Department of Cell Biology, Rostock University Medical Center, Rostock, Germany.
Department of Sports Medicine, Charité University Medicine Berlin, Berlin, Germany.
Am J Sports Med. 2020 Jun;48(7):1727-1734. doi: 10.1177/0363546520911035. Epub 2020 Apr 13.
Platelet-rich plasma (PRP) is widely used in sports medicine. However, neither preparation nor parameters for clinical application, such as concentration, timing, and number of applications, are standardized, making research and clinical utilization challenging.
To investigate the effect of varying doses of PRP powder in terms of different concentrations, timing, and number of applications on human chondrocytes in a reproducible cell culture model.
Controlled laboratory study.
A standardized lyophilized platelet growth factor preparation (PRP powder) was used to stimulate human chondrocytes. Chondrocytes were cultivated for 2 weeks with different stimulation frequencies (2×, 3×, 6×) and different concentrations of PRP powders (0.5%, 1%, 5%). Cell proliferation and metabolic cell activity were analyzed on days 7 and 14. Phenotypic changes were visualized through live-dead staining. Chondrogenic differentiation was quantified with enzyme-linked immunosorbent assay to assess the synthesis of procollagen types 1 and 2. Furthermore, sulfated proteoglycans and glycosaminoglycans were analyzed.
Human chondrocytes exhibited a significant dose- and time-dependent increase after 14 days in cell number (1% and 5% PRP powder vs unstimulated control: 7.95- and 15.45-fold increase, respectively; 2× vs 6× stimulation with 5% PRP powder: 4.00-fold increase) and metabolic cell activity (1% and 5% PRP powder vs unstimulated control: 3.27-fold and 3.58-fold change, respectively). Furthermore, cells revealed a significant increase in the amount of bone-specific procollagen type 1 (14 days, 1.94-fold) and sulfated glycosaminoglycans (14 days, 2.69-fold); however, no significant change was observed in the amount of cartilage-specific collagen type 2.
We showed that chondrocytes exhibit a significant dose- and time-dependent increase in cell number and metabolic cell activity. The standardized use of growth factor concentrates in cell culture models can contribute to clinical knowledge in terms of dosage and timing of PRP applications.
Problems with PRP, such as the absence of standardization, lack of consistency among studies, and unknown dosage, could be solved by using characterized PRP powder made by pooling and lyophilizing multiple platelet concentrates. The innovative PRP powder generates new possibilities for PRP research, as well as for the treatment of patients.
富血小板血浆（PRP）在运动医学中被广泛应用。然而，其制备方法和临床应用参数（如浓度、时间和应用次数）尚未标准化，这使得研究和临床应用具有挑战性。
在可重复的细胞培养模型中，研究不同浓度、时间和次数的 PRP 粉末剂量对人软骨细胞的影响。
对照实验室研究。
使用标准化的冻干血小板生长因子制剂（PRP 粉末）刺激人软骨细胞。软骨细胞在不同的刺激频率（2×、3×、6×）和不同浓度的 PRP 粉末（0.5%、1%、5%）下培养 2 周。在第 7 天和第 14 天分析细胞增殖和代谢细胞活性。通过死活染色观察表型变化。通过酶联免疫吸附试验定量评估前胶原蛋白 1 和 2 的合成，以评估软骨形成分化。此外，还分析了硫酸化蛋白聚糖和糖胺聚糖。
在第 14 天，细胞数量（1%和 5% PRP 粉末与未刺激对照相比：分别增加了 7.95 倍和 15.45 倍；5% PRP 粉末 2×与 6×刺激相比：增加了 4.00 倍）和代谢细胞活性（1%和 5% PRP 粉末与未刺激对照相比：分别增加了 3.27 倍和 3.58 倍）均呈现出显著的剂量和时间依赖性增加。此外，细胞中骨特异性前胶原蛋白 1 的含量（14 天，增加 1.94 倍）和硫酸化糖胺聚糖（14 天，增加 2.69 倍）也显著增加，而软骨特异性胶原蛋白 2 的含量没有明显变化。
我们表明，软骨细胞的细胞数量和代谢细胞活性均呈现出显著的剂量和时间依赖性增加。在细胞培养模型中标准化使用生长因子浓缩物可以为 PRP 应用的剂量和时间提供临床知识。
PRP 存在标准化缺失、研究间一致性差以及剂量未知等问题，使用汇集和冻干多个血小板浓缩物制成的有特征的 PRP 粉末可以解决这些问题。创新的 PRP 粉末为 PRP 研究以及患者治疗提供了新的可能性。
