Mussel adhesive Protein-conjugated Vitronectin (fp-151-VT) Induces Anti-inflammatory Activity on LPS-stimulated Macrophages and UVB-irradiated Keratinocytes

Immunol Invest. 2019 Apr;48(3):242-254. doi: 10.1080/08820139.2018.1506476. Epub 2018 Sep 6.

Abstract

Background: Skin inflammation and dermal injuries are a major clinical problem because current therapies are limited to treating established scars, and there is a poor understanding of healing mechanisms. Mussel adhesive proteins (MAPs) have great potential in many tissue engineering and biomedical applications. It has been successfully demonstrated that the redesigned hybrid type MAP (fp-151) can be utilized as a promising adhesive biomaterial. The aim of this study was to develop a novel recombinant protein using fp-151 and vitronectin (VT) and to elucidate the anti-inflammatory effects of this recombinant protein on macrophages and keratinocytes.

Methods: Lipopolysaccharide (LPS) was used to stimulate macrophages and UVB was used to stimulate keratinocytes. Inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 were analyzed by Western Blot. Inflammatory cytokines and NO and ROS production were analyzed.

Result: In macrophages stimulated by LPS, expression of the inflammatory factors iNOS, COX-2, and NO production increased, while the r-fp-151-VT-treated groups had suppressed expression of iNOS, COX-2, and NO production in a dose-dependent manner. In addition, keratinocytes stimulated by UVB and treated with r-fp-151-VT had reduced expression of iNOS and COX-2. Interestingly, in UVB-irradiated keratinocytes, inflammatory cytokines, such as interleukin (IL)-1b, IL-6, and tumor necrosis factor (TNF)-a, were significantly reduced by r-fp-151-VT treatment.

Conclusions: These results suggest that the anti-inflammatory activity of r-fp-151-VT was more effective in keratinocytes, suggesting that it can be used as a therapeutic agent to treat skin inflammation.

Keywords: Mussel adhesive protein; Vitronectin; anti-inflammatory activity.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / isolation & purification
  • Anti-Inflammatory Agents / pharmacology*
  • Anti-Inflammatory Agents / therapeutic use
  • Cell Line
  • Dermatitis / drug therapy
  • Dermatitis / immunology
  • Humans
  • Keratinocytes / drug effects
  • Keratinocytes / immunology
  • Keratinocytes / radiation effects
  • Lipopolysaccharides / immunology
  • Macrophages / drug effects
  • Macrophages / immunology
  • Mice
  • Proteins / genetics
  • Proteins / isolation & purification
  • Proteins / pharmacology*
  • Proteins / therapeutic use
  • RAW 264.7 Cells
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / pharmacology*
  • Recombinant Fusion Proteins / therapeutic use
  • Ultraviolet Rays / adverse effects
  • Vitronectin / genetics
  • Vitronectin / isolation & purification
  • Vitronectin / pharmacology*
  • Vitronectin / therapeutic use

Substances

  • Anti-Inflammatory Agents
  • Lipopolysaccharides
  • Proteins
  • Recombinant Fusion Proteins
  • Vitronectin
  • adhesive protein, mussel