial-like cells in vitro 被引量:1
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作者 Min Yao Jian Chen +4 位作者 Xiao-Xi Yang Xiao-Ling Zhang Qing-Shan Ji Qing Zhou Jin-Tang Xu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2013年第5期564-572,共9页
·AIM: To explore the feasibility that human amniotic epithelial cells (hAECs) have the potential to differentiate into corneal epithelial -like cells under the microenvironment replicated by spontaneously immorta... ·AIM: To explore the feasibility that human amniotic epithelial cells (hAECs) have the potential to differentiate into corneal epithelial -like cells under the microenvironment replicated by spontaneously immortalized human corneal epithelial cells (S-ihCECs). ·METHODS: hAECs were isolated by enzyme digestion, and flow cytometry was used to analysis the expression of CD29/90/166/73/34 and HLA -DR. Recovered and cultured S -ihCECs, immunocytochemistry was used to detect the expression of CK3/12. The proliferation of S - ihCECs handled by different concentrations of mitomycin was detected by CCK -8. The proliferation of hAECs cultured by S-ihCECs culture media collected at different time was analyzed by CCK -8. After filtered out the optimal conditions, we collected S-ihCECs culture media for 5 days, then prepared conditioned medium to incubate hAECs, inverted phase contrast microscope and scanning electron microscope were used to observe the change of morphology in hAECs. Quantitative real -time reverse transcription -polymerase chain reaction (QRT - PCR) was carried out to evaluate the expression of Oct - 4, NANOG, PAX6, and CK12 in the differentiation period. Immunocytochemistry and western bloting were used to detect the expression of CK3/12. ·RESULTS: The culture media collected every 12h, from 20μg/mL mitomycin pretreatment S -ihCECs could significantly promote the proliferation of hAECs. In the period of differentiation, the morphology of differentiated hAECs was obviously different compared with the control group, and the distinctive CK3/12 for corneal epithelial cells was detected.·CONCLUSION: This study showed that hAECs can differentiate into corneal epithelial -like cells by replication of the corneal epithelial microenvironment, using the culture media collected from S -ihCECs, and it is possible that S -ihCECs culture media could be used in corneal tissue engineering. · 展开更多
关键词 human amniotic epithelial cells spontaneously immortalized human corneal epithelial cells mytomicin MICROENVIRONMENT tissue engineering
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Corneal endothelial cells and acoustic cavitation in phacoemulsification 被引量:1
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作者 Kai Chen Wen-Ya Xu +1 位作者 Si-Si Sun Hong-Wei Zhou 《World Journal of Clinical Cases》 SCIE 2023年第8期1712-1718,共7页
Postoperative complications of phacoemulsification,such as corneal edema caused by human corneal endothelial cell(CEC)injury,are still a matter of concern.Although several factors are known to cause CEC damage,the inf... Postoperative complications of phacoemulsification,such as corneal edema caused by human corneal endothelial cell(CEC)injury,are still a matter of concern.Although several factors are known to cause CEC damage,the influence of ultrasound on the formation of free radicals during surgery should be considered.Ultrasound in aqueous humor induces cavitation and promotes the formation of hydroxyl radicals or reactive oxygen species(ROS).ROS-induced apoptosis and autophagy in phacoemulsification have been suggested to significantly promote CEC injury.CEC cannot regenerate after injury,and measures must be taken to prevent the loss of CEC after phacoemulsification or other CEC injuries.Antioxidants can reduce the oxidative stress injury of CEC during phacoemulsification.Evidence from rabbit eye studies shows that ascorbic acid infusion during operation or local application of ascorbic acid during phacoemulsification has a protective effect by scavenging free radicals or reducing oxidative stress.Both in experiments and clinical practice,hydrogen dissolved in the irrigating solution can also prevent CEC damage during phacoemulsification surgery.Astaxanthin(AST)can inhibit oxidative damage,thereby protecting different cells from most pathological conditions,such as myocardial cells,luteinized granulosa cells of the ovary,umbilical vascular endothelial cells,and human retina pigment epithelium cell line(ARPE-19).However,existing research has not focused on the application of AST to prevent oxidative stress during phacoemulsification,and the related mechanisms need to be studied.The Rho related helical coil kinase inhibitor Y-27632 can inhibit CEC apoptosis after phacoemulsification.Rigorous experiments are required to confirm whether its effect is realized through improving the ROS clearance ability of CEC. 展开更多
关键词 CATARACT PHACOEMULSIFICATION corneal endothelial cells ULTRASOUND Acoustic cavitation Oxidative stress ANTIOXIDANT
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Corneal subepithelial nerve fibers in type 2 diabetes:potential biomarker of diabetic neuropathy
