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富血小板血浆和间充质干细胞对肌腱细胞人工伤口愈合的影响:一项比较新鲜和冷冻富血小板血浆的体外研究

PRP and MSCs on tenocytes artificial wound healing: an in vitro study comparing fresh and frozen PRP.

作者信息

Veronesi Francesca, Pagani Stefania, Torricelli Paola, Filardo Giuseppe, Cavallo Carola, Grigolo Brunella, Fini Milena

机构信息

Laboratory of Preclinical and Surgical Studies, IRCCS - Instituto Ortopedico Rizzoli, Bologna, Italy.

Applied Translational Research (ATR) Center, IRCCS - Instituto Ortopedico Rizzoli, Bologna, Italy.

出版信息

Histol Histopathol. 2018 Dec;33(12):1323-1334. doi: 10.14670/HH-18-018. Epub 2018 Jul 3.

Abstract

Tendon tissue has poor regenerative capacity due to its low vascularization, cell density and extracellular matrix (ECM) production. Therefore, tendon injuries are an increasing clinical problem because of the formation of scar tissue with traditional therapies. Regenerative medicine aims at triggering a healing response through the use of biological treatments such as mesenchymal stromal cells (MSCs) and growth factors (GFs). MSCs show several advantages in tendon clinical setting, while platelet rich plasma (PRP) has gained popularity because of its high GF concentration, although its applications in the tendon clinical setting are still controversial. The aim of the present study was to evaluate a combined treatment of MSCs and PRP in an in vitro microwound model of tendon injuries. In addition, fresh and frozen PRP were compared. Single human tenocytes cultures or co-cultures with bone marrow derived MSCs (BMSCs) were set up with or without human PRP, fresh or frozen. After 24 hours of culture, it was observed that MSCs alone significantly increased tenocyte migration speed, microwound healing rate, fibronectin, collagen I and aggrecan production. These effects were enhanced by the combination with PRP, fresh being more effective than frozen PRP. In addition, the number of MSCs and tenocytes inside the microwound was significantly increased, especially with fresh PRP. In conclusion, the combination of MSCs and PRP, especially the fresh one, increases tenocytes and MSC migration speed, as well as ECM protein production compared to the use of MSCs alone.

摘要

由于肌腱组织血管化程度低、细胞密度低以及细胞外基质(ECM)生成能力差,其再生能力较弱。因此,由于传统治疗会形成瘢痕组织,肌腱损伤成为一个日益严重的临床问题。再生医学旨在通过使用间充质基质细胞(MSCs)和生长因子(GFs)等生物治疗方法来引发愈合反应。MSCs在肌腱临床治疗中显示出多种优势,而富含血小板血浆(PRP)因其高GF浓度而受到欢迎,尽管其在肌腱临床治疗中的应用仍存在争议。本研究的目的是在肌腱损伤的体外微伤口模型中评估MSCs和PRP的联合治疗效果。此外,还比较了新鲜PRP和冷冻PRP。建立了单独的人肌腱细胞培养物或与骨髓来源的MSCs(BMSCs)的共培养物,添加或不添加人PRP,新鲜的或冷冻的。培养24小时后,观察到单独使用MSCs可显著提高肌腱细胞迁移速度、微伤口愈合率、纤连蛋白、I型胶原蛋白和聚集蛋白聚糖的产生。与PRP联合使用可增强这些效果,新鲜PRP比冷冻PRP更有效。此外,微伤口内的MSCs和肌腱细胞数量显著增加,尤其是使用新鲜PRP时。总之,与单独使用MSCs相比,MSCs和PRP的联合使用,尤其是新鲜PRP,可提高肌腱细胞和MSCs的迁移速度以及ECM蛋白的产生。

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