【作者】 李强；

【导师】 张朝跃；

【作者基本信息】 中南大学 ， 临床医学， 2013， 硕士

【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(P>0.05)。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(P<0.01),在第1、3、7天各浓度之间均无差异性(P>0.05),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。更多还原

【Abstract】 OBJECTIVE:To evaluate the biological compatibility of stone needle via animal implantation experiments, cytotoxicity experiments in vitro, and animal skin irritation experiments.METHODS:According to GB/T1688and ISO/10993standards, stone needle from Sibin area is used to conduct animal implantation experiments, cytotoxicity experiments in vitro, and animal skin irritation experiments.1. Animal implantation experiments:①Experimental materials:stone needles from Sibin area and T-shaped silicone stents。②Experimental groups:Sterile stone needle and silica module are embedded between the superficial fascia and muscle in the rat’s back respectively. Rats implanted with stone needle is the experimental group, Rats implanted with T-shaped silicone stent the control group and Rats cut open and then sutured on waist the blank group. Kill the Rats under narcotism on the7th,30th and90th days after surgery respectively, and then take out the embedded modules together with a few of surrounding tissues. Observe the inflammatory cell responses and changes of surrounding tissues of the implanted materials through microscope.2. cytotoxicity experiments in vitro:Fibroblast (L929) of mice were fostered with1640culture medium adding leach liquors of different concentrations from stone needle and silica gel respectively to assess cytotoxicity of stone needle through MTT staining method on the1th,3th, and7th days.3. Animal skin irritation experiments:remain the pasters on the surface of skin for at least4hours. Stone needle grinded into powders is the experimental group, sterile desensitization medical gauze the control group. Record and grade the condition of contact area respectively in1h,24h,48h and72h after removing the paster till the14th day.RESULTS1. Animal implantation experiments:7days after implantation, increased capillary dilatation, congestion, neutrophils, and lymphocytes infiltrating surrounding capillaries are found in the surrounding tissues of two groups and cyst wall formation is not found.30days after implantation, a moderate quantity of new capillaries, a less than moderate quantity of inflammatory cells (including lymphs and neutrophils) and a small quantity of loose cyst wall structures are found in surrounding tissues.90days after implantation, dense, thin-walled and transparent cyst wall structures are found in the surrounding tissues. There is no significant difference between the two kinds of materials (P>0.05).2. cytotoxicity experiments in vitro:Leach liquors of different concentrations from stone needle contribute to the proliferation of L929cells (P<0.01). There are no significant differences among various concentrations on the1th,3th and7th days (P>0.05), and the cytotoxicity of stone needle is rated grade0-1.3. Animal skin irritation experiments:1h,24h,48h and72h after being contacted with two groups of material for5hours, no chromatosis, no erythema, no eschar and edema are found in the surface of skin through the naked eyes and microscope (5times). After being observed for14days continuously, primary irritating index (PII) of the skin is0, and there is no significant difference between two groups (P>0.05).CONCLUSION:Stone needle has good biological compatibility, which offers definite experimental basis to clinical application.更多还原