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Bone regeneration with osteogenic matrix cell sheet and tricalcium phosphate:An experimental study in sheep

World Journal of Orthopedics 2017年第10期8卷 754-760页

作者： Tsutomu Kira Manabu Akahane Shohei Omokawa Takamasa Shimizu Kenji Kawate tadanobu onishi Yasuhito Tanaka Department of Orthopedic Surgery Nara Medical University Department of Public Health Health Management and PolicyNara Medical University School of Medicine Department of Hand Surgery Nara Medical University Department of Artificial Joint and Regenerative Medicine for Bone and Cartilage Nara Medical University

AIM To determine the effects of a cell sheet created from sheep bone marrow and tricalcium phosphate(TCP) on *** Bone marrow cells were harvested from a sheep and cultured in a minimal essential medium(MEM) containing ascorbic acid phosphate(AscP) and dexamethasone(Dex). After 2 wk, the formed osteogenic matrix cell sheet was lifted from the culture dish using a scraper. Additionally, harvested bone marrow cells were cultured in MEM only as a negative control group, and in MEM with AscP, Dex, and β-glycerophosphate as a positive control group. For in vitro evaluation, we measured the alkaline phosphatase(ALP) activity and osteocalcin(OC) content in the media of the cultured cells from each group. For in vivo analysis, a porous TCP ceramic was used as a scaffold. We prepared an experimental group comprising TCP scaffolds wrapped with the osteogenic matrix cell sheets and a control group consisting of the TCP scaffold only. The constructs wereimplanted subcutaneously into athymic rats and the cell donor sheep, and bone formation was confirmed by histology after 4 *** In the in vitro part, the mean ALP activity was 0.39 ± 0.03 mg/well in the negative control group, 0.67 ± 0.04 mg/well in the sheet group, and 0.65 ± 0.07 mg/well in the positive control group. The mean OC levels were 1.46 ± 0.33 ng/well in the negative control group, 3.92 ± 0.16 ng/well in the sheet group, and 4.4 ± 0.47 ng/well in the positive control group, respectively. The ALP activity and OC levels were significantly higher in the cell sheet and positive control groups than in the negative control group(P 0.05). TCP constructs wrapped with cell sheets prior to implantation showed bone formation, in contrast to TCP scaffolds alone, which exhibited poor bone formation when implanted, in the subcutaneous layer both in athymic rats and in the s

关键词： Cell sheet Osteogenesis Sheep Bone marrow Mesenchymal stromal cell

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