Macrophage Response to Allogeneic Adipose Tissue-Derived Stromal Cells in Hyaluronan-Based Hydrogel in a Porcine Vocal Fold Injury Model

Ann Otol Rhinol Laryngol. 2017 Jun;126(6):463-477. doi: 10.1177/0003489417702923. Epub 2017 Apr 6.

Abstract

Objective: Adipose tissue-derived stromal cells (ASC) embedded in hyaluronan scaffold is a beneficial prophylactic treatment for vocal fold (VF) surgical scar. Here, we investigated the macrophage inflammatory response to allogeneic ASC-constructs and identified changes in lamina propria extracellular matrix.

Method: Pig ASC were characterized and transfected with GFP+ lentivirus. Thirty-three pigs underwent VF biopsies, and after 3 days, gel alone, gel+pASC, placebo, or pASC alone was injected into wound bed. Animals were sacrificed 3, 7, or 26 days post-injection. Flow cytometry; qPCR for NF-α, TGFβ, IL-10, IL-4, IFNγ, IL-12, FGF2, Col1A1, and HGF; and immunohistochemistry for collagen, elastin, HA, and fibronectin were performed to characterize macrophage phenotype, quantify cytokine transcription, analyze extracellular matrix remodeling, and track GFP+ cells.

Results: No significant differences were found in SWC3+/SWC9+ phenotype or mRNA expression between cells+gel, gel, or placebo. The ASC alone exhibited significantly greater collagen, gel alone resulted in significantly less hyaluronan, and gel+pASC significantly more fibronectin (all P < .05). The pASC-GFP+ were detected 26 days post-injection.

Conclusions: The ASC-constructs were biocompatible; they did not influence the macrophage inflammatory response or provoke increases in collagen expression. Long-term engraftment was confirmed.

Keywords: adipose; hyaluronic acid; inflammation; injury; macrophages; mesenchymal stem/stromal cells; surgical injury; vocal fold; wound healing.

MeSH terms

  • Adipose Tissue / cytology*
  • Animals
  • Cicatrix / pathology*
  • Cytokines / metabolism
  • Disease Models, Animal
  • Extracellular Matrix / metabolism
  • Female
  • Humans
  • Hyaluronic Acid
  • Hydrogel, Polyethylene Glycol Dimethacrylate
  • Inflammation / pathology
  • Inflammation / physiopathology
  • Macrophages / physiology*
  • Mucous Membrane / metabolism
  • Neutrophils / physiology
  • Stromal Cells / physiology*
  • Swine
  • Tissue Scaffolds*
  • Vocal Cords / pathology*
  • Vocal Cords / physiopathology

Substances

  • Cytokines
  • Hydrogel, Polyethylene Glycol Dimethacrylate
  • Hyaluronic Acid