Terapia fotodinâmica com ftalocianina de zinco tetracarboxi-N-metilglucamina na doença periodontal induzida em ratos
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Data
2017-12-07
Autores
Corbi, Sâmara Cruz Tfaile
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Universidade Estadual Paulista (Unesp)
Resumo
A Terapia Fotodinâmica Antimicrobiana (Antimicrobial Photodynamic Therapy – aPDT), tem sido utilizada como um tratamento complementar na doença periodontal (DP). Ela combina um fotossensibilizador (FS) com uma fonte de luz que induz a produção de espécies reativas de oxigênio que elimina células microbianas. O objetivo deste trabalho foi avaliar, in vivo, os efeitos da aPDT (com o FS ftalocianina de zinco tetracarboxi-N-metilglucamina – 10μg/mL e luz LED vermelho – 655mn, 0,45W de potência), coadjuvante a Raspagem e Alisamento Radicular (RAR) e como monoterapia, além de verificar as respostas e alterações teciduais da DP induzida em ratos, pelas avaliações: microtomográfica, histométrica, estereométrica e histológica. Ligaduras foram inseridas nos sulcos dos segundos molares superiores para indução da DP. No Estudo 1, as ligaduras permaneceram por 15 dias e foram removidas para aplicar os tratamentos e no Estudo 2, as ligaduras foram colocadas por 7 dias e continuaram em posição por todo o experimento. 40 animais foram utilizados no Estudo 1 e distribuídos em 4 grupos: DP (Somente indução da doença, sem tratamento); RAR (Indução e tratamento básico periodontal); aPDT (Indução e aplicação da aPDT – FS ftalocianina de zinco tetracarboxi-N-metilglucamina e luz LED vermelho); RAR+aPDT (Indução, tratamento básico periodontal e aplicação da aPDT). 42 animais foram utilizados no Estudo 2 e divididos também em 4 grupos: FS (Tratamento somente com a ftalocianina de zinco tetracarboxi-N-metilglucamina); Luz (Tratamento somente com irradiação de luz LED vermelha); aPDT (Tratamento com a terapia fotodinâmica – FS+Luz) E DP (Indução da doença, sem tratamento). No Estudo 1, um dia após a remoção das ligaduras, foi aplicado os tratamentos e os animais foram eutanasiados nos períodos de 7 e 30 dias. No estudo 2, os animais receberam a aplicação das terapias e foram eutanasiados nos períodos de 7 e 15 dias. Para ambos os estudos, foi aplicado o teste paramétrico ANOVA one way, seguida do pós-teste de Tukey. No Estudo 1: na histometria, não foram encontradas diferenças estatísticas; a análise microtomográfica mostrou diferenças significantes nos dois períodos entre o grupo DP e RAR+aPDT e em 7 dias para DP e aPDT na região de furca. Nas proximais não houve diferenças significantes e na histologia, mostrou que não houve danos aos tecidos. No Estudo 2, na análise histométrica e radiográfica tridimensional, os resultados não mostraram diferenças estatisticamente significantes na região de furca e nas regiões interproximais, o perfil inflamatório demonstrou uma tendência a apresentar menor quantidade de células inflamatórias no grupo aPDT em 7 dias e na análise histológica não houve diferenças entre os grupos, indicando também que as terapias não causaram danos aos tecidos. Pode-se concluir que a aPDTcom ftalocianina de zinco tetracarboxi-N- metilglucamina foi efetiva no controle de perda óssea em DP induzida em ratos e que a aplicação da aPDT (com os componentes como monoterapias) e com a preservação da ligadura, favoreceu a permanência das bactérias no local e inibiu a ação dos tratamentos.
