Key Lab for Oral Biomedicine of Ministry of Education and Department of Endodontics, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei Province, People's Republic of China.
J Biomed Mater Res A. 2020 Dec;108(12):2519-2526. doi: 10.1002/jbm.a.34064. Epub 2012 Feb 18.
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.
在这项研究中,通过冷冻干燥工艺制备了多孔壳聚糖/胶原支架,并负载了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功效,将人牙髓干细胞(DPSCs)接种在该支架上进行体外和体内研究。体外结果表明,负载的质粒可成功转染细胞,并持续分泌 BMP-7 至第 24 天。通过 DNA 含量、ALP 活性、钙含量、SEM 和实时 PCR 的评估表明,与纯支架上的细胞相比,基因激活支架上的细胞表现出更好的增殖特性和成牙本质分化行为。然后,将这些细胞-支架复合物皮下植入,并在 4 周后取出进行组织学评估。体内结果表明,基因激活支架组在第 4 周仍能追踪到转染细胞的存在,与纯支架组相比,DSPP 的表达上调。基于我们的结果,壳聚糖/胶原负载 BMP-7 DNA 似乎是一种有效的基因传递候选底物,确实能增强 DPSCs 在体外和体内向成牙本质细胞样表型的分化。©2012 Wiley Periodicals, Inc. J 生物材料 Res 部分 A:,2012。