Paracrine factors from fibroblast aggregates in a fibrin-matrix carrier enhance keratinocyte viability and migration

J Biomed Mater Res A. 2010 Nov;95(2):658-64. doi: 10.1002/jbm.a.32881.

Abstract

Efficient re-epithelialization of skin lesions is dependent on paracrine support from connective tissue fibroblasts. In deep skin defects, the supporting growth factor incentive is lacking. Current methods of keratinocyte transplantation with compromised attachment, spread, and cell proliferation warrant improvement and refinement. We describe here how human keratinocytes can be stimulated by matrix-embedded factors from a novel process of fibroblast activation: nemosis. Interestingly, the unique set of mediators released in this process also plays a key role in normal wound healing. To develop a system for targeted delivery of nemosis-derived paracrine effectors, herein designated as Finectra, we combined them with fibrin to establish a controlled-release gel. Keratinocytes seeded to cover this active matrix showed better adherence, outgrowth, and viability than did cells on control matrix. The matrix incorporating Finectra supported viability of both primary keratinocytes and green fluorescent protein (GFP)-labeled HaCaT cells, as evaluated by MTT assay and persistence of GFP-fluorescence. The fibrin-Finectra matrix promoted migration of keratinocytes to cover a larger area on the matrix, suggesting better wound coverage on transplantation. An inhibitor of EGFR/c-Met receptor tyrosine kinases abolished keratinocyte responses to fibrin-Finectra matrix. This matrix can thus deliver biologically relevant synergistic stimuli to keratinocytes and hasten re-epithelialization.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Biocompatible Materials / chemistry
  • Biocompatible Materials / metabolism
  • Cell Line
  • Cell Movement / drug effects*
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects*
  • Collagen / chemistry
  • Delayed-Action Preparations
  • Drug Carriers / chemistry
  • Drug Carriers / metabolism
  • Extracellular Matrix / chemistry
  • Female
  • Fibrin / chemistry*
  • Fibroblasts / cytology
  • Fibroblasts / metabolism*
  • Humans
  • Intercellular Signaling Peptides and Proteins / pharmacology*
  • Keratinocytes* / cytology
  • Keratinocytes* / drug effects
  • Keratinocytes* / physiology
  • Materials Testing
  • Paracrine Communication*
  • Skin / cytology
  • Skin / drug effects
  • Skin / pathology
  • Wound Healing / drug effects

Substances

  • Biocompatible Materials
  • Delayed-Action Preparations
  • Drug Carriers
  • Intercellular Signaling Peptides and Proteins
  • Fibrin
  • Collagen