Ito Y, Fitzsimmons J S, Sanyal A, Mello M A, Mukherjee N, O'Driscoll S W
Cartilage and Connective Tissue Research Laboratory, Mayo Clinic, Mayo Foundation, Rochester, MN 55905, USA.
Osteoarthritis Cartilage. 2001 Apr;9(3):215-23. doi: 10.1053/joca.2000.0378.
Periosteal chondrogenesis is relevant to cartilage repair and fracture healing. Periosteum contains two distinct layers: a thick, outer fibrous layer and a thin, inner cambium layer which is adjacent to the bone. Specific chondrocyte precursors are known to exist in periosteum but have not yet been identified. In this study, the location of the chondrocyte precursors in periosteum was determined.
One hundred and twenty periosteal explants from 30 2-month-old NZ rabbits were cultured for up to 42 days. Histomorphological changes and spatio-temporal localization of Col. II mRNA and protein were analysed.
On day 7, chondrocyte differentiation appeared in the most juxtaosseous region in the cambium layer. Col. II mRNA and protein were also evident in the same region. By day 14, chondrocyte differentiation progressed further into the juxtaosseous cambium layer, as did Col. II mRNA and protein. With growth of the neocartilage, the cambium layer gradually diminished to the extent that by 21-28 days it was no longer evident. Cartilage growth was significant and followed an appositional pattern, growing away from the fibrous layer. The fibrous layer remained essentially unchanged from 0-42 days, without evidence of hypertrophy or atrophy. Col. II mRNA expression was never seen in the fibrous layer.
From these data, three conclusions can be drawn concerning chondrogenesis from periosteum: (1) the chondrocyte precursors are located in the cambium layer of periosteum; (2) chondrogenesis commences in the juxtaosseous area in the cambium layer and progresses from the juxtaosseous region to the juxtafibrous region of the cambium layer; (3) neocartilage growth is appositional, which displaces the fibrous layer away from the cartilage already formed, as new cartilage is formed between these two layers. These findings suggest that the least differentiated (stem or reserve) cells are located in the cambium layer furthest from the bone.
These findings show that the chondrocyte precursors are located in the cambium layer of periosteum. Preservation of this layer is essential for chondrogenesis. As neocartilage growth is appositional, away from the fibrous layer, it can be expected that the new cartilage deposited in and adjacent to a periosteal graft would be expected to be located on the side of the cambium layer, rather than on the side of the fibrous layer of the graft.
骨膜软骨形成与软骨修复及骨折愈合相关。骨膜包含两层不同的结构:外层较厚的纤维层和与骨相邻的内层较薄的生发层。已知骨膜中存在特定的软骨细胞前体,但尚未被鉴定出来。在本研究中,确定了骨膜中软骨细胞前体的位置。
从30只2月龄新西兰兔获取120个骨膜外植体,培养长达42天。分析组织形态学变化以及Ⅱ型胶原mRNA和蛋白的时空定位。
第7天,软骨细胞分化出现在生发层最靠近骨的区域。Ⅱ型胶原mRNA和蛋白在同一区域也很明显。到第14天,软骨细胞分化进一步向靠近骨的生发层发展,Ⅱ型胶原mRNA和蛋白也是如此。随着新软骨的生长,生发层逐渐减少,到21 - 28天时不再明显。软骨生长显著,呈表面生长模式,远离纤维层生长。纤维层在0 - 42天基本保持不变,没有肥大或萎缩的迹象。纤维层中从未观察到Ⅱ型胶原mRNA表达。
根据这些数据,关于骨膜软骨形成可得出三个结论:(1)软骨细胞前体位于骨膜的生发层;(2)软骨形成始于生发层靠近骨的区域,并从靠近骨的区域向生发层靠近纤维的区域发展;(3)新软骨生长是表面生长,随着新软骨在这两层之间形成,将纤维层从已形成的软骨上推开。这些发现表明,分化程度最低(干细胞或储备细胞)的细胞位于离骨最远的生发层。
这些发现表明软骨细胞前体位于骨膜的生发层。保留这一层对软骨形成至关重要。由于新软骨生长是表面生长,远离纤维层,可以预期,植入骨膜移植物内及相邻部位沉积的新软骨将位于生发层一侧,而非移植物纤维层一侧。