Human Adipose-Derived Stem Cell Conditioned Media and Exosomes Containing MALAT1 Promote Human Dermal Fibroblast Migration and Ischemic Wound Healing.

Denise R Cooper,Andrea Trujillo,Rehka Patel,Mack H Wu,Chunyan Wang,Jamie Prather,Lisa J Gould,Niketa A Patel

doi:10.1089/wound.2017.0775

Abstract

Objective: Chronically ill patients heal recalcitrant ulcerative wounds more slowly. Human adipose-derived stem cells (hADSCs) play an important role in tissue regeneration and exosomes secreted by hADSC contribute to their paracrine signaling. In addition to cytokines, lipids and growth factors, hADSC secrete mRNA, miRNA, and long noncoding (lnc) RNA into exosomes. In this study we examined the role of lncRNA MALAT1 (metastasis-associated lung adenocarcinoma transcript 1), an abundant lncRNA in exosomes from conditioned media (CM), on cell migration and ischemic wound healing.Approach: CM and isolated exosomes from hADSC were applied to human dermal fibroblast (HDF) in scratch assays and electric cell-substrate impedance sensing (ECIS) assays. CM was also applied to a rat model of ischemic wound healing and wound closure was followed.Results: CM stimulated cell migration of HDFs in vitro by 48%. CM stimulated the closure of ischemic wounds in a rat model 50% faster than unconditioned media. The depletion of MALAT1 in adipose-derived stem cell (ADSC) CM significantly reduced cell migration. Since MALAT1 is secreted into exosomes, a purified population of exosomes was applied to HDF where they enhanced cell migration in a similar manner to FGF-2 or basic fibroblast growth factor (bFGF) in ECIS wound healing assays. The uptake of exosomes by HDF was shown using dynasore, an inhibitor that blocks clathrin- and caveolin-dependent endocytosis. Depletion of MALAT1 in hADSC with antisense oligonucleotides resulted in exosomes without MALAT1. These exosomes had an effect similar to the unconditioned, control media in ECIS assays.Innovation: Exosomes contain lncRNA MALAT1 and other factors that have the potential to stimulate HDF cell migration and angiogenesis involved in wound healing without applying stem cells to wounds.Conclusion: Our results show the potential of using topically applied ADSC-derived exosomes containing MALAT1 for treating ischemic wounds. This allows for harnessing the power of stem cell paracrine signaling capabilities without applying the cells.

Highlights

Ill patients are faced with challenges when they develop recalcitrant wounds
We demonstrate that adipose-derived stem cell (ADSC)-derived exosomes increase wound healing by stimulating the migration of dermal fibroblasts, and that the lncRNA, MALAT1, is largely responsible for increased cell migration in vitro
CLINICAL PROBLEM ADDRESSED This study focuses on exosomes, extracellular vesicles secreted by stem cells, which, when isolated from conditioned media (CM), can alter the response of wounded cells, and increase in vivo wound healing

Summary

Introduction

Ill patients are faced with challenges when they develop recalcitrant wounds. A number of local and systemic factors impact wound healing including oxygenation, infection, age, sex hormones, stress, diabetes, obesity, medications such as chemotherapy, alcoholism, smoking, and nutrition.[1] The wound healing process consists of four integrated and overlapping phases: hemostasis, inflammation, proliferation, and tissue remodeling.[2] These must occur rapidly and appropriately in the proper sequence and continue for a specific duration for wound healing to occur successfully.[3] Investigations and clinical studies have provided a wealth of information about normal and impaired wound healing. Incomplete removal of bacteria and endotoxins can lead to infections at the injured skin surface, lengthening the inflammatory phase. Prolongation and escalation of the inflammatory phase contributes to a failure to heal. Systemic factors contribute to impaired wound healing.[1]

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Conclusion