Funayama Atsushi, Niki Yasuo, Matsumoto Hideo, Maeno Shinichi, Yatabe Taku, Morioka Hideo, Yanagimoto Shigeru, Taguchi Tetsushi, Tanaka Junzo, Toyama Yoshiaki
Department of Orthopedic Surgery, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo, Japan.
J Orthop Sci. 2008 May;13(3):225-32. doi: 10.1007/s00776-008-1220-z. Epub 2008 Jun 6.
Recently, tissue-engineered chondrocyte transplantation has been tried to treat full-thickness cartilage defects. We developed an injectable type II collagen gel scaffold by chemically reacting type II collagen with polyethylene glycol crosslinker. This type II collagen was prepared from the nasal septa of cattle. In the present study, chondrocytes embedded in type II collagen gel were injected into rabbit full-thickness cartilage defects without a periosteal graft, and the feasibility for clinical application of the gel was evaluated.
Chondrocytes were isolated from 1-kg New Zealand white rabbits. A full-thickness articular cartilage defect (5 mm diameter, 4 mm depth) was created on the patellar groove of the femur of 16 male 3-kg New Zealand white rabbits. A type II collagen solution of mixed chondrocytes at a density of 1 x 10(7) cells/ml was injected and transplanted into the defect in the right knee. The controls were the defect only in the left knee. At 4, 8, 12, and 24 weeks after operation, four cases from each group were evaluated macroscopically and histologically.
After injection into the cartilage defect, the gel bonded to the adjacent cartilage and bone within several minutes. Macroscopic examination revealed that the surface of the transplanted area was smooth and exhibited similar coloration and good integration with the surrounding cartilage at 12 and 24 weeks after transplantation. Histological examination at 8 weeks revealed favorable hyaline cartilage regeneration with good chondrocyte morphology. At 12 and 24 weeks, reparative cartilage remained rich in type II collagen. According to O'Driscoll histological scores, significant differences between the transplanted and control groups were apparent at 12 and 24 weeks. Immunohistochemical staining indicated sufficient type II collagen synthesis in regenerated cartilage 8 weeks after transplantation, and it was maintained until 24 weeks.
These results indicate that type II collagen gel is suitable for injection into cartilage defects without any covering of a graft and offers a useful scaffold during chondrocyte transplantation.
最近,组织工程软骨细胞移植已被尝试用于治疗全层软骨缺损。我们通过使II型胶原蛋白与聚乙二醇交联剂发生化学反应,开发出一种可注射的II型胶原蛋白凝胶支架。这种II型胶原蛋白是从牛鼻中隔制备的。在本研究中,将包埋于II型胶原蛋白凝胶中的软骨细胞注入兔全层软骨缺损处,不进行骨膜移植,并评估该凝胶临床应用的可行性。
从1千克重的新西兰白兔中分离软骨细胞。在16只3千克重的雄性新西兰白兔的股骨髌沟处制造一个全层关节软骨缺损(直径5毫米,深度4毫米)。将密度为1×10⁷个细胞/毫升的混合软骨细胞的II型胶原蛋白溶液注入并移植到右膝的缺损处。对照组为仅左膝有缺损。在术后4、8、12和24周,每组取4只动物进行大体和组织学评估。
注入软骨缺损后,凝胶在几分钟内与相邻软骨和骨结合。大体检查显示,移植区域表面光滑,在移植后12周和24周时颜色与周围软骨相似且融合良好。8周时的组织学检查显示有良好的透明软骨再生,软骨细胞形态良好。在12周和24周时,修复性软骨中II型胶原蛋白含量仍然丰富。根据奥德里斯科尔组织学评分,移植组和对照组在12周和24周时有明显差异。免疫组织化学染色表明,移植后8周再生软骨中有足够的II型胶原蛋白合成,并且一直维持到24周。
这些结果表明,II型胶原蛋白凝胶适用于在不进行任何移植覆盖的情况下注入软骨缺损处,并在软骨细胞移植过程中提供有用的支架。