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作者 Ling-Rui Meng Hua Chen +4 位作者 Wen-Qian Chen Yi Gao Zi-Wei Li Zi Ye Zhao-Hui Li 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2024年第11期2060-2066,共7页
AIM:To observe the changes in corneal subepithelial nerve fibers(CNFs)and Langerhans cells(LCs)in patients with type 2 diabetes using corneal laser confocal microscopy(CLCM).METHODS:A total of 60 patients(64 eyes),inc... AIM:To observe the changes in corneal subepithelial nerve fibers(CNFs)and Langerhans cells(LCs)in patients with type 2 diabetes using corneal laser confocal microscopy(CLCM).METHODS:A total of 60 patients(64 eyes),including 40 patients with type 2 diabetes(DM group)and 20 subjects without diabetes(control group)were included with CLCM.Neuron J plugin of Image J software were used for quantitative analysis of CNF length(CNFL),CNF density(CNFD),corneal nerve branch fiber density(CNBD),main branch length density,branch length density,corneal nerve fiber tortuosity(NT)score,and LCs density.An independent samples t-test to analyze the variability between the two groups was performed,and Pearson correlation analysis was used to analyze the relationships between CNF and multiple biochemical indicators in the DM group.The predictive power of CNF for type 2 diabetes was assessed using the receiver operating characteristic(ROC)curve.RESULTS:There were significant differences in the CNFL,CNFD,and main branch length density between two groups.The results of Pearson correlation analysis showed a significant negative correlation between CNFD and the duration of diabetes as well as triglyceride levels and total cholesterol,and a significant positive correlation between CNFD and serum albumin.In addition,the NT score showed a positive correlation and urea nitrogen,similar to the positive correlation observed between LC density and glycosylated hemoglobin(HbA1c)levels.CNFD showed the highest area under the curve(AUC of ROC)value,followed by main branch length density and CNFL.The AUC of the ROC curve under the logistic regression model also demonstrated good predictive values.The cut-off values of CNFD,CNFL,and main branch length density for diabetes showed 31.25,18.85,and 12.56,respectively.CONCLUSION:In patients with type 2 diabetes,there is a notable reduction in both CNFL and CNFD.These measurements can be influenced by various blood biochemical factors.However,the compromised nerve fibers can serve as valuable indicators for predicting the onset of type 2 diabetes and also as biomarkers for detecting diabetic neuropathy and its related complications. 展开更多
关键词 corneal subepithelial nerve DIABETES glycated hemoglobin Langerhans cells diabetic neuropathy
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Limbal stem cells: Central concepts of corneal epithelial homeostasis 被引量:12
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作者 Jinny J Yoon Salim Ismail Trevor Sherwin 《World Journal of Stem Cells》 SCIE CAS 2014年第4期391-403,共13页
A strong cohort of evidence exists that supports the localisation of corneal stem cells at the limbus. The distinguishing characteristics of limbal cells as stem cells include slow cycling properties, high proliferati... A strong cohort of evidence exists that supports the localisation of corneal stem cells at the limbus. The distinguishing characteristics of limbal cells as stem cells include slow cycling properties, high proliferative potential when required, clonogenicity, absence of differentiation marker expression coupled with positive expression of progenitor markers, multipotency, centripetal migration, requirement for a distinct niche environment and the ability of transplanted limbal cells to regenerate the entire corneal epithelium. The existence of limbal stem cells supports the prevailing theory of corneal homeostasis, known as the XYZ hypothesis where X represents proliferation and stratification of limbal basal cells, Y centripetal migration of basal cells and Z desquamation of superficial cells. To maintain the mass of cornea, the sum of X and Y must equal Z and very elegant cell tracking experiments provide strong evidence in support of this theory. However, several recent stud-ies have suggested the existence of oligopotent stem cells capable of corneal maintenance outside of the limbus. This review presents a summary of data which led to the current concepts of corneal epithelial homeostasis and discusses areas of controversy surrounding the existence of a secondary stem cell reservoir on the corneal 展开更多
关键词 Limbal stem cell corneal EPITHELIUM XYZ HYPOTHESIS corneal HOMEOSTASIS corneal wound repair
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Establishment of an untransfected human corneal epithelial cell line and its biocompatibility with denuded amniotic membrane 被引量:22
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作者 Ting-Jun Fan Bin Xu +3 位作者 Jun Zhao Hong-Shou Yang Rui-Xin Wang and Xiu-Zhong Hu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2011年第3期228-234,共7页
