异种关节软骨脱细胞基质支架的免疫反应研究

Research of immunoreaction of heterogeneic articular cartilage extracellular matrix derived scaffold

[目的]观察异种脱细胞软骨基质支架的免疫排斥反应,探讨其用于组织工程软骨支架的可行性。[方法]分别获取猪和新西兰大白兔的关节软骨,制作成:①猪脱细胞软骨支架;②猪未脱细胞软骨支架;③兔脱细胞软骨支架。将9只新西兰大白兔随机分成三组,分别在兔背部深筋膜与肌膜之间埋入:①异种(猪)脱细胞软骨支架;②异种(猪)未脱细胞软骨支架;③同种(兔)异体的脱细胞软骨支架,术后分组饲养。于术后1、2、4周分别取支架埋藏部位组织,进行普通病理观察及CD4+、CD8+T淋巴细胞免疫组织化学检测,并在术前和术后1周,2周,4周抽取外周血,分离检测血清中对支架产生的抗体的浓度(ELISA法)。[结果]所有新西兰大白兔无不良反应,细胞免疫显示异种(猪)和同种(兔)的脱细胞软骨支架及周围埋藏部位组织无明显淋巴细胞浸润,而异种未脱细胞组支架在埋入第4周时有明显淋巴细胞浸润,体液免疫显示术前后抗体浓度无明显差异(P>0.05)。[结论]猪来源的脱细胞软骨支架用于异种移植时无免疫反应,从免疫学方面为异种脱细胞软骨支架的临床应用提供了可行性。

[ Objective] To observe the immune rejection of xenogenic acellular cartilage matrix scaffold, and to investigate its possibility for cartilage tissue engineering. [ Methods] Articular cartilage was obtained from pigs and New Zealand white rabbits, respectively, then it was used to produce porcine acellular cartilage scaffold , porcine undecellularized cartilage scaffold, and rabbit acellular cartilage scaffold. Nine New Zealand white rabbits were randomly divided into three groups , then the three types of cartilage scaffold were buried into the back of rabbits, between the deep fascia and the sarcolemma respectively. At the 1, 2 and 4 weeks after surgery , the tissue where the scaffold was buried was obtained for pathological testing and immunohistochemical analysis of CD4＋ and CD8＋ T - cell. And peripheral blood samples were collected to detect serum antibody concentration using ELISA. [ Results] There were no adverse reactions in any of New Zealand white rabbits. Cell - mediated immunity showed the acellular cartilage scaffold and tissue around the burial site had no significant lymphocytic infiltration. In the xenogenic undercellularized group, there were marked lymphocytic infiltration at the 4th week after surgery. Humoral immunity showed there was no significant difference in antibody concentration before and after surgery （ P 〉0. 05） . [ Conclusion] Porcine articular cartilage extracellular matrix derived scaffold used in heterogenic transplantation or allograft have no immunologic rejection. This research provides a feasibility for application of xenogenic acellular cartilage to clinic.