title: Keloid pathogenesis via Drosophila similar to mothers against decapentaplegic (SMAD) signaling in a primary epithelial-mesenchymal in vitro model treated with biomedical-grade chitosan porous skin regenerating template.
abstract: The effects of locally produced chitosan (CPSRT-NC-bicarbonate) in the intervention of keloid pathogenesis were investigated in vitro. A human keratinocyte-fibroblast co-culture model was established to investigate the protein levels of human collagen type-I, III and V in a western blotting analysis, the secreted transforming growth factor-β1 (TGF-β1) in an enzyme-linked immunosorbent assay (ELISA) and the mRNA levels of TGF-β1's intracellular signaling molecules (SMAD2, 3, 4 and 7) in a real-time PCR analysis. Keratinocyte-fibroblast co-cultures were maintained in DKSFM:DMEM:F12 (2:2:1) medium. Collagen type-I was found to be the dominant form in primary normal human dermal fibroblast (pNHDF) co-cultures, whereas collagen type-III was more abundant in primary keloid-derived human dermal fibroblast (pKHDF) co-cultures. Collagen type-V was present as a minor component in the skin. TGF-β1, SMAD2 and SMAD4 were expressed more in the pKHDF than the pNHDF co-cultures. Co-cultures with normal keratinocytes suppressed collagen type-III, SMAD2, SMAD4 and TGF-β1 expressions and CPSRT-NC-bicarbonate enhanced this effect. In conclusion, the CPSRT-NC-bicarbonate in association with normal-derived keratinocytes demonstrated an ability to reduce TGF-β1, SMAD2 and SMAD4 expressions in keloid-derived fibroblast cultures, which may be useful in keloid intervention.
authors.affiliation: Department of Orthopaedic Surgery, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Chin Keong
authors.initials: CK
authors.lastname: Lim
authors.forename: Ahmad Sukari
authors.initials: AS
authors.lastname: Halim
authors.forename: Nik Soriani
authors.initials: NS
authors.lastname: Yaacob
authors.forename: Ismail
authors.initials: I
authors.lastname: Zainol
authors.forename: Kartini
authors.initials: K
authors.lastname: Noorsal
mesh_terms: D001672:Biocompatible Materials / Q000494:pharmacology*; D002478:Cells, Cultured; D048271:Chitosan / Q000494:pharmacology*; D018920:Coculture Techniques; D024042:Collagen Type I / Q000378:metabolism; D024061:Collagen Type III / Q000378:metabolism; D024062:Collagen Type V / Q000378:metabolism; D020405:Dermis / Q000166:cytology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007627:Keloid / Q000378:metabolism*; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology; D008954:Models, Biological; D012038:Regeneration; D015398:Signal Transduction / Q000187:drug effects; D012879:Skin Physiological Phenomena; D051785:Smad Proteins / Q000235:genetics / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D001672:Biocompatible Materials; D024042:Collagen Type I; D024061:Collagen Type III; D024062:Collagen Type V; D051785:Smad Proteins; D053773:Transforming Growth Factor beta1; D048271:Chitosan