论文部分内容阅读
目的研究脱细胞的细胞外基质(ECM)材料植入小鼠后对其固有免疫(包括巨噬细胞和中性粒细胞)的影响,初步建立一套以小鼠为实验动物的动物源材料免疫原性去除效果的测试评价方法。方法将新鲜膜材料和新工艺制备的脱细胞和除DNA的ECM材料移植到小鼠腹腔中,7 d后,检测脾脏指数的变化,采用BD绝对计数管检测小鼠外周血中性粒细胞和腹腔液巨噬细胞的绝对数量,采用小鼠Th1、Th2、Th17细胞因子流式微球分析技术检测小鼠血浆和腹腔液白细胞介素2(IL-2)、IL-4、IL-6、γ干扰素(IFN-γ)、肿瘤坏死因子α(TNF-α)、IL-17A和IL-10分泌。结果使用新鲜膜材料的小鼠体质量不增加、但脾脏指数增加,小鼠外周血中性粒细胞和腹腔液巨噬细胞的数量明显增加,IL-6和IFN-γ的分泌水平增加。而使用脱细胞的ECM材料小鼠以上参数仅有微弱的影响或无影响。结论脱细胞、除DNA的ECM材料对小鼠的固有免疫应答无明显影响。
Objective To study the effects of decellularized extracellular matrix (ECM) material on innate immunity (including macrophages and neutrophils) after implantation in mice and to establish a set of animal-derived materials Test evaluation method of the original removal effect. Methods The decellularized and DNA-removed ECM materials prepared by fresh membrane material and new technology were transplanted into the peritoneal cavity of mice. The changes of spleen index were detected after 7 days. The numbers of peripheral blood neutrophils and The amount of peritoneal macrophages was determined by the method of flow cytometry of mouse Th1, Th2 and Th17 cytokines. The levels of interleukin 2 (IL-2), IL-4, IL-6, Interferon (IFN-γ), tumor necrosis factor α (TNF-α), IL-17A and IL-10 secretion. Results The body mass of mice with fresh membrane material did not increase, but the spleen index increased, the number of neutrophils and peritoneal macrophages in peripheral blood of mice increased obviously, and the secretion of IL-6 and IFN-γ increased. The use of acellular ECM material mouse above parameters only weakly or no effect. Conclusion The decellularization and DNA removal of ECM did not affect the innate immune response in mice.