TGF‐β1‐activated type 2 dendritic cells promote wound healing and induce fibroblasts to express tenascin c following corneal full‐thickness hydrogel transplantation

Christine Moelzer, Sucharita P Shankar, Vlad Masalski, May Griffith, Lucia Kuffova (Corresponding Author), John V Forrester (Corresponding Author)

Research output: Contribution to journalArticle

Abstract

We showed previously that 1‐ethyl‐3‐(3‐dimethylamino‐propyl)‐carbodiimide hydrochloride (EDC) cross‐linked recombinant human collagen III hydrogels promoted stable regeneration of the human cornea (continued nerve and stromal cell repopulation) for over 4 years. However, as EDC cross linking kinetics were difficult to control, we additionally tested a sterically bulky carbodiimide. Here, we compared the effects of two carbodiimide cross linkers—bulky, aromatic N‐cyclohexyl‐N0‐(2‐morpholinoethyl)‐carbodiimide (CMC), and nonbulky EDC—in a mouse corneal graft model. Murine corneas undergoing full‐thickness implantation with these gels became opaque due to dense retro‐corneal membranes (RCM). Corneal epithelial cytokeratin 12 and alpha smooth muscle actin indicative of functional tissue regeneration and wound contraction were observed in RCM surrounding both hydrogel types. However, quantitatively different levels of infiltrating CD11c+ dendritic cells (DC) were found, suggesting a hydrogel‐specific innate immune response. More DC infiltrated the stroma surrounding EDC‐N‐hydroxysuccinimide (NHS) hydrogels concurrently with higher fibrosis‐associated tenascin c expression. The opposite was true for CMC‐NHS gels that had previously been shown to be more tolerising to DC. In vitro studies showed that DC cultured with transforming growth factor β1 (TGF‐β1) induced fibroblasts to secrete more tenascin c than those cultured with lipopolysaccharide and this effect was blocked by TGF‐β1 neutralisation. Furthermore, tenascin c staining was found in 40‐ to 50μm long membrane nanotubes formed in fibroblast/DC cocultures. We suggest that TGF‐β1 alternatively activated (tolerising) DC regulate fibroblast‐mediated tenascin c secretion, possibly via local production of TGF‐β1 in early wound contraction, and that this is indirectly modulated by different hydrogel chemistries.
Original languageEnglish
Pages (from-to)1507-1517
Number of pages11
JournalJournal of Tissue Engineering and Regenerative Medicine
Volume13
Issue number9
Early online date25 Jul 2019
DOIs
Publication statusPublished - Sep 2019

Fingerprint

Tenascin
Hydrogel
Wound Healing
Dendritic Cells
Fibroblasts
Transplantation
Transforming Growth Factors
Carbodiimides
Hydrogels
Cornea
Membranes
Keratin-12
Regeneration
Gels
Nanotubes
Wounds and Injuries
Stromal Cells
Coculture Techniques
Innate Immunity
Smooth Muscle

Keywords

  • biomaterial
  • corneal transplantation
  • tissue repair
  • transforming growth factor beta 1
  • membrane nanotubes
  • tectonic graft
  • MIGRATION
  • REGENERATION
  • INNATE
  • IMPLANTS
  • MATURATION
  • RESPONSES
  • EPITHELIUM
  • transforming growth factor β1, membrane nanotubes, tectonic graft

ASJC Scopus subject areas

  • Biomedical Engineering
  • Medicine (miscellaneous)
  • Biomaterials

Cite this

TGF‐β1‐activated type 2 dendritic cells promote wound healing and induce fibroblasts to express tenascin c following corneal full‐thickness hydrogel transplantation. / Moelzer, Christine; Shankar, Sucharita P; Masalski, Vlad; Griffith, May; Kuffova, Lucia (Corresponding Author); Forrester, John V (Corresponding Author).

In: Journal of Tissue Engineering and Regenerative Medicine, Vol. 13, No. 9, 09.2019, p. 1507-1517.

Research output: Contribution to journalArticle

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abstract = "We showed previously that 1‐ethyl‐3‐(3‐dimethylamino‐propyl)‐carbodiimide hydrochloride (EDC) cross‐linked recombinant human collagen III hydrogels promoted stable regeneration of the human cornea (continued nerve and stromal cell repopulation) for over 4 years. However, as EDC cross linking kinetics were difficult to control, we additionally tested a sterically bulky carbodiimide. Here, we compared the effects of two carbodiimide cross linkers—bulky, aromatic N‐cyclohexyl‐N0‐(2‐morpholinoethyl)‐carbodiimide (CMC), and nonbulky EDC—in a mouse corneal graft model. Murine corneas undergoing full‐thickness implantation with these gels became opaque due to dense retro‐corneal membranes (RCM). Corneal epithelial cytokeratin 12 and alpha smooth muscle actin indicative of functional tissue regeneration and wound contraction were observed in RCM surrounding both hydrogel types. However, quantitatively different levels of infiltrating CD11c+ dendritic cells (DC) were found, suggesting a hydrogel‐specific innate immune response. More DC infiltrated the stroma surrounding EDC‐N‐hydroxysuccinimide (NHS) hydrogels concurrently with higher fibrosis‐associated tenascin c expression. The opposite was true for CMC‐NHS gels that had previously been shown to be more tolerising to DC. In vitro studies showed that DC cultured with transforming growth factor β1 (TGF‐β1) induced fibroblasts to secrete more tenascin c than those cultured with lipopolysaccharide and this effect was blocked by TGF‐β1 neutralisation. Furthermore, tenascin c staining was found in 40‐ to 50μm long membrane nanotubes formed in fibroblast/DC cocultures. We suggest that TGF‐β1 alternatively activated (tolerising) DC regulate fibroblast‐mediated tenascin c secretion, possibly via local production of TGF‐β1 in early wound contraction, and that this is indirectly modulated by different hydrogel chemistries.",
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note = "ACKNOWLEDGEMENTS We are grateful to the Iain Fraser Flow Cytometry Centre, the Microscopy and Histology Facility, the Quantitative PCR Facility, and the Medical Research Facility at the University of Aberdeen. This work was supported by the Royal College of Surgeons of Edinburgh, UK and Saving Sight in Grampian/Development Trust of the University of Aberdeen, UK. Funding Information: Saving Sight in Grampian/Development Trust of the University of Aberdeen, UK Royal College of Surgeons of Edinburgh, UK",
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