title: A Co-Culture Model of Fibroblasts and Adipose Tissue-Derived Stem Cells Reveals New Insights into Impaired Wound Healing After Radiotherapy.
abstract: External radiation seems to be associated with increased amounts of cytokines and other cellular modulators. Impaired microcirculation and fibrosis are examples of typical long term damage caused by radiotherapy. Adipose tissue-derived stem cells (ASC) are discussed to enhance wound healing, but their role in wounds due to radiotherapy is poorly understood. Normal human fibroblasts (NHF) and ASCs were co-cultured and external radiation with doses from 2-12 Gray (Gy) was delivered. Cell proliferation and mRNA levels of matrix metalloproteinases (MMP1, MMP2 and MMP13) were determined 48 h after irradiation of the co-cultures by qPCR. Additionally, tissue inhibitors of matrix metalloproteinases (TIMP1, TIMP2) were determined by enzyme-linked immunosorbent assay (ELISA). There was a reduction of cell proliferation after external radiation in mono-cultures of NHFs and ASCs compared to controls without irradiation. The co-culture of ASCs and NHFs showed reduced impairment of cell proliferation after external radiation. Gene expression of MMP1 and MMP13 was reduced after external irradiation in NHF. MMP2 expression of irradiated NHFs was increased. In the co-culture setting, MMP1 and MMP2 gene expression levels were upregulated. TIMP1 and TIMP2 protein expression was increased after irradiation in NHFs and their co-cultures with ASCs. ASCs seem to stimulate cell proliferation of NHFs and modulate relevant soluble mediators as well as proteinases after external radiation.
authors.affiliation: Department of Otorhinolaryngology, Division of Facial Plastic Surgery, University Medical Center, Regensburg 93053, Germany. frank.haubner@ukr.de.
authors.forename: Frank
authors.initials: F
authors.lastname: Haubner
authors.affiliation: Department of Otorhinolaryngology, Division of Facial Plastic Surgery, University Medical Center, Regensburg 93053, Germany. dominique.muschter@ukr.de.
authors.forename: Dominique
authors.initials: D
authors.lastname: Muschter
authors.affiliation: Department of Radiotherapy, University Medical Center, Regensburg 93053, Germany. fabian.pohl@ukr.de.
authors.forename: Fabian
authors.initials: F
authors.lastname: Pohl
authors.affiliation: Department of Dermatology, University Medical Center, Regensburg 93053, Germany. stephan.schreml@ukr.de.
authors.forename: Stephan
authors.initials: S
authors.lastname: Schreml
authors.affiliation: Department of Plastic and Reconstructive Surgery, University Medical Center, Regensburg 93053, Germany. lukas.prantl@ukr.de.
authors.forename: Lukas
authors.initials: L
authors.lastname: Prantl
authors.affiliation: Department of Otorhinolaryngology, Division of Facial Plastic Surgery, University Medical Center, Regensburg 93053, Germany. holger.gassner@ukr.de.
authors.forename: Holger G
authors.initials: HG
authors.lastname: Gassner
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D049109:Cell Proliferation / Q000528:radiation effects; D018920:Coculture Techniques; D005347:Fibroblasts / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D011832:Radiation Injuries; D011878:Radiotherapy / Q000009:adverse effects; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000378:metabolism; D014945:Wound Healing
chemical_list: D016212:Transforming Growth Factor beta; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1
keywords: adipose derived stem cells; external radiation; fibroblasts; matrix metalloproteinases; tissue inhibitors of matrix metalloproteinases