AIM: To establish an untransfected human corneal epithelial (HCEP) cell line and characterize its biocompatibility with denuded amniotic membrane (dAM). METHODS: The torn HCEP pieces were primarily cultured in DMEM/F1... AIM: To establish an untransfected human corneal epithelial (HCEP) cell line and characterize its biocompatibility with denuded amniotic membrane (dAM). METHODS: The torn HCEP pieces were primarily cultured in DMEM/F12 media (pH 7.2) supplemented with 20% fetal bovine serum and other necessary factors, yielding an HCEP cell line which was its growth performance, chromosome morphology, tumorigenicity and expression of marker proteins analyzed. In addition, the biocompatibility of HCEP cells with dAM was evaluated through histological and immunocytochemistry analyses and with light, electron and slit-lamp microscopies. RESULTS: HCEP cells proliferated to confluence in 3 weeks, which have been subcultured to passage 160. A continuous untransfected HCEP cell line, designated as utHCEPC01, was established with a population doubling time of 45.42 hours as was determined at passage 100. The cells retained HCEP cell properties as were approved by chromosomal morphology and the expression of keratin 3. They, with no tumorigenicity, formed a multilayer epithelium-like structure on dAMs through proliferation and differentiation during air-liquid interface culture, maintained expression of marker proteins including keratin 3 and integrin p 1 and attached tightly to dAMs. The reconstructed HCEP was highly transparent and morphologically and structurally similar to the original. CONCLUSION: An untransfected and non-tumorigenic HCEP cell line was established in this study. The cells maintained expression of marker proteins. The cell line was biocompatible with dAM. It holds the potential of being used for in vitro reconstruction of tissue-engineered HCEP, promising for the treatment of diseases caused by corneal epithelial disorders. 展开更多
关键词 human corneal epithelial cell cell line untransfected BIOCOMPATIBILITY denuded amniotic membrane
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Nerve Growth Factor Modulate Proliferation of Cultured Rabbit Corneal Endothelial Cells and Epithelial Cells 被引量:9
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作者 李新宇 李中国 +2 位作者 邱良秀 赵长松 胡竹林 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2005年第5期575-577,共3页
Summary: In order to investigate the effect of nerve growth factor (NGF) on the proliferation of rabbit corneal endothelial cells and epithelial cells, the in vitro cultured rabbit corneal endothelial cells and epi... Summary: In order to investigate the effect of nerve growth factor (NGF) on the proliferation of rabbit corneal endothelial cells and epithelial cells, the in vitro cultured rabbit corneal endothelial cells and epithelial cells were treated with different concentrations of NGF.MTT assay was used to examine the clonal growth and proliferation of the cells by determining the absorbency values at 570 nm. The results showed that NGF with three concentrations ranging from 5 U/mL to 500 U/mL enhanced the proliferation of rabbit corneal endothelial cells in a concentration-dependent manner. 50 U/mI. and 500 U/mI. NGF got more increase of proliferation than that of 5 U/mL NGF did. Meanwhile, 50 U/mL and 500 U/mL NGF could promote the proliferation of the rabbit corneal epithelial cells significantly in a concentration-dependent manner. However, 5 U/mL NGF did not enhance the proliferation of epithelial cells. It was suggested that exogenous NGF can stimulate the proliferation of both rabbit corneal endothelial and epithelial cells, but the extent of modulation is different. 展开更多
关键词 nerve growth factor corneal endothelial cells corneal epithelial cells PROLIFERATION
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Acellular porcine corneal matrix as a carrier scaffold for cultivating human corneal epithelial cells and fibroblasts in vitro 被引量:7
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作者 ju zhang Can-Wei Zhang +1 位作者 Li-Qun Du Xin-Yi Wu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2016年第1期1-8,共8页
AIM:To investigate the feasibility of corneal anterior lamellar reconstruction with human corneal epithelial cells and fibroblasts,and an acellular porcine cornea matrix(APCM) in vitro.·METHODS:The scaffold w... AIM:To investigate the feasibility of corneal anterior lamellar reconstruction with human corneal epithelial cells and fibroblasts,and an acellular porcine cornea matrix(APCM) in vitro.·METHODS:The scaffold was prepared from fresh porcine corneas which were treated with 0.5%sodium dodecyl sulfate(SDS)solution and the complete removal of corneal cells was confirmed by hematoxylin-eosin(HE)staining and 4’,6-diamidino-2-phenylindole(DAPI)staining.Human corneal fibroblasts and epithelial cells were cultured with leaching liquid extracted from APCM,and then cell proliferative ability was evaluated by MTT assay.To construct a human corneal anterior lamellar replacement,corneal fibroblasts were injected into the APCM and cultured for 3d,followed by culturing corneal epithelial cells on the stroma construction surface for another 10d.The corneal replacement was analyzed by HE staining,and immunofluorescence staining.