Cell population kinetics of collagen scaffolds in ex vivo oral wound repair

PLoS One. 2014 Nov 14;9(11):e112680. doi: 10.1371/journal.pone.0112680. eCollection 2014.

Abstract

Biodegradable collagen scaffolds are used clinically for oral soft tissue augmentation to support wound healing. This study sought to provide a novel ex vivo model for analyzing healing kinetics and gene expression of primary human gingival fibroblasts (hGF) within collagen scaffolds. Sponge type and gel type scaffolds with and without platelet-derived growth factor-BB (PDGF) were assessed in an hGF containing matrix. Morphology was evaluated with scanning electron microscopy, and hGF metabolic activity using MTT. We quantitated the population kinetics within the scaffolds based on cell density and distance from the scaffold border of DiI-labled hGFs over a two-week observation period. Gene expression was evaluated with gene array and qPCR. The sponge type scaffolds showed a porous morphology. Absolute cell number and distance was higher in sponge type scaffolds when compared to gel type scaffolds, in particular during the first week of observation. PDGF incorporated scaffolds increased cell numbers, distance, and formazan formation in the MTT assay. Gene expression dynamics revealed the induction of key genes associated with the generation of oral tissue. DKK1, CYR61, CTGF, TGFBR1 levels were increased and integrin ITGA2 levels were decreased in the sponge type scaffolds compared to the gel type scaffold. The results suggest that this novel model of oral wound healing provides insights into population kinetics and gene expression dynamics of biodegradable scaffolds.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Becaplermin
  • Cell Count
  • Collagen / physiology*
  • Collagen / ultrastructure
  • Connective Tissue Growth Factor / metabolism
  • Cysteine-Rich Protein 61 / metabolism
  • Fibroblasts / metabolism
  • Gene Expression Profiling / methods
  • Gene Expression Regulation / physiology*
  • Gingiva / cytology*
  • Gingiva / injuries*
  • Humans
  • In Vitro Techniques
  • Integrin alpha2 / metabolism
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Microscopy, Electron, Scanning
  • Polymerase Chain Reaction
  • Protein-Serine-Threonine Kinases / metabolism
  • Proto-Oncogene Proteins c-sis / metabolism*
  • Receptor, Transforming Growth Factor-beta Type I
  • Receptors, Transforming Growth Factor beta / metabolism
  • Tetrazolium Salts
  • Thiazoles
  • Tissue Scaffolds*
  • Wound Healing / physiology*

Substances

  • CCN1 protein, human
  • CCN2 protein, human
  • Cysteine-Rich Protein 61
  • DKK1 protein, human
  • ITGA2B protein, human
  • Integrin alpha2
  • Intercellular Signaling Peptides and Proteins
  • Proto-Oncogene Proteins c-sis
  • Receptors, Transforming Growth Factor beta
  • Tetrazolium Salts
  • Thiazoles
  • Connective Tissue Growth Factor
  • Becaplermin
  • Collagen
  • Protein-Serine-Threonine Kinases
  • Receptor, Transforming Growth Factor-beta Type I
  • TGFBR1 protein, human
  • thiazolyl blue

Associated data

  • GEO/GSE61314
  • GEO/GSM1501968
  • GEO/GSM1501969
  • GEO/GSM1501970
  • GEO/GSM1501971

Grant support

This study was supported by Geistlich Pharma AG (www.geistlich-pharma.com, Wolhusen, Switzerland). Christoph Görlach is employed by Geistlich Pharma AG. The authors confirm that the funder provided support in the form of salary for Christoph Görlach, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of this author are articulated in the ‘author contributions’ section. Hermann Agis received the Erwin Schrödinger Fellowship from the Austrian Science Fund (FWF, fwf.ac.at): J3379-B19. The Austrian Science Fund had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.