do Amaral Ronaldo Jfc, Matsiko Amos, Tomazette Marcel Rp, Rocha Wanessa Kr, Cordeiro-Spinetti Eric, Levingstone Tanya J, Farina Marcos, O'Brien Fergal J, El-Cheikh Marcia C, Balduino Alex
Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brasil ; Excellion Serviços Biomédicos, Amil/UnitedHealth Group, Petrópolis, Brasil.
Tissue Engineering Research Group, Royal College of Surgeons in Ireland (RCSI), Dublin, Ireland ; Trinity Centre for Bioengineering, Trinity College Dublin (TCD), Dublin, Ireland ; Advanced Materials and Bioengineering Research (AMBER) Centre, RCSI & TCD, Dublin, Ireland.
J Tissue Eng. 2015 Jul 7;6:2041731415594127. doi: 10.1177/2041731415594127. eCollection 2015 Jan-Dec.
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.
富含血小板血浆已被用于治疗关节软骨缺损,以期获得合成代谢和抗炎作用。然而,其在细胞软骨生成或纤维生成趋向方面的作用仍存在争议。在此,我们在体外评估了富含血小板血浆激活后获得的产物——富含血小板血浆释放物在细胞向软骨生成谱系趋向中的作用。分别使用人鼻中隔软骨生成细胞和人骨髓间充质基质细胞作为已趋向软骨生成谱系的细胞类型和未分化细胞,与常用的胎牛血清相比,测试了不同浓度的富含血小板血浆释放物。对于这两种细胞类型,低浓度的富含血小板血浆释放物(2.5%)与10%胎牛血清相比,对细胞生长具有相似的作用。在三维培养系统中,单独的富含血小板血浆释放物不会诱导完全鼻中隔软骨生成细胞形成软骨样微球。尽管如此,富含血小板血浆释放物在细胞趋向中发挥了重要作用,因为高代次鼻中隔软骨生成细胞仅在富含血小板血浆释放物而非胎牛血清存在的情况下扩增时才产生软骨样微球。组织学分析和微球面积测量表明,即使是低浓度的富含血小板血浆释放物也足以防止鼻中隔软骨生成细胞因体外扩增而丧失软骨生成潜能,从而促进其重新趋向。在软骨生成培养基中添加低浓度的富含血小板血浆释放物也增加了接种在胶原-透明质酸支架上的间充质基质细胞的软骨生成潜能,体外培养后软骨生成相关基因表达增加、硫酸化糖胺聚糖产生增加以及压缩模量增加即可观察到这一点。相反,较高浓度的富含血小板血浆释放物(10%)则阻碍了其中一些特征。总之,富含血小板血浆释放物能够防止细胞软骨生成能力丧失,诱导其表型恢复,并调节细胞趋向。我们的数据支持富含血小板血浆具有软骨生成潜能的假说,当在特定细胞类型和培养阶段需要软骨生成趋向时,在培养基中特定浓度下允许用富含血小板血浆释放物替代胎牛血清。