·R ESULTS:Histological examination indicated that there were no cells in the APCM by HE staining,and DAPI staining did not detect any residual DNA.The leaching liquid from APCM had little influence on the proliferation ability of human corneal fibroblasts and epithelial cells.At 10d,a continuous 3 to 5 layers of human corneal epithelial cells covering the surface of the APCM was observed,and the injected corneal fibroblasts distributed within the scaffold.The phenotype of the construction was similar to normal human corneas,with high expression of cytokeratin 12 in the epithelial cell layer and high expression of Vimentin in the stroma.·CONCLUSION:Corneal anterior lamellar replacement can be reconstructed in vitro by cultivating human corneal epithelial cells and fibroblasts with an acellular porcine cornea matrix.This laid the foundation for the further transplantation in vitro. 展开更多
关键词 corneal epithelial cells corneal keratocytes acellular porcine cornea matrix corneal tissue engineering limbal epithelial cells
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The roles of surfactant protein D during Aspergillus fumigatus infection in human corneal epithelial cells 被引量:13
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作者 Cheng-Ye Che Wen-Yan Jia +6 位作者 Qiang Xu Na Li Li-Ting Hu Nan Jiang Jing Lin Qing Wang and Gui-Qiu Zhao 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2012年第1期13-17,共5页
AIM: To investigate roles of surfactant protein D (SP-D) and relative cytokines in human corneal epithelial (HCE) cells Exposed to aspergillus fumigatus (AF) antigens. METHODS: HCE cells cultured 47 in vitro with AF a... AIM: To investigate roles of surfactant protein D (SP-D) and relative cytokines in human corneal epithelial (HCE) cells Exposed to aspergillus fumigatus (AF) antigens. METHODS: HCE cells cultured 47 in vitro with AF antigens and sampled at 0, 0.5, 1 hour, 2, 4, 6 and 8 hours. The Expression of SP-D mRNA was evaluated by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).The expression of SP-D protein was shown by ELISA and immunocytochemistry SP methods. The expression of NF-kappa B and relative downstream cytokines such as TNF-alpha, IL-1 beta, IL-8 and IL-10 in supernatant fluid were measured by ELISA. RESULTS: SP-D mRNA and protein were detected in untreated HCE cells. The expression of SP-D and the relative downstream cytokines rose after being stimulated with AF antigens. SP-D mRNA began to rise at 0.5 hour and the most significantly peak was in 2 hours. The protein of SP-D in supernatant fluid had the same trend with mRNA. Immunocytochemistry of SP-D showed positive expression and gradually increased to 6 hours, and then the expression began to decline. NF-kappa B was activated after treated by AF antigens and the changes had correlation with SP-D. TNF-alpha, IL-1 beta, IL-8 and IL-10 began to rise after given AF antigens 1 hour and were 1.82, 1.43, 1.12 and 1.28 times higher than the untreated HCE cells separately. The expression of TNF-alpha and IL-1 beta reached the peak at 2 hours, separately 2.80 and 2.86 times than the untreated. The expression of IL-8 and IL-10 gradually increased with a time-dependent manner. ' CONCLUSION: HCE cells exists SP-D and it may play a significant role in pathogenesis of keratomycosis. AF may induce human corneal epithelial cells to express inflammatory cytokines via SP-D and NF-kappa B pathway. SP-D possibly mediates the recognition to AF mycelium. 展开更多
关键词 corneal epithelial cells aspergillus fumigatus surfactant protein D innate immune
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Mesenchymal stem cells: Potential role in corneal wound repair and transplantation 被引量:8
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作者 Fei Li Shao-Zhen Zhao 《World Journal of Stem Cells》 SCIE CAS 2014年第3期296-304,共9页
Corneal diseases are a major cause of blindness in the world. Although great progress has been achieved in the treatment of corneal diseases, wound healing after severe corneal damage and immunosuppressive therapy aft... Corneal diseases are a major cause of blindness in the world. Although great progress has been achieved in the treatment of corneal diseases, wound healing after severe corneal damage and immunosuppressive therapy after corneal transplantation remain prob-lematic. Mesenchymal stem cells(MSCs) derived from bone marrow or other adult tissues can differentiate into various types of mesenchymal lineages, such as osteocytes, adipocytes, and chondrocytes, both in vivo and in vitro. These cells can further differentiate into specific cell types under specific conditions. MSCs migrate to injury sites and promote wound healing by secreting anti-inflammatory and growth factors. In ad-dition, MSCs interact with innate and acquired immune cells and modulate the immune response through their powerful paracrine function. Over the last decade, MSCs have drawn considerable attention because of their beneficial properties and promising therapeutic prospective. Furthermore, MSCs have been applied to various studies related to wound healing, autoim-mune diseases, and organ transplantation. This review discusses the potential functions of MSCs in protecting corneal tissue and their possible mechanisms in corneal wound healing and corneal transplantation. 展开更多
