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Chitosan/collagen scaffold containing bone morphogenetic protein‐7 DNA supports dental pulp stem cell differentiation in vitro and in vivo
Journal of Biomedical Materials Research Part A ( IF 4.9 ) Pub Date : 2012-02-18 , DOI: 10.1002/jbm.a.34064
Xuechao Yang 1 , Guangli Han 1 , Xin Pang 1 , Mingwen Fan 1
Affiliation  

In this study, porous chitosan/collagen scaffolds were prepared through a freeze‐drying process, and loaded with the plasmid vector encoding human bone morphogenetic protein‐7 (BMP‐7) gene. To investigate the feasibility and efficacy of this gene‐activated scaffold on dental tissue engineering, human dental pulp stem cells (DPSCs) were seeded in this scaffold for in vitro and in vivo study. In vitro results indicated that cells can be transfected successfully by loaded plasmid and secrete BMP‐7 until day 24. Evaluation of DNA content, ALP activity, calcium content, SEM, and real‐time PCR revealed that cells on gene‐activated scaffold showed better proliferation properties and odontoblastic differentiation behaviors than cells on pure scaffolds. Then, these cell–scaffold complexes were implanted subcutaneously and retrieved after 4 weeks for histology evaluation. In vivo results that gene‐activated scaffold group could still trace the existence of tranfected cells at week 4 and showed the upregulated expression of DSPP compared to pure scaffold groups. On the basis of our results, chitosan/collagen‐loaded BMP‐7 DNA appears to be an effective substrate candidate for gene delivery and indeed enhanced DPSCs differentiation toward an odontoblast‐like phenotype in vitro and in vivo. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A:, 2012.

中文翻译:

含有骨形态发生蛋白-7 DNA 的壳聚糖/胶原支架支持体外和体内牙髓干细胞分化

在这项研究中,通过冷冻干燥过程制备多孔壳聚糖/胶原支架,并加载编码人骨形态发生蛋白-7(BMP-7)基因的质粒载体。为了研究这种基因激活支架在牙科组织工程中的可行性和有效性,将人牙髓干细胞 (DPSCs) 接种到该支架中进行体外体内研究。体外结果表明细胞可以通过加载的质粒成功转染并分泌 BMP-7 直到第 24 天。 DNA 含量、ALP 活性、钙含量、SEM 和实时 PCR 的评估表明基因激活支架上的细胞显示出更好的增殖特性和成牙本质细胞的分化行为与纯支架上的细胞相比。然后,将这些细胞 - 支架复合物植入皮下并在 4 周后取出用于组织学评估。体内结果表明,基因激活支架组在第 4 周仍可以追踪转染细胞的存在,并且与纯支架组相比,DSPP 的表达上调。根据我们的结果,装载壳聚糖/胶原蛋白的 BMP-7 DNA 似乎是基因传递的有效底物候选物,并且确实在体外体内增强了 DPSCs 向成牙本质细胞样表型的分化。© 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A:,2012。
更新日期:2019-12-11
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