成骨细胞异种移植的免疫观察

IMMUNOLOGICAL RESEARCH OF RAT OSTEOBLAST XENOTRANSPLANTATION

目的 探讨异种成骨细胞移植及组织工程骨构建的可能性。 方法 用二步酶消化 -组织块联合接种法行 SD大鼠骨细胞原代培养 ,传代后用碱性磷酸酶 (AL P)染色法进行细胞鉴定 ;再将 2 8只大耳白兔分为 3组 :自体移植组 (A组 )、异种移植组 (B组 )、正常对照组 (C组 ) ,分别于腹直肌袋内注入自体软骨细胞悬液、SD大鼠成骨细胞及生理盐水 ,于 1、2、4及 8周行免疫学及组织形态学检测。 结果 细胞原代接种后 5天成单层 ,经鉴定为成骨细胞 ;实验兔各组移植后受体无明显全身及局部排异反应 ;B组移植后 2、4周检测到特异性抗体 ,2周检测到细胞介导的细胞毒反应 ,4周达高峰 ,8周开始减退 ,HE染色见大量炎性细胞浸润 ,至 8周无明显异位成骨现象。 A、C两组移植后未见明显体液及细胞免疫反应。 结论 单纯异种成骨细胞不能作为细胞移植及组织工程骨构建的可靠细胞来源。

Objective To detect the immunoreaction after osteoblast xenotransplantation and to investigate the possibility of heterogenic osteocyte transplantation and tissue engineered bone reconstruction. Methods Rat osteoblasts were isolated by two-part bony digestion/elements inculturing, and incubated in vitro at 37℃, 5% carbon dioxide for 5 days until they multiplied and formed a monolayer on the bottom of dish. Twenty-eight rabbits were divided into 3 groups. Autograft of osteoblasts(group A), xenograft of osteoblasts(group B) and normal saline(group C) were implanted into the rectus abdominus muscle. The immunological and histological observations were performed after 1,2,4 and 8 weeks of transplantation. Results Cultured cells reached confluence within 5 days and was identified as osteoblasts by ALP staining and Bon kossa staining. The result of host versus graft reaction was negative. In group B, specific antibody reaction was detected 2 weeks and 4 weeks after transplantation. Cell mediated cytotoxicity was detected after 2 weeks, reached the peak value 4 weeks later, and then began to decline 8 weeks later. HE staining showed mass inflammatory cells and no ectopic ossification after 8 weeks. Conclusion Heterogenic osteoblast transplantation will lead to an obvious change in host humoral and cellular immunity and lost the ability of bone formation . So, it can not be used for the reliable cell sources for osteocyte transplantation or tissue-engineering bone reconstruction.