title: The role of interleukin-10 and hyaluronan in murine fetal fibroblast function in vitro: implications for recapitulating fetal regenerative wound healing.
abstract: BACKGROUND
  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 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.
  
  
  RESULTS
  To 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
  Our 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.
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Swathi
authors.initials: S
authors.lastname: Balaji
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Alice
authors.initials: A
authors.lastname: King
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Emily
authors.initials: E
authors.lastname: Marsh
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Maria
authors.initials: M
authors.lastname: LeSaint
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Sukanta S
authors.initials: SS
authors.lastname: Bhattacharya
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Nathaniel
authors.initials: N
authors.lastname: Han
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Yashu
authors.initials: Y
authors.lastname: Dhamija
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Rajeev
authors.initials: R
authors.lastname: Ranjan
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Louis D
authors.initials: LD
authors.lastname: Le
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Stanford University School of Medicine, Palo Alto, California, United States of America.
authors.forename: Paul L
authors.initials: PL
authors.lastname: Bollyky
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America; Center for Children's Surgery, Children's Hospital Colorado and The University of Colorado School of Medicine, Aurora, Colorado, United States of America.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Crombleholme
authors.affiliation: Laboratory for Regenerative Wound Healing, Division of Pediatric, General, Thoracic and Fetal Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America; Division of Pediatric General and Thoracic Surgery, Baylor College of Medicine and Texas Children's Hospital, Houston, Texas, United States of America.
authors.forename: Sundeep G
authors.initials: SG
authors.lastname: Keswani
mesh_terms: D000818:Animals; D017209:Apoptosis; D002465:Cell Movement; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism; D047109:Fetal Development / Q000502:physiology*; D005347:Fibroblasts / Q000502:physiology*; D006820:Hyaluronic Acid / Q000378:metabolism*; D066298:In Vitro Techniques; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D014945:Wound Healing / Q000502:physiology*
chemical_list: C556062:IL10 protein, mouse; D016753:Interleukin-10; D006820:Hyaluronic Acid