ROLE OF DIFFERENTIATION FACTORS IN CARTILAGE AND BONE FORMATION AND REGENERATION

本项目的目标是研究软骨和骨 诱导因子，并确定它们在胚胎发生和 在组织形成和疾病中的作用。诸如组织 再生概括了胚胎组织的发育顺序 形成，可以想象，了解作用机制 可溶性分化因子的合成是生物学上 骨骼组织的受控再生。 这将产生 对治疗先天性和/或后天性 骨骼疾病如大的骨缺损，受损的骨折愈合， 骨关节炎、骨质疏松症和牙周炎。 该项目的重点是 软骨和骨诱导分子的进一步表征， 他们的生物活动。 含软骨和骨的蛋白质组分 在体内诱导活性，已经从关节软骨中分离 并纯化至同质;来自胰蛋白酶的蛋白质测序数据 已获得肽;数据库搜索未显示同源性 与任何已知的序列。新的组织特异性定位 蛋白制备物通过使用多克隆抗体的免疫定位显示， 针对N-末端序列的抗体。进一步表征 cDNA克隆正在进行中。 重组瞬时表达 分离的cDNA克隆正在进行中。 使用分子探针，我们 研究它们各自对软骨、软骨内分泌和 膜状骨形成。 免疫组织化学定位和原位 软骨和骨诱导蛋白的杂交，以及研究 在体外，表明这些分子的选择性贡献 软骨细胞的启动、增强、维持和成熟 和成骨细胞表型。 这项工作有助于基本的 了解从头软骨和骨形成，并设置阶段 为了正确指示和使用这些可溶性分化 临床环境中的因素。

The objectives of this project are to study the cartilage and bone inducing factors and to define their role in embryogenesis and in postnatal life, both in tissue formation and in disease. As tissue regeneration recapitulates the developmental sequence of embryonic tissue formation, it is conceivable that understanding the mechanisms of action of the soluble differentiation factors is a key step towards biologically controlled regeneration of skeletal tissues. This will have a significant impact on the treatment of congenital and/or acquired skeletal diseases such as large bone defects, impaired fracture healing, osteoarthritis, osteoporosis and periodontitis. This project focuses on the further characterization of cartilage and bone inducing molecules and their biological activities. Protein fractions with cartilage and bone inducing activity in vivo, have been isolated from articular cartilage and purified to homogeneity; protein sequencing data from tryptic peptides have been obtained; databank searches did not reveal homology with any known sequences. The tissue specific localization of the new protein preparation was shown by immunolocalization using a polyclonal antibody against the N-terminal sequence. Further characterization by cDNA cloning is in progress. Recombinant transient expression of isolated cDNA clones is ongoing. Using the molecular probes, we are studying their respective contribution to cartilage, endochondral and membranous bone formation. Immunohistochemical localization and in situ hybridization of cartilage and bone inducing proteins, as well as studies in vitro, indicate the selective contribution of these molecules to initiation, enhancement, maintenance and maturation of the chondrocytic and osteoblastic phenotype. This work contributes to the basic understanding of de novo cartilage and bone formation, and sets the stage for the proper indication and use of these soluble differentiation factors in a clinical setting.