Department of Stomatology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China.
Chin Med J (Engl). 2011 Dec;124(23):4022-8.
The seed cell is a core problem in bone tissue engineering research. Recent research indicates that human dental pulp stem cells (hDPSCs) can differentiate into osteoblasts in vitro, which suggests that they may become a new kind of seed cells for bone tissue engineering. The aim of this study was to evaluate the osteogenic differentiation of hDPSCs in vitro and bone-like tissue formation when transplanted with three-dimensional gelatin scaffolds in vivo, and hDPSCs may become appropriate seed cells for bone tissue engineering.
We have utilized enzymatic digestion to obtain hDPSCs from dental pulp tissue extracted during orthodontic treatment. After culturing and expansion to three passages, the cells were seeded in 6-well plates or on three-dimensional gelatin scaffolds and cultured in osteogenic medium. After 14 days in culture, the three-dimensional gelatin scaffolds were implanted subcutaneously in nude mice for 4 weeks. In 6-well plate culture, osteogenesis was assessed by alkaline phosphatase staining, Von Kossa staining, and reverse transcription-polymerase chain reaction (RT-PCR) analysis of the osteogenesis-specific genes type I collagen (COL I), bone sialoprotein (BSP), osteocalcin (OCN), RUNX2, and osterix (OSX). In three-dimensional gelatin scaffold culture, X-rays, hematoxylin/eosin staining, and immunohistochemical staining were used to examine bone formation.
In vitro studies revealed that hDPSCs do possess osteogenic differentiation potential. In vivo studies revealed that hDPSCs seeded on gelatin scaffolds can form bone structures in heterotopic sites of nude mice.
These findings suggested that hDPSCs may be valuable as seed cells for bone tissue engineering. As a special stem cell source, hDPSCs may blaze a new path for bone tissue engineering.
种子细胞是骨组织工程研究的核心问题。最近的研究表明,人牙髓干细胞(hDPSCs)在体外可分化为成骨细胞,这表明它们可能成为骨组织工程的一种新型种子细胞。本研究旨在评估 hDPSCs 在体外的成骨分化能力,以及在体内与三维明胶支架共移植后形成类骨组织的能力,从而推断 hDPSCs 可能成为合适的骨组织工程种子细胞。
我们采用酶消化法从正畸治疗中提取的牙髓组织中获得 hDPSCs。培养和扩增至三代后,将细胞接种于 6 孔板或三维明胶支架上,并在成骨培养基中培养。培养 14 天后,将三维明胶支架皮下植入裸鼠体内 4 周。在 6 孔板培养中,通过碱性磷酸酶染色、Von Kossa 染色和骨形成特异性基因Ⅰ型胶原(COL I)、骨涎蛋白(BSP)、骨钙素(OCN)、RUNX2 和骨形成蛋白 2(OSX)的逆转录聚合酶链反应(RT-PCR)分析评估成骨作用。在三维明胶支架培养中,通过 X 射线、苏木精/伊红染色和免疫组织化学染色来检测骨形成。
体外研究表明 hDPSCs 具有成骨分化潜能。体内研究表明,接种在明胶支架上的 hDPSCs 可在裸鼠异位部位形成骨结构。
这些发现表明 hDPSCs 可能是骨组织工程中有价值的种子细胞。作为一种特殊的干细胞来源,hDPSCs 可能为骨组织工程开辟新途径。