Elder Steven, Thomason John
Department of Agricultural & Biological Engineering, Bagley College of Engineering, Mississippi State University, Starkville, MS, USA.
Department of Clinical Sciences, College of Veterinary Medicine, Mississippi State University, Starkville, MS, USA.
Open Orthop J. 2014 Apr 4;8:78-84. doi: 10.2174/1874325001408010078. eCollection 2014.
Platelet-rich plasma (PRP) may have the potential to enhance articular cartilage regeneration through release of growth factors including transforming growth factor isoforms. The purpose of this study was to investigate the potential for PRP to stimulate chondrogenic differentiation in three-dimensional PRP hydrogel constructs. Allogenic PRP was prepared using a double centrifugation protocol which resulted in a platelet concentration approximately 250% above baseline. Canine marrow stromal cells were encapsulated at 6.8×10(6) cells/ml in either 2% sodium alginate or in a 3:1 mixture of freshly prepared PRP and 2% alginate. PRP and alginate beads were cultured in chemically defined chondrogenic medium with and without 10 ng/ml TGF-β3. PRP cultures were additionally supplemented with frozen-thawed PRP. In the absence of TGF-β3, PRP had a mild stimulatory effect on cell proliferation. PRP did not stimulate cell proliferation in the presence of TGF-β3. Cells exposed to TGF-β3 accumulated significantly more GAG/DNA than those which were not, but there was not a statistically significant difference between alginate and PRP. Total collagen content was greater in PRP than in alginate, regardless of TGF-β3. Chondrogenesis in PRP was qualitatively and spatially different than that which occurred in conventional alginate beads and was characterized by isolated centers of intense chondrogenesis. Overall the results demonstrate that PRP alone weakly promotes chondroinduction of marrow stromal cells, and the effect is greatly augmented by TGF-β3.
富含血小板血浆（PRP）可能具有通过释放包括转化生长因子亚型在内的生长因子来增强关节软骨再生的潜力。本研究的目的是探讨PRP在三维PRP水凝胶构建物中刺激软骨形成分化的潜力。使用双重离心方案制备同种异体PRP，其血小板浓度比基线高出约250%。将犬骨髓基质细胞以6.8×10⁶个细胞/毫升的浓度包封在2%海藻酸钠中，或包封在新鲜制备的PRP与2%海藻酸钠的3:1混合物中。将PRP和海藻酸钠珠在化学限定的软骨形成培养基中培养，添加或不添加10 ng/ml的转化生长因子-β3（TGF-β3）。PRP培养物还额外添加了冻融的PRP。在没有TGF-β3的情况下，PRP对细胞增殖有轻度刺激作用。在有TGF-β3的情况下，PRP不刺激细胞增殖。暴露于TGF-β3的细胞比未暴露的细胞积累的糖胺聚糖/DNA显著更多，但海藻酸钠和PRP之间没有统计学上的显著差异。无论是否存在TGF-β3，PRP中的总胶原蛋白含量均高于海藻酸钠。PRP中的软骨形成在质量和空间上与传统海藻酸钠珠中的不同，其特征是有孤立的强烈软骨形成中心。总体而言，结果表明单独的PRP对骨髓基质细胞的软骨诱导作用较弱，而TGF-β3可大大增强这种作用。
