The role of interleukin-10 and hyaluronan in murine fetal fibroblast function in vitro: implications for recapitulating fetal regenerative wound healing.

Swathi Balaji,Sukanta S Bhattacharya,Nathaniel Han,Louis D Le,Maria Lesaint,Alice King,Paul L Bollyky,Timothy M Crombleholme,Yashu Dhamija,Rajeev Ranjan,Sundeep G Keswani,Emily Marsh,Kandiah Jeyaseelan

doi:10.1371/journal.pone.0124302

Abstract

BackgroundMid-gestation fetal cutaneous wounds heal scarlessly and this has been attributed in part to abundant hyaluronan (HA) in the extracellular matrix (ECM) and a unique fibroblast phenotype. We recently reported a novel role for interleukin 10 (IL-10) as a regulator of HA synthesis in the fetal ECM, as well as the ability of the fetal fibroblast to produce an HA-rich pericellular matrix (PCM). We hypothesized that IL-10-mediated HA synthesis was essential to the fetal fibroblast functional phenotype and, moreover, that this phenotype could be recapitulated in adult fibroblasts via supplementation with IL-10 via an HA dependent process.Methodology/Principal FindingsTo evaluate the differences in functional profile, we compared metabolism (MTS assay), apoptosis (caspase-3 staining), migration (scratch wound assay) and invasion (transwell assay) between C57Bl/6J murine fetal (E14.5) and adult (8 weeks) fibroblasts. We found that fetal fibroblasts have lower rates of metabolism and apoptosis, and an increased ability to migrate and invade compared to adult fibroblasts, and that these effects were dependent on IL-10 and HA synthase activity. Further, addition of IL-10 to adult fibroblasts resulted in increased fibroblast migration and invasion and recapitulated the fetal phenotype in an HA-dependent manner.Conclusions/SignificanceOur data demonstrates the functional differences between fetal and adult fibroblasts, and that IL-10 mediated HA synthesis is essential for the fetal fibroblasts' enhanced invasion and migration properties. Moreover, IL-10 via an HA-dependent mechanism can recapitulate this aspect of the fetal phenotype in adult fibroblasts, suggesting a novel mechanism of IL-10 in regenerative wound healing.

Highlights

Cutaneous wound repair occurs in a highly orchestrated sequence of events that begins with hemostasis, proceeds to inflammation and proliferation, and concludes with dermal remodeling
We recently reported a novel role for interleukin 10 (IL-10) as a regulator of HA synthesis in the fetal extracellular matrix (ECM), as well as the ability of the fetal fibroblast to produce an HA-rich pericellular matrix (PCM)
Our results suggest that IL-10 mediated HA synthesis is essential to the fetal fibroblast functional profile which can be recapitulated in adult fibroblasts (AFB) by addition of IL-10

Summary

Introduction

Cutaneous wound repair occurs in a highly orchestrated sequence of events that begins with hemostasis, proceeds to inflammation and proliferation, and concludes with dermal remodeling. The synthesis and remodeling of the ECM is primarily regulated by dermal fetal fibroblast (FFB) and it is believed to be an integral contributor to the fetal regenerative phenotype [18]. Previous reports have described functional differences between adult fibroblasts (AFB) and fetal fibroblasts (FFB) including differences in migratory phenotype [20,21,22], proliferation [23], differentiation and cytokine/ ECM synthesis [22, 24, 25] [26]. Mid-gestation fetal cutaneous wounds heal scarlessly and this has been attributed in part to abundant hyaluronan (HA) in the extracellular matrix (ECM) and a unique fibroblast phenotype. We hypothesized that IL-10-mediated HA synthesis was essential to the fetal fibroblast functional phenotype and, that this phenotype could be recapitulated in adult fibroblasts via supplementation with IL-10 via an HA dependent process

Methods

Results

Discussion

Conclusion