527

TO DEVELOP AN IN-VITRO (EFT-300) SKIN MODEL FOR THE STUDY OF WOUND HEALING.

Singh1, M., Mallampati1, R., Patlolla1, R.R., Vashi1, P., Hayden1, P., Klausner1, M. 1College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, FL 32307. 2MatTek Corporation, 200 Homer Avenue, Ashland, MA 01721.
Abstract

This study by scientists at Florida A&M University College of Pharmacy and Pharmaceutical Sciences and MatTek Corp. demonstrated that MatTek’s EpiDermFT full-thickness in vitro human skin tissue equivalent is an excellent in vitro model for studying the human skin wound healing process. Wound healing is a natural process, which involves the regeneration of epidermal and dermal tissue. The initiation of wound healing involves sequential completion of coagulation, inflammation and removal of damaged matrix components, followed by cellular proliferation and migration, angiogenesis, matrix synthesis and deposition, re-epithelisation and remodeling. Epiderm full thickness tissue (EFT-300) consists of Normal human derived Epidermal keratinocytes (NHEK) and Dermal Fibroblasts (NHFB). Epiderm full thickness is an in vitro model has both epidermis and dermis which closely resembles to human skin.> Histological studies clearly showed that there was a regeneration of keratinocytes in epidermal layers when treated with 32N KOH and concentrated H2SO4 and regeneration of fibroblasts, collagen in the dermis when treated with acrylic acid. Our results showed up regulation of IL-1alpha, IL-6 and IL-8 on earlier days followed by their subsequent down regulation by the sixth day. Immunohistochemistry studies demonstrated expression of Collagen IV (in dermis with acrylic acid treatment), and EGF in epidermis (H2SO4) Western blotting clearly showed expression of Collagen IV, which increased as a function of time. EGF expression was in the order; AA > H2SO4 > KOH. Expression of EGF was less in 32N KOH treated cultures, indicate that only upper epidermal layers and stratum corneum was damaged. This could possibly be due to the increased permeation of acrylic acid compared to H2SO4 and KOH. CONCLUSION: Our results strongly support that the EFT-300 cultures could be a good in-vitro model for studying wound healing process.

Keywords

Biomarkers, EFT-300, EGF Collagen, EpiDerm-FT, EpiDermFT, Harsh chemicals, Histological sections, IL-1a, IL-6, IL-8, Western blotting, Wound healing

Materials Tested

Acrylic acid, Potassium hydroxide (KOH), Sulfuric acid (H2SO4)

Request a copy of this paper, click here.