Pure Platelet-Rich Plasma promotes Semaphorin3A expression: a novel insight to ameliorate intervertebral disc degeneration in vitro

Abstract

Abstract Introduction: Platelet-rich plasma(PRP) has been proven to have therapeutic potential for intervertebral disc degeneration (IVDD). Pure PRP (P-PRP) with the exclusion of leukocytes has been proved to be a better choice for mitigating IVDD, while the potential mechanism is unclear. Sema3A, an inhibitor of innervation and angiogenesis, plays a vital role in maintaining the homeostasis of IVDD. However, it was seldom studied whether PRP prevents IVDD by modulating Semaphorin3A (Sema3A). The purpose of this study is to elucidate the effect of P-PRP on Sema3A in the progress of IVDD in vitro. Methods: Nucleus pulposus cells (NPCs) isolated from 8-week-old male Sprague-Dawle rats were exposed to 10ng/ml IL-1β, and then treated with P-PRP or leukocyte platelet-rich plasma (L-PRP) in vitro, followed by measuring cell proliferation, apoptosis and microstructures, inflammatory gene and Sema3A expression, as well as anabolic and catabolic protein expression by immunostaining, quantitative real-time polymerase chain reaction (qPCR), Western blot, and enzyme-linked immunosorbent assay (ELISA). Results: The concentration of growth factors in P-PRP was higher than that of L-PRP, while the concentration of inflammatory elements was lower. The proliferation of NPCs was enhanced by P-PRP and the apoptosis level was alleviated after the intervention of IL-1β. The expression levels of anabolic genes and aggrecan, collagen II were elevated. On the contrary, the expression levels of catabolic or inflammatory genes including MMP-3, ADAMTS-4 were decreased. The Sema3A activity was promoted after intervention of P-PRP, while the expression levels of CD31 and NF200 were down regulated. Conclusions: P-PRP improved the function of NPCs in IVDD by modulating the NF-κB signaling pathway and promoting Sema3A expression, which may provide a new insight for IVDD treatment. The translational potential of this article: The study elucidates the potential mechanism of PRP reveals novel insights into the role of Sema3A in the progression of IVDD and provides a new therapeutic target for the treatment of IVDD.