接骨丹介导成骨细胞增殖促进骨折愈合的观察

Bone- Joining Elixir mediated osteoblast proliferation could promote fracture healing

[目的]观察接骨丹对新西兰乳兔成骨细胞增殖的影响,探讨接骨丹促进骨折愈合的机制。[方法]普通级新西兰大白乳兔,耳缘静脉空气栓塞处死,无菌条件下取颅盖骨,Ⅰ型胶原酶消化法获取成骨细胞并进行体外培养、鉴定及传代。将鉴定后的第二代成骨细胞随机分为正常对照组及实验组,正常对照组应用正常兔血清培养,实验组应用接骨丹含药血清培养,MTT法观测两组细胞的生长曲线;不同血清干预48 h后,倒置显微镜观测接骨丹对成骨细胞碱性磷酸酶及Ⅰ型胶原表达的影响,Elisa法观测接骨丹对成骨细胞培养上清液中骨钙素表达的影响。[结果]Ⅰ型胶原酶消化法成功获取并建立成骨细胞体外培养体系,MTT观测显示实验组能够显著促进成骨细胞的增殖,倒置显微镜观测实验组中碱性磷酸酶及Ⅰ型胶原的表达要优于正常对照组,并能够促进成骨细胞分泌骨钙素。[结论]接骨丹能够促进骨折的愈合,其机制可能与促进成骨细胞中碱性磷酸酶、Ⅰ型胶原及骨钙素的表达进而促进成骨细胞增殖和分化有关。

[ Objective] To observe the Bone -Joining Elixir influence on osteoblasts proliferation of New Zealand newborn rabbit,and to explore the mechanism of Bone -Joining Elixir promoting fracture healing. [ Method] Conventional grade New Zealand white newborn rabbit were sacrificed by ear venous air embolism, craniums was taken in aseptic conditions, and osteo- blasts was got by type I collagenase digestion method for in vitro culture,identification of osteoblasts and and passages. After i- dentification,the second generation of osteoblasts were randomly divided into normal control group and experimental group. In the normal control group, normal rabbit serum was used for culturing, while in the experimental group Bone -Joining Elixir medicated serum was applied for culturing, MTT method were determined to observe the cell growth curve of the two groups. After 48 h intervention of different serum, the influence of Bone - Joining Elixir on osteoblast alkaline phosphatase was observed and the expression of collagen type l was surveyed by inverted microscope,and osteocalcin expression of Bone -Joining Elixir on osteo- blast culture supernatant was detected by ELISA. [ Result] Type I collagenase digestion method successfully established the os- teoblasts cuhrue system in vitro, MTT observation showed the experiment could significantly promote the proliferation of osteo- blast, the inverted microscope observed alkaline phosphatase and the expression of collagen type I in the experimental group were superior to that of the normal control group, and could promote osteoblast secrete osteocalcin. [ Conclusion] The Bone - Joining Elixir can promote the healing of fracture, the mechanism may be related to promoting of the expression of ostenblast al- kaline phosphatase,collagen type I and osteocalcin,resulting in promoting proliferation and differentiation of osteoblasts.