ABSTRACT
Abstract The aim of this study was to evaluate the expression of MMP2 and MMP9 during apical periodontitis (AP) progression in TLR2 (TLR2 KO) and in MyD88 (MyD88 KO) knockout mice compared to wild type (WT) mice. AP was induced in mandibular first molars of TLR2 KO (n= 18), MyD88 KO (n= 18), and WT mice (n= 18). After 7, 21, and 42 days, the animals were euthanized and the jaws were dissected and subjected to histotechnical processing. Subsequent sections were stained by immunohistochemistry and evaluated for detection of MMP2 and MMP9. Statistical analysis of the semi-quantitative analysis of immunohistochemistry was performed using chi-square test (α = 0.05). In the initial periods of AP progression, an increased expression of MMP9 in the TLR2 KO and MyD88 KO mice was observed. In the final periods of AP progression, a reduction of MMP2 expression and an increase of MMP9 expression in the TLR2 KO mice were observed. MMP2 and MMP9 production was modulated for TLR2 and MyD88 during apical periodontitis progression.
Resumo O objetivo deste estudo foi avaliar a expressão de MMP2 e MMP9 durante a progressão da periodontite apical (AP) em camundongos knockout para TLR2 (TLR2 KO) e MyD88 (MyD88 KO) comparados aos camundongos wild type (WT). A AP foi induzida nos primeiros molares inferiores dos camundongos TLR2 KO (n = 18), MyD88 KO (n = 18) e WT (n = 18). Após 7, 21 e 42 dias, os animais foram eutanaziados e as mandíbulas foram dissecadas e submetidas a processamento histotécnico. As lâminas foram coradas por imuno-histoquímica e analisadas para a detecção de MMP2 e MMP9. A análise estatística semi-quantitativa da imuno-histoquímica foi realizada pelo teste qui-quadrado (α = 0,05). Nos períodos iniciais de progressão AP, foi observada uma expressão aumentada de MMP9 nos camundongos TLR2 KO e MyD88 KO. Nos períodos finais de progressão AP, observou-se uma redução da expressão de MMP2 e um aumento da expressão de MMP9 nos camundongos TLR2 KO. A produção de MMP2 e MMP9 foi modulada por TLR2 e MyD88 durante a progressão da periodontite apical.
Subject(s)
Animals , Mice , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Myeloid Differentiation Factor 88/physiology , Periapical Periodontitis/enzymology , Toll-Like Receptor 2/physiology , Disease Progression , Immunohistochemistry , Mice, Inbred C57BL , Mice, Knockout , Myeloid Differentiation Factor 88/genetics , Periapical Periodontitis/metabolism , Periapical Periodontitis/pathologyABSTRACT
OBJETIVO: Los procesos destructivos del periodonto apical están en su mayoría mediados por proteasas específicas. Existe evidencia de que los niveles de metaloproteinasa de matriz extracelular-9 (MMP-9) en el fluido crevicular gingival (FCG) podrían reflejar la presencia de periodontitis apical asintomática (PAA). El objetivo de este estudio fue evaluar la actividad de MMP-9 durante la respuesta reparativa periapical en controles postendodóncicos entre una semana y 6 meses y en controles sanos. MATERIALES Y MÉTODOS: En este estudio prospectivo se incluyeron 28 pacientes con periodontitis apical asintomática, y se tomaron muestras de FCG a partir de dientes con PAA en línea base y controles postendodóncicos de una semana, uno, 3 y 6 meses. Adicionalmente se incluyeron controles contralaterales sanos. Las muestras se eluyeron y analizaron mediante gelatinogramas y densitometría. RESULTADOS: En pacientes con periodontitis apical asintomática se identificaron tanto la proforma como la forma activa de la MMP-9. Estas presentaron aumentos significativos en controles postendodóncicos de 3 y 6 meses con signos de reparación periapical. CONCLUSIONES: Los niveles de MMP-9 aumentaron significativamente en dientes con diagnóstico de PAA durante la fase reparativa. Estos resultados sugieren que MMP-9 también cumpliría un papel fisiológico durante la reparación periapical, que es susceptible de ser evaluado a través del análisis del FCG.
