Citation: Sz˝ ucs, D.; Miklós, V.; Monostori, T.; Guba, M.; Kun-Varga, A.; Póliska, S.; Kis, E.; Bende, B.; Kemény, L.; Veréb, Z. Effect of Inflammatory Microenvironment on the Regenerative Capacity of Adipose-Derived Mesenchymal Stem Cells. Cells 2023, 12, 1966. https:// doi.org/10.3390/cells12151966 Received: 26 May 2023 Revised: 12 July 2023 Accepted: 22 July 2023 Published: 29 July 2023 Copyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). cells Article Effect of Inflammatory Microenvironment on the Regenerative Capacity of Adipose-Derived Mesenchymal Stem Cells Diána Sz ˝ ucs 1,2,3 , Vanda Miklós 4 , Tamás Monostori 1,3 , Melinda Guba 1,3 , Anikó Kun-Varga 1 , Szilárd Póliska 5 , Erika Kis 6 , Balázs Bende 6 , Lajos Kemény 1,3,7 and Zoltán Veréb 1,3, * 1 Regenerative Medicine and Cellular Pharmacology Laboratory, Department of Dermatology and Allergology, University of Szeged, 6720 Szeged, Hungary; szucs.diana@med.u-szeged.hu (D.S.); monostori.tamas.bence@med.u-szeged.hu (T.M.); guba.melinda@med.u-szeged.hu (M.G.); kun-varga.aniko@med.u-szeged.hu(A.K.-V.); kemeny.lajos@med.u-szeged.hu (L.K.) 2 Doctoral School of Clinical Medicine, University of Szeged, 6720 Szeged, Hungary 3 Centre of Excellence for Interdisciplinary Research, Development and Innovation, University of Szeged, 6720 Szeged, Hungary 4 Biobank, University of Szeged, 6720 Szeged, Hungary; miklos.vanda@szte.hu 5 Genomic Medicine and Bioinformatics Core Facility, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, H-4032 Debrecen, Hungary; poliska@med.unideb.hu 6 Dermatosurgery and Plastic Surgery, Department of Dermatology and Allergology, University of Szeged, 6720 Szeged, Hungary; kis.erika.gabriella@med.u-szeged.hu (E.K.); bende.balazs@med.u-szeged.hu (B.B.) 7 Hungarian Centre of Excellence for Molecular Medicine-USz Skin Research Group, University of Szeged, 6720 Szeged, Hungary * Correspondence: vereb.zoltan@med.u-szeged.hu Abstract: Adipose-derived mesenchymal stem cells are increasingly being used in regenerative medicine as cell therapy targets, including in the treatment of burns and ulcers. The regenerative potential of AD-MSCs and some of their immunological properties are known from in vitro studies; however, in clinical applications, cells are used in non-ideal conditions and can behave differently in inflammatory environments, affecting the efficacy and outcome of therapy. Our aim was to investigate and map the pathways that the inflammatory microenvironment can induce in these cells. High-throughput gene expression assays were performed on AD-MSCs activated with LPS and TNFα. Analysis of RNA-Seq data showed that control, LPS-treated and TNFα-treated samples exhibited distinct gene expression patterns. LPS treatment increased the expression of 926 genes and decreased the expression of 770 genes involved in cell division, DNA repair, the cell cycle, and several metabolic processes. TNFα treatment increased the expression of 174 genes and decreased the expression of 383 genes, which are related to cell division, the immune response, cell proliferation, and differentiation. We also map the biological pathways by further investigating the most altered genes using the Gene Ontology and KEGG databases. Secreted cytokines, which are important in the immunological response, were also examined at the protein level, and a functional assay was performed to assess wound healing. Activated AD-MSC increased the secretion of IL-6, IL-8 and CXCL-10, and also the closure of wounds. AD-MSCs presented accelerated wound healing under inflammation conditions, suggesting that we could use this cell in clinical application. Keywords: adipose-derived mesenchymal stem cells; lipopolysaccharide; tumor necrosis factor α; inflammation; regenerative medicine 1. Introduction Adipose tissue can be found in several locations in the human body, such as subcuta- neous and visceral sites, intra-articular, intramuscular, intra-hepatic depots, and in bone marrow. Adipose tissue is not only an energy reservoir but also an endocrine organ since it produces mediators of metabolism and cell function. Adipocytes and nonadipocytes secrete adipokines (leptin, adiponectin, omentin, and resistin), pro- and anti-inflammatory Cells 2023, 12, 1966. https://doi.org/10.3390/cells12151966 https://www.mdpi.com/journal/cells