Nicoletti Giovanni, Saler Marco, Villani Laura, Rumolo Agnese, Tresoldi Marco Mario, Faga Angela
Plastic and Reconstructive Surgery, Department of Clinical Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Pavia, Italy.
Advanced Technologies for Regenerative Medicine and Inductive Surgery Research Center, University of Pavia, Pavia, Italy.
Front Bioeng Biotechnol. 2019 Jan 22;7:2. doi: 10.3389/fbioe.2019.00002. eCollection 2019.
This study reports on the development of an original, wounded skin culture protocol using autologous Platelet Rich Plasma (PRP) and enriched Dulbecco's Modified Eagle's Medium (DMEM). Human skin samples obtained from specimens harvested during reduction mammoplasty procedures, were injured in their central portion-to create a standard wound-and cultured under three different conditions: - enriched DMEM with saline solution in the central wound (control)- enriched DMEM with the same medium in the central wound- enriched DMEM plus 2.5% autologous PRP, with the same PRP added medium in the central wound. Morphological analysis was carried out at 0 h (T) and on days 1, 3, 5 and 10 (T-T-T-T) using Hematoxylin and Eosin; Masson's trichrome staining; Weigert staining and Ki-67 staining to identify the skin histological features in the different experimental conditions. The combination of DMEM and PRP allowed a favorable modulation of the epithelial cells and fibroblasts proliferation, and a relevant anti-inflammatory action. PRP also demonstrated an inhibitory effect on both the collagen and elastic fibers' de-structuration and a favorable modulation of the re-organization of these fibers. The step by step histological and immune-histo-chemical regenerative effects of PRP on human skin wound repair and regeneration process was observed over a period of 10 days.
本研究报告了一种使用自体富血小板血浆（PRP）和富集的杜氏改良伊格尔培养基（DMEM）的原创性伤口皮肤培养方案的开发情况。从缩乳手术中获取的标本中获得的人类皮肤样本，在其中心部分造成损伤以形成标准伤口，并在三种不同条件下进行培养：- 在中心伤口处用生理盐水的富集DMEM（对照）- 在中心伤口处用相同培养基的富集DMEM- 在中心伤口处用富集DMEM加2.5%自体PRP，且在中心伤口处添加相同的含PRP培养基。在0小时（T）以及第1、3、5和10天（T1、T2、T3、T4）使用苏木精和伊红、马松三色染色、魏格特染色和Ki-67染色进行形态学分析，以识别不同实验条件下的皮肤组织学特征。DMEM和PRP的组合有利于调节上皮细胞和成纤维细胞的增殖，并具有显著的抗炎作用。PRP还对胶原蛋白和弹性纤维的解构表现出抑制作用，并对这些纤维的重组起到有利的调节作用。在10天的时间里观察到PRP对人类皮肤伤口修复和再生过程逐步产生的组织学和免疫组织化学再生效果。
