Liu Keng-Fan, Chen Rong-Fu, Li Yun-Ting, Lin Yun-Nan, Hsieh Dar-Jen, Periasamy Srinivasan, Lin Sin-Daw, Kuo Yur-Ren
Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital, Kaohsiung 80756, Taiwan.
R&D Center, ACRO Biomedical Co., Ltd., Kaohsiung 82151, Taiwan.
Biomedicines. 2021 Dec 3;9(12):1825. doi: 10.3390/biomedicines9121825.
Large bone fractures with segmental defects are a vital phase to accelerate bone integration. The present study examined the role of supercritical carbon dioxide (scCO) decellularized bone matrix (scDBM) seeded with allogeneic adipose-derived mesenchymal stem cells (ADSC) as bio-scaffold for bone regeneration. Bio-scaffold produced by seeding ADSC to scDBM was evaluated by scanning electron microscopy (SEM). Rat segmental femoral defect model was used as a non-union model to investigate the callus formation in vivo. Histological analysis and osteotomy gap closure in the defect area were analyzed at 12 and 24 weeks post-surgery. Immunohistochemical expression of Ki-67, BMP-2 and osteocalcin was evaluated to assess the ability of new bone formation scDBM. ADSC was found to attach firmly to scDBM bioscaffold as evidenced from SEM images in a dose-dependent manner. Callus formation was observed using X-ray bone imaging in the group with scDBM seeded with 2 × 10 and 5 × 10 ASCs group at the same time-periods. H&E staining revealed ASCs accelerated bone formation. IHC staining depicted the expression of Ki-67, BMP-2, and osteocalcin was elevated in scDBM seeded with 5 × 10 ASCs group at 12 weeks after surgery, relative to other experimental groups. To conclude, scDBM is an excellent scaffold that enhanced the attachment and recruitment of mesenchymal stem cells. scDBM seeded with ASCs accelerated new bone formation.
伴有节段性骨缺损的大型骨折是加速骨整合的关键阶段。本研究探讨了接种异体脂肪来源间充质干细胞（ADSC）的超临界二氧化碳（scCO）脱细胞骨基质（scDBM）作为骨再生生物支架的作用。通过扫描电子显微镜（SEM）评估将ADSC接种到scDBM上产生的生物支架。大鼠节段性股骨缺损模型用作骨不连模型，以研究体内骨痂形成情况。在术后12周和24周分析缺损区域的组织学分析和截骨间隙闭合情况。评估Ki-67、BMP-2和骨钙素的免疫组织化学表达，以评估新骨形成scDBM的能力。从SEM图像可以看出，ADSC以剂量依赖性方式牢固地附着在scDBM生物支架上。在同一时间段，使用X射线骨成像观察到接种2×10和5×10个ASC的scDBM组有骨痂形成。苏木精-伊红（H&E）染色显示ASC加速了骨形成。免疫组织化学（IHC）染色显示，术后12周，接种5×10个ASC的scDBM组中Ki-67、BMP-2和骨钙素的表达相对于其他实验组有所升高。总之，scDBM是一种出色的支架，可增强间充质干细胞的附着和募集。接种ASC的scDBM加速了新骨形成。
