Department of Orthopedics, Shuang Ho Hospital, Taipei Medical University, New Taipei City 23561, Taiwan.
Department of Orthopedics, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan.
Int J Mol Sci. 2020 Jan 16;21(2):577. doi: 10.3390/ijms21020577.
The main aim of this study is to develop a one-stage method to combine platelet-rich fibrin (PRF) and autologous cartilage autografts for porcine articular cartilage repair. The porcine chondrocytes were treated with different concentrations of PRF-conditioned media and were evaluated for their cell viability and extracellular glycosaminoglycan (GAG) synthesis during six day cultivation. The chemotactic effects of PRF on chondrocytes on undigested cartilage autografts were revealed in explant cultures. For the in vivo part, porcine chondral defects were created at the medial femoral condyles of which were (1) left untreated, (2) implanted with PRF combined with hand-diced cartilage grafts, or (3) implanted with PRF combined with device-diced cartilage grafts. After six months, gross grades, histological, and immunohistochemical analyses were compared. The results showed that PRF promotes the viability and GAG expression of the cultured chondrocytes. Additionally, the PRF-conditioned media induce significant cellular migration and outgrowth of chondrocytes from undigested cartilage grafts. In the in vivo study, gross grading and histological scores showed significantly better outcomes in the treatment groups as compared with controls. Moreover, both treatment groups showed significantly more type II collagen staining and minimal type I collagen staining as compared with controls, indicating more hyaline-like cartilage and less fibrous tissue. In conclusion, PRF enhances the viability, differentiation, and migration of chondrocytes, thus, showing an appealing capacity for cartilage repair. The data altogether provide evidences to confirm the feasibility of a one-stage, culture-free method of combining PRF and cartilage autografts for repairing articular cartilage defects. From translational standpoints, these advantages benefit clinical applications by simplifying and potentiating the efficacy of cartilage autograft transplants.
本研究的主要目的是开发一种将富含血小板的纤维蛋白 (PRF) 和自体软骨移植物结合起来用于猪关节软骨修复的一步法。将猪软骨细胞用不同浓度的 PRF 条件培养基处理，并在 6 天培养过程中评估其细胞活力和细胞外糖胺聚糖 (GAG) 合成。在 explant 培养中揭示了 PRF 对未消化软骨移植物上软骨细胞的趋化作用。在体内部分，在股骨内侧髁处创建猪软骨缺损，其中（1）未处理，（2）植入 PRF 与手工切碎软骨移植物结合，或（3）植入 PRF 与器械切碎软骨移植物结合。6 个月后，比较大体评分、组织学和免疫组织化学分析。结果表明，PRF 促进了培养的软骨细胞的活力和 GAG 表达。此外，PRF 条件培养基诱导未消化软骨移植物中的软骨细胞显著迁移和生长。在体内研究中，与对照组相比，治疗组的大体分级和组织学评分显示出明显更好的结果。此外，与对照组相比，两组治疗组均显示出明显更多的 II 型胶原染色和最小的 I 型胶原染色，表明更类似于透明软骨和更少的纤维组织。总之，PRF 增强了软骨细胞的活力、分化和迁移，从而显示出对软骨修复的吸引力。这些数据提供了证据，证实了将 PRF 与软骨移植物结合用于修复关节软骨缺损的无细胞一步法的可行性。从转化的角度来看，这些优势通过简化和增强软骨移植物移植的疗效，使临床应用受益。
