Use of Cryopreserved Osteogenic Matrix Cell Sheets for Bone Reconstruction  

作　　者：Tomohiko Kura Manabu Akahane Takamasa Shimizu Yoshinobu Uchihara Yasuaki Tohma Yusuke Morita Munehisa Koizumi Kenji Kawate Yasuhito Tanaka Tomohiko Kura;Manabu Akahane;Takamasa Shimizu;Yoshinobu Uchihara;Yasuaki Tohma;Yusuke Morita;Munehisa Koizumi;Kenji Kawate;Yasuhito Tanaka(Department of Orthopedic Surgery, Nara Medical University School of Medicine, Kashihara, Japan;Department of Public Health, Health Management and Policy, Nara Medical University School of Medicine, Kashihara, Japan;Departmentof Biomedical Engineering, Doshisha University, Kyotanabe, Japan;Department of Artificial Joint and Regenerative Medicine, Nara Medical University, Kashihara, Japan)

机构地区：[1]Department of Orthopedic Surgery, Nara Medical University School of Medicine, Kashihara, Japan [2]Department of Public Health, Health Management and Policy, Nara Medical University School of Medicine, Kashihara, Japan [3]Departmentof Biomedical Engineering, Doshisha University, Kyotanabe, Japan [4]Department of Artificial Joint and Regenerative Medicine, Nara Medical University, Kashihara, Japan

出　　处：《Stem Cell Discovery》2016年第1期13-23,共11页干细胞探索（英文）

摘　　要：Skeletal diseases, such as nonunion and osteonecrosis, are now treatable with tissue engineering techniques. Single cell sheets called osteogenic matrix cell sheets (OMCSs) grown from cultured bone marrow-derived mesenchymal stem cells show high osteogenic potential;however, long preparation times currently limit their clinical application. Here, we report a cryopreservation OMCS transplantation method that shortens OMCS preparation time. Cryopreserved rat OMCSs were prepared using slow- and rapid-freezing methods, thawed, and subsequently injected scaffold-free into subcutaneous sites. Rapid- and slow-frozen OMCSs were also transplanted directly to the femur bone at sites of injury. Slow-freezing resulted in higher cell viability than rapid freezing, yet all two cryopreservation methods yielded OMCSs that survived and formed bone tissue. In the rapid- and slow-freezing groups, cortical gaps were repaired and bone continuity was observed within 6 weeks of OMCS transplantation. Moreover, while no significant difference was found in osteocalcin expression between the three experimental groups, the biomechanical strength of femurs treated with slow-frozen OMCSs was significantly greater than those of non-transplant at 6 weeks post-injury. Collectively, these data suggest that slow-frozen OMCSs have superior osteogenic potential and are better suited to produce a mineralized matrix and repair sites of bone injury.Skeletal diseases, such as nonunion and osteonecrosis, are now treatable with tissue engineering techniques. Single cell sheets called osteogenic matrix cell sheets (OMCSs) grown from cultured bone marrow-derived mesenchymal stem cells show high osteogenic potential;however, long preparation times currently limit their clinical application. Here, we report a cryopreservation OMCS transplantation method that shortens OMCS preparation time. Cryopreserved rat OMCSs were prepared using slow- and rapid-freezing methods, thawed, and subsequently injected scaffold-free into subcutaneous sites. Rapid- and slow-frozen OMCSs were also transplanted directly to the femur bone at sites of injury. Slow-freezing resulted in higher cell viability than rapid freezing, yet all two cryopreservation methods yielded OMCSs that survived and formed bone tissue. In the rapid- and slow-freezing groups, cortical gaps were repaired and bone continuity was observed within 6 weeks of OMCS transplantation. Moreover, while no significant difference was found in osteocalcin expression between the three experimental groups, the biomechanical strength of femurs treated with slow-frozen OMCSs was significantly greater than those of non-transplant at 6 weeks post-injury. Collectively, these data suggest that slow-frozen OMCSs have superior osteogenic potential and are better suited to produce a mineralized matrix and repair sites of bone injury.

关 键 词：Bone Marrow Stromal Cell CRYOPRESERVATION Cell Sheet Injectable Bone Bone Reconstruction 

分 类 号：R32[医药卫生—人体解剖和组织胚胎学]