利用脱细胞角膜基质作为羟基磷灰石义眼座包裹材料的实验研究

发布时间：2018-04-19 14:55

本文选题：脱细胞角膜基质 + 包裹材料　；参考：《南昌大学》2014年硕士论文

【摘要】：目的：利用反复冻融+电泳法制备脱细胞角膜基质,同时初步观察猪脱细胞角膜基质(APCS)作为羟基磷灰石(HA)义眼台包裹材料的可行性，为其在眼整形方面的临床应用提供理论和实验基础。方法： 1、采用反复冻融+电泳法去除猪角膜细胞成分，制备脱细胞角膜基质（APCS），利用病理切片染色方法检查其细胞去除和细胞外基质结构保留情况，同时利用皮下植入法检测其生物相容性； 2、将30只新西兰白兔随机分为A、B2组,每组15只，各组均将HA义眼台植入兔左眼眶内。A组为对照组，植入自体巩膜包裹的HA义眼台，B组植入APCS包裹的HA义眼台。术后每天肉眼和裂隙灯显微镜检查术眼，观测眼睑、结膜囊愈合情况及义眼台有无暴露，并分别于术后1周、2周、3周、4周、5周对每组兔均进行双眼增强磁共振扫描，同时两组各随机取出1只义眼台植入物行组织病理学检查，比较植入义眼台纤维血管化程度的差异。结果： 1、新鲜猪角膜经过脱细胞进程处理，基质细胞已全部移除，细胞外基质构架保存良好（含前弹力层及前2/3基质层），免疫反应实验结果显示：皮下植入后第2周，APCS组织周边炎性细胞聚集明显，以中性粒细胞为主，植入后4周，实验组与对照组均未发现明显排斥反应， APCS组织中未见中性粒细胞及淋巴细胞浸润，周围包裹纤维结缔组织，，与周边形成明显界限。 2、术后1周，A、 B组均开始有不同程度的血管化，B组血管化速度缓慢，两组磁共振扫描结果有统计学差异（A、B组间p＜0.05）。术后2周，两组血管化程度都提高，但均未完全血管化，B组血管化程度稍低于A组，两组之间血管化差异有统计学意义（A、B组间p＜0.05）。术后3周，两组植入物血管化程度加深，较前有明显提高，但未完全血管化，两组之间的血管化无统计学差异（A、B组间p＞0.05）。术后4周及以后， A、B两组均达到完全血管化，两组间无统计学差异。结论： 1、利用反复冻融+电泳法能完全去除猪角膜基质细胞,同时良好保留细胞外基质,使其结构不受破坏。 2、APCS对早期的HA义眼台血管化无显著影响，并未延缓羟基磷灰石义眼台完全血管化进程。 3、利用APCS作为HA义眼台植入的包裹材料具有一定可行性。
[Abstract]:Objective:Acellular corneal stroma was prepared by repeated freeze-thaw electrophoresis, and the feasibility of APCS as hydroxyapatite (HA) implantation material was preliminarily observed.To provide theoretical and experimental basis for its clinical application in ocular plastic surgery.Methods:1. The acellular corneal stroma (APCS) was prepared by repeated freeze-thaw electrophoresis, and the cell removal and extracellular matrix structure retention were examined by pathological section staining.At the same time, the biocompatibility was detected by subcutaneous implantation.2. Thirty New Zealand white rabbits were randomly divided into two groups: group A (n = 15), group A (n = 15), and group A (n = 15): the HA prosthesis was implanted into the left orbit of rabbits as a control group.The healing of eyelid and conjunctival sac and the exposure of prosthetic ocular platform were observed every day after operation with slit lamp microscope. The enhanced magnetic resonance imaging (enhanced magnetic resonance imaging) was performed in each group at 1 week, 2 weeks, 3 weeks, 4 weeks and 5 weeks, respectively.At the same time, one implant was randomly removed from each group for histopathological examination to compare the degree of fibrous vascularization.Results:1. After the fresh porcine cornea was treated with acellular process, all the stromal cells were removed.The extracellular matrix structure was well preserved (including the anterior elastic layer and the first 2 / 3 matrix layer). The results of immunoreaction showed that the inflammatory cells around the tissue of APCS were mainly neutrophils at 2 weeks after subcutaneous implantation, and were mainly neutrophils at 4 weeks after implantation.No significant rejection was found in the experimental group and control group, no neutrophil and lymphocyte infiltration was found in the APCS tissue, and the surrounding fibrous connective tissue formed a clear boundary with the peripheral tissue.(2) at 1 week after operation, the velocity of vascularization in group B began to be slow with different degrees of vascularization. There was a significant difference between the two groups in the results of magnetic resonance imaging (P < 0.05).Two weeks after operation, the degree of vascularization increased in both groups, but the degree of vascularization in group B was slightly lower than that in group A, and the difference between the two groups was statistically significant (p < 0.05).At 3 weeks after operation, the degree of vascularization of the implants in the two groups was deepened and increased obviously, but not completely vascularized. There was no significant difference in vascularization between the two groups (p > 0.05).After 4 weeks and after operation, the two groups reached complete vascularization, and there was no statistical difference between the two groups.Conclusion:1. The porcine corneal stromal cells can be completely removed by repeated freeze-thaw electrophoresis, and the extracellular matrix can be preserved so that its structure can not be destroyed.(2) APCS had no significant effect on the vascularization of HA prosthesis in early stage, and did not delay the complete vascularization of hydroxyapatite prosthesis.3. It is feasible to use APCS as the encapsulation material for HA implant.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R779.6

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