Evaluation of How Methacrylate Gelatin Hydrogel Loaded with Ximenia americana L. Extract (Steam Bark) Effects Bone Repair Activity Using Rats as Models.

The use of bioactive materials, such as Ximenia americana L., to stimulate the bone repair process has already been studied; however, the synergistic effects of its association with light emitting diode (LED) have not been reported. The present work aims to evaluate the effect of its stem bark extract incorporated into methacrylate gelatin hydrogel (GelMA) on the bone repair process using pure hydrogel and hydrogel associated with LED therapy. For this purpose, the GelMA hydrogel loaded with Ximenia americana L. extract (steam bark) was produced, characterized and applied in animal experiments. The tests were performed using 50 male Wistar rats (divided into 5 groups) submitted to an induced tibia diaphyseal fracture. The therapy effects were verified for a period of 15 and 30 days of treatment using histological analysis and Raman spectroscopy. After 15 days of induced lesion/treatment, the new bone formation was significantly higher in the GXG (GelMA + X. americana L.) group compared to the control group (p < 0.0001). After 30 days, a statistically significant difference was observed when comparing the GXLEDG (GelMA + X. americana L. + LED) and the control group (p < 0.0001), the GXG and the control group (p < 0.001), and when comparing the GG, GXG (p < 0.005) and GXLEDG (p < 0.001) groups. The results shows that the Ximenia americana L. stem extract incorporated into GelMA hydrogel associated with LED therapy is a potentiator for animal bone repair.

Identification of bioactive compounds and cytogenotoxicity of the essential oil from the leaves of Croton heliotropiifolius Kunth.

Croton heliotropiifolius Kunth, popularly known as "quince" and "velame," contains a high concentration of volatile oils in the leaves, and widely used in folk medicine as an antiseptic, analgesic, sedative, anti-inflammatory, spasmolytic and local anesthetic. The objectives of this investigation were to (1) identify the phytochemical compounds and (2) assess the cytogenotoxicity of the essential oil extracted from the leaves of C. heliotropiifolius Kunth. The oil was extracted utilizing hydrodistillation and phytochemical profile determined using gas chromatography and mass spectrometry (GCMS). In the toxicogenetics analysis, Allium cepa roots were exposed to 1% dimethylsulfoxide or methylmethanesulfonate (MMS, 10 µg/ml) negative and positive controls, respectively, and to C. heliotropiifolius oil at 6 concentrations (0.32; 1.6; 8; 40; 200 or 1000 µg/ml). The phytochemical profile exhibited 40 chromatographic bands, and 33 compounds identified. α-pinene (16.7%) and 1,8-cineole (13.81%) were identified as the major compounds. Some of these identified secondary metabolites displayed biological and pharmacological activities previously reported including antiseptic, analgesic, sedative, anti-inflammatory as well insecticidal, antiviral, anti-fungal actions. In the A. cepa test, C. heliotropiifolius leaves oil induced cytotoxicity at concentrations of 0.32, 1.6 or 200 µg/ml and genotoxicity at 200 or 1000 µg/ml as evidenced by increased presence of micronuclei and significant chromosomal losses. Based upon our observations data demonstrated that the essential oil of C. heliotropiifolius leaves contain monoterpene hydrocarbons, and oxygenated monoterpenes, sesquiterpenes, and oxygenated sesquiterpenes which are associated with cytotoxic and genotoxic responses noted in on A. cepa cells.

Cecropia pachystachya Trécul: identification, isolation of secondary metabolites, in silico study of toxicological evaluation and interaction with the enzymes 5-LOX and α-1-antitrypsin.

