Masaoka Tomokazu, Yoshii Toshitaka, Yuasa Masato, Yamada Tsuyoshi, Taniyama Takashi, Torigoe Ichiro, Shinomiya Kenichi, Okawa Atsushi, Morita Sadao, Sotome Shinichi
Department of Rehabilitation Medicine, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan; Department of Orthopaedic and Spinal Surgery, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.
Department of Orthopaedic and Spinal Surgery, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.
Open Biomed Eng J. 2016 Mar 18;10:2-11. doi: 10.2174/1874120701610010002. eCollection 2016.
Reconstruction of large bone defects is a great challenge in orthopedic research. In the present study, we prepared composites of bone marrow-derived stromal cells (BMSCs) and β-tricalcium phosphate (β-TCP) with three novel aspects: proliferation of BMSCs with continuous dexamethasone treatment, cell loading under low pressure, and use of autologous plasma as the cell loading medium. The effectiveness of the resulting composite for large bone-defect reconstruction was tested in a non-human primate model, and the bone union capability of the regenerated bones was examined.
Primary surgery: Bone defects (5 cm long) were created in the left femurs of nine cynomolgus monkeys with resection of the periosteum (five cases) or without resection (four cases), and porous β-TCP blocks were transplanted into the defects. Secondary surgery: Bone marrow aspirates harvested from seven of the nine monkeys were cultured with dexamethasone, and BMSCs were obtained. BMSCs were suspended in autologous plasma and introduced into a porous β-TCP block under low-pressure conditions. The BMSC/β-TCP composites were transplanted into bone defects created at the same sites as the primary surgery. Bone union evaluation: Five regenerated femurs were shortened by osteotomy surgery 8 to 15 months after transplantation of the β-TCP/BMSC composites, and bone union was evaluated radiographically.
After the primary surgery and treatment with β-TCP alone, one of the five periosteum-resected monkeys and two of the four periosteum-preserved monkeys exhibited successful bone reconstruction. In contrast, five of the seven cases treated with the β-TCP/MSC composite showed successful bone regeneration. In four of the five osteotomy cases, bone union was confirmed.
We validated the effectiveness of a novel β-TCP/BMSC composite for large bone defect regeneration and confirmed the bone union capability of the regenerated bone.
大骨缺损的修复是骨科研究中的一项巨大挑战。在本研究中，我们制备了骨髓源性基质细胞（BMSCs）与β-磷酸三钙（β-TCP）的复合材料，该复合材料具有三个新特点：通过持续地塞米松处理使BMSCs增殖、在低压下进行细胞接种以及使用自体血浆作为细胞接种介质。在非人灵长类动物模型中测试了所得复合材料用于大骨缺损修复的有效性，并检查了再生骨的骨愈合能力。
一期手术：在9只食蟹猴的左股骨上制造骨缺损（长5厘米），其中5例切除骨膜，4例未切除骨膜，将多孔β-TCP块植入缺损处。二期手术：从9只猴子中的7只采集骨髓抽吸物，用地塞米松进行培养，获得BMSCs。将BMSCs悬浮于自体血浆中，并在低压条件下引入多孔β-TCP块中。将BMSC/β-TCP复合材料植入与一期手术相同部位制造的骨缺损处。骨愈合评估：在β-TCP/BMSC复合材料移植后8至15个月，对5根再生股骨进行截骨手术使其缩短，并通过X线片评估骨愈合情况。
一期手术后仅用β-TCP治疗时，5例切除骨膜的猴子中有1例、4例保留骨膜的猴子中有2例实现了成功的骨重建。相比之下，7例接受β-TCP/MSC复合材料治疗的病例中有5例显示出成功的骨再生。在5例截骨病例中的4例中，确认了骨愈合。
我们验证了一种新型β-TCP/BMSC复合材料用于大骨缺损再生的有效性，并证实了再生骨的骨愈合能力。
