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

Hermann Agis; Amy Collins; Andrei D Taut; Qiming Jin; Laura Kruger; Christoph Görlach; William V Giannobile

doi:10.1371/journal.pone.0112680

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.

Authors

Hermann Agis¹, Amy Collins², Andrei D Taut², Qiming Jin³, Laura Kruger², Christoph Görlach⁴, William V Giannobile⁵

Affiliations

¹ Periodontics and Oral Medicine, School of Dentistry, University of Michigan, Ann Arbor, Michigan, United States of America; Department of Conservative Dentistry and Periodontology, Medical University of Vienna, Vienna, Austria; Austrian Cluster for Tissue Regeneration, Vienna, Austria.
² Periodontics and Oral Medicine, School of Dentistry, University of Michigan, Ann Arbor, Michigan, United States of America.
³ Department of Cariology, Restorative Sciences, and Endodontics, School of Dentistry, University of Michigan, Ann Arbor, Michigan, United States of America.
⁴ Geistlich Pharma AG, Wolhusen, Switzerland.
⁵ Periodontics and Oral Medicine, School of Dentistry, University of Michigan, Ann Arbor, Michigan, United States of America; Department of Biomedical Engineering, College of Engineering, University of Michigan, Ann Arbor, Michigan, United States of America.

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

Grants and funding

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.