Cecropia pachystachya Tréc., popularly known as embaúba, belongs to the Cecropiaceae family and is used by the native population in the treatment of bronchitis, asthma, high blood pressure, fever, and as a diuretic. The pharmacological actions including anti-inflammatory, antioxidant, cardiotonic and sedative were previously reported. The objective of this study was to (1) isolate and identify bioactive compounds extracted from the ethanolic extract of C. pachystachya roots (ERCP), as well as (2) verify the affinity of these metabolites with the enzymes 5-lipoxygenase (5-LOX) and α-1-antitrypsin through in silico tests. Isolation and/or identification were performed using GC-MS, HPLC, Infrared (IR), and nuclear magnetic resonance (NMR) techniques. After isolation and identification of the active compounds, these substances were subjected to the in silico investigation that proceeded by performing PreADMET simulations and molecular docking calculations. The bioactive compounds identified were 1-(+)-ascorbic acid 2,6-dihexadecanoate, ethyl hexadecanoate, ethyl (9E,12E)-octadec-9,12-dienoate, ethyl (Z)-octadec-9-enoate and ethyl octadecanoate by GC-MS; chlorogenic acid, catechin, epicatechin, syringaldehyde by HPLC; ß-sitosterol, sitostenone, beccaridiol, tormentic acid, lupeol, α- and ß-amyrin by classical chromatography, IR, 1H and 13C NMR techniques. The ADMET (absorption, distribution, metabolism, excretion, and toxicity) properties were determined for each bioactive compound. Tormentic acid demonstrated a greater affinity for 5-LOX enzyme while sitostenone demonstrated a higher affinity for the α-1-antitrypsin enzyme. Our findings demonstrated a diverse range of secondary metabolites isolated from C. pachystachya that showed relevant interactions with the enzymes 5-LOX and α-1-antitrypsin. Thus, "embaúba" may be employed in in vivo experimental studies as a form of alternative treatment for chronic lung diseases.Abbreviations: ADT: Autodock Tools; BBB: Blood-brain barrier; CaCo2: Human colonic adenocarcinoma cells; CC: Classic/open Column; TLC: Thin Layer Chromatography; CD40: Differentiation Cluster 40; CENAUREMN: Centro Nordestino de Aplicação e Uso da Ressonância Magnética Nuclear; GC-MS: Gas Chromatography coupled to mass spectrometry; HPLC: High-Perfomance Liquid Chromatography; CYP2C9, CYP2C19, CYP2D6 and CYP3A4: Cytochrome P450 isoenzymes; COPD: Chronic Obstructive Pulmonary Disease; DRX-500: X-Ray Diffraction - 500; ERCP: Ethanolic extract of the roots of C. pachystachya; FAPEPI: Fundação de Amparo à Pesquisa do Piauí; HIA: Human Intestinal Absorption; IR: Infrared; Ki: Inhibition constant; 5-LOX: 5-Lipoxygenase; mM: miliMolar; nM: nanoMolar; OECD423: acute toxic class method; PDB: Protein Data Bank; P-gP: P-glycoprotein; PM2,5: Small inhalable particles 2,5; PPB: Plasm Protein Binding; PreADMET: Prediction Absorption, Distribution, Metabolization, Excretion and Toxicity; NMR: Nuclear Magnetic Resonance; +S9: with metabolic activation; -S9: no metabolic activation; SisGen: Sistema Nacional de Gestão de Patrimônio Genético e do Conhecimento Tradicional Associado; RT: Retention time; TA100: Ames test with TA100 cells line; TA1535: Ames test with cells of the TA1535 cell line; UESPI: State University of Piauí; V79: lung fibroblast cells; ΔG: Gibbs free energy (Kcal/mol); µM: microMolar.

Phytochemical screening, phenolic and flavonoid contents, antioxidant and cytogenotoxicity activities of Combretum leprosum Mart. (Combretaceae).

Combretum leprosum Mart. (Combretaceae), a shrub popularly known as mofumbo, is used in folk medicine for treatment of uterine bleeding, pertussis, gastric pain, and as a sedative. The aim of this study was to (1) determine the phytochemical profile,(2) identify chemical constituents and (3) examine antioxidant and cytogenotoxic activity of ethanolic extracts and fractions of stem bark and leaves. The plant material (leaf and stem bark) was submitted to extraction with ethanol, followed by partition using hexane, chloroform, and ethyl acetate. It was possible to identify and quantify the epicatechin in the ethanolic stem bark extract (0.065 mg/g extract) and rutin in the leaf extract (3.33 mg/g extract). Based upon in vitro tests a significant relationship was noted between findings from antioxidant tests and levels of total phenolic and flavonoid. Comparing all samples (extracts and fractions), the ethyl acetate fractions of stem bark (411.40 ± 15.38 GAE/g) and leaves (225.49 ± 9.47 GAE/g) exhibited higher phenolic content, whereas hexanic fraction of stem bark (124.28 ± 56 mg/g sample) and ethyl acetate fraction of leaves (238.91 ± 1.73 mg/g sample) demonstrated a higher content of flavonoids. Among the antioxidant tests, the intermediate fraction of stem bark (28.5 ± 0.60 µg/ml) and ethyl acetate fraction of leaves (40 ± 0.56 µg/ml) displayed a higher % inhibition of free radical DPPH activity, whereas intermediate fraction of stem bark (27.5 ± 0.9 µg/ml) and hydromethanol fraction of leaves (81 ± 1.4 µg/ml) demonstrated inhibition of the free radical ABTS. In biological tests (Allium cepa and micronucleus in peripheral blood), data showed that none of the tested concentrations of ethanolic extracts of leaves and stem bark produced significant cytotoxicity, genotoxicity, and mutagenic activity.Abbreviations AA%: percentage of antioxidant activity; ABTS: 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid); CEUA: Ethics Committee in the Use of Animals; TLC: Thin Layer Chromatography; DNA: deoxyribonucleic acid; DPPH: 1,1-diphenyl-2-picrylhydrazyl; ROS: Reactive oxygen species; EEB: ethanol extract of the stem bark; HFB: Hexanic fraction of stem bark; IFB: Intermediate fraction of stem bark; CFB: Chloroform fraction of stem bark; EAFB: Ethyl acetate fraction of stem bark; HMFB: Hydromethanol fraction of the stem bark; EEL: Ethanol extract from leaves; HFL: Hexane fraction of leaves; CFL: Chloroform fraction of leaves; EAFL: Ethyl acetate fraction of leaves; HMFL: Hydromethanol fraction of leaves; GAE: Gallic Acid Equivalent; IC50: 50% inhibition concentration; HCOOH: Formic acid; HCl: hydrochloric acid; HPLC: High-performance liquid chromatography; MN: micronucleus; WHO: World Health Organization; UFLC: Ultra-Fast Liquid Chromatography; UESPI: State University of Piauí.

