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| 1 | Local anesthetic lidocaine induces apoptosis in human corneal stromal cells in vitro显示文摘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. | Xin Zhou Yi-Han Li Hao-Ze Yu Rui-Xin Wang Ting-Jun Fan | 2013 | International Journal of Ophthalmology(English edition)2013，6，6： | 4 |
| 2 | Using bovine pituitary extract to increase proliferation of keratocytes and maintain their phenotype in vitro显示文摘AIM:To investigate the effects of bovine pituitary extract on the proliferation of keratocytes and maintaining the keratocyte phenotype/n v/tro.METHODS:Single keratocytes were isolated by enzyme digestion for in vitro culture.Three groups were designed according to the different culture media:a bovine pituitary extract(BPE)group,a fetal bovine serum(FBS)group and the control group.The phenotypes and proliferation of cultured cells were evaluated by morphology,immunofluorescent staining and mRNA expression of CD34,Lumican,VSX1,α-SMA and proliferating cell nuclear antigen(PCNA).in the BPE group,cells underwent serial subcultivation,and their phenotypes were identified by immunofluorescent staining.To analyze the proliferation of keratocytes in different concentrations of BPE,six different concentrations were designed to ascertain the most appropriate amount.RESULTS:In the BPE group,the cells spread out and presented dendritic morphology,and their dendrites connected to one another to form networks.On the third passage,most cells maintained their phenotype.In the FBS group,the cells exhibited a dendritic appearance in early cultured stages,but their morphology subsequently changed into a fibroblast-like shape.The number ofdendritic cells in BPE group was more than FBS and control groups.Immunofluorescent staining and realtime polymerase chain reaction(PCR)confirmed that few keratocytes underwent fibroblastic transformation in the BPE and control groups,and that proliferation was higher in the BPE group than in the control group.Although the proliferation was higher in the FBS group,many keratocytes underwent fibroblastic transformation.The analysis of cell morphology and mRNA expressions of CD34,PCNA and VSX1 in six group showed that different concentrations of BPE affected the proliferation obviously but didn't affect the keratocyte phenotype,and the concentration of 40μg/mL was the most appropriate one.CONCLUSION:BPE can improve the proliferation of keratocytes and maintain their phenotype in vitro.Many keratocytes can be harvested rapidly and provide seeds for the construction of corneal stroma. | Zhong-Zhong Xu Zhi-Jie Li Lian-Xin Du Jing Li Li-Ya Wang | 2013 | International Journal of Ophthalmology(English edition)2013，6，6： | 1 |
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