去白细胞优化富血小板血浆促进肩袖腱-骨愈合的机制研究

Multiple studies have been published to demonstrate the inspiring outcomes of autologous platelet-rich plasma (PRP) in reducing rotator cuff re-tear to enhance the prognosis of rotator cuff tear by promoting tendon-to-bone healing. However, most PRP formulations used in clinical practice contain concentrated leukocytes, and our preliminary studies indicated that leukocytes in PRP might release pro-inflammatory cytokines to activate NF-κB signaling pathway to induce harmful effect on tendon-to-bone healing, and weaken the therapeutic effect of PRP on rotator cuff tear. Therefore, the objective of the current study is to elucidated the effect and mechanism of leukocytes in PRP on tendon-to-bone healing in rotator cuff tear, in an attempt to enhance the effects of platelet-rich plasma on rotator cuff tear by leukocyte depletion. Firstly, we are planning to compare the effects of leukocyte-rich PRP (L-PRP) and pure PRP (P-PRP) on dermal fibroblasts and osteoblasts in vitro, and tendon-to-bone healing in rat chronic rotator cuff tear model. Secondly, the effects of L-PRP and P-PRP on NF-κB signaling pathway will be investigated. Then, the effects of specific inhibitor of NF-κB signaling pathway on the in vitro and in vivo effects of L-PRP and P-PRP will be explored. Finally, the effect and mechanism of leukocytes in PRP on tendon-to-bone healing in rotator cuff tear will be elucidated. These findings will provide substantial proofs for the optimization of PRP therapy in the treatment of rotator cuff tear in clinical practice.

研究证实富血小板血浆（PRP）可以促进肩袖腱-骨愈合，减少肩袖术后再断裂，提高肩袖修复手术疗效。但是临床使用的大部分PRP中含有大量白细胞，我们的前期研究提示白细胞可能可以通过释放炎症因子激活NF-κB通路产生不利于腱-骨愈合的作用，从而减弱PRP对肩袖损伤的疗效。因此，本项研究拟探讨通过去白细胞来优化PRP，避免激活NF-κB通路，从而进一步提高PRP对肩袖损伤的疗效。我们拟首先制备去白细胞PRP（P-PRP），然后从细胞水平比较富白细胞PRP（L-PRP）和P-PRP对成纤维细胞和成骨细胞的作用，并从动物水平在大鼠陈旧性肩袖损伤模型上验证；同时，通过观察L-PRP和P-PRP对NF-κB通路的作用并比较NF-κB通路阻断剂对L-PRP和P-PRP对细胞和动物作用的影响探讨相关机制；最终，阐明PRP中白细胞对肩袖腱-骨愈合的作用及其机制，为临床进一步优化治疗肩袖损伤的PRP疗法提供依据。