关键词 MESENCHYMAL stem cells corneal injury WOUND repair IMMUNE modulation TRANSPLANTATION
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Evaluating alternative stem cell hypotheses for adult corneal epithelial maintenance 被引量:6
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作者 John D West Natalie J Dorà J Martin Collinson 《World Journal of Stem Cells》 SCIE CAS 2015年第2期281-299,共19页
In this review we evaluate evidence for three different hypotheses that explain how the corneal epithelium is maintained. The limbal epithelial stem cell(LESC)hypothesis is most widely accepted. This proposes that ste... In this review we evaluate evidence for three different hypotheses that explain how the corneal epithelium is maintained. The limbal epithelial stem cell(LESC)hypothesis is most widely accepted. This proposes that stem cells in the basal layer of the limbal epithelium, at the periphery of the cornea, maintain themselves and also produce transient(or transit) amplifying cells(TACs). TACs then move centripetally to the centre of the cornea in the basal layer of the corneal epithelium and also replenish cells in the overlying suprabasal layers. The LESCs maintain the corneal epithelium during normal homeostasis and become more active to repair significant wounds. Second, the corneal epithelial stem cell(CESC) hypothesis postulates that, during normal homeostasis, stem cells distributed throughout the basal corneal epithelium, maintain the tissue. According to this hypothesis, LESCs are present in the limbus but are only active during wound healing. We also consider a third possibility, that the corneal epithelium is maintained during normal homeostasis by proliferation of basal corneal epithelial cells without any input from stem cells. After reviewing the published evidence, we conclude that the LESC and CESC hypotheses are consistent with more of the evidence than the third hypothesis, so we do not consider this further. The LESC and CESC hypotheses each have difficulty accounting for one main type of evidence so we evaluate the two key lines of evidence that discriminate between them. Finally, we discuss how lineage-tracing experiments have begun to resolve the debate in favour of the LESC hypothesis. Nevertheless, it also seems likely that some basal corneal epithelial cells can act as long-term progenitors if limbal stem cell function is compromised. Thus, this aspect of the CESC hypothesis may have a lasting impact on our understanding of corneal epithelial maintenance, even if it is eventually shown that stem cells are restricted to the limbus as proposed by the LESC hypothesis. 展开更多
关键词 Eye CORNEA corneal EPITHELIUM Limbalepithelium Stem cell LINEAGE tracing
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Establishment of an untransfected human corneal stromal cell line and its biocompatibility to acellular porcine corneal stroma 被引量:5
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作者 Ting-Jun Fan Xiu-Zhong Hu +4 位作者 Jun Zhao Ying Niu Wen-Zhuo Zhao Miao-Miao Yu and Yuan Ge 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2012年第3期286-292,共7页
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 o... 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. 展开更多
关键词 human corneal stromal cells cell line untransfected BIOCOMPATIBILITY acellular porcine corneal stroma
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Evaluation of corneal cell growth on tissue engineering materials as artificial cornea scaffolds 被引量:8
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作者 Hai-Yan Wang Rui-Hua Wei Shao-Zhen Zhao 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2013年第6期873-878,共6页
The keratoprosthesis(KPro;artificial cornea)is a special refractive device to replace human cornea by using heterogeneous forming materials for the implantation into the damaged eyes in order to obtain a certain visio... The keratoprosthesis(KPro;artificial cornea)is a special refractive device to replace human cornea by using heterogeneous forming materials for the implantation into the damaged eyes in order to obtain a certain vision.The main problems of artificial cornea are the biocompatibility and stability of the tissue particularly in penetrating keratoplasty.The current studies of tissue-engineered scaffold materials through comprising composites of natural and synthetic biopolymers together have developed a new way to artificial cornea.Although a wide agreement that the long-term stability of these devices would be greatly improved by the presence of cornea cells,modification of keratoprosthesis to support cornea cells remains elusive.Most of the studies on corneal substrate materials and surface modification of composites have tried to improve the growth and biocompatibility of cornea cells which can not only reduce the stimulus of heterogeneous materials,but also more importantly continuous and stable cornea cells can prevent the destruction of collagenase.The necrosis of stroma and spontaneous extrusion of the device,allow for maintenance of a precorneal tear layer,and play the role of ensuring a good optical surface and resisting bacterial infection.As a result,improvement in corneal cells has been the main aim of several recent investigations;some effort has focused on biomaterial for its well biological properties such as promoting the growth of cornea cells.The purpose of this review is to summary the growth status of the corneal cells after the implantation of several artificial corneas. 展开更多
