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携带头蛋白小干扰RNA的胶原蛋白/生物玻璃复合材料的成骨作用

[Osteogenic effect of collagen/bioglass composites carrying noggin siRNA].

作者信息

Chen Yanling, Chen Liangjiao, Li Zhengmao, Lan Zedong

机构信息

Department of Orthodontics, Stomatological Hospital Affiliated to Fujian Medical University, Fuzhou 350004, China.

Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou 510140, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2018 Jan 30;38(1):106-111. doi: 10.3969/j.issn.1673-4254.2018.01.17.

Abstract

OBJECTIVE

To investigate osteogenic effect of collagen/bioglass composites loaded with a small interfering RNA (siRNA) targeting noggin.

METHODS

The collagen/bioglass composites loaded with the negative control siRNA or noggin siRNA were prepared by freeze-drying method. CCK8 test was used to evaluate the proliferation of MC3T3 cells exposed to the aqueous extracts of collagen/bioglass composites and the siRNA-loaded collagen/bioglass composites. ALP activity assay, quantitative real-time PCR and Alizarin Red staining were used to assess the effect of the 3 composites on mineralization in MC3T3 cells.

RESULTS

MC3T3 cells cultured for 3 and 5 days in the presence of the extracts of the 3 composites all showed significantly more active proliferation than the blank control cells ( < 0.05). Compared with the cells seeded on the scaffold without siRNA, MC3T3 cells seeded on collagen/bioglass scaffold loaded with noggin siRNA showed a significantly higher ALP activity at 14 days after seeding ( < 0.05) with significantly increased expression of , and BSP mRNAs ( < 0.05). Alizarin Red staining showed that the cells seeded on the noggin siRNA-loading collagen/bioglass scaffold contained significantly more mineralized nodules than the other cells ( < 0.05).

CONCLUSIONS

The collagen/bioglass composites loaded with noggin siRNA have a good biocompatibility, and the collagen/bioglass composites and noggin siRNA show a synergistic effect in promoting osteogenesis.

摘要

目的

研究负载靶向头蛋白的小干扰RNA(siRNA)的胶原蛋白/生物玻璃复合材料的成骨作用。

方法

采用冷冻干燥法制备负载阴性对照siRNA或头蛋白siRNA的胶原蛋白/生物玻璃复合材料。使用CCK8试验评估暴露于胶原蛋白/生物玻璃复合材料及负载siRNA的胶原蛋白/生物玻璃复合材料水提取物中的MC3T3细胞的增殖情况。采用碱性磷酸酶(ALP)活性测定、定量实时聚合酶链反应(PCR)和茜素红染色来评估这3种复合材料对MC3T3细胞矿化的影响。

结果

在3种复合材料提取物存在的情况下培养3天和5天的MC3T3细胞,其增殖活性均显著高于空白对照细胞(P<0.05)。与接种在未负载siRNA支架上的细胞相比,接种在负载头蛋白siRNA的胶原蛋白/生物玻璃支架上的MC3T3细胞在接种后14天显示出显著更高的ALP活性(P<0.05),且Runx、Ocn和骨涎蛋白(BSP)信使核糖核酸(mRNAs)的表达显著增加(P<0.05)。茜素红染色显示,接种在负载头蛋白siRNA的胶原蛋白/生物玻璃支架上的细胞比其他细胞含有显著更多的矿化结节(P<0.05)。

结论

负载头蛋白siRNA的胶原蛋白/生物玻璃复合材料具有良好的生物相容性,且胶原蛋白/生物玻璃复合材料与头蛋白siRNA在促进成骨方面显示出协同作用。

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