| 陈 骥,王君俊,张 旻,程百祥,李德华,张 茜.minTBP-1/IGF-1融合蛋白钛涂层对高糖高脂环境下成骨细胞分化活性的影响[J].,2017,17(29):5611-5617 |
| minTBP-1/IGF-1融合蛋白钛涂层对高糖高脂环境下成骨细胞分化活性的影响 |
| Enhanced Osteoblast Differentiation of Directional minTBP-1/IGF-1 Coatings Titanium under High-Glucose/ High-Fat Microenvironment |
| 投稿时间：2017-02-25 修订日期：2017-03-18 |
| DOI：10.13241/j.cnki.pmb.2017.29.003 |
| 中文关键词: 高糖高脂环境 钛 minTBP-1 胰岛素样生长因子-1 成骨分化 |
| 英文关键词: High-glucose / high-fat environment Titanium minTBP-1 IGF-1 Osteogenic differentiation |
| 基金项目:国家自然科学基金项目（81300910）；全军医学科技青年培育计划（16QNP109） |
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| 中文摘要: |
| 摘要 目的：评价自行设计的minTBP-1与IGF-1融合蛋白涂层修饰的纯钛表面对高糖高脂环境下成骨相关基因及蛋白表达的影响。方法：借助互联网ProtParam工具设计并筛选出亲水性较高的minTBP-1与IGF-1融合蛋白，用于修饰钛表面，分别将正常成骨细胞和高糖高脂环境下培养的成骨细胞接种于其表面，以钛表面无涂层的正常成骨细胞为对照组，通过碱性磷酸酶（ALP）、茜素红S染色及定量，比较成骨细胞的生物学特性；1周、2周时，Real-time PCR 检测成骨细胞ALP、Ι型胶原、Runx2和OCN基因表达水平的变化；1周、2周和3周时，Western Blot分别检测成骨细胞ALP、OCN蛋白的表达量。结果：高糖高脂环境下培养的成骨细胞扁平、呈不规则多边形，其ALP的表达、钙化结节形成均低于正常成骨细胞（P＜0.05）。高糖高脂环境下，在目标融合蛋白修饰的钛表面接种成骨细胞1周时，ALP基因及蛋白均有明显表达，与正常成骨细胞相比无统计学差异（P＞0.05）；2周时Ι型胶原、Runx2以及OCN的基因明显高表达，第3周时OCN蛋白显著表达，与正常成骨细胞相比无统计学差异（P＞0.05）。结论：minTBP-1与IGF-1融合蛋白涂层修饰的纯钛表面有利于改善高糖高脂环境下成骨细胞的生物学活性、促进了成骨相关基因及蛋白的表达，有望为2型糖尿病条件下钛-骨结合的种植体表面改性提供新策略。 |
| 英文摘要: |
| ABSTRACT Objective: To evaluate the osteoblast differentiation activity of osteoblasts cultured in high-glucose/high -fat environ- ment by titanium coatings directional minTBP-1/IGF-1. Methods: The hydrophilic minTBP-1/IGF-1 protein was designed by ProtParam tool in Internet. The healthy or osteoblasts in high-glucose/high -fat environment were cultured onto titanium, respectively. The activity of osteoblast differentiation was detected by following parameters, such as ALP, alizarin red S and cell cloning. The relative expression of ALP mRNA (protein), Col1a1 mRNA, Runx2 mRNA and Osteocalcin mRNA (protein) were investigated. Results: After analysis of the ALP activity, numbers of calcified nodules and cell cloning ability, we found the osteoblast cultured in high-glucose/high -fat environ- ment were inferior to the normal osteoblast. Compared to the normal osteoblasts cultured on the titanium disks, there was no significant difference in osteogenesis related genes (ALP, Col1a1, Runx2 and OCN) and protein(ALP and OCN) expression of morbid osteoblasts incubated by three kinds of fusion protein (P＞0.05). Conclusion: The enhanced osteoblast differentiation ability by regulating the titani- um-bone response of high-glucose/high -fat microenvironment through directional minTBP-1/IGF-1 may be of great value for in vivo ap- plications of T2DM osseointegration. |
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