Urist M R, Iwata H, Ceccotti P L, Dorfman R L, Boyd S D, McDowell R M, Chien C
Proc Natl Acad Sci U S A. 1973 Dec;70(12):3511-5. doi: 10.1073/pnas.70.12.3511.
Insoluble bone gelatin with inclusions of insoluble noncollagenous protein produces new bone when implanted in muscle in allogeneic rats. The implanted residue provides the milieu for expression of bone morphogenetic potential of migratory mesenchymal cells. Neutral buffer solutions activate endogenous enzymes that degrade components essential for cell interactions and differentiation of bone. Chloroform-methanol either denatures or extracts constituents responsible for degradation. Insoluble bone gelatin produces new bone after extraction at 2 degrees with neutral salts, 0.5 M EDTA, 0.1 M Tris.HCl, 4 M urea, 0.5 M hydroxylamine, and 10 M KCNS, as well as after limited digestion with pepsin or collagenase, but not after extraction with 5 M guanidine, 7 M urea, water saturated with phenol, or after alkali hydrolysis with 0.1 N NaOH. The specific activity of cell populations interacting with insoluble bone gelatin suggests that a chemical bond between collagen and a noncollagenous protein or part of a protein, cleaved by a neutral proteinase, controls the bone morphogenetic reaction.
含有不溶性非胶原蛋白包涵体的不溶性骨明胶,植入同种异体大鼠肌肉后可生成新骨。植入的残余物为迁移的间充质细胞表达骨形态发生潜能提供了环境。中性缓冲溶液激活内源性酶,这些酶可降解细胞相互作用和骨分化所必需的成分。氯仿 - 甲醇可使负责降解的成分变性或提取出来。不溶性骨明胶在与中性盐、0.5M乙二胺四乙酸(EDTA)、0.1M三羟甲基氨基甲烷盐酸盐(Tris.HCl)、4M尿素、0.5M羟胺和10M硫氰酸钾在2℃下提取后,以及用胃蛋白酶或胶原酶进行有限消化后,均可产生新骨,但在用5M胍、7M尿素、酚饱和水提取后或用0.1N氢氧化钠进行碱水解后则不能产生新骨。与不溶性骨明胶相互作用的细胞群体的比活性表明,胶原与一种非胶原蛋白或蛋白质的一部分之间的化学键(被中性蛋白酶裂解)控制着骨形态发生反应。