生长分化因子-5及碱性成纤维细胞生长因子诱导家兔椎间盘髓核细胞成骨作用的研究

Effects of growth differentiation factor-5 and basic fibroblast growth factor on bone formation in rabbit disc nucleus pulposus cells

目的建立家兔椎间盘髓核细胞的体外培养模型,研究重组人生长分化因子-5(recombinant human growth differentiation factor-5,rhGDF-5)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)对髓核细胞成骨潜能的激发作用。方法将诱导剂rhGDF-5和bFGF分别及联合加入体外培养的髓核细胞中,观察髓核细胞的成骨表型表达和细胞学特性的变化。结果 rhGDF-5和bFGF均能促进钙盐沉积形成钙结节。rhGDF-5抑制髓核细胞增殖同时增加骨钙素表达;bFGF促进髓核细胞增殖及Ⅰ型胶原表达,但对骨钙素表达无显著影响。联合使用rhGDF-5和bFGF对髓核细胞成骨潜能(促进髓核细胞增殖、Ⅰ型胶原及骨钙素表达和钙盐沉积)的激发作用均优于单独使用其中任一细胞因子。结论 rhGDF-5诱导髓核细胞向成骨细胞分化,bFGF加强该诱导作用,联合使用rhGDF-5和bFGF能充分激发髓核细胞的成骨潜能。

Obj ective To research the effect of growth differentiation factor-5 (rhGDF-5 )and basic fibroblast growth factor (bFGF)on the osteogenesis potential excitation of nucleus pulposus cells ,while establishing a model of observing cytological and biological properties of rabbit nucleus pulposus cells in vitro.Methods The inducers,rhGDF-5 and bFGF were administerred to the nucleus pulposus cells respec-tively and simultaneously,then the phenotype and cytological characteristics of bone-formation were ob-served.Results Both rhGDF-5 and bFGF promoted the deposition of calcium salt to form calcium nodes. RhGDF-5 restrained cell proliferation,but enhanced the expression of osteocalcin of nucleus pulposus cells;bFGF promoted cell proliferation and up-regulated the expression of type I collagen,but it had no significant effect on the osteocalcin expression of nucleus pulposus cells.Moreover,the osteogenesis po-tential excitation (promoting cells proliferation,enhancing the expression of type I collagen or osteocalcin and improving the deposition of calcium salt)was better by co-administering rhGDF-5 and bFGF than u-sing either of them alone to nucleus pulposus cells.Conclusion RhGDF-5 can stably induce ossification of nucleus pulposus cells in vitro,and bFGF can enhance its effect.Co-administering rhGDF-5 and bFGF can sufficiently excite the osteogenesis potential of nucleus pulposus cells .