砭石生物相容性的实验研究

发布时间：2018-06-07 00:39

本文选题：砭石 + 生物相容性　；参考：《中南大学》2013年硕士论文

【摘要】：目的：通过动物体内植入、体外细胞毒性及动物皮肤刺激实验评估砭石的生物相容性。方法：根据GB/T1688.和IS0/10993.标准规定的方法对泗滨砭石进行动物体内植入、体外细胞毒性及动物皮肤刺激实验。 1.动物体内植入实验：①实验材料选取泗滨砭石,硅胶T型支架。②分组：植入砭石为实验组,植入硅胶T型支架为对照组,大鼠于腰部切开后缝合,作为空白组。分别于手术后7、30、90天麻醉后处死大鼠,取出埋植模块及周围少量包裹组织。HE染色光镜下观察埋入材料周围组织、细胞反应和变化。 2.细胞毒性实验：培养小鼠成纤维细胞(L929),采用不同浓度的砭石浸提液、硅胶浸提液、1640培养基分别培养细胞,在第1、3、7天通过MTT法染色,对砭石的细胞毒性进行评价。 3.动物皮肤刺激实验：砭石研磨成粉状为实验组,无菌脱敏医用纱布为对照组,皮肤固定敷贴片至少4h。单次接触试验时,在除去敷贴片后1h,24h,48h、72h,观察至14d,记录各接触部位情况并评分。结果： 1.动物体内植入实验：植入后7天,两组周围组织均见毛细血管增多,扩张充血,中性粒细胞、淋巴细胞增多浸润围绕毛细血管,材料周围组织未发现囊壁形成。植入后30天周围组织见中等量新生毛细血管、中等偏少炎性细胞(包括淋巴、中性粒细胞),见少量疏松的纤维囊壁结构。植入后90天周围组织见致密、质薄、透明纤维囊壁结构,两种材料之间差异无显著性意义(P0.05)。 2.体外细胞毒性实验：不同浓度砭石浸提液对L929细胞均有促进增殖作用(P0.01),在第1、3、7天各浓度之间均无差异性(P0.05),砭石的细胞毒性等级评分为0级-1级。 3.动物皮肤刺激实验：两组材料接触5h后,除去敷贴片后1h、24h、48h和72h在直视及5倍放大镜下观察,皮肤无色素沉着、无红斑、焦痂形成,无水肿。术后连续观察14d,原发性刺激指数(P工I)0分,两组无明显差异。结论：砭石具有良好的生物相容性,为临床应用提供了一定的实验依据。
[Abstract]:Objective: Biocompatibility of Bian-stone was evaluated by in vivo implantation, in vitro cytotoxicity and skin stimulation. Methods: According to GB / T 1688. And IS0 / 10993. The standard method was used to implant Sibin stone in vivo and to test in vitro cytotoxicity and skin irritation. 1. Sibin stone and silica gel T stent were divided into two groups: the experimental group was implanted with silica gel T stent, the control group was implanted with silica gel T stent, and the rats were sutured after lumbar incision. The rats were anesthetized 7 days after operation for 30 days. The implanted modules and a small amount of surrounding tissue were taken out. The tissue around the implanted material was observed under light microscope and the cell reaction and changes were observed under light microscope. 2. Cytotoxicity test: mouse fibroblasts were cultured in different concentrations of Bian stone extract and silica gel extract on the medium of 1640. The cytotoxicity of Bian stone was evaluated by MTT staining on the 3rd day of the first day. 3. Animal skin irritation test: the stone was ground into powder as experimental group, sterile desensitized medical gauze as control group, and skin fixation application for at least 4 hours. During the single contact test, the contact sites were recorded and evaluated at 1 h, 24 h, 48 h and 72 h after the application was removed. Results: 1. In vivo implantation experiment: 7 days after implantation, capillary increased, dilated and hyperemia, neutrophil and lymphocyte infiltration were observed in the surrounding tissues of both groups, and no cyst wall was found in the surrounding tissues. 30 days after implantation, a moderate amount of neocapillaries was found in the surrounding tissues, and a few loose fibrous cystic wall structures were observed in the medium less inflammatory cells (including lymph nodes, neutrophils and neutrophilic granulocytes). The surrounding tissue was dense, thin and transparent fiber wall structure was found 90 days after implantation. There was no significant difference between the two materials (P 0.05). 2. In vitro cytotoxicity test: different concentrations of Bian stone extract could promote the proliferation of L929 cells. There was no difference between different concentrations on the 1st day 3 ~ (th) day. The cytotoxicity grade of Bian stone was grade 0 ~ (-1). 3. Animal skin stimulation test: after 5 hours of contact with the two groups of materials, the skin was observed under direct vision and 5 times magnifying glass at 24 h, 48 h and 72 h after application. There was no pigmentation, no erythema, no scab formation and no edema. After 14 days, there was no significant difference between the two groups. Conclusion: Bian stone has good biocompatibility and provides some experimental basis for clinical application.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R318.08

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