Uchihara Yoshinobu, Akahane Manabu, Okuda Akinori, Shimizu Takamasa, Masuda Keisuke, Kira Tsutomu, Kawate Kenji, Tanaka Yasuhito
Department of Orthopedic Surgery, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8522, Japan.
Department of Public Health, Health Management and Policy, Nara Medical University School of Medicine, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan.
J Orthop Sci. 2018 May;23(3):578-584. doi: 10.1016/j.jos.2018.01.015. Epub 2018 Mar 1.
To evaluate whether osteogenic matrix cell sheets can supply osteogenesis to dead bone.
Femur bone fragments (5 mm in length) were obtained from Fisher 344 rats and irradiated by a single exposure of 60 Gy to produce bones that were no longer viable. Osteogenic matrix cell sheets were created from rat bone marrow-derived stromal cells (BMSCs). After wrapping the dead bone with an osteogenic matrix cell sheet, it was subcutaneously transplanted into the back of a rat and harvested after 4 weeks. Bone formation around the dead bone was evaluated by X-ray imaging and histology. Alkaline phosphatase (ALP) and osteocalcin (OC) mRNA expression levels were measured to confirm osteogenesis of the transplanted bone. The contribution of donor cells to bone formation was assessed using the Sry gene and PKH26.
After the cell sheet was transplanted together with dead bone, X-ray images showed abundant calcification around the dead bone. In contrast, no newly formed bone was seen in samples that were transplanted without the cell sheet. Histological sections also showed newly formed bone around dead bone in samples transplanted with the cell sheet, whereas many empty lacunae and no newly formed bone were observed in samples transplanted without the cell sheet. ALP and OC mRNA expression levels were significantly higher in dead bones transplanted with cell sheets than in those without a cell sheet (P < 0.01). Sry gene expression and cells derived from cell sheets labeled with PKH26 were detected in samples transplanted with a cell sheet, indicating survival of donor cells after transplantation.
Our study indicates that osteogenic matrix cell sheet transplantation can supply osteogenesis to dead bone.
评估成骨基质细胞片能否为死骨提供成骨作用。
从Fisher 344大鼠获取股骨骨碎片（长度5毫米），单次接受60 Gy照射以产生不再具有活力的骨。从大鼠骨髓来源的基质细胞（BMSCs）制备成骨基质细胞片。用成骨基质细胞片包裹死骨后，将其皮下移植到大鼠背部，4周后收获。通过X线成像和组织学评估死骨周围的骨形成。测量碱性磷酸酶（ALP）和骨钙素（OC）mRNA表达水平以确认移植骨的成骨情况。使用Sry基因和PKH26评估供体细胞对骨形成的贡献。
细胞片与死骨一起移植后，X线图像显示死骨周围有大量钙化。相比之下，未移植细胞片的样本中未见新形成的骨。组织学切片也显示，移植了细胞片的样本中死骨周围有新形成的骨，而未移植细胞片的样本中观察到许多空陷窝且无新形成的骨。移植了细胞片的死骨中ALP和OC mRNA表达水平显著高于未移植细胞片的死骨（P < 0.01）。在移植了细胞片的样本中检测到Sry基因表达以及用PKH26标记的细胞片来源的细胞，表明移植后供体细胞存活。
我们的研究表明，成骨基质细胞片移植可为死骨提供成骨作用。
