Relevant Degree Programs
Affiliations to Research Centres, Institutes & Clusters
Wound healing, cell communication, fibroblast biology, cell therapy
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Graduate Student Supervision
Doctoral Student Supervision (Jan 2008 - Nov 2020)
Wound healing in human oral mucosal gingiva is faster and results in significantly reduced scar formation as compared to similar skin wounds. Fibroblasts are the most abundant group of connective tissue cells that play a key role in wound healing and scar formation. It is possible that differential healing outcomes in skin and gingiva may relate to the distinct phenotypic features of fibroblasts residing in these tissues. In fibroblasts, cells-to-cell communication occurs partly through connexin (Cx) hemichannels (HCs) and gap junctions (GJs). Findings from previous studies have shown that functional blockage of connexin 43 (Cx43), the most ubiquitous Cx in skin (SFBLs) and gingival fibroblasts (GFBLs), accelerates wound closure in skin and may alleviate scarring, but the mechanisms are poorly understood and may involve modulation of Cx43 function in fibroblasts. In the present dissertation, we show that (1) Cx43 was the most abundant Cx present in cultured human SFBLs and GFBLs. (2) Its abundance was potently downregulated at the early stage of human gingival wound healing. (3) Cx43 assembled into GJ and HC plaques in skin and gingival epithelium and connective tissue fibroblasts, although its distribution into GJs or HCs was markedly different in these two tissues. (4) Cx43 mainly assembled into HCs in GFBLs while in SFBLs only a few HCs were present in vivo and in vitro. (5) Using an in vivo-like 3D culture model and Cx43 mimetic peptides to block its function, we showed that the GJ, HC, and channel-independent functions of Cx43 distinctly upregulate anti-fibrotic and downregulate profibrotic wound healing-related genes in GFBLs and SFBLs. (6) In GFBLs this response was mainly mediated by activation of ERK1/2 pathway via Cx43 HC blockage. Thus, Cx43 assembly into GJs and HCs and its function are distinct in SFBLs and GFBLs, which may contribute to the different wound healing outcomes in these tissues. Furthermore, specific blockage of Cx43 HC functions may provide a novel target to promote wound healing and alleviate scar formation.
No abstract available.
Master's Student Supervision (2010 - 2018)
Scar formation as a result of wound healing in skin is associated with increased deposition of extracellular matrix (ECM) and reduced ECM turnover by fibroblasts. Remarkably, wound healing in the human oral mucosa results in scarless healing, while wound healing in the skin can often result in scarring. Therefore comparing fibroblast phenotype and interactions with their ECM niche in the gingiva and skin may provide novel information about the factors that regulate scar formation. To this end, a novel 3D cell culture model was utilized to yield a cellular microenvironment (niche) that closely mimics the in vivo situation and primary gingival (GFBL) and skin fibroblasts (SFBL) phenotype was characterized. Furthermore, fibroblasts were reseeded on cell-free 3D ECM derived from GFBL and SFBL and the effects of the 3D ECM on cell phenotype were analyzed. Interestingly, SFBL in 3D cultures had greater expression of ECM deposition associated genes, including collagens, matricellular proteins, SLRPs, TGF-β1 and CTGF, intracellular ECM degradation and myofibroblast differentiation and function-associated genes, while GFBL had a greater expression of matrix remodeling associated genes (MMPs). We also found that the 3D cultures showed a significant difference in expression of certain genes (MMPs and myofibroblast function-associated genes) between GFBL and SFBL compared to cells reseeded on the 3D ECM or 2D control substrate. Thus, the 3D culture conditions may differentially regulate expression of a subset of genes in these cells. Interestingly, SFBL had a greater expression of matrix deposition associated genes (collagens, SLRPs, tenascins) irrespective of the culture conditions, suggesting that expression of these genes is inherently distinct between GFBL and SFBL. This was associated with greater autogenous TGF-β expression and SMAD3 phosphorylation in SFBL than GFBL, which may partly explain the innate difference in gene expression. In addition, there was greater ERK1/2 phosphorylation in fibroblasts when seeded on 3D ECM compared to 2D substrate. Greater ERK1/2 phosphorylation may have promoted greater expression of AP-1-dependent MMPs seen in SFBL and GFBL on 3D ECM. In conclusion, reduced expression of matrix deposition associated genes and greater expression of matrix remodeling genes in GFBL may contribute to scarless healing in gingiva.
Objectives: Wound healing in skin often results in scar formation, whereas wound healing in oral palatal mucosa is fast and rarely results in scarring. Understanding the mechanisms that promote oral scarless wound healing may provide novel approaches to prevent scar formation in skin. The goal of the study was to compare the abundance of the major collagenases MMP-1 and MMP-13 and gelatinases MMP-2 and MMP-9 in normal unwounded oral mucosa and skin and in experimental excisional wounds in skin (healing results to scar formation) and oral mucosa (wounds heal with minimal scar formation) at various time points post-wounding at the protein level. We hypothesized that the abundance of MMPs will be higher in scarless oral mucosal wound healing, compared to skin wound healing.Methods: Experimental wounds were created in oral palatal mucosa and dorsal skin of red Duroc pigs. Wound biopsies were collected before wounding and at various time points after wounding. The abundance of MMPs at the protein level was assessed by Western blotting and zymography. Results: All studied MMPs showed a significantly increased accumulation in the wound tissue already at day three post-wounding. Their abundance remained high until day 28 when MMP-9 and MMP-13 returned to the level of unwounded tissue, while MMP-1 and MMP-2 remained significantly elevated. Oral mucosal wounds showed in general a robust early up-regulation of MMP-1, MMP-2 and MMP-9 as compared to skin wounds already at day 3 after wounding. In contrast, the peak abundance of these MMPs occurred at day 14 in skin wounds. Unwounded oral mucosa showed significantly higher abundance of total MMP-2 and active MMP-9 as compared to unwounded skin. Thus, MMPs needed for early wound healing response are already present in higher abundance in oral mucosa as compared to skin before tissue injury possibly allowing a fast wound healing response. Conclusions: Results suggest that oral mucosal wound healing is associated with fast and robust regulation of MMPs. Rapid controlled processing of wound extracellular matrix may play a key role in scarless palatal wound healing. In addition, MMPs may regulate inflammatory reaction that plays a central role in scar formation.