目的

以简便、有效、安全的方法获得富血小板血浆（platelet-rich plasma，PRP），探讨PRP 来源的富生长因子血清（serum rich in growth factors，SRGF）对体外原代培养的人成骨细胞增殖的影响。

方法

改进传统二次离心方法制备得到PRP，进行血细胞计数分析和ELISA 法定量分析TGF-β1 浓度。 原代培养并传代人成骨细胞，取第四代人成骨细胞分为3 组，分别加入5%SRGF、5%SPGF 或无血清细胞培养基， 在添加因素后24、48、72 和96 h 用MTT 法检测成骨细胞的增殖。

结果

改良法制备得到PRP，其血小板浓度（P＝0.003）、血小板富集系数（P＝0.004）和血小板获得率（P＝0.005）TGF-β1 浓度（P＝0.041）、TGF-β1 富集系数（P＝0.028）均高于传统法。 SRGF 组吸光度值（A 值）与SPGF 组同时点数值进行两两比较，其差异在48、72 和96 h 均有统计学意义（P ＜0.05）；与无血清对照组同时点A 值两两比较，其差异在24、48、72 和96 h 均有统计学意义（P ＜0.05）。

结论

改良法可获得高质量的PRP，PRP 来源的SRGF 具有促人成骨细胞增殖的作用。

Objective

To improve the original tube method for isolating platelet-rich plasma（PRP）， and study the effect of serum rich in growth factors （SRGF） derived from platelet-rich plasma（PRP） on the biological function of human osteoblast.

Methods

Whole blood was collected from 8 healthy donors and then PRP was separated with both the tube method and the syringe method.Samples were evaluated for platelet count and activated to get serum rich-in growth factors （SRGF）.The level of TGF-β1 in SRGF were assayed with enzyme-linked immunoassay （ELISA）. Human osteoblast were cultured and identified. Treated with 5% SRGF， 5% SPGF and serum-free Dulbecco's modified Eagle medium （DMEM）， the cellular mitogenic activity was evaluated by thiazoly blue（MTT） colorimetric assay at 24， 48， 72 and 96 hour.

Results

The end product of syringe method showed both a higher platelet count in PRP （P＝0.003） and a higher level of TGF-β1 in SRGF （P＝0.041）. The proliferation of human osteoblast were promoted after treated with SRGF. The differences between SPGF group and SRGF group were significant at 48， 72 and 96 hour （P ＜0.05）. The proliferation of osteoblasts almost stoped in serum-free medium， and the differences in absorbency，compared with other groups，were significant （P ＜0.05）.

Conclusion

High quality of PRP can be achieved by the improved method and SRGF is capable of up-regulating the proliferation of human osteoblast.