Efecto in vitro de una solución estabilizada de ácido hipocloroso (HOCl) para uso odontológico sobre la expresión de factores de crecimiento y el ciclo celular en fibroblastos orales humanos

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dc.contributor.advisorPerdomo, Sandra Janeth
dc.contributor.advisorLafaurie Villamil, Gloria Ines
dc.contributor.authorCalderón Mendoza, Justo Leonardo
dc.date.accessioned2023-03-03T14:58:56Z
dc.date.available2023-03-03T14:58:56Z
dc.date.issued2016
dc.description.abstractLa reparación de los tejidos de la mucosa oral después de una lesión requiere un balance entre la producción de citocinas y factores de crecimiento permitiendo la reparación tisular. En algunas circunstancias clínicas, se utilizan enjuagues que favorecen el proceso de reparación de los tejidos. Sin embargo, las alternativas disponibles en el mercado a pesar de usarse a bajas concentraciones, tienen un alto grado de citotoxicidad. El HOCl es una solución antimicrobiana utilizada por más de 10 años en Colombia, en el tratamiento de heridas de piel y mucosas. Su eficacia antimicrobiana y efectos sobre la reparación tisular demostrados en el uso médico, lo hacen una importante alternativa terapéutica para ser utilizada para el tratamiento de heridas de la mucosa oral. En este trabajo se evaluó el efecto de una solución estabilizada de HOCl pH 5.6 (+/-0.2) diseñada para uso odontológico sobre la línea celular de fibroblastos gingivales humanos (HGnF 2620), con el fin de elucidar mecanismos celulares y moleculares que puedan proponer al HOCl, como una alternativa terapéutica para la reparación de heridas de cavidad oral. Para ello, se evaluó la citotoxicidad, los niveles en la expresión de los genes TGFΒ, FGF, NFkB, VEGF, EGF como marcadores de reparación y BAX, p53, BCL2 y E2F como marcadores de apoptosis y ciclo celular. Se evaluó el efecto del HOCl sobre el contenido de ADN por citometría de flujo y la expresión de proteínas IP3k/AKT, JNK, ERK y P38 MAPK en FGH expuestos a tratamientos con HOCl por western blot. . La expresión de los genes en función a la concentración y tiempo de exposición a HOCl se analizó por ANOVA de dos vías. Los resultados muestran que el HOCl tiene efectos sobre los FGH dependientes de concentración y pH. A concentraciones de ≤47,5µM detiene el ciclo celular de los FGH e induce aneuploidia. A concentraciones entre 95µM y 190µM el HOCl estimula la expresión de los factores de crecimiento TGF-β, FGF y VEGF y activa proteínas de la familia MAPK (ERK). En conclusión el HOCl está involucrando en importantes vías de señalización de reparación tisular dependiendo de concentración y pH, hallazgos que permiten considerar su uso como posible alternativa terapéutica en el tratamiento de lesiones en cavidad oral.spa
dc.description.abstractenglishMucosal wound healing after an injury requires coordination between growth factors and cytokines production enabling tissue repair. In many clinical circumstances, it is required the use of agents to promote tissue repair processes. However, the available alternatives in the market even at low concentrations have a high degree of cytotoxicity. HOCl is an antimicrobial solution used in Colombia for over 10 years in the treatment of wounds in skin and mucosa. Its antimicrobial efficacy on pathogens and its effects on tissue repair demonstrated in the medical setting, make it an important therapeutic alternative to be used during the treatment of wounds in the oral cavity. The effect of a stable solution of HOCl to be used in oral health at pH 5.2 (+/-0.2) and 5.6 (+/-0.2) on the human gingival fibroblast (HGnF 2620) cell line was evaluated, with the objective to elucidate cellular and molecular mechanisms which may introduce the HOCl as a novel therapeutic for the treatment and repair of wounds in oral cavity. Briefly, cytotoxicity and changes in the expression of TGF-β, FGF, NFkB, VEGF, EGF genes as markers of tissue repair and BAX, p53, E2F, BCL2 as markers of cell cycle and apoptosis in HGF was evaluated. The HOCl effect over DNA content in the HGF was evaluated by flow cytometry and the protein expression of IP3k/AKT, JNK, ERK and P38 MAPK in HGF after treatment with HOCl was evaluated by western blot. Gene expression in function to the concentration and exposure time to HOCl was analyzed using a two ways ANOVA. Results suggest HOCl has effects on the HGF depending on concentration and pH. HOCl Concentrations of ≤47.5um could induce cell cycle arrest in HGF and aneuploidy. At Concentrations between 95µM and 190µM the HOCl stimulates the expression of growth factors such as TGF-β, FGF and VEGF; and activates MAPK signaling pathway (ERK). In conclusion, the HOCl is involved in important tissue repair signaling pathways, depending on its concentration and pH. Important findings to consider its use as a possible therapeutic alternative in the treatment of oral wounds.eng
dc.description.degreelevelMaestríaspa
dc.description.degreenameMagíster en Ciencias Básicas Biomédicasspa
dc.format.mimetypeapplication/pdf
dc.identifier.instnameinstname:Universidad El Bosquespa
dc.identifier.reponamereponame:Repositorio Institucional Universidad El Bosquespa
dc.identifier.repourlrepourl:https://repositorio.unbosque.edu.co
dc.identifier.urihttps://hdl.handle.net/20.500.12495/10098
dc.language.isospa
dc.publisher.facultyFacultad de Medicinaspa
dc.publisher.grantorUniversidad El Bosquespa
dc.publisher.programMaestría en Ciencias Básicas Biomédicasspa
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dc.rights.accessrightsinfo:eu-repo/semantics/closedAccess
dc.rights.accessrightshttps://purl.org/coar/access_right/c_14cb
dc.rights.localAcceso cerradospa
dc.subjectEfecto in vitrospa
dc.subjectÁcido hipocloroso (HOCl)spa
dc.subjectFibroblastos orales humanosspa
dc.subject.keywordsIn vitro effectspa
dc.subject.keywordsHypochlorous acid (HOCl)spa
dc.subject.keywordsHuman oral fibroblastsspa
dc.subject.nlmW 50
dc.titleEfecto in vitro de una solución estabilizada de ácido hipocloroso (HOCl) para uso odontológico sobre la expresión de factores de crecimiento y el ciclo celular en fibroblastos orales humanosspa
dc.title.translatedIn vitro effect of a stabilized hypochlorous acid (HOCl) solution for dental use on the expression of growth factors and cell cycle in human oral fibroblasts.spa
dc.type.coarhttps://purl.org/coar/resource_type/c_bdcc
dc.type.coarversionhttps://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.driverinfo:eu-repo/semantics/masterThesis
dc.type.hasversioninfo:eu-repo/semantics/acceptedVersion
dc.type.localTesis/Trabajo de grado - Monografía - Maestríaspa

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Efecto in vitro de una solución estabilizada de ácido hipocloroso (HOCl) para uso odontológico sobre la expresión de factores de crecimiento y el ciclo celular en fibroblastos orales humanos
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Maestría en Ciencias Básicas Biomédicas