hyaluronan fragments improve wound healing on in vitro cutaneous model through p2x7 purinoreceptor basal activation role of molecular weight透明质酸的碎片改善伤口愈合体外皮肤模型通过p2x7 purinoreceptor基底分子量的激活作用.pdfVIP

Hyaluronan Fragments Improve Wound Healing on In Vitro Cutaneous Model through P2X7 Purinoreceptor Basal Activation: Role of Molecular Weight Kamelia Ghazi, Uriell Deng-Pichon, Jean-Michel Warnet, Patrice Rat* ´ ´ ´ Chimie-Toxicologie Analytique et Cellulaire (EA 4463), Universite Paris Descartes, Sorbonne Paris Cite, Faculte de Pharmacie, Paris, France Abstract Background: hyaluronan biopolymer is used in dermatology but the underlying mechanism and the impact of its molecular weight have not yet been investigated in skin wound healing. The aim of our work was to study the role of HA molecular weight in the proliferative phase of wound healing and to understand how this physiological biopolymer acts to promote wound healing on a human keratinocyte in vitro model. Methodology and Findings: wound healing closure was evaluated using scratch test assay, cell proliferation by counting cell with haemocytometer, expression of CD44 and ZO-1 (protein present in tight junctions specific of epithelia) using flow cytometry, and P2X7 receptor activation on living using a cytoflurometric method. Our study showed that medium hyaluronan fragment (MMW-HA, between 100 and 300 kDa) induced a significant increase in wound closure, increased ZO-1 protein expression and induced a slight activation of P2X7 receptor, contrary to high (between 1000 and 1400 kDa) and low (between 5 and 20 kDa) molecular hyaluronan fragments that had no healing effects. Basal activation of P2X7 receptor is already known to stimulate cell proliferation and this activation in our model plays a pivotal role in MMW-HA-induced wound healing. Indeed, we showed that use of BBG, a specific inhibitor of P2X7 receptor, blocked completely the beneficial effects of MMW-HA on wo