多肽P-15对大鼠成骨细胞附着增殖分化的影响

Effects of Polypeptide P-15 on Attachment,Proliferation and Differentiation of Cultured Osteoblast on Bone Mineral Particles

目的研究多肽P-15对小牛骨表面生长的大鼠成骨细胞附着、增殖、分化的影响。方法对照组和实验组分别采用单纯无机小牛骨粉(anorganic bone mineral,ABM)和复合P-15的无机小牛骨粉接种大鼠成骨细胞进行体外培养。四甲基偶氮唑蓝[3-(4,5-dimehyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazoliumbromide,MTT]比色法检测大鼠成骨细胞在2组骨粉表面附着、增殖的速度,倒置相差显微镜观察成骨细胞在2组骨粉表面及周围的增殖情况,用对硝基苯磷酸二钠盐(p-nitrophenyl phosphate,PNPP)法检测2组成骨细胞碱性磷酸酶(alkaline phosphatase,ALP)活性。结果成骨细胞体外培养24h,实验组与对照组MTT比色法检测的光密度值分别为1.597±0.035、1.239±0.063,差异有统计学意义(P<0.01)。培养72h,实验组和对照组PNPP法检测的ALP活性分别为(35.533±1.051)金氏单位/100mL和(34.645±1.187)金氏单位/100mL,实验组略高于对照组,但差异无统计学意义(P>0.05)。结论多肽P-15加快了大鼠成骨细胞在骨粉表面粘附、增殖的速度,但对大鼠成骨细胞的分化无明显促进作用。

Objective To study the effects of peptide P-15 on attachment, proliferation and differentiation of rat osteoblast cultured on the anorganic bone mineral. Methods The control group and experimental group were used anorganic bone mineral （ABM） and the compound P-15 ABM （P-15＋ABM） inoculated rat osteoblasts cultured in vitro. Cell functions such as attachment, spreading, proliferation and differentiation of rat osteoblast those cultured on the PepGenP-15TM or on the OstenGraf/N-300 were investigated and compared by phase-contrast microscope, MTT assay and ALP staining. Results After 24 hours’culturing, in the experimental and control group MTT （OD values） were 1.597±0.035, 1.239±0.063 respectively, and the difference was statistically significant （P0.01）. Peptide P-15 significantly speeded the rat osteoblast attachment. After 72h culturing, in experimental group, ALP activity was 35.533±1.051, slightly higher than 34.645±1.187 in control group, but P 0.05, difference was not statistically significant. Peptide P-15 had no significant role in promoting rat osteoblast differentiation. Conclusion Compound P-15 ABM is a good biological activity of bone-induced scaffold materials.