Department of Orthopedics, Guangdong Key Laboratory of Orthopaedic Technology and Implant Materials, The First Affiliated Hospital of 26468Guangzhou Medical University, Guangzhou, China.
J Biomater Appl. 2021 Feb;35(7):814-822. doi: 10.1177/0885328220948030. Epub 2020 Aug 10.
Critical size bone defects are one of the most serious complications in orthopedics due to the lack of effective osteogenesis treatment. We fabricated carboxymethyl cellulose with phenol moieties (CMC-ph) microcapsules loaded with gene-modified rat bone mesenchymal stem cells (rBMSCs) that secrete hBMP2 following doxycycline (DOX) induction. The results showed that the morphology of microcapsules was spherical, and their diameters have equally distributed in the range of 100-150 μm; the viability of rBMSCs was unchanged over time. Through real-time PCR and Western blot analyses, the rBMSCs in microcapsules were found to secrete hBMP2 and to have upregulated mRNA and protein expression of osteogenesis-related genes in vitro and in vivo. Furthermore, the in vivo results suggested that the group with the middle concentration of cells expressed the highest amount of osteogenic protein over time. In this study, we showed that gene-modified rBMSCs in CMC-ph microcapsules had good morphology and viability. The BMP2-BMSCs/CMC-Ph microcapsule system could upregulate osteogenic mRNA and protein in vitro and in vivo. Further analysis demonstrated that the medium concentration of cells had a suitable density for transplantation in nude mice. Therefore, BMP2-BMSCs/CMC-Ph microcapsule constructs have potential for bone regeneration in vivo.
临界尺寸骨缺损是骨科中最严重的并发症之一,因为缺乏有效的成骨治疗。我们制备了载有基因修饰的大鼠骨髓间充质干细胞(rBMSCs)的羧甲基纤维素与苯酚部分(CMC-ph)微胶囊,该细胞在强力霉素(DOX)诱导下分泌 hBMP2。结果表明,微胶囊的形态为球形,其直径在 100-150μm 范围内均匀分布;rBMSCs 的活力随时间的推移而保持不变。通过实时 PCR 和 Western blot 分析,发现微胶囊中的 rBMSCs 分泌 hBMP2,并在体外和体内上调与成骨相关基因的 mRNA 和蛋白表达。此外,体内结果表明,细胞中浓度适中的组随时间推移表达出最高量的成骨蛋白。在这项研究中,我们表明载有基因修饰的 rBMSCs 的 CMC-ph 微胶囊具有良好的形态和活力。BMP2-BMSCs/CMC-Ph 微胶囊系统可在体外和体内上调成骨 mRNA 和蛋白。进一步的分析表明,细胞的中等浓度具有适合在裸鼠中移植的合适密度。因此,BMP2-BMSCs/CMC-Ph 微胶囊构建物具有体内骨再生的潜力。