OBJECTIVE: Apical destructive periodontal processes are largely mediated by specific proteases. Evidence supports that the levels of extracellular matrix metalloproteinase-9 (MMP-9) could reflect the presence of asymptomatic apical periodontitis (AAP) in gingival crevicular fluid (GCF). The aim of this study was to evaluate the activity of MMP-9 during reparative response in periapical post endodontic controls between 1 week and 6 months later and in healthy controls. MATERIALS AND METHODS: A prospective study was performed on 28 patients with a diagnosis of AAP. GCF samples were taken from AAP teeth at baseline and post-endodontic controls at 1 week, 1, 3 and 6 months. Additional healthy contralateral controls were obtained, and samples were eluted and analyzed by densitometric scanning and gelatin zymography. RESULTS: In patients with asymptomatic apical periodontitis, both the pro form and the active form of MMP-9 were identified. These showed significant increases in post-endodontic controls at three and six months, with signs of periapical repairing. CONCLUSIONS: MMP-9 levels significantly increased in teeth diagnosed with AAP during the reparative phase. These results suggest that MMP-9 might be involved in the healing of apical tissues that might be reflected in GCF.
Subject(s)
Humans , Male , Adolescent , Adult , Periapical Periodontitis/enzymology , Periapical Periodontitis/therapy , Gingival Crevicular Fluid/enzymology , Matrix Metalloproteinase 9/metabolismABSTRACT
La periodontitis apical asintomática (PAa) es una patología infecciosa caracterizada por destrucción ósea perirradicular asociada a un proceso inflamatorio crónico y producción de mediadores inflamatorios, entre los cuales se encuentran las metaloproteinasas de matriz extracelular (MMPs). Entre éstas, las MMPs-13, -14, -2 y -9, son producidas por el tejido óseo y degradan sinérgicamente el colágeno tipo I, principal componente de los tejidos periodontales, y gelatina, producto de la degradación y desnaturación del colágeno. El objetivo de este estudio fue determinar el patrón de expresión de las MMPs-2, -9, -13 y -14 en granulomas periapicales (GPAs), quistes radiculares inflamatorios (QRIs) y ligamento periodontal sano (LS). Materiales y Métodos: Se seleccionaron 12 pacientes con diagnóstico clínico de PAa e indicación de exodoncia a partir de los cuales se obtuvieron biopsias de lesiones periapicales (LPAs). Como controles, se seleccionaron 7 individuos con indicación de exodoncia de premolares por ortodoncia, obteniéndose biopsias de LS. Se efectuó el diagnóstico anátomo-patológico de los especímenes y se caracterizó la expresión de las MMPs en estudio mediante inmunohistoquímica. Resultados: Las MMPs en estudio sólo se detectaron en GPAs y QRIs, y se inmunolocalizaron principalmente en el infiltrado inflamatorio de éstos. Adicionalmente, la MMP-2 se identificó en fibroblastos del tejido conectivo. Conclusiones: MMPs-2, -9, -13 y -14 se expresan predominantemente en el infiltrado inflamatorio de las LPAs y no en LS, y por tanto se sugiere la participación de estos mediadores en la patogénesis de la PAa.
Asymptomatic apical periodontitis (aAP) is an infectious disease characterized by perirradicular bone destruction associated with chronic inflammation and release of inflammatory mediators, such as matrix metalloproteinases (MMPs). MMPs-13, -14 and -2, -9 are bone-expressed enzymes that can synergistically degrade collagen I, the main component of periodontal extracellular matrix, and gelatin, the product of degradation and denaturation of collagen. The aim of this study was to characterize the expression pattern of MMPs-2, -9, -13, and -14 in periapical granulomas (PGs), radicular cysts (RCs) and healthy periodontal ligament (PDL). Materials and Methods: Individuals with clinical diagnosis of aAP and indication of extraction were selected (N=12), and biopsies of periapical lesions (PLs) were obtained. For controls, 7 subjects with indication of premolar extraction for orthodontic reasons were selected, and PDL biopsies were obtained. Samples were diagnosed by anatomopathological examination and immunohistochemical staining was carried out to characterize MMPs expression. Results: MMPs-2, -9, -13 and -14 detection was limited to PLs and were localized mainly to inflammatory infiltrate on both, PGs and RCs. Additionally, MMP-2 was immunolocalized to fibroblasts from the connective tissue. Conclusions: Whereas MMPs-2, -9, -13 and -14 were not detected in healthy periodontal ligament, they were highly expressed on inflammatory infiltrate from PGs and RCs, suggesting a role of these mediators in aAP pathogenesis.