O'Driscoll S W, Recklies A D, Poole A R
Cartilage and Connective Tissue Research Laboratory, St. Michael's Hospital, University of Toronto, Canada.
J Bone Joint Surg Am. 1994 Jul;76(7):1042-51. doi: 10.2106/00004623-199407000-00013.
Periosteal grafts have chondrogenic potential and have been used to repair defects in articular cartilage. We studied the effects of the culture conditions and of transforming growth factor-beta 1 on chondrogenesis in rabbit periosteal explants that were cultured in vitro. A total of 390 periosteal explants were obtained from the anteromedial sides of the proximal parts of the tibiae of eleven rabbits that were two weeks, two months, or six months old. The culture medium (alpha minimum essential medium or Dulbecco minimum essential medium) contained fetal calf serum, with or without transforming growth factor-beta 1, at a concentration of one or ten nanograms per milliliter for the first two weeks of culture. Three hundred and twenty-one explants were submerged in liquid medium and sixty-nine were suspended in an agarose gel; they were then evaluated histochemically, histomorphometrically, and by collagen-typing. In the media without agarose, in the presence of ten nanograms of transforming growth factor-beta 1 per milliliter, chondrogenesis was commonly seen after two to four weeks with use of safranin-O staining and histomorphometry. In the agarose gels, chondrogenesis from the periosteum was observed at four and six weeks and was enhanced by the presence of one or ten nanograms of transforming growth factor-beta 1 per milliliter. The combination of agarose with transforming growth factor-beta 1 most favored the formation of cartilage, which was maximum at six weeks in the presence of ten nanograms of transforming growth factor-beta 1 per milliliter. Under these conditions, chondrogenesis occurred in almost every explant, with 50 +/- 30 per cent of the tissue being composed of cartilage. Type-II collagen was present in the explants that had undergone chondrogenesis.
骨膜移植物具有软骨形成潜能，已被用于修复关节软骨缺损。我们研究了培养条件和转化生长因子-β1对体外培养的兔骨膜外植体软骨形成的影响。从11只2周、2个月或6个月大的兔子胫骨近端前内侧获取了总共390个骨膜外植体。培养基（α-最低必需培养基或杜尔贝科最低必需培养基）含有胎牛血清，在培养的前两周添加或不添加浓度为每毫升1或10纳克的转化生长因子-β1。321个外植体浸没在液体培养基中，69个悬浮在琼脂糖凝胶中；然后对它们进行组织化学、组织形态计量学和胶原分型评估。在不含琼脂糖的培养基中，每毫升含有10纳克转化生长因子-β1时，在2至4周后通过番红O染色和组织形态计量学通常可见软骨形成。在琼脂糖凝胶中，在4周和6周时观察到骨膜的软骨形成，并且每毫升含有1或10纳克转化生长因子-β1可增强软骨形成。琼脂糖与转化生长因子-β1的组合最有利于软骨形成，在每毫升含有10纳克转化生长因子-β1时，6周时软骨形成达到最大值。在这些条件下，几乎每个外植体都发生了软骨形成，其中50±30%的组织由软骨组成。发生软骨形成的外植体中存在II型胶原。
