Ripamonti U, Ma S S, Cunningham N S, Yeates L, Reddi A H
Medical Research Council/University of the Witwatersrand, Johannesburg, South Africa.
Plast Reconstr Surg. 1993 Jan;91(1):27-36. doi: 10.1097/00006534-199301000-00005.
Information concerning the efficacy of osteogenin, a bone morphogenetic protein, and demineralized bone matrix in orthotopic sites in nonhuman primates is a prerequisite for potential clinical application in humans. After exposure of the calvaria, 84 cranial defects, 25 mm in diameter, were prepared in 26 adult male baboons (Papio ursinus). Defects were implanted with insoluble collagenous bone matrix (ICBM, the inactive collagenous residue after dissociative extraction of bone matrix with 4 M guanidine hydrochloride) reconstituted with osteogenin fractions isolated from baboon bone matrix by chromatography on heparin-Sepharose and hydroxyapatite-Ultrogel (Og Hep-HA) or osteogenin further purified using Sephacryl S-200 gel filtration chromatography (Og S-200). Baboon osteogenin with the highest biologic activity in a rodent bioassay, as determined by alkaline phosphatase activity, calcium content, and histologic analysis, was used for orthotopic implantation in baboons. Additional defects were implanted with baboon demineralized bone matrix (DBM) or ICBM without osteogenin as control. Defects also were grafted with corticocancellous bone harvested from the iliac crest or left ungrafted to monitor the spontaneous regeneration potential of the adult baboon calvaria. Undecalcified bone sections at 7 microns were prepared from the harvested specimens 30 and 90 days after surgery. Histomorphometry demonstrated that Og S-200 induced copious amounts of bone and osteoid as early as day 30 (P < 0.01 versus ICBM, autogenous grafts and untreated defects). At day 90, in implants of Og S-200, Og Hep-HA, and DBM, bone and marrow formation was extensive, culminating in complete regeneration of the craniotomies. In implants of DBM, bone formed with an intervening phase of cartilage development. This provides the phenotypic evidence of endochondral bone differentiation by induction in defects of membranous calvarial bone in adult primates. These results establish the potential therapeutic application of osteogenin and demineralized bone matrix for the architectural reconstruction of the bone-bone marrow organ in humans.
骨形态发生蛋白骨生成素及脱矿骨基质在非人灵长类动物原位部位的疗效信息是其在人类中潜在临床应用的前提条件。在暴露颅骨后，于26只成年雄性狒狒（山魈）身上制备了84个直径25毫米的颅骨缺损。缺损处植入用通过肝素 - 琼脂糖和羟基磷灰石 - 优他胶（骨生成素肝素 - 羟基磷灰石）层析从狒狒骨基质中分离出的骨生成素组分重构的不溶性胶原骨基质（ICBM，即经4M盐酸胍解离提取骨基质后的无活性胶原残渣）或经Sephacryl S - 200凝胶过滤层析进一步纯化的骨生成素（骨生成素S - 200）。通过碱性磷酸酶活性、钙含量和组织学分析确定在啮齿动物生物测定中具有最高生物活性的狒狒骨生成素用于狒狒的原位植入。另外的缺损处植入狒狒脱矿骨基质（DBM）或不含骨生成素的ICBM作为对照。缺损处还用取自髂嵴的皮质松质骨移植或不进行移植以监测成年狒狒颅骨的自发再生潜力。术后30天和90天从收获的标本制备7微米厚的未脱钙骨切片。组织形态计量学表明，骨生成素S - 200早在第30天就诱导了大量的骨和类骨质形成（与ICBM、自体移植物和未处理的缺损相比，P < 0.01）。在第90天，在骨生成素S - 200、骨生成素肝素 - 羟基磷灰石和DBM的植入物中，骨和骨髓形成广泛，最终导致颅骨切开术完全再生。在DBM植入物中，骨形成伴有软骨发育的中间阶段。这为成年灵长类动物膜性颅骨骨缺损中通过诱导实现软骨内骨分化提供了表型证据。这些结果确立了骨生成素和脱矿骨基质在人类骨 - 骨髓器官结构重建方面的潜在治疗应用。
