Anitua E, Sánchez M, Zalduendo M M, de la Fuente M, Prado R, Orive G, Andía I
Biotechnology Institute, BTI IMASD, Vitoria-Gasteiz, Spain.
Cell Prolif. 2009 Apr;42(2):162-70. doi: 10.1111/j.1365-2184.2009.00583.x. Epub 2009 Feb 24.
Preparations rich in growth factors (PRGF) release them plus bioactive proteins at localized sites, with the aim of triggering healing and regenerative processes. The prevailing paradigm suggests that their influence on proliferation, angiogenesis and the extracellular matrix synthesis is minimal. However, variations in their composition and impact on different cell phenotypes have not been examined.
Sixteen fibroblast cultures obtained from three different anatomical sites (skin, synovium and tendon) of 16 donors were exposed to the molecular pool released from PRGF scaffolds, with increasing amounts of platelets. We evaluated cell proliferation, secretion of angiogenic growth factors (VEGF and HGF), synthesis of type I collagen and hyaluronic acid (HA), considering platelet dose and anatomical origin of the cells. Activity of transforming growth factor-beta (TGF-beta) in type I procollagen and HA synthesis was examined by adding exogenous TGF-beta to plasma preparations.
All plasma preparations induced a significant proliferative response compared to non-stimulated cells (P < 0.05). Maximum proliferation rate was obtained with PRGF with 2-fold or 4-fold platelet concentration. Exposure to PRGF stimulated VEGF synthesis exclusively in tendon cells (P < 0.05), which also exhibited a different pattern of HGF production (P < 0.05). PRGF enhanced HA synthesis (P < 0.05), but did not alter collagen I production. Platelet-secreted TGF-beta may be involved in HA, but not in type I procollagen synthesis.
Optimizing composition and use of platelet-rich products is crucial to enhancing the therapeutic potential of this technology. Our data show that the biological effects of PRGF may depend on concentration of platelets and on the anatomical source of the cells.
富含生长因子的制剂(PRGF)在局部位点释放生长因子及生物活性蛋白,旨在引发愈合和再生过程。普遍观点认为,它们对增殖、血管生成和细胞外基质合成的影响极小。然而,其组成的变化及其对不同细胞表型的影响尚未得到研究。
从16名供体的三个不同解剖部位(皮肤、滑膜和肌腱)获取的16种成纤维细胞培养物,暴露于PRGF支架释放的分子库中,血小板含量逐渐增加。我们评估了细胞增殖、血管生成生长因子(VEGF和HGF)的分泌、I型胶原蛋白和透明质酸(HA)的合成,同时考虑了血小板剂量和细胞的解剖来源。通过向血浆制剂中添加外源性TGF-β,检测I型前胶原蛋白和HA合成中转化生长因子-β(TGF-β)的活性。
与未刺激的细胞相比,所有血浆制剂均诱导了显著的增殖反应(P < 0.05)。血小板浓度为2倍或4倍的PRGF获得了最大增殖率。暴露于PRGF仅刺激肌腱细胞中的VEGF合成(P < 0.05),其HGF产生模式也有所不同(P < 0.05)。PRGF增强了HA合成(P < 0.05),但未改变I型胶原蛋白的产生。血小板分泌的TGF-β可能参与HA合成,但不参与I型前胶原蛋白合成。
优化富含血小板产品的组成和使用对于提高该技术的治疗潜力至关重要。我们的数据表明,PRGF的生物学效应可能取决于血小板浓度和细胞的解剖来源。