Phytochemical study, molecular docking, genotoxicity and therapeutic efficacy of the aqueous extract of the stem bark of Ximenia americana L. in the treatment of experimental COPD in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Ximenia americana L. is popularly known as yellow plum, brave plum or tallow wood. All the parts of this plant are used in popular medicine. Its reddish and smooth bark are used to treat skin infections, inflammation of the mucous membranes and in the wound healing process. OBJECTIVE: Verification of phytochemical profile, the molecular interaction between flavonoid, (-) epi-catechin and 5-LOX enzyme, by means of in silico study, the genotoxic effect and to investigate the pharmacological action of the aqueous extract of the stem bark of X. americana in pulmonary alterations caused by experimental COPD in Rattus norvegicus. MATERIALS AND METHODS: The identification of secondary metabolites was carried out by TLC and HPLC chromatographic methods, molecular anchoring tests were applied to analyze the interaction of flavonoid present in the extract with the enzyme involved in pulmonary inflammation process and the genotoxic effect was assessed by comet assay and micronucleus test. For induction of COPD, male rats were distributed in seven groups. The control group was exposed only to ambient air and six were subjected to passive smoke inhalations for 20 min/day for 60 days. One of the groups exposed to cigarette smoke did not receive treatment. The others were treated by inhalation with beclomethasone dipropionate (400 mcg/kg) and aqueous and lyophilized extracts of X. americana (500 mg/kg) separately or in combination for a period of 15 days. The structural and inflammatory pulmonary alterations were evaluated by histological examination. Additional morphometric analyses were performed, including the alveolar diameter and the thickness of the right ventricle wall. RESULTS: The results showed that the aqueous extract of the bark of X. americana possesses (-) epi -catechin, in silico studies with 5-LOX indicate that the EpiC ligand showed better affinity parameters than the AracA ligand, which is in accordance with the results obtained in vivo studies. Genotoxity was not observed at the dose tested and the extract was able to stagnate the alveolar enlargement caused by the destruction of the interalveolar septa, attenuation of mucus production and decrease the presence of collagen fibers in the bronchi of animals submitted to cigarette smoke. CONCLUSION: Altogether, the results proved that the aqueous extract of X. americana presents itself as a new option of therapeutic approach in the treatment of COPD.

EFICÁCIA DA FONOFORESE COM XIMENIA AMERICANA L. NA INFLAMAÇÃO DE TENDÃO DE RATOS / PHONOPHORESIS EFFECTIVENESS WITH XIMENIA AMERICANA L. IN RATS TENDON INFLAMMATION / EFICACIA DE LA FONOFORESIS CON XIMENIA AMERICANA L. EN LA INFLAMACIÓN DE TENDÓN DE RATAS

RESUMO Introdução: Diversos recursos terapêuticos, como laser e ultrassom isolado, combinados com fármacos e fonoforese têm sido utilizados em casos de inflamação e reparo de tendão, sendo o ultrassom pulsado bastante eficaz em tendinites crônicas. Objetivo: Analisar a eficácia do uso da fonoforese com o extrato etanólico das cascas do caule da Ximenia americana L. na resolução do processo inflamatório crônico em Rattus norvegicus. Métodos: A amostra consistiu em 120 animais para análise da resposta inflamatória, utilizando-se como variáveis edema, força biomecânica, número de fibroblastos e análise histológica. Resultados: Nos 7° e no 14° dia, verificou-se diferença significativa entre o grupo ultrassom com gel do caule de Ximenia americana L. e o grupo ultrassom com placebo (p < 0,05) quanto à redução de edema, aumento da força máxima de ruptura, redução extremamente significativa da deformação máxima (p < 0,001), além de aumento dos fibroblastos. Na análise histológica, houve melhora do processo inflamatório inicial e aceleração do reparo tendíneo, com redução de células inflamatórias e com deposição de colágeno organizado com matriz extracelular densa. Conclusão: O ultrassom pulsado combinado com o gel do caule de Ximenia americana L. é uma forma terapêutica eficaz para a resolução do processo inflamatório crônico.

