羊脱细胞真皮基质的体内外毒性实验

发布时间：2018-05-21 04:44

  本文选题：羊皮 + 脱细胞真皮基质　； 参考：《中南大学学报(医学版)》2016年10期

【摘要】：目的:利用溶血反应、热原反应和细胞毒性反应实验了解羊脱细胞真皮基质(acellular dermal matrix,ADM)在动物体内外的毒性反应强度。方法:溶血反应:分别制作交联型和非交联型羊ADM的浸提液,为交联组和非交联组;并设置阳性对照组10 m L灭菌注射用水,阴性对照组10 m L 0.9%氯化钠溶液。将新鲜抗凝稀释兔血加入实验组和对照组试管中,离心后取上清液测量吸光度值,计算溶血度。热原实验:按照条件入选16只新西兰兔,分为初试A,B两组各3只和复试C,D两组各5只。分别将交联与未交联的两种ADM浸提液注射入两组新西兰兔体内,注射后每隔0.5 h测量一次体温,共测量6次,以6次中最高一次减去正常体温即为该兔的体温升高度数。据此判断是否符合生物制品热原实验的评价标准。MTT实验:收集对数期的小鼠成纤维细胞细胞,种植于含有不同浓度羊ADM浸提液的培养基,测量吸光度值进行细胞活力测定。结果:实验A,B组溶血度均小于5%,热原反应测得新西兰兔体温升高之和低于1.8℃。MTT实验显示10%~90%羊ADM浸提液培养基对体外细胞的毒性为1级,100%的羊浸提液对体外细胞的毒性为2级,交联型与非交联型组的浸提液对体外细胞的毒性的无显著性差异(P0.05)。均对体外细胞的增殖影响轻微。结论:羊ADM植入新西兰兔体内引起的溶血反应和热原反应在评价标准之内,在体外对细胞的增殖和活力影响不明显。
[Abstract]:Objective: to investigate the intensity of acellular dermal adm in vivo and in vitro by hemolytic reaction pyrogen reaction and cytotoxicity test. Methods: hemolysis reaction: the extract of cross-linked and non-cross-linked sheep ADM was made into cross-linked group and non-cross-linked group respectively, and 10 mL sterilizing water was set up in the positive control group and 0.9% sodium chloride solution in the negative control group. The fresh anticoagulant diluted rabbit blood was added to the test tube of the experimental group and the control group. The absorbance of the supernatant was measured after centrifugation and the hemolysis was calculated. Pyrogen test: sixteen New Zealand rabbits were selected according to the conditions and divided into two groups: group A (n = 3) and group C (n = 5). The cross-linked and uncrosslinked ADM extractions were injected into the two groups of New Zealand rabbits respectively. The body temperature was measured every 0.5 h after injection. According to the results, the mouse fibroblasts in logarithmic phase were collected and planted in the medium containing different concentrations of sheep ADM extract, and the cell viability was measured by measuring the absorbance value. Results: the hemolysis degree of group A and B was less than 5. The sum of the elevated body temperature of New Zealand rabbits was less than 1.8 鈩，

本文编号：1917824