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.
Highlights
A widely used strategy in oral reconstructive surgical procedures is soft tissue augmentation [1,2,3]
Gel type scaffolds are composed of rat tail collagen type I (BD Biosciences, Bedford, MA, USA) at 2.5 mg/ml supplemented with Dulbecco’s modified Eagle’s medium (DMEM) with 1% fetal bovine serum (FBS), penicillin (100 units/mL; GIBCOBRL) with and without platelet-derived growth factor-BB (PDGF) (R&D Systems) at a concentration of 250 ng/mL. pH was neutralized with NaOH
Cell population kinetics within the scaffolds Fluorescence microscopy showed DiI-labeled human gingival fibroblasts, coming from the ‘‘active zone’’ and populating sponge type and gel type scaffolds within one day and increasing throughout the 14-day observation period (Figure 2)
Summary
A widely used strategy in oral reconstructive surgical procedures is soft tissue augmentation [1,2,3] Autologous grafts such as subepithelial connective tissue grafts are successfully applied to increase soft tissue volume, these techniques are associated with several disadvantages. These include increased patient’s morbidity due to a second surgical procedure and limited graft material due to anatomical reasons. To overcome these drawbacks of autologous grafts, several strategies using allogeneic and synthetic devices have been developed [4,5,6,7,8,9,10,11]. To extend the structural properties and mechanical integrity, cross-linking of collagen has been developed [9,13,16,17]
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