Platelet Rich Plasma Enhancement of Skin Regeneration in an ex-vivo Human Experimental Model.

Giovanni Nicoletti,Angela Faga,Agnese Rumolo,Laura Villani,Marco Mario Tresoldi,Marco Saler

doi:10.3389/fbioe.2019.00002

Abstract

This study reports on the development of an original, ex-vivo 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 (T0) and on days 1, 3, 5 and 10 (T1-T3-T5-T10) 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.

Highlights

Several experimental models are currently available for the investigation of complex skin physiology.The in-vitro models are usually made of a single skin cell type—fibroblasts or keratinocytes— cultured in the appropriate medium (Johnen et al, 2008)
The aim of this study is the development of an original, exvivo human wounded skin culture protocol by adding autologous Platelet Rich Plasma (PRP) to a conventional culture medium, in order to enhance the tissue regeneration process
A few individual fibroblasts were observed up to T5 whereas at T10 their amount significantly increased with a homogeneous distribution in the dermis, with the exception of the sub-epithelial dermal layer

Summary

Introduction

Several experimental models are currently available for the investigation of complex skin physiology.The in-vitro models are usually made of a single skin cell type—fibroblasts or keratinocytes— cultured in the appropriate medium (Johnen et al, 2008). Several experimental models are currently available for the investigation of complex skin physiology. The disadvantages are the fine differences in skin physiology between humans and animals and the ethical issues that strongly limit this model. The ex-vivo organotypic model (3D) allows the best approximation to living, human skin. The bioengineered exvivo organotypic model allows for control of the keratinocytes’ differentiation, it does not include all of the different cell types of living skin. It lacks, for example, the skin adnexa and, its manufacturing process is time and money consuming. The model which best approximates living human skin, is the organotypic model as it enables immediate and short-term evaluation of a particular effect on cells and surrounding tissue components, it is seldom used due to its technical complexity and the limited availability of full thickness human skin samples (Safferling et al, 2013; Mori et al, 2016)

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