The Dynamics of Subcutaneous Tissue Response to Microorganisms Associated with the Extract of Araçá (Psidium cattleianum): An Edemogenic and Microscopic Analysis / A Dinâmica da Resposta do Tecido Subcutâneo para Associação de Microorganismos com o Extrato de Araçá (Psidium cattleianum): Uma Análise Microscópica e Edemogênica

Objective: To evaluate in vivo tissue reaction to the extract of araçá (Psidium cattleianum) associated with inactivated microorganisms. Material and Methods: A 0.1 mL suspension was used containing Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Enterococcus faecalis, Peptostreptococcus micros, and Porphyromonas endodontalis, which were inactivated by heat and mixed into a 1.0 mL saline (control group), an aqueous solution, or a hydroalcoholic extract of araçá. Eighteen male rats (Rattus norvegiccus) under general anesthesia received 0.2 mL of 1% intravenous Evans blue. Thirty minutes later, 0.1 mL of one of the associations was injected into the animals' dorsal region. The animals were euthanized after 3 and 6 hours, and the materials obtained were placed in formamide for 72 hours then analyzed in a spectrophotometer (λ=630 ηm). For the morphological analysis, 30 rats received polyethylene tubes implants with the extracts or the saline with the associations in the dorsal region and euthanized after 7 and 30 days to be analyzed according to an inflammation cell score. Results: No significant difference (p>0.05) was observed in the edema among groups. The optical microscopy results showed a repair in the 30-day-period, which was higher when compared to the 7-day-period (p< 0.0001). Nevertheless, in the 7-day-period, the hydroalcoholic extract presented a significant response compared to the aqueous extract (p=0.05) and a trend for better results than the control group. Conclusion: The aqueous and hydroalcoholic araçá extracts associated with inactivated microorganisms showed similar responses to control, indicating no interference on the toxic effects of the bacterial components in tissue repair. (AU)

Objetivo: Avaliar in vivo a reação tecidual do extrato de araçá (Psidium cattleianum) associado com microorganismos inativados. Material e Métodos: Uma suspensão de 0.1mL foi usada contendo Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Enterococcus faecalis, Peptostreptococcus micros e Porphyromonas endodontalis dos quais foram inativos por aquecimento e misturados a 1,0 mL de soro fisiológico (grupo controle), uma solução aquosa ou hidroalcoólica de araçá. Dezoito ratos machos (Rattus norvegiccus) sob anestesia geral receberam 0,2mL de Azul de Evans a 1% intravenoso. Após trinta minutos, 0,1mL de um dos extratos (associado com microorganismos inativos) foi injetado nos animais na região dorsal. Os animais foram eutanasiados após 3 e 6 horas, e os materiais obtidos colocados em formamida por 72 horas para análise em espectrofotômetro (λ=630 ηm). Para análise morfológica, 30 ratos receberam implante subcutâneo de tubo de polietileno com as associações na região dorsal, eutanasiados após 7 e 30 dias para serem analisados de acordo com um escore de células inflamatórias. Resultados: Não houve diferença significativa (p>0,05) no edema entre os grupos. Os resultados obtidos em microscópio óptico apontaram reparo em 30 dias superior ao de 7 dias (p< 0,0001). No período de 7 dias a solução hidroalcoólica apresentou resposta superior a solução aquosa (p=0,05) e uma tendência de melhor resultado que o controle. Conclusão: A solução aquosa e hidroalcoólica de extrato de araçá associadas a microrganismos inativados apresentaram respostas biológicas semelhantes ao controle, indicando que não há interferência sobre os efeitos tóxicos advindos dos componentes bacterianos, no sentido de favorecer o reparo(AU)

Viability of Human Gingival Fibroblast (FGH) Treated with Ethanolic "Aroeira" Extract (Myracrodruon urundeuva Allemão)

ABSTRACT The aim of this study was to evaluate the effect of ethanolic "aroeira" (Myracrodruon urundeuva) extract on the viability of human gingival fibroblast. For this, fibroblasts (2x103 cells/well) were plated in a 96-well plate and incubated for 24 h; the medium (Eagle's medium modified by Dulbecco - DMEM) supplemented with 10% fetal bovine serum was replaced by DMEM with different ethanolic extract concentration (0, 0.1, 1, 10, 100, and 1000μg / mL). The fibroblast viability was analyzed after 48,72, and 96 h by the neutral red capture test and violet crystal. The "aroeira" extract, at high concentrations (100 and 1000 µg/mL) caused decrease in both cellular viability tests (p<0.05). However, dilutions between 0.1 and 10 µg/mL did not affect the viability of the cells. It was concluded that "aroeira" extract was able to change the gingival fibroblast viability, and this effect was concentration dependent.

