Thakkar, Dhwani (2013) In vitro characterization of oral irritation and healing. (MSc(R) thesis), Kingston University, .

Abstract

Wound healing is an important physiological process whereby several cellular and extra-cellular matrix components (ECM) interact with each other to regain integrity of the wounded tissue. The aim of this study was to establish in-vitro models for oral wound healing to characterize this process and to investigate agents that could promote or stimulate wound healing in the gingival tissues. The experimental models considered were: (1) Scratch model, and (2) 3D tissue model. The monolayer scratch model with Human Gingival Fibroblasts (HGF) showed wound closure within 96 hours post scratch while the co-culture model with HGF and Human Gingival Epithelial progenitors (HGEp, precursors of human oral keratinocytes) healed more quickly, within 48 hours. To investigate the effectiveness of agents that could promote healing in the oral compartment, calcium chloride (0.014%), phosphoryl oligosaccharide of calcium (POs Ca; 0.1%) and human collagen type IV (0.01% and 0.1%) were tested using the co-culture scratch model. There was no evidence that these reagents improved gingival healing using this model. SPARC (secreted protein acidic and rich in cysteine) can delay wound closure in dermal fibroblasts. Using siRNA knockdown to inhibit SPARC expression in gingival fibroblasts the rate of healing was enhanced, similar to the scenario in dermal fibroblasts. However, a statistically significant conclusion could not be reached within the timeframe of this project (p.0.07). Using the GIN-100 tissue 3D model, it was observed that recovery from chemical-induced damage (overnight treatment with 0.1%Sodium Lauryl Sulphate or SLS) could be observed over a 5 day period specifically when the GIN-100-ASY medium was used. These models provide the basis for the development of easy and reliable tools for oral wound healing studies.

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