Department of Immunology, School of Basic Medical Sciences, NHC Key Laboratory of Medical Immunology, Peking University, Beijing 100191, China.
Beijing Advanced Innovation Center for Biomedical Engineering, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100083, China.
Biomed Environ Sci. 2018 Nov;31(11):829-842. doi: 10.3967/bes2018.110.
Graft rejection, with the possibility of a violent immune response, may be severe and life threatening. Our aim was to thoroughly investigate the biocompatibility and immunotoxicology of collagen-based dermal matrix (DM) before assessment in clinical trials.
DM was subcutaneously implanted in BALB/c mice in two doses to induce a potential immune response. The spleen and lymph nodes were assessed for shape, cell number, cell phenotype via flow cytometry, cell activation via CCK8 kit, Annexin V kit, and Ki67 immunostaining. Serum samples were used to measure antibody concentration by enzyme-linked immunosorbent assay. Local inflammation was analyzed by histology and immunohistochemistry staining. Data analysis was performed by one-way ANOVA and non-parametric tests.
Our data illustrate that the spleen and lymph node sizes were similar between the negative control mice and mice implanted with DM. However, in the high-dose DM (DM-H) group, the total cell populations in the spleen and lymph nodes, T cells and B cells in the spleen had slight increases in prophase, and the low-dose DM (DM-L) group did not display gross abnormities. Moreover, DM-H initiated moderate cell activation and proliferation in the early phase post-immunization, whereas DM-L did not. Neither DM-H nor DM-L implantation noticeably increased IgM and IgG serum concentrations. Examination of the local cellular response revealed only benign cell infiltration and TNF-α expression in slides of DM in the early phase.
Overall, DM-H may have induced a benign temporary acute immune response post-implantation, whereas DM-L had quite low immunogenicity. Thus, this DM can be regarded as a safe product.
移植物排斥反应可能伴有剧烈的免疫反应，严重时会危及生命。我们的目的是在临床试验评估之前，彻底研究基于胶原蛋白的真皮基质（DM）的生物相容性和免疫毒性。
DM 以两种剂量皮下植入 BALB/c 小鼠，以诱导潜在的免疫反应。通过流式细胞术评估脾和淋巴结的形状、细胞数量、细胞表型，通过 CCK8 试剂盒、Annexin V 试剂盒和 Ki67 免疫染色评估细胞激活，使用血清样本通过酶联免疫吸附试验测量抗体浓度。通过组织学和免疫组织化学染色分析局部炎症。数据分析采用单因素方差分析和非参数检验。
我们的数据表明，阴性对照组和 DM 植入组的小鼠脾脏和淋巴结大小相似。然而，在高剂量 DM（DM-H）组中，脾和淋巴结中的总细胞群体、脾中的 T 细胞和 B 细胞在前中期略有增加，低剂量 DM（DM-L）组未显示明显异常。此外，DM-H 在免疫接种早期引发中度细胞激活和增殖，而 DM-L 则没有。DM-H 和 DM-L 的植入均未明显增加 IgM 和 IgG 血清浓度。早期 DM 切片的局部细胞反应检查仅显示良性细胞浸润和 TNF-α 表达。
总体而言，DM-H 可能在植入后引发良性短暂的急性免疫反应，而 DM-L 的免疫原性较低。因此，这种 DM 可以被视为一种安全的产品。
