Yoshioka Tomokazu, Mishima Hajime, Ohyabu Yoshimi, Sakai Shinsuke, Akaogi Hiroshi, Ishii Tomoo, Kojima Hiroko, Tanaka Junzo, Ochiai Naoyuki, Uemura Toshimasa
Department of Orthopaedic Surgery, Control of Musculoskeletal System, Advanced Biomedical Applications, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
J Orthop Res. 2007 Oct;25(10):1291-8. doi: 10.1002/jor.20426.
Our objective was to examine the technique of regenerating cartilage tissue from bone marrow-derived cells by three-dimensional (3D) culture using the rotating wall vessel (RWV) bioreactor. Three-dimensional and cylindrical aggregates of allogeneic cartilage with dimensions of 10 x 5 mm (height x diameter) formed by the RWV bioreactor were transplanted into osteochondral defects of Japanese white rabbits (Group T, n = 15). For the control, some osteochondral defects were left empty (Group C, n = 18). At 4, 8, and 12 weeks postimplantation, the reparative tissues were evaluated macroscopically, histologically, and biochemically. In Group T at as early as 4 weeks, histological observation, especially via safranin-O staining, suggested that the reparative tissues resembled hyaline cartilage. And we observed no fibrous tissues between reparative tissue and adjacent normal tissues. In the deeper portion of the bony compartment, the osseous tissues were well remodeled. At 4 and 8 weeks postimplantation, the mean histological score of Group T was significantly better than that of Group C (p < 0.05). The glycosaminoglycans (GAG)/DNA ratio in both groups increased gradually from 4 to 8 weeks and then decreased from 8 to 12 weeks. We herein report the first successful regeneration of cartilage in osteochondral defects in vivo using allogeneic cartilaginous aggregates derived from bone marrow-derived cells by 3D culture using the RWV bioreactor.
我们的目标是研究使用旋转壁式生物反应器(RWV)通过三维(3D)培养从骨髓来源的细胞再生软骨组织的技术。将由RWV生物反应器形成的尺寸为10×5毫米(高×直径)的同种异体软骨的三维圆柱形聚集体移植到日本白兔的骨软骨缺损处(T组,n = 15)。作为对照,一些骨软骨缺损处保持空置(C组,n = 18)。在植入后4、8和12周,对修复组织进行宏观、组织学和生化评估。在T组中,早在4周时,组织学观察,尤其是通过番红O染色表明,修复组织类似于透明软骨。并且我们在修复组织和相邻正常组织之间未观察到纤维组织。在骨腔的较深部分,骨组织得到了良好的重塑。在植入后4周和8周,T组的平均组织学评分显著优于C组(p < 0.05)。两组中的糖胺聚糖(GAG)/DNA比值从4周到8周逐渐增加,然后从8周到12周下降。我们在此报告首次成功地在体内使用通过RWV生物反应器进行3D培养从骨髓来源的细胞获得的同种异体软骨聚集体再生骨软骨缺损处的软骨。