Caracterização do extrato de aroeira (Myracrodruon urundeuva) e seu efeito sobre a viabilidade de fibroblastos gengivais humanos / Characterization of extract of mastic (M. urundeuva) and its effect on the viability of human gingival fibroblasts

A aroeira (Myracrodruon urundeuva) é uma árvore cujas folhas vem sendo estudadas com fins terapêuticos na medicina e odontologia por apresentar potencial antiinflamatório e antimicrobiano, além de favorecer o processo de reparo e cicatrização. O objetivo do trabalho foi caracterizar quimicamente o extrato de aroeira e avaliar seu efeito sobre fibroblastos gengivais humanos. Foi realizado a caracterização química da aroeira por meio do processo de triagem cromatográfica que consistiu na coleta das folhas sadias, maceração em metanol (MeOH) 80%, partição entre os solventes hexano, acetato de etila (AcOEt) e n-butanol (n-(BuOH)) Em seguida, realizaram-se análises dos compostos químicos em cromatografia liquida de alta eficiência acoplado a detector de arranjo de fotodiodo HPLC-PAD, análises por espectrometria de massas MS e cromatografia liquida acoplada a espectrometria de massas (LC-MS). Posteriormente foi realizada a análise de citotoxicidade do extrato hidroalcoólico de aroeira em fibroblastos gengivais humanos (linhagem FGH). Para a realização dos experimentos foram plaqueadas 2x103 células/poço em placas de 96 poços. O meio de cultivo foi substituído por meio de cultura de Eagle modificado por Dullbecco complementado por 10% soro fetal bovino (SFB) em diferentes concentrações do extrato hidroalcoólico de aroeira (extrato bruto; 1:10; 1:100; 1:1000; 1:10000 e controle). As análises de viabilidade foram feitas nos tempos experimentais de 24, 48, 72 e 96 horas, por meio dos testes de redução do MTT (brometo de 3-(4,5-dimetiltiazol-2-yl)-2,5-difeniltetrazólio), captação do vermelho neutro e coloração pelo cristal violeta. A estatística foi realizada em análise de variância de 2 critérios seguido de uma análise de teste Tukey (p<0,05). Os compostos encontrados no extrato foram os derivados de ácido gálico, galotaninos, além de flavonoides. Para a redução do MTT os resultados mostraram uma ligeira oscilação da absorbância em todos os grupos...

Aroeira (M. urundeuva) is a tree whose leaves have been studied with therapeutic purpose in dentistry and medicine, due it antimicrobial and antinflamatory potencial protect, besides the repair and cicatrization processes. The aim of the study was characterize the aroeiras extract and evaluate effect over human gingival fibroblast. The aroeiras characterization was realized by means of chromatography screenuy, process that consists in healthy leaves maceration in 80% MeOH, partition in the BuOH, AcOEt and hexanes solvents. Next, the chemicals compounds were analysed in High Performance Liquide Chromatography-Photo Diode Array (HPLC-PAD), Liquide Chromatography Mass Spectrometry (LCMS) and Mass Spectrometry (MSs). Subsequently was realized the citotoxicitys analysis from the aroeira hidroalcooholic extract in human gingival fibroblast (FGH lineage). For the experiments realization was platted 2x103 cels/well in 96 wells plate. The cultives mean was substituted for Eagles means culture changed for complemented Dullbeco for 10% bovine fetal serum in aroeiras hidroalcooholic extracts concentrations different (brute extract, 1:10, 1:100, 1:1000, 1:10000). The viability analysis was carried out in experimental times 24, 48, 72 and 96 hours, by means of MTT reductions test (brometo3-(4,5-dimetiltiazol-2-yl)-2,5-difeniltetrazolio), neutral red captation and violet crystal). The statistics were performed for analysis of variance in 2 criteria continue with a Tukey test analysis (p<0.05). The founded compounds in the extract was the galic acid, gallotanine linked to glucose, besides flavonoids. For the MTTs reduction the results display an absorbance oscillation lightness in all the groups in experimental periods. Distinction for the control group and 1:100, 1:1000 and 1:10000 dilution, that present, by and large, for all the periods, the biggest valuable, while the brute extract group and 1:10 dilution present valuable lower to the others groups (p<0.05)...

Evaluation of tissue reaction to Aroeira (Myracrodruon urundeuva) extracts: a histologic and edemogenic study

OBJECTIVES: This study evaluated subcutaneous tissue response to Aroeira (Myracrodruon urundeuva) extract employing edemogenic and histological analyses. MATERIAL AND METHODS: Test groups consisted of aqueous and ethanolic Aroeira extracts and saline (control). For edema quantification, 18 rats received an intravenous injection of Evan's Blue. After 30 min, the extracts and saline were injected on the dorsum of the rats, which were then sacrificed after 3 and 6 h. Readings were performed in a spectrophotometer. For subcutaneous implantation, 30 rats received a polyethylene tube containing the extracts on their dorsum and then they were killed after 7 and 28 days. The samples were processed for histological analysis and evaluated with a light microscope. The inflammatory infiltrate was quantified. RESULTS: There were no statistically significant differences between aqueous extract and saline groups in relation to edema quantification in the different periods (p>0.05). Ethanolic solution resulted in more edema independently of the experimental period (p<0.05). Histological analysis showed similar results on the 7-day period for the 3 groups. There was a notable reduction on inflammatory cell number for saline and aqueous extract groups at 28 days. CONCLUSION: The aqueous extract showed biocompatible properties similar to those of saline.