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摘要:
免疫应答反应一直是限制可植入生物材料应用的关键因素之一。实验评估了2种用于骨修复的胶原膜在体内的免疫反应，希望为其临床试验提供依据。在将2种膜皮下植入到BALB/c小鼠后的第14天，与阴性对照（NC，未植入材料）相比，脾和淋巴结没有明显肿大，淋巴结细胞数几乎没差异，而脾细胞数大约是NC的2倍。流式细胞术分析显示植入胶原膜1导致脾中T细胞比例减少了约13%，但是没有影响T细胞亚群，而植入胶原膜2对小鼠的脾细胞组成没有明显影响；2种胶原膜都激活了一定的B细胞，激活率大约为NC小鼠的2倍。淋巴细胞体外增殖实验显示与NC组无显著差异。酶联免疫吸附测试表明胶原膜1导致了第14天血清中的抗体浓度升高至NC小鼠的2倍。局部H&E染色显示2种材料都引起了轻微的细胞浸润。这2种胶原膜引起的免疫反应很微弱，可以被应用于临床试验。
Abstract:Immune response is one of the critical factors to limit implantable biomaterials' application. In this study, two kinds of collagen membranes for bone regeneration were evaluated in terms of immunological response in vivo, which is hoped to lay foundations for clinical trials. At day 14 following subcutaneous implantation of the two membranes in BALB/c mice, compared with negative control (NC, no implant), spleen and lymph nodes showed no obvious swelling, and normal lymph nodes'cell population as well as one-fold more splenic cell population was observed. Flow cytometry analysis demonstrated that in mice with collagen membrane 1, splenic T lymphocytes percentage decreased by about 13%, but no significant change was discovered in T lymphocyte subsets; collagen membrane 2 did not trigger apparent splenic cells composition alteration; the two membranes activated one-fold more B lymphocytes than NC. Enzyme linked immunosorbent assay indicated that one-fold higher IgG concentration than NC was detected in mice with collagen membrane 1 in day 14. Lymphocytes proliferation assay in vitro did not show significant difference from NC. H&E staining of local tissues exhibited slight cell infiltration around the two membranes. The two collagen membranes have engendered mild immune response and can be applied to clinical trials.
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Key words:
- collagen membrane /
- bone regeneration /
- immune response /
- flow cytometry /
- inflammation
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图 2 脾和淋巴结的免疫细胞计数结果(***p < 0.005)
Figure 2. Immune cell population of spleen and lymph nodes (***p < 0.005)
图 4 CD4+T细胞、CD8+T细胞和CD19+细胞的Ki67表达情况
Figure 4. Ki67 expression of CD4+T lymphocytes, CD8+T lymphocytes and CD19+ lymphocytes
图 5 脾和淋巴结细胞体外增殖情况(**p < 0.01)
Figure 5. Proliferation in vitro of splenic cells and lymph nodes cells (**p < 0.01)
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