ABSTRACT Introduction: Several therapeutic resources such as laser and isolated ultrasound combined with drugs and phonophoresis have been used in cases of inflammation and tendon repair, and the pulsed ultrasound is quite effective in chronic tendinitis. Objective: To analyze the efficacy of phonophoresis with the ethanol extract of the stem bark of Ximenia americana L. to manage the chronic inflammatory process in Rattus norvegicus. Methods: The sample consisted of 120 animals for analysis of the inflammatory response using edema, biomechanical strength, number of fibroblasts and histological analysis as variables. Results: On the 7th and 14th day, there was a significant difference between the group ultrasound with Ximenia americana L. gel and the ultrasound with placebo group (p<0.05) for edema reduction, increased maximum rupture strength, highly significant reduction of the maximum deformation (p<0.001), in addition to an increase in fibroblasts. In the histological analysis, there was improvement in the inflammatory process and acceleration of tendon repair with reduction of inflammatory cells and deposition of organized collagen with dense extracellular matrix. Conclusion: The pulsed ultrasound combined with gel of Ximenia americana L. is an effective therapy to manage the chronic inflammatory process.

RESUMEN Introducción: Varios recursos terapéuticos, tales como láser y ultrasonido aislado, en combinación con fármacos y la fonoforesis se han utilizado en casos de inflamación y reparación del tendón, y el ultrasonido pulsado ha sido muy eficaz en la tendinitis crónica. Objetivo: Analizar la eficacia de la fonoforesis con el extracto de etanol de la corteza del tallo de Ximenia americana L. en la resolución del proceso inflamatorio crónico en Rattus norvegicus. Métodos: La muestra fue de 120 animales para analizar la respuesta inflamatoria, utilizando como variables el edema, la resistencia biomecánica, el número de fibroblastos y el análisis histológico. Resultados: En el 7° y en el 14° día, se observó una diferencia significativa entre el grupo de ultrasonido con gel de Ximenia americana L. y el grupo ultrasonido con placebo (p < 0,05) en la reducción del edema, el aumento de la resistencia máxima a la rotura, la reducción altamente significativa de la deformación máxima (p < 0,001), además del aumento de los fibroblastos. En el análisis histológico hubo una mejoría en el proceso inflamatorio y la aceleración de la reparación del tendón, con reducción de células inflamatorias y deposición de colágeno organizado con matriz extracelular densa. Conclusión: La eficacia del ultrasonido pulsado combinado con gel de Ximenia americana L. es una forma eficaz para la resolución del proceso inflamatorio crónico.

Análise da ação do gel da Bryophyllum calycinum Salisb (folha santa) associado ao ultrassom na cicatrização de feridas cutâneas / Analysis of the action of Bryophyllum calycinum Salisb (holy sheet) gel associated with ultrasound in skin wound healing

Introdução: A cicatrização é uma cascata de eventos celulares que interagem para que ocorra a reconstituição do tecido. Objetivo: Analisar a ação do gel da Bryophyllum calycinum Salisb, associado ao ultrassom pulsado, na cicatrização de feridas. Métodos: Vinte ratos foram feridos, divididos em quatro grupos, tratados e observados diariamente. O Grupo1 não recebeu nenhum tratamento; o 2 foi tratado com ultrassom (modo pulsado 10%, frequência de 1 MHz, intensidade de 0,5 W/cm2, método de acoplamento direto com movimentos oscilatórios numa ERA de 1 cm2, durante dois minutos) + gel da B. calycinum Salisb a 2%; o 3, com gel da B. calycinum Salisb a 2%; o 4, com ultrassom. Resultados: O Grupo 2 teve maior redução das áreas feridas, diminuição do colágeno tipo III e aumento do tipo I, quando comparado ao controle. Conclusões: O gel da folha santa associado ao ultrassom é capaz de acelerar o processo de cicatrização tecidual.

Introduction: Healing is a cascade of cellular events that interact to occur replenish the tissue. Objective: To analyze the action of the Bryophyllum calycinum Salisb gel associated with the pulsed ultrasound in wound healing. Methods: Twenty rats were wounded and divided into four groups. After this, they were treated and observed daily. Group 1 did not receive any treatment; Group 2 was treated with ultrasound (10% pulsed mode, frequency of 1 MHz intensity of 0.5 W / cm2, direct coupling method with an oscillatory motion was 1 cm2 for two minutes) + Gel B. calycinum Salisb 2%; Group 3, with B. calycinum Salisb gel 2%; and Group 4, with ultrasound. Results: Group 2 had a greater reduction in wound areas, a decrease of type III collagen and an increase of the type I when compared to the control group. Conclusions: The gel of the holy sheet associated with ultrasound is able to accelerate the wound healing process.