【作者】 李波；

【导师】 徐大宝；

【作者基本信息】 中南大学 ， 妇产科学， 2011， 硕士

【摘要】 目的：通过动物埋植实验及体外细胞毒性实验评估宫腔粘连治疗装置(自主专利产品)的生物相容性。方法：根据GB/T1688.和ISO10993.标准规定的方法对自主专利产品宫腔粘连治疗装置进行动物埋植实验及体外细胞毒性试验。动物埋植实验：①实验材料：将自行研制的专利产品硅橡胶(以下简称：硅胶)宫腔粘连治疗装置、已认证的T-CU环的聚乙烯混硫酸钡T型支架,统一制成直径为1.5mm,长5mm的规则光滑圆柱体。②实验分组：按照随机数字表法将9只健康的SD雌性大鼠编号,分别在大鼠的左右后腿浅筋膜与肌肉层之间植入已消毒的硅胶和聚乙烯模块,植入硅胶的为实验组,植入聚乙烯模块的为对照组。右前腿切开后缝合,不做特殊处理,作为空白组。为防止系统误差,将硅胶植入编号为单数的大鼠左后腿和编号为双数大鼠的右后腿；将聚乙烯模块植入编号为单数的大鼠右后腿和编号为双数大鼠的左后腿。完成手术后继续饲养大鼠,分别于术后7、30、90天麻醉后处死1-3号、4-6号、7-9号大鼠,取出埋植模块周围组织,制做石蜡切片,苏木精-伊红染色。实验评估：光镜下观察埋入材料周围组织、细胞的反应和变化,判断两种材料的组织相容性差异。细胞毒性实验：利用体外培养的小鼠成纤维细胞(L929),采用不同浓度的宫腔粘连治疗装置浸提液(100%、50%、25%)为实验组、已认证的T型环聚乙烯支架浸提液为对照组、1640培养基为空白组分别培养细胞,于培养后的第1、3、7天通过MTT法染色,在酶标490nm波长处测定吸光值,计算相对增值率,逐日用倒置显微镜观察比较各组细胞形态学发生的改变,对宫腔粘连治疗装置的细胞毒性进行评价。结果：动物埋植实验：所有大鼠安全经受手术,术后未出现感染,未使用抗生素,未出现排异反应。所有大鼠切口正常愈合。①植入后7天,两种材料周围见大量中性粒细胞浸润,并伴有明显血管充血,材料周围无囊壁形成。②植入后30天,材料周围可见少量疏松的囊壁组织,周围见中量中性粒细胞,周边组织无明显渗血。③植入后90天,两种材料周围均可见透明、致密、质薄囊壁形成,厚度明显薄于植入后30天,两种材料之间差异无显著性意义(P>0.05)。细胞毒性实验：随着时间的推移,不同浓度宫腔粘连治疗装置浸提液对L929细胞均有促进增殖作用(P<0.01),在第1、3、7天各浓度之间无明显差异性(P>0.05),提示L929细胞增殖与浸提液的浓度无明显相关,实验组细胞数量及形态与阴性对照组、空白组无明显差异,评分宫腔粘连治疗装置的细胞毒性等级为0级-1级。结论：宫腔粘连治疗装置具有良好的生物相容性,无细胞毒性,具有良好的细胞相容性,符合临床使用要求。更多还原

【Abstract】 Objective To evaluate the biocompatibility of a sample of an IUD for the treatment of IUAs,as a new patent by animal embedding experiment and cytotoxicity in vitro.Methods According to GB/T16886 and ISO10993 as a standard of the animal embedding experiment and cytotoxicity in vitro were using a sample of an IUD for the treatment of IUAs.The animal of the embedding experiment:①The sample of an IUD for the treatment of IUAs with silicone rubber and T-Cu IUD of polythene,they were make a uniform velvet cylinder for diameter 1.5mm and height 5mm.②Thirty healthy Sprague Dawley rats were selected and numbered randomly to placed silica and polythene module between the superficial fascia and muscle of both left and right leg, which served as a experimental group and control group. To dissect and suture the front legs, which serves as a blank group. For preventing of systematic error, the silicone rubber was implanted into the left leg of animals numbered in odd and the right leg of animals numbered in dual, and the polythene into the right leg of odd number of the animals and left leg of dual number of the animals. No.1-3, 4-6, and 7-9 SD rats were executed under anesthesia on days 7,30, and 90 after operation respectively. The modules and surrounding tissues were taken out from the animals for paraffin section, and HE staining. The reaction and changes of cell and surrounding tissues were observed under inverted microscope to compare their tissue-compatibility. The cytotoxicity experiment:L929 cells were cultured with different concentration to extracts the sample of an IUD for the treatment of IUAs (100%,50%,25%, V/V),the T-Cu IUD of polythene bracketed and RPMI-1640, which served as a experimental group,control group and blank group in vitro,and MTT assay was performed to evaluate the cell proliferation on days1,4,and 7.The morphologic changes of cells were observed by inverted microscope to evaluate the cytotoxicity.Results The animal embeding experiment:None of the SD rats was infected after operation and no antibiotics were used and no rejection were encountered.All of the SD rats’s wound were healed.①seven days after implantation, there were a great amount of infiltrated heterophilic granulocytg were accompanied by obvious congestion around the modules, but no cystwall formed.②Thirty days later, there were medium-amount of infiltrated heterophilic granulocytes and around two kinds of modules have loose cystwall formed. There were no capillary hemorrhage around the tissues.③Ninety days late, around two kinds of modules have pellucidly,compactly and sleazy cystwall formed, but the thickness were much thinner than after implantation, these two kinds of modules have no statistics discrepancy(P>0.5). The cytotoxicity experiment:MTT assay revealed to extracts of the sample of an IUD for the treatment of IUAs, enhanced the cell proliferation (P<0.01), but not a dose-dependent manner (P>0.05).In the experiment group, the cell proliferation and morphologic have no discrepancy compared with the control group and blank group. The cytotoxicity grade of the sample of an IUD for the treatment of IUAs was 0 or 1, suggesting that the material was almost free of cytotoxicity.Conclusion The sample of an IUD for the treatment of IUAs possesses ideal biocompatibility, free of cytotoxicity, according to the standard of clinical use, and has a great potential.更多还原