海藻酸钠壳聚糖支架复合成骨细胞特异性多肽修复兔颅骨缺损

发布时间：2018-05-03 09:26

本文选题：成骨细胞特异性识别多肽 + 颅骨缺损　；参考：《安徽医科大学》2017年硕士论文

【摘要】：成骨细胞特异性识别多肽是以人牙龈成纤维细胞为阴性吸附细胞,人颅骨成骨细胞为阳性靶细胞,对噬菌体展示十二肽库进行全细胞差减筛选,获得的与人成骨细胞有高度亲和性的多肽分子。其氨基酸序列为VLAVPWSEPGYL(缬氨酸-亮氨酸-丙氨酸-缬氨酸-脯氨酸-色氨酸-丝氨酸-谷氨酸-脯氨酸-甘氨酸-络氨酸-亮氨酸)。本实验依据组织工程学的原理,采用壳聚糖、海藻酸钠为支架,成骨细胞特异性识别多肽为信号因子,将二者复合植入兔颅骨极限骨缺损模型中,以评价该特异性多肽的促成骨效应。为临床上解决颅颌面部骨缺损及口腔种植中普遍存在的骨质、骨量不足问题提供新的研究思路和治疗方法。具体内容如下:第一部分:海藻酸钠/壳聚糖/多肽复合凝胶的制备在无菌条件下将海藻酸钠/多肽混悬液与壳聚糖溶液混合,维持二者为非凝胶化的溶液状态,并通过搅拌反应使得多肽均匀分布于混合液中,调节多肽到目的终浓度160μg/ml,在一定温度条件下制备成海藻酸钠/壳聚糖/多肽复合凝胶。同时制备不加载成骨细胞特异性识别多肽的复合凝胶,在实验中用作对照组。第二部分:制备兔颅骨极限骨缺损动物模型选取健康新西兰兔12只,制备兔颅骨双侧15 mm直径圆形极限骨缺损。实验组缺损处植入海藻酸钠/壳聚糖/多肽水凝胶,对照组植入海藻酸钠/壳聚糖复合水凝胶,空白组不植入任何载体及药物,随机分组。术后8周活体行CBCT扫描、高频钼靶X线扫描、HE染色检测成骨效果。结果表明:影像学及组织形态学结果可见,实验组缺损区成骨量明显多于对照组及空白组,差异有统计学意义(P0.05)。结论:成骨细胞特异性识别多肽可促进兔颅骨缺损的骨修复。
[Abstract]:Human gingival fibroblasts were used as negative adsorption cells and human skull osteoblasts as positive target cells. The phage display dodecapeptide library was screened by whole cell subtractive screening. Polypeptide molecules with high affinity to human osteoblasts were obtained. Its amino acid sequence is VLAVPWSEPGYL (valine-leucine-alanine -valine-proline-tryptophan-serine-glutamate-proline-glycine-complex acid-leucine). According to the principle of tissue engineering, chitosan and sodium alginate were used as scaffolds, and osteoblast specific recognition polypeptide was used as signal factor. To evaluate the bone-promoting effect of the specific polypeptide. To provide a new way of thinking and treatment for the clinical solution of craniomaxillofacial bone defects and oral implants. The main contents are as follows: the preparation of sodium alginate / chitosan / polypeptide composite gel in aseptic condition mixed sodium alginate / polypeptide suspensions with chitosan solution to maintain a non-gelatinized solution state. The polypeptide was evenly distributed in the mixed solution by stirring reaction, and the final concentration of the polypeptide was adjusted to 160 渭 g / ml. The sodium alginate / chitosan / polypeptide composite gel was prepared under certain temperature. At the same time, the composite gel without loading osteoblast specific recognition polypeptide was prepared and used as control group. In the second part, 12 healthy New Zealand rabbits were selected to establish the animal model of rabbit cranial limit bone defect. The bilateral 15mm diameter circular limited bone defect of rabbit skull was prepared. Sodium alginate / chitosan / polypeptide hydrogel was implanted in the defect of the experimental group, and sodium alginate / chitosan hydrogel was implanted in the control group. 8 weeks after operation, CBCT scanning was performed in vivo, and high frequency molybdenum target X-ray scanning was used to detect the osteogenic effect by HE staining. The results showed that the amount of osteogenesis in the experimental group was significantly higher than that in the control group and the blank group, and the difference was statistically significant (P 0.05). Conclusion: osteoblast specific recognition peptide can promote bone repair of rabbit skull defect.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R782

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