羧基化的聚酰胺—胺/无定形磷酸钙诱导Ⅰ型胶原纤维矿化的研究

Study on mineralization of type-I collagen fibrils induced by carboxylated polyamide-amine/amorphous calcium phosphate

  • 摘要: 目的:利用羧基化的聚酰胺—胺稳定矿化物前驱相无定形磷酸钙,合成并表征同时具有有机和无机材料特性的仿生矿化复合物(ACP/CPAMAM),并观察其诱导胶原纤维仿生矿化的效果。方法:制备ACP/CPAMAM纳米复合物,并通过透射电子显微镜、选区电子衍射、粒径分析、电位分析等对其表征和验证。最终体外实验应用牛跟腱胶原纤维,借助透射电镜—能谱分析观察胶原纤维内部及周围晶体形成情况。结果:ACP/CPAMAM 纳米复合物成功合成并且为无定形相,平均粒径为25.13 nm。透射电子显微镜及选区电子衍射显示,ACP的表面出现10 nm厚的低衬度包裹层,证实了ACP/CPAMAM的成功合成且其结构为非晶相。透射电镜—能谱分析显示,应用ACP/CPAMAM矿化肌腱Ⅰ型胶原纤维后可见高密度晶体,胶原纤维周围可见致密、连续的晶体形成,证实该纳米复合物可连续诱导胶原纤维矿化。结论:ACP/CPAMAM作为一种新型的纳米复合生物材料具有良好的生物活性和仿生矿化特性,能够诱导胶原纤维形成矿化产物,提示ACP/CPAMAM可能成为一种新型诱导胶原纤维仿生再矿化的纳米材料。

     

    Abstract: Objective:To synthesize and characterize a biomimetic mineralization complex material (ACP/CPAMAM) with both organic and inorganic properties using carboxylated polyamide-amine to stabilize the precursor of mineralized amorphous calcium phosphate, and to observe its biomimetic mineralization effect on collagen fibrils.Methods:ACP/CPAMAM nanocomplex was prepared, characterized and verified by transmission electron microscopy(TEM), electron diffraction(SAED), particle size analysis and potential analysis.Collagen fibrils of bovine tendon were applied in vitro experiment, and the crystal formation in and around collagen fibrils was observed by TEM and the energy spectrum analysis.Results:ACP/CPAMAM nanocomplex was successfully synthesized and was amorphous with an average size of 25.13 nm.TEM and SAED showed that the surface of ACP had a low-contrast coating with a thickness of 10 nm, which confirmed the successful synthesis of ACP/CPAMAM was amorphous.TEM and energy spectrum analysis showed that high density crystals were observed after the application of ACP/CPAMAMmineralized tendon type-Ⅰcollagen fibers, and dense and continuous crystal formation was observed around the collagen fibers, which confirmed the nanocomplex could continuously induce collagen fiber mineralization.Conclusion:ACP/CPAMAM, as a novel nanocomplex, has good biological activity and biomimetic mineralization characteristics.It can induce the type-I collagen fibrils to form mineralized products, suggesting that ACP/CPAMAM may become a novel nanomaterial to induce biomimetic re-mineralization of collagen fibers.

     

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