ingest cdrApp 2017-07-05T20:36:58.756Z d36eae88-cb6b-42c1-ba08-197eadfa9868 modifyDatastreamByValue RELS-EXT fedoraAdmin 2017-07-05T21:22:12.855Z Setting exclusive relation modifyDatastreamByValue RELS-EXT fedoraAdmin 2017-07-05T21:22:21.114Z Setting exclusive relation addDatastream MD_TECHNICAL fedoraAdmin 2017-07-05T21:22:28.996Z Adding technical metadata derived by FITS modifyDatastreamByValue RELS-EXT fedoraAdmin 2017-07-05T21:22:45.001Z Setting exclusive relation addDatastream MD_FULL_TEXT fedoraAdmin 2017-07-05T21:22:53.422Z Adding full text metadata extracted by Apache Tika modifyDatastreamByValue RELS-EXT fedoraAdmin 2017-07-05T21:23:09.395Z Setting exclusive relation modifyDatastreamByValue RELS-EXT cdrApp 2017-07-06T11:39:26.208Z Setting exclusive relation modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2017-09-11T17:30:15.528Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-01-25T01:44:18.956Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-01-27T02:30:43.275Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-03-13T22:06:46.999Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-05-16T19:52:34.401Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-07-10T20:39:41.015Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-07-17T16:54:48.036Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-08-08T16:21:15.849Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-08-14T20:46:36.897Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-08-16T16:30:41.357Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-09-21T14:05:04.449Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-09-26T16:59:55.546Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-10-10T17:19:46.170Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2018-10-11T17:52:12.858Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2019-02-28T01:01:51.862Z modifyDatastreamByValue MD_DESCRIPTIVE cdrApp 2019-03-19T20:18:56.912Z Eman Hefni Author School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. Spring 2017 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text Eman Hefni Author School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. Spring 2017 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. Spring 2017 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. Spring 2017 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017-05 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 University of North Carolina at Chapel Hill Degree granting institution Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1; DNA methylation; Epigenetic; epithelial resistance; Periodontal Disease; PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 University of North Carolina at Chapel Hill Degree granting institution Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1, DNA methylation, Epigenetic, epithelial resistance, Periodontal Disease, PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1; DNA methylation; Epigenetic; epithelial resistance; Periodontal Disease; PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Dentistry (Oral and Craniofacial Medicine) Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Eman Hefni Creator School of Dentistry Epigenetics and Transcriptional Dynamics in Periodontal Disease Objectives: Several studies have shown the involvement of epigenetics with periodontal disease. Since functional dissociation of Paracellular permeability is expected during bacterial infection, we hypothesize that the methylation of host oral epithelial DNA represents an important element in the disruption of barrier function and pathogenesis of periodontal diseases. With this In vitro study, we aimed to assess whether there is altered epithelial permeability measuring trans epithelial resistance after Porphyromonas gingivalis (P. gingivalis), Campylobacter rectus (C. rectus) and Fusobacterium nucleatum (F. nucleatum) infection. Plakophilin2 (PKP2) methylation status and expression levels were also investigated. In addition, investigate the potential effects of DNA methyltransferase (DNMT) inhibitors on epithelial barrier function in response to infection with periodontal pathogen in human gingival epithelial cells. Methods: Primary human gingival epithelial cells (HGEPs) were stimulated with P. gingivalis, strain, C. rectus and F. nucleatum (MOI 50) either in the presence or absence of DNMT inhibitors (10 μM of RG108 or EGCG). CellTiter-Blue® Cell Viability Assay (Promega) was used to determine an optimum cell density and maximum inhibitor concentration at which cell viability is maintained. Transepithelial electrical resistance (TER) at various time points were performed using an EVOM® electrical resistance system. DNA methylation was quantified by qPCR using EpiTect Methyl II PCR Primer Assays for PKP2. Immunofluorescence analysis was performed using PKP2 antibody and analysis performed using Zeiss710 confocal microscope. Results: Exposure of HGEPs to P. gingivalis resulted in decreased TER (P=<0.001) associated with increased cell permeability. Methylation assays showed increased methylation levels of the PKP2 in comparison to non-infected controls (P=<0.001) and an associated PKP2 down- regulation (P=<0.005). For infected cells treated with DNMT inhibitors, PKP2 mRNA expression was increased (P=<0.001) and TER values similar to non-infected cells. Comparatively, immunofluorescent staining of the PKP2 protein showed reduced protein expression in infected cells not treated with DNMT inhibitors. Conclusion: DNA methylation levels of PKP2 can affect epithelial barrier function potentially conferring increased susceptibility to infection. DNMT inhibitors can affect cell adhesion dissociation in response to infection minimizing the disturbance to the barrier function. 2017 Dentistry CDH1; DNA methylation; Epigenetic; epithelial resistance; Periodontal Disease; PKP2 eng Master of Science Masters Thesis University of North Carolina at Chapel Hill Graduate School Degree granting institution Silvana Barros Thesis advisor Steven Offenbacher Thesis advisor Ramiro Mura Thesis advisor text 2017-05 Hefni_unc_0153M_16773.pdf uuid:97f7290a-5d44-4d0e-97bd-c8e4c74eb627 2019-07-05T00:00:00 2017-04-12T17:30:58Z proquest application/pdf 5670098 yes