关键词 artificial cornea KERATOPROSTHESIS tissue-engineered scaffold corneal cells collagen FIBRIN amniotic membrane biomaterial
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Niche regulation of corneal epithelial stem cells at the limbus 被引量:22
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作者 YasutakaHayashida SchefferCGTseng 《Cell Research》 SCIE CAS CSCD 2007年第1期26-36,共11页
Among all adult somatic stem cells, those of the corneal epithelium are unique in their exclusive location in a defined limbal structure termed Palisades of Vogt. As a result, surgical engraftment of limbal epithelial... Among all adult somatic stem cells, those of the corneal epithelium are unique in their exclusive location in a defined limbal structure termed Palisades of Vogt. As a result, surgical engraftment of limbal epithelial stem cells with or without ex vivo expansion has long been practiced to restore sights in patients inflicted with limbal stem cell deficiency. Neverthe- less, compared to other stem cell examples, relatively little is known about the limbal niche, which is believed to play a pivotal role in regulating self-renewal and fate decision oflimbal epithelial stem cells. This review summarizes relevant literature and formulates several key questions to guide future research into better understanding of the pathogenesis of limbal stem cell deficiency and further improvement of the tissue engineering of the corneal epithelium by focusing on the limbal niche. 展开更多
关键词 corneal epithelium stem cell NICHE
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Expression of dectin-1 during fungus infection in human corneal epithelial cells 被引量:6
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作者 Cui Li Gui-Qiu Zhao +6 位作者 Cheng-Ye Che Na Li Jing Lin Qiang Xu Qian Wang Ying Liu Sheng Qiu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2014年第1期34-37,共4页
AIM: To evaluate the expression of dendritic cell-associated C-type lectin-1(dectin-1) in human corneal epithelial(HCE) cells infected by fungus. · METHODS: A total of 20 cases of healthy donor corneas were group... AIM: To evaluate the expression of dendritic cell-associated C-type lectin-1(dectin-1) in human corneal epithelial(HCE) cells infected by fungus. · METHODS: A total of 20 cases of healthy donor corneas were group A,and 20 patients(20 eyes) suffered from fungal keratitis(FK) composed group B. Real-time qPCR and immunohistochemistry were applied to detect dectin-1 expression in corneal epithelium of both groups. HCE cells were cultured with aspergillus fumigatus(AF) antigens in vitro. The expression of dectin-1 mRNA was measured by real-time qPCR at the stimulation of 0,4,8 and 24h separately. Dectin-1 protein was detected by immunocytochemistry at 0 and 24h separately. ·RESULTS: Dectin-1 expressed in corneal epithelium of normal persons and FK patients. Vitro cellular experiment showed that the expression of dectin-1 mRNA in HCE cells began to increase after stimulation of AF antigens at 4h,and dectin-1 protein expression increased after stimulation at 24h. · CONCLUSION: Dectin-1 expressed in corneal epithelium of normal persons. AF antigens stimulation can elevate the expression of dectin-1 in HCE cells in vitro. 展开更多
关键词 DECTIN-1 corneal epithelial cells fungal keratitis HUMAN
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Transplantation of tissue-engineered human corneal epithelium in limbal stem cell deficiency rabbit models 被引量:4
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作者 Bin Xu Ting-Jun Fan +6 位作者 Jun Zhao Ai Sun Rui-Xin Wang Xiu-Zhong Hu Hao-Ze Yu Xian-Yuan Fan and Xiao-Hui Xu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2012年第4期424-429,共6页
AIM: To evaluate the biological functions of tissue-engineered human corneal epithelium (TE-HCEP) by corneal transplantation in limbal stem cell deficiency (LSCD) rabbit models. METHODS: TE-HCEPs were reconstructed wi... AIM: To evaluate the biological functions of tissue-engineered human corneal epithelium (TE-HCEP) by corneal transplantation in limbal stem cell deficiency (LSCD) rabbit models. METHODS: TE-HCEPs were reconstructed with DiI-labeled untransfected HCEP cells and denuded amniotic membrane (dAM) in air-liquid interface culture, and their morphology and structure were characterized by hematoxylin-eosin (HE) staining of paraffin-sections, immunohistochemistry and electron microscopy. LSCD models were established by mechanical and alcohol treatment of the left eyes of New Zealand white rabbits, and their eyes were transplanted with TE-HCEPs with dAM surface outside by lamellar keratoplasty (LKP). Corneal transparency, neovascularization, thickness, and epithelial integrality of both traumatic