Effect of kindlin-1 deficiency on inflammatory cytokine expression in gingival keratinocytes in the presence of oral biofilm
department or school: Dentistry, Faculty of
data provider: DSpace
degree campus: UBCV
creator: Kehler, Angela
issued: 2014-07-23T14:57:47Z
issued: 2014
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: Kindler syndrome (KS) is an autosomal recessive skin disorder of unknown etiology resulting in congenital skin blisters, photosensitivity, generalized progressive poikiloderma, and mucosal alterations. Early onset and severe periodontal disease has been noted as a common clinical finding. Kindlin-1 is an intracellular focal adhesion protein that functions in cellular adhesion in the epidermis and mucosal tissues through interaction and activation of integrins. Kindlin-1 deficiency, as occurs in KS, results in impaired adhesion of the junctional epithelium to the basement membrane and may contribute to severe periodontal disease. We hypothesize that kindlin-1 deficiency may affect other aspects of keratinocyte behavior, including expression of cytokines and mediators involved in inflammation and repair. Expression of kindlin-1 was down-regulated in human gingival keratinocytes (HGK) and confirmed via Western blot analysis and RT-PCR. HGKs were exposed to varying concentrations of a native and heat-inactivated oral biofilm extract as well as a 3-week live oral biofilm. mRNA was isolated from control and kindlin-1 deficient HGKs and RT-PCR was used to assess relative gene expression of the following gene products: β6 integrin, fibronectin ED-A and ED-B, IL-1α, IL-1β, IL-6, IL-8, TNFα, TGFβ-1, TGFβ-3, MMP-1, MMP-2, MMP-9, and tenascin C. Kindlin-1 deficiency in HGKs was found to significantly up-regulate the expression of genes encoding for β6 integrin, IL-1α, and IL-1β. β6 integrin expression was increased in K1-deficient cells in the absence of biofilm treatment and when exposed to native biofilm extract. The pro-inflammatory cytokines IL-1α and IL-1β were significantly up-regulated in K1-deficient cells as compared to control cells in keratinocytes exposed to native oral biofilm extract, and in control cells exposed to heat- inactivated biofilm. ii MMP-2, MMP-9, IL-8, tenascin C, ED-B fibronectin, and TGFβ-3 were differentially regulated by the presence of biofilm ex…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/48473
note: EFFECT OF KINLDIN-1 DEFICIENCY ON INFLAMMATORY CYTOKINE EXPRESSION IN GINGIVAL KERATINOCYTES IN THE PRESENCE OF ORAL BIOFILM by Angela Kehler D.M.D., The University of Manitoba, 2010 B.Sc., The University of Manitoba, 2006 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Craniofacial Science) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) July 2014 © Angela Kehler, 2014 ii Abstract Kindler syndrome (KS) is an autosomal recessive skin disorder of unknown etiology resulting in congenital skin blisters, photosensitivity, generalized progressive poikiloderma, and mucosal alterations. Early onset and severe periodontal disease has been noted as a common clinical finding. Kindlin-1 is an intracellular focal adhesion protein that functions in cellular adhesion in the epidermis and mucosal tissues through interaction and activation of integrins. Kindlin-1 deficiency, as occurs in KS, results in impaired adhesion of the junctional epithelium to the basement membrane and may contribute to severe periodontal disease. We hypothesize that kindlin-1 deficiency may affect other aspects of keratinocyte behavior, including expression of cytokines and mediators involved in inflammation and repair. Expression of kindlin-1 was down-regulated in human gingival keratinocytes (HGK) and confirmed via Western blot analysis and RT-PCR. HGKs were exposed to varying concentrations of a native and heat-inactivated oral biofilm extract as well as a 3-week live oral biofilm. mRNA was isolated from control and kindlin-1 deficient HGKs and RT-PCR was used to assess relative gene expression of the following gene products: β6 integrin, fibronectin ED-A and ED-B, IL-1α, IL-1β, IL-6, IL-8, TNFα, TGFβ-1, TGFβ-3, MMP-1, MMP-2, MMP-9, and tenascin C. Kindlin-1 deficiency in HGKs was found to significantly up-regulate the expression of genes encoding for β6 integrin, IL-1α, and IL-1β. β6 integrin expressio…
has type: Thesis/Dissertation
date issued: 2014-09
is shown at: 10.14288/1.0167556
language: eng
degree discipline: Craniofacial Science
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivs 2.5 Canada
rights URI: http://creativecommons.org/licenses/by-nc-nd/2.5/ca/
scholar level: Graduate
type: Text
identifier URI: http://hdl.handle.net/2429/48473