Alice King1, Swathi Balaji1, Louis D Le1, Emily Marsh1, Timothy M Crombleholme2, Sundeep G Keswani1. 1. Division of Pediatric, Thoracic and General Surgery, The Center for Molecular Fetal Therapy, Cincinnati Children's Hospital and the University of Cincinnati College of Medicine, Cincinnati, OH. 2. Division of Pediatric Surgery, The Children's Hospital Colorado, Aurora, CO.
Abstract
BACKGROUND/ PURPOSE: Mid-gestational (E14.5) fetal wounds heal regeneratively with attenuated inflammation and high levels of hyaluronan (HA) in their extracellular matrix (ECM), whereas late-gestational (E18.5) fetal wounds heal with scarring. IL-10 plays an essential role in the fetal regenerative phenotype and is shown to recapitulate scarless wound healing postnatally. We hypothesize a novel role of IL-10 as a regulator of HA in the ECM. METHODS: Murine fetal fibroblasts (FFb) from C57Bl/6 and IL-10-/- mice were evaluated in vitro. Pericellular matrix (PCM) and HA synthesis were quantified using a particle exclusion assay and ELISA. The effects of hyaluronidase and hyaluronan synthase (HAS) inhibitor (4-methylumbelliferone[4-MU]) were evaluated. An ex vivo fetal forearm culture incisional wound model comparing mid-gestation and late-gestation fetuses was used to evaluate IL-10's effect on HA-rich ECM production with pentachrome and immunohistochemistry. RESULTS: FFb produce a robust HA-rich PCM which is IL-10 dependent and attenuated with hyaluronidase and HAS inhibition. Mid-gestation fetal wounds produce more ground substance and HA than late-gestation fetal wounds. IL-10 in late-gestation fetal wounds results in elevated ground substance levels and HA staining. CONCLUSIONS: Our data demonstrate that IL-10 regulates an HA-rich ECM deposition, suggesting a novel non-immunoregulatory mechanism of IL-10 in mediating regenerative wound healing.
BACKGROUND/ PURPOSE: Mid-gestational (E14.5) fetal wounds heal regeneratively with attenuated inflammation and high levels of hyaluronan (HA) in their extracellular matrix (ECM), whereas late-gestational (E18.5) fetal wounds heal with scarring. IL-10 plays an essential role in the fetal regenerative phenotype and is shown to recapitulate scarless wound healing postnatally. We hypothesize a novel role of IL-10 as a regulator of HA in the ECM. METHODS:Murine fetal fibroblasts (FFb) from C57Bl/6 and IL-10-/- mice were evaluated in vitro. Pericellular matrix (PCM) and HA synthesis were quantified using a particle exclusion assay and ELISA. The effects of hyaluronidase and hyaluronan synthase (HAS) inhibitor (4-methylumbelliferone[4-MU]) were evaluated. An ex vivo fetal forearm culture incisional wound model comparing mid-gestation and late-gestation fetuses was used to evaluate IL-10's effect on HA-rich ECM production with pentachrome and immunohistochemistry. RESULTS: FFb produce a robust HA-rich PCM which is IL-10 dependent and attenuated with hyaluronidase and HAS inhibition. Mid-gestation fetal wounds produce more ground substance and HA than late-gestation fetal wounds. IL-10 in late-gestation fetal wounds results in elevated ground substance levels and HA staining. CONCLUSIONS: Our data demonstrate that IL-10 regulates an HA-rich ECM deposition, suggesting a novel non-immunoregulatory mechanism of IL-10 in mediating regenerative wound healing.
Authors: William H Peranteau; Liping Zhang; Nidal Muvarak; Andrea T Badillo; Antoneta Radu; Philip W Zoltick; Kenneth W Liechty Journal: J Invest Dermatol Date: 2008-01-17 Impact factor: 8.551
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Authors: Swathi Balaji; Chad M Moles; Sukanta S Bhattacharya; Maria LeSaint; Yashu Dhamija; Louis D Le; Alice King; Mykia Kidd; Muhammad F Bouso; Aimen Shaaban; Timothy M Crombleholme; Paul Bollyky; Sundeep G Keswani Journal: J Surg Res Date: 2014-02-22 Impact factor: 2.192
Authors: Miguel Quiros; Hikaru Nishio; Philipp A Neumann; Dorothee Siuda; Jennifer C Brazil; Veronica Azcutia; Roland Hilgarth; Monique N O'Leary; Vicky Garcia-Hernandez; Giovanna Leoni; Mingli Feng; Gabriela Bernal; Holly Williams; Priya H Dedhia; Christian Gerner-Smidt; Jason Spence; Charles A Parkos; Timothy L Denning; Asma Nusrat Journal: J Clin Invest Date: 2017-08-07 Impact factor: 19.456
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Authors: Swathi Balaji; Xinyi Wang; Alice King; Louis D Le; Sukanta S Bhattacharya; Chad M Moles; Manish J Butte; Vinicio A de Jesus Perez; Kenneth W Liechty; Thomas N Wight; Timothy M Crombleholme; Paul L Bollyky; Sundeep G Keswani Journal: FASEB J Date: 2016-11-30 Impact factor: 5.191
Authors: Kavya L Singampalli; Swathi Balaji; Xinyi Wang; Umang M Parikh; Aditya Kaul; Jamie Gilley; Ravi K Birla; Paul L Bollyky; Sundeep G Keswani Journal: Front Cell Dev Biol Date: 2020-07-17
Authors: Emily H Steen; Xinyi Wang; Swathi Balaji; Manish J Butte; Paul L Bollyky; Sundeep G Keswani Journal: Adv Wound Care (New Rochelle) Date: 2020-02-07 Impact factor: 4.947