and post transplantation eyes were checked once a week by slit-lamp corneal microscopy, a corneal pachymeter, and periodic acid-Schiff (PAS) staining. At day 120 post surgery, the rabbits in each group were sacrificed and their corneas were examined by DiI label observation, HE staining, immunohistochemistry and electron microscopy. RESULTS: After cultured for 5 days on dAM, HCEP cells, maintaining keratin 3 expression, reconstructed a 6-7 layer TE-HCEP with normal morphology and structure. The traumatic rabbit corneas, entirely opaque, conjunctivalized and with invaded blood vessels, were used as LSCD models for TE-HCEP transplantation. After transplantation, obvious edema was not found in TE-HCEP-transplanted corneas which became more and more transparent, the invaded blood vessels reduced gradually throughout the monitoring period. The corneas decreased to normal thickness on day 25, while those of dAM eyes were over 575 mu m in thickness during the monitoring period. A 45 layer of epithelium consisting of TE-HCEP originated cells attached tightly to the anterior surface of stroma was reconstructed 120 days after TE-HCEP transplantation, which was similar to the normal control eye in morphology and structure. In contrast, intense corneal edema, turbid, invaded blood vessels were found in dAM eyes, and no multilayer epithelium was found but only a few scattered conjunctiva-like cells appeared. CONCLUSION: The TE-HCEP, with similar morphology and structure to those of innate HCEP, could reconstruct a multilayer corneal epithelium with normal functions in restoring corneal transparency and thickness of LSCD rabbits after transplantation. It may be a promising HCEP equivalent for clinical therapy of corneal epithelial disorders. 展开更多
关键词 tissue-engineered human corneal epithelium limbal stem cell deficiency rabbit lamellar keratoplasty human corneal epithelial cells denuded amniotic membrane RECONSTRUCTION
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Construction of a full-thickness human corneal substitute from anterior acellular porcine corneal matrix and human corneal cells 被引量:3
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作者 Kai Zhang Xiao-Xiao Ren +2 位作者 Ping Li Kun-Peng Pang Hong Wang 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2019年第3期351-362,共12页
AIM: To construct functional human full-thickness corneal replacements.METHODS: Acellular porcine corneal matrix(APCM) was developed from porcine cornea by decellulariztion. The biomechanical properties of anterior-AP... AIM: To construct functional human full-thickness corneal replacements.METHODS: Acellular porcine corneal matrix(APCM) was developed from porcine cornea by decellulariztion. The biomechanical properties of anterior-APCM(AAPCM) and posterior-APCM(PAPCM) were checked using uniaxial tensile testing. Human corneal cells were obtained by cell culture. Suspending ring was designed by deformation of an acupuncture needle. MTT cytotoxicity assay was used to check the cytotoxicity of suspending ring soaking solutions. A new three-dimensional organ culture system was established by combination of suspending ring, 48-well plate and medium together. A human full-thickness corneal substitute was constructed from human corneal cells with AAPCM in an organ coculture system. Biochemical marker expression of the construct was measured by immunofluorescent staining and morphological structures were observed using scanning electron microscopy. Pump function and biophysical properties were examined by penetrating keratoplasty and follow-up clinical observations.RESULTS: There were no cells in the AAPCM or PAPCM, whereas collagen fibers, Bowman's membrane, and Descemet's membrane were retained. The biomechanical property of AAPCM was better than PAPCM. Human corneal cells grew better on the AAPCM than on the PAPCM.There was no cytotoxicity for the suspending ring soaking solutions. For the constructed full-depth human corneal replacements keratocytes scattered uniformly throughout the AAPCM and expressed vimentin. The epithelial layer was located on the surface of Bowman's membrane and composed of three or four layers of epithelial cells expressing cytokeratin 3. One layer of endothelial cells covered the stromal surface of AAPCM, expressed Na+/K+ATPase and formed the endothelial layer. The construct was similar to normal human corneas, with many microvilli on the epithelial cell surface, stromal cells with a long shuttle shape, and zonula occludens on the interface of endothelial cells. The construct withstood surgical procedures during penetrating keratoplasty. The corneal transparency increased gradually and was almost completely restored 7 d after surgery.CONCLUSION: AAPCM is an ideal scaffold for constructing full-thickness corneal replacement, and functional human full-thickness corneal replacements are successfully constructed using AAPCM and human corneal cells. 展开更多
关键词 full-thickness human corneal SUBSTITUTE anterior-acellular PORCINE corneal MATRIX posterior-acellular PORCINE corneal MATRIX human corneal cells
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Cytotoxic effects of betaxolol on healthy corneal endothelial cells both in vitro and in vivo 被引量:3
