Li Huimin, Xia Ting, Zeng Hao, Qiu Yun, Wei Yan, Cheng Yihong, Wang Yulan, Zhang Xiaoxin, Ke Jin, Miron Richard, He Qing
State Key Laboratory of Oral and Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
Front Bioeng Biotechnol. 2023 Dec 7;11:1301430. doi: 10.3389/fbioe.2023.1301430. eCollection 2023.
Recently, liquid platelet-rich fibrin (PRF), a rich source of concentrated platelets and growth factors, has emerged as a promising agent for stimulating tissue regeneration. However, its specific efficacy in chondrocyte proliferation and cartilage regeneration remains underexplored. To address this question, we investigated liquid PRF obtained through horizontal centrifugation and compared its effects with hyaluronic acid (HA), a high molecular weight glucosamine supplement widely used in clinical practice to safeguard against chondral damage. Liquid PRF, produced using horizontal centrifugation (liquid H-PRF) at 500 g for 8 min, served as our experimental agent. We conducted cell viability and proliferation assays using PRF-conditioned medium. We assessed the chondrocyte phenotype of ATDC5 cells through toluidine blue and alcian blue staining, real-time polymerase chain reaction (RT-PCR), Western blotting, and immunofluorescence staining. Furthermore, we examined the expression of genes involved in inflammation through RT-PCR and Western blot analysis. Liquid H-PRF exerted notable effects on chondrocytes, influencing proliferation, inflammatory responses, and chondrogenic differentiation. The H-PRF group displayed significantly higher expression of chondrogenic markers, including Col2a1, compared to HA-treated cells, whereas aggrecan expression was significantly higher in the HA group. PRF also demonstrated the ability to reduce inflammatory levels in chondrogenic ATDC5 cells, and this effect was further enhanced when PRF from the buffy coat zone was added. In comparison, chondrocytes cultured in the HA group produced significantly fewer inflammatory factors than those in the PRF group, as confirmed qualitatively by Western blot analysis. Liquid H-PRF emerged as a potent stimulator for chondrogenesis and a regulator of the inflammatory response, achieving levels similar to HA. Moreover, liquid H-PRF exhibited strong potential for enhancing the production of cartilage extracellular matrix and promoting chondrogenic regeneration with notably increased Col2a1 levels. Future research should encompass animal studies and human trials to further evaluate the comparative effectiveness of liquid PRF HA, potentially as an alternative or complementary strategy for future clinical applications.
最近，富含浓缩血小板和生长因子的液体富血小板纤维蛋白（PRF）已成为一种有前景的促进组织再生的制剂。然而，其在软骨细胞增殖和软骨再生方面的具体功效仍未得到充分研究。为了解决这个问题，我们研究了通过水平离心获得的液体PRF，并将其效果与透明质酸（HA）进行比较，HA是一种在临床实践中广泛用于预防软骨损伤的高分子量氨基葡萄糖补充剂。使用水平离心（液体H-PRF）在500g下离心8分钟产生的液体PRF作为我们的实验制剂。我们使用PRF条件培养基进行细胞活力和增殖测定。我们通过甲苯胺蓝和阿尔辛蓝染色、实时聚合酶链反应（RT-PCR）、蛋白质印迹和免疫荧光染色评估了ATDC5细胞的软骨细胞表型。此外，我们通过RT-PCR和蛋白质印迹分析检查了炎症相关基因的表达。液体H-PRF对软骨细胞有显著影响，影响其增殖、炎症反应和软骨形成分化。与HA处理的细胞相比，H-PRF组中包括Col2a1在内的软骨形成标志物的表达显著更高，而聚集蛋白聚糖的表达在HA组中显著更高。PRF还显示出降低软骨形成的ATDC5细胞中炎症水平的能力，当添加来自血沉棕黄层区域的PRF时，这种效果进一步增强。相比之下，如蛋白质印迹分析定性证实的那样，在HA组中培养的软骨细胞产生的炎症因子明显少于PRF组中的软骨细胞。液体H-PRF成为软骨形成的有效刺激剂和炎症反应的调节剂，达到了与HA相似的水平。此外，液体H-PRF在提高软骨细胞外基质产生和促进软骨形成再生方面具有强大潜力，Col2a1水平显著增加。未来的研究应包括动物研究和人体试验，以进一步评估液体PRF与HA的比较效果，可能作为未来临床应用的替代或补充策略。
