ABSTRACT
Background Excessive elongation of axis and expansion of sclera is one of the hot topics in the study of the pathogenesis of high myopia.To establish a human scleral fibroblasts (HSFs)-collagen matrix culture model is helpful for understanding the reciprocal and adaptive interactions between HSFs and the collagen matrix in tissue.Objective The aim of this study was to establish a HSFs-seeded collagen three-dimension culture system that may mimic the sclera remolding in myopia.Methods HSFs were isolated and cultured from donor eyes by explant culture and purified by passages culture in vitro.The expressions of vimentin and keratin in the cells were detected by immunofluorescence technique to identify the cells.Rat tail tendon was obtained from 8-week-old SPF SD rats to prepare the collagen matrix.The mixed solution of 400 μl collagen matrix and 1 100 μl PBS,200 μ1 nutrient medium,50 μ1 NaOH and HSF suspension were mixed to prepare the collagen gel three-demension culture system.The growth and morphology of the cells in the culture system were observed under the inverted phase contrast microscope,and IPP-5 software was used to measure the contraction area of collagen gel,and the mechanical creep properties of the HSFs-seeded collagen matrix were measured by a biomechanics test instrument.Results HSFs emigrated from tissue 7 days after culture and passage could be performed 14 days after culture.The expression of keratin was absent in HSFs,while vimentin was positively expressed.The free-cell collagen gel was clear and unchanged in the experimental duration.However,the cells were obviously increased on the three-demension culture system and showed a tissue-like structure of net-like arrangement on dozens of layers.In 7-14 days after culture,the collagen gel area in a three-demension collagen matrix revealed a decrease of 90％.Duotriode-like and fusiform cells were seen 24 hours after culture.The biomechanical creep curve of HSFs-seeded collagen matrix consisted of the nonlinear section (0-100 seconds) and linear section (100-600 seconds),and the former appeared to be an elastic change of the gel under the temporal stress,and the latter was the creeping of the gel with the time.Conclusions Rat tail collagen appears to have a good biocompatibility to HSFs.HSFs-seeded collagen matrix can retain the mechanical creep properties,and it may be a good tool for the study on the relationship between HSFs and extracellular matrix or intercellular biological behaviour for scleral remodeling.
ABSTRACT
OBJECTIVE:To investigate the clinical efficacy of levodopa combined with comprehensive therapy for ametropic amblyopia children and teenagers and its effeets on pattern visual evoked potential(P-VEP)and visual function. METHODS:70 pa-tients(80 eyes)with ametropic amblyopia were randomly divided into observation group(35 cases,40 eyes)and control group(35 cases,38 eyes). Control group received comprehensive therapy as wearing correctional glasses,occlusion therapy and visual stimu-lation training;observation group additionally received levodopa 0.125(d1-3)-0.25 g/time,bid,on the basis of control group for 6 months. Therapeutic efficacy,P-VEP,Visual sensitivity and visual function were compared between two groups,and the occur-rence of ADR was recorded. RESULTS:The effective rate of observation group(92.50%)was significantly higher than that of con-trol group(76.32%),with statistical significance(P0.05). CONCLUSIONS:Levodopa combined with comprehensive therapy can improve the function of visual central neurons,and improve visual acuity and binocular stereo vision functions so as to improve therapy efficacy.