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作者 Ying Miao Qian Sun +4 位作者 Qian Wen Yue Qiu Yuan Ge Miao-Miao Yu Ting-Jun Fan 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2014年第1期14-21,共8页
AIM: To demonstrate the cytotoxic effect of betaxolol and its underlying mechanism on human corneal endothelial cells(HCE cells) in vitro and cat corneal endothelial cells(CCE cells) in vivo,providing experimental bas... AIM: To demonstrate the cytotoxic effect of betaxolol and its underlying mechanism on human corneal endothelial cells(HCE cells) in vitro and cat corneal endothelial cells(CCE cells) in vivo,providing experimental basis for safety anti-glaucoma drug usage in clinic of ophthalmology. ·METHODS: In vivo and in vitro experiments were conducted to explore whether and how betaxolol participates in corneal endothelial cell injury. The in vitro morphology,growth status,plasma membrane permeability,DNA fragmentation,and ultrastructure of HCE cells treated with 0.021875-0.28g/L betaxolol were examined by light microscope,3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide(MTT) assay,acridine orange(AO)/ethidium bromide(EB) double-fluorescent staining,DNA agarose gel electrophoresis,and transmission electron microscope(TEM). The in vivo density,morphology,and ultrastructure of CCE cells,corneal thickness,and eye pressure of cat eyes treated with 0.28g/L betaxolol were investigated by specular microscopy,applanation tonometer,alizarin red staining,scanning electron microscope(SEM),and TEM. · RESULTS: Exposure to betaxolol at doses from 0.0875g/L to 2.8g/L induced morphological and ultrastructural changes of in vitro cultured HCE cells such as cytoplasmic vacuolation,cellular shrinkage,structural disorganization,chromatin condensation,and apoptotic body appearance. Simultaneously,betaxolol elevated plasma membrane permeability and induced DNA fragmentation of these cells in a dose-dependent manner in AO/EB staining. Furthermore,betaxolol at adose of 2.8g/L also induced decrease of density of CCE cells in vivo,and non-hexagonal and shrunk apoptotic cells were also found in betaxolol-treated cat corneal endothelia. ·CONCLUSION: Betaxolol has significant cytotoxicity on HCE cells in vitro by inducing apoptosis of these cells,and induced apoptosis of CCE cells in vivo as well. The findings help provide new insight into the apoptosis-inducing effect of anti-glaucoma drugs in eye clinic. 展开更多
关键词 BETAXOLOL CYTOTOXICITY APOPTOSIS human corneal endothelial cells cat corneal endothelial cells
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Local anesthetic lidocaine induces apoptosis in human corneal stromal cells in vitro 被引量:4
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作者 Xin Zhou Yi-Han Li +2 位作者 Hao-Ze Yu Rui-Xin Wang Ting-Jun Fan 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2013年第6期766-771,共6页
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 ... 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. 展开更多
关键词 LIDOCAINE apoptosis-inducing effect apoptotic body DNA fragmentation human corneal stromal cell
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Transforming growth factor-β2 induces morphological alteration of human corneal endothelial cells in vitro 被引量:5
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作者 Jing Wang Ting-Jun Fan +1 位作者 Xiu-Xia Yang Shi-Min Chang 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2014年第5期759-763,共5页
AIM:To investigate the morphological altering effect of transforming growth factor-β2(TGF-β2) on untransfected human corneal endothelial cells(HCECs)in vitro.METHODS:After untransfected HCECs were treated with TGF-... AIM:To investigate the morphological altering effect of transforming growth factor-β2(TGF-β2) on untransfected human corneal endothelial cells(HCECs)in vitro.METHODS:After untransfected HCECs were treated with TGF-β2 at different concentrations, the morphology,cytoskeleton distribution, and type IV collagen expression of the cells were examined with inverted contrast light microscopy, fluorescence microscopy,immunofluorescence or Western Blot.RESULTS:TGF-β2 at the concentration of 3-15 μg/L had obviously alterative effects on HCECs morphology in dose and time-dependent manner, and 9 μg/L was the peak concentration. TGF-β2(9 μg/L) altered HCE cell morphology after treatment for 36 h, increased the mean optical density(P 【0.01) and the length of F-actin,reduced the mean optical density(P 【0.01) of the collagen type IV in extracellular matrix(ECM) and induced the rearrangement of F-actin, microtubule in cytoplasm and collagen type IV in ECM after treatment for 72 h.·CONCLUTION: TGF-β2 has obviously alterative effect on the morphology of HCECs from polygonal phenotype to enlarged spindle-shaped phenotype, in dose and time-dependence manner by inducing more, elongation and alignment of F-actin, rearrangement of microtubule and larger spread area of collagen type IV. 展开更多
关键词 human corneal endothelial cell transforming growth factor-β 2 F-ACTIN MICROTUBULE collagen type IV
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