Antimicrobial Photodynamic Therapy (PDT) is a minimally invasive method consisting in the application of a photosensitive dye, which is subsequently stimulated by a light source and reacts with oxygen, producing reactive species. The aim of this study was to evaluate in vivo, the aPDT effects (with the PS zinc tetracarboxy-N-metylglucamine phthalocyanine 10μg/mL, and red LED light with 655nm), as adjuvant treatment to Scaling Root and Planing (SRP) and as monotherapy and verify the responses and tissue changes after aPDT application in PD-induced rats by microtomographic, histometric, stereometric and histological evaluations. Ligatures were placed around the second maxillary molars for PD induction. In Study 1, the ligatures were placed for 15 days and then they were removed. On the following day the treatments were performed. In Study 2, the ligatures were placed for 7 days and remained in position throughout the periods. Forty animals were used in Study 1 and they were divided into 4 groups: PD (disease induction only, without treatment); SRP (induction and basic periodontal treatment); PDT (Induction and application of photodynamic therapy); SRP+PDT (induction, application of photodynamic therapy and basic periodontal treatment). Forty-two animals were used in Study 2 and they divided into 4 groups: PS (Treatment with zinc tetracarboxy-N-metylglucamine phthalocyanine only); Light (Treatment with red LED light irradiation only); aPDT (Treatment with photodynamic therapy – PS + Light) and PD (Periodontal disease induction, without treatment). In Study 1, the animals were euthanized after 7 and 30 days of treatment. In Study 2, the therapies were applied at zero period and the animals were euthanized at 7 and 15 days. For both studies, one- way ANOVA parametric test was applied, followed by Tukey’s post-test. In Study 1, concerning histometry data, no statistical differences were observed between groups. The microtomographic analysis indicated significant differences in the two periods for the PD and SRP+PDT groups and, at 7 days, for the PD and PDT groups in the furcation area. No significant differences the interproximal regions were observed. Regarding the histological analyses, no tissue damage was observed. In Study 2, the three-dimensional radiographic and histometric analyses revealed no statistically different results for the furcation and interproximal regions. The inflammatory profile presented a trend of lower amounts of inflammatory cells in the aPDT group at 7 days, while the histological analysis indicated no significant differences between the groups, indicating that the therapies did not cause tissue damage. Thus, these data indicate that PDT using zinc tetracarboxy-N-metylglucamine phthalocyanine was effective in maintaining bone loss in PD-induced in rats, although further studies are required to further elucidate PDT effects and the application of aPDT and its components as monotherapies in PD-induced in rats with the preservation of the ligatures, favored in situ bacteria permanence and inhibited treatment action.
Antimicrobial Photodynamic Therapy (PDT) is a minimally invasive method consisting in the application of a photosensitive dye, which is subsequently stimulated by a light source and reacts with oxygen, producing reactive species. The aim of this study was to evaluate in vivo, the aPDT effects (with the PS zinc tetracarboxy-N-metylglucamine phthalocyanine 10μg/mL, and red LED light with 655nm), as adjuvant treatment to Scaling Root and Planing (SRP) and as monotherapy and verify the responses and tissue changes after aPDT application in PD-induced rats by microtomographic, histometric, stereometric and histological evaluations. Ligatures were placed around the second maxillary molars for PD induction. In Study 1, the ligatures were placed for 15 days and then they were removed. On the following day the treatments were performed. In Study 2, the ligatures were placed for 7 days and remained in position throughout the periods. Forty animals were used in Study 1 and they were divided into 4 groups: PD (disease induction only, without treatment); SRP (induction and basic periodontal treatment); PDT (Induction and application of photodynamic therapy); SRP+PDT (induction, application of photodynamic therapy and basic periodontal treatment). Forty-two animals were used in Study 2 and they divided into 4 groups: PS (Treatment with zinc tetracarboxy-N-metylglucamine phthalocyanine only); Light (Treatment with red LED light irradiation only); aPDT (Treatment with photodynamic therapy – PS + Light) and PD (Periodontal disease induction, without treatment). In Study 1, the animals were euthanized after 7 and 30 days of treatment. In Study 2, the therapies were applied at zero period and the animals were euthanized at 7 and 15 days. For both studies, one- way ANOVA parametric test was applied, followed by Tukey’s post-test. In Study 1, concerning histometry data, no statistical differences were observed between groups. The microtomographic analysis indicated significant differences in the two periods for the PD and SRP+PDT groups and, at 7 days, for the PD and PDT groups in the furcation area. No significant differences the interproximal regions were observed. Regarding the histological analyses, no tissue damage was observed. In Study 2, the three-dimensional radiographic and histometric analyses revealed no statistically different results for the furcation and interproximal regions. The inflammatory profile presented a trend of lower amounts of inflammatory cells in the aPDT group at 7 days, while the histological analysis indicated no significant differences between the groups, indicating that the therapies did not cause tissue damage. Thus, these data indicate that PDT using zinc tetracarboxy-N-metylglucamine phthalocyanine was effective in maintaining bone loss in PD-induced in rats, although further studies are required to further elucidate PDT effects and the application of aPDT and its components as monotherapies in PD-induced in rats with the preservation of the ligatures, favored in situ bacteria permanence and inhibited treatment action.
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Palavras-chave
Doenças periodontais, Fotoquimioterapia, Microtomografia por raio- X, Periodontal disease, Photochemotherapy, Computadorized microtomography