Platelet-rich plasma releasate differently stimulates cellular commitment toward the chondrogenic lineage according to concentration.
Platelet-rich plasma releasate differently stimulates cellular commitment toward the chondrogenic lineage according to concentration.
复制标题
DOI：
10.1177/2041731415594127
复制
发表时间：
2015-01
影响因子：
8.2
通讯作者：
Balduino A
中科院分区：
文献类型：
--
作者：
do Amaral RJ;Matsiko A;Tomazette MR;Rocha WK;Cordeiro-Spinetti E;Levingstone TJ;Farina M;O'Brien FJ;El-Cheikh MC;Balduino A
富血小板血浆已被用于治疗关节软骨缺损，期望具有合成代谢和抗炎作用。然而，其在细胞软骨形成或纤维形成中的作用仍存在争议。本文在体外评估了富血小板血浆释放（富血小板血浆激活后获得的产物）对细胞向软骨细胞谱系的作用。人鼻中隔软骨细胞和人骨髓间充质间质细胞分别被用作已经形成软骨细胞谱系的细胞类型和未分化细胞类型，因为与常用的胎牛血清相比，测试了不同浓度的富血小板血浆释放。对于两种细胞类型，低浓度富血小板血浆释放物（2.5%）与10%胎牛血清相比，对细胞生长的影响相似。在三维培养系统中，仅富血小板血浆释放不能诱导完整的鼻中隔软骨细胞软骨样颗粒形成。尽管如此，富血小板血浆释放在细胞活动中发挥了重要作用，因为高传代鼻中隔软骨细胞只有在富血小板血浆释放而不是胎牛血清存在下扩张时才形成软骨样微球。组织学分析和颗粒面积测量表明，即使是低浓度的富血小板血浆释放也足以防止鼻中隔软骨细胞因体外扩张而失去其成软骨潜能，从而促进其再沉积。低浓度的富血小板血浆释放物在成软骨培养基中补充，也增加了胶原-透明质酸支架上的间充质基质细胞的成软骨潜能，这是通过在体外培养后软骨相关基因表达、硫酸糖胺聚糖产生和压缩模量的增加观察到的。相反，高浓度的富血小板血浆释放（10%）阻碍了这些特征。总之，富血小板血浆释放能够防止细胞软骨生成能力丧失，诱导其表型恢复，并调节细胞承诺。我们的数据支持富血小板血浆成软骨潜能的假设，当特定细胞类型和培养时刻需要成软骨时，允许胎牛血清替代培养基中特定浓度的富血小板血浆释放。
Platelet-rich plasma has been used to treat articular cartilage defects, with the expectations of anabolic and anti-inflammatory effects. However, its role on cellular chondrogenic or fibrogenic commitment is still a controversy. Herein, the role of platelet-rich plasma releasate, the product obtained following platelet-rich plasma activation, on cellular commitment toward the chondrogenic lineage was evaluated in vitro. Human nasoseptal chondrogenic cells and human bone marrow mesenchymal stromal cells were used as cell types already committed to the chondrogenic lineage and undifferentiated cells, respectively, as different concentrations of platelet-rich plasma releasate were tested in comparison to commonly used fetal bovine serum. Low concentration of platelet-rich plasma releasate (2.5%) presented similar effects on cellular growth compared to 10% fetal bovine serum, for both cell types. In a three-dimensional culture system, platelet-rich plasma releasate alone did not induce full nasoseptal chondrogenic cells cartilage-like pellet formation. Nonetheless, platelet-rich plasma releasate played a significant role on cell commitment as high-passage nasoseptal chondrogenic cells only originated cartilage-like pellets when expanded in the presence of platelet-rich plasma releasate rather than fetal bovine serum. Histological analyses and measurements of pellet area demonstrated that even low concentrations of platelet-rich plasma releasate were enough to prevent nasoseptal chondrogenic cells from losing their chondrogenic potential due to in vitro expansion thereby promoting their recommitment. Low concentration of platelet-rich plasma releasate supplemented in chondrogenic medium also increased the chondrogenic potential of mesenchymal stromal cells seeded on collagen-hyaluronic acid scaffolds, as observed by an increase in chondrogenic-related gene expression, sulfated glycosaminoglycan production, and compressive modulus following in vitro culture. On the contrary, higher concentration of platelet-rich plasma releasate (10%) hampered some of these features. In conclusion, platelet-rich plasma releasate was able to prevent cellular chondrogenic capacity loss, inducing regain of their phenotype, and modulate cell commitment. Our data support the hypothesis of platelet-rich plasma chondrogenic potential, allowing fetal bovine serum substitution for platelet-rich plasma releasate at specific concentrations in culture medium when chondrogenic commitment is desired on specific cell types and moments of culture.