Effect of the use of antibiotics on the skin repair process in animal model

Efeito do uso de antibióticos no processo de reparo cutâneo em modelo animal. Orientadora: Reggiani Vilela Gonçalves. Coorientadores: Rômulo Dias Novaes, Mariáurea Matias Sarandy Souza e Sirlene Souza Rodrigues Sartori. A busca por um tratamento eficaz que promova o fechamento rápido de feridas cutâneas tem como objetivo garantir ao paciente menor sofrimento e reduzir custos. Uma ferida pode demorar a cicatrizar devido a falha na progressão entre as fases da cicatrização. A cronificação do processo geralmente está associada a uma fase inflamatória prolongada. O objetivo desse trabalho foi investigar o uso de antibióticos, em especial da doxiciclina (Dx), na cicatrização de feridas cutâneas não infectadas em animais. Primeiro, realizamos uma revisão sistemática seguindo as diretrizes PRISMA. A plataforma SYRCLE’s Risk of Bias foi utilizada para análise da qualidade metodológica dos estudos. Em nossa revisão, vinte e sete estudos foram selecionados para a extração de dados e análise de viés metodológico. A compilação dos dados mostrou que o tratamento antibiótico pode promover redução do infiltrado inflamatório, aumento do número de fibroblastos e dos constituintes da matriz extracelular, reepitelização e aumento resistência mecânica do tecido neoformado. No entanto, observamos grande viés metodológico entre os estudos, o que compromete a validade interna e externa dos mesmos. Após os achados promissores encontrados nesta revisão, partimos para investigação da ação do antibiótico doxiciclina, in vitro na viabilidade celular de macrófagos e in vivo no reparo cutâneo. Ratos Wistar machos (n=15) foram alojados em gaiolas individuais com comida e água ad libitum (número de registo 72/2017). Após anestesia, três feridas circulares (12 mm de diâmetro) foram feitas no dorso de cada animal. Estes foram divididos em três grupos e receberam o tratamento durante 21 dias, via gavagem: C (controle - água); Dx1 (Dx 10 mg/kg/dia) e Dx2 (Dx 30 mg/kg/dia). A área e a taxa de contração das feridas foram avaliadas e os tecidos neoformados foram coletadas para análise histologia e bioquímica. Animais tratados com Dx apresentaram maior proporção de células (macrófagos e mastócitos) e dos componentes da matriz extracelular (colágeno e fibra elástica), aumento inicial seguido de redução a proporção de vasos sanguíneos e reduziu o estresse oxidativo e os níveis de metaloproteinase de matriz-10 (MMP-10) do tecido cicatricial. Além disso, resultou em maior contração da ferida e melhor cicatrização em relação ao controle. Em conclusão, os antibióticos, que são drogas com ampla utilização na população, podem interferir no reparo de feridas 6 cutâneas não decorrente de infecção, evidenciando a eficácia do antibiótico Dx no processo de cicatrização de feridas de segunda intenção em ratos Wistar. Palavras-chave: Antibioticoterapia. Revisão sistemática. In vitro e in vivo análises. Cicatrização de ferida.The search for an effective treatment that promotes rapid closure of skin wounds aims to ensure the patient less suffering and reduce costs. A wound may take time to heal due to failure to progress between healing phases. Chronification of the process is usually associated with a prolonged inflammatory phase. The aim of this study was to investigate the use of antibiotics, especially doxycycline (Dx), in the healing of uninfected skin wounds in animals. First, we performed a systematic review following the PRISMA guidelines. The SYRCLE's Risk of Bias platform was used to analyze the methodological quality of the studies. In our review, twenty- seven studies were selected for data extraction and methodological bias analysis. The compilation of data showed that antibiotic treatment can promote reduction of inflammatory infiltrate, increase in the number of fibroblasts and extracellular matrix constituents, reepithelization and increased mechanical resistance of neoformed tissue. However, we observed a great methodological bias between the studies, which compromises their internal and external validity. After the promising findings found in this review, we set out to investigate the action of doxycycline antibiotic, in vitro on macrophage cell viability and in vivo on skin repair. Male Wistar rats (n = 15) were housed in individual cages with food and water ad libitum (registration number 72/2017). After anesthesia, three circular wounds (12 mm in diameter) were made on the back of each animal. These were divided into three groups and received treatment for 21 days via gavage: C (control - water); Dx1 (Dx 10 mg / kg / day) and Dx2 (Dx 30 mg / kg / day). Area and rate of wound contraction were evaluated, and newly formed tissues were collected for histological and biochemical analysis. Dx-treated animals had a higher proportion of cells (macrophages and mast cells) and extracellular matrix components (collagen and elastic fiber), an initial increase followed by a reduction in the proportion of blood vessels and reduced oxidative stress and matrix-metalloproteinase levels. 10 (MMP-10) of scar tissue. In addition, it resulted in greater wound contraction and better healing compared to control. In conclusion, antibiotics, which are widely used drugs in the population, may interfere with the repair of non-infection skin wounds, showing the efficacy of the antibiotic Dx in the healing process of secondary intention wounds in Wistar rats. 8 Keywords: Antibiotic therapy. Systematic review. In vitro and in vivo analyzes. Wound healing.Coordenação de Aperfeiçoamento de Pessoal de Nível Superio

Controle alternativo de Colletotrichum musae com extrato de mil folhas Alternative control of Colletotrichum musae with extract of yarrow

Made available in DSpace on 2018-02-15T18:27:44Z (GMT). No. of bitstreams: 2 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) yasmim_freitas_et_all.pdf: 87751 bytes, checksum: 972daf626011e5fc044deb5481b925bc (MD5) Previous issue date: 9Universidade Federal de Lavras. Programa de Pós-graduação em Fitopatologia. Departamento de Fitopatologia. Lavras, MG, Brasil.Universidade Federal de Viçosa. Programa de Pós-graduação em Biologia Celular e Estrutural. Departamento de Biologia Geral. Viçosa, MG, Brasil.Universidade Federal do Espírito Santo. Centro Universitário Norte do Espírito Santo. Departamento de Ciências Agrárias e Biológica. Laboratório de Fitopatologia. São Mateus, ES, Brasil.Empresa Usina Magdalena S.A. Departamento de Manejo Integrado de Plagas. Laboratório de Microbiologia. Escuintla, Guatelama.Universidade Federal do Espírito Santo. Centro Universitário Norte do Espírito Santo. Departamento de Ciências Agrárias e Biológica. Laboratório de Fitopatologia. São Mateus, ES, Brasil.Extratos de planta de mil folhas (Achillea millefolium L.), podem auxiliar no controle de doenças de pós-colheita da banana (Musa sp.), como a antracnose (Colletotrichum musae B. & M.A.C.). O objetivo deste trabalho foi avaliar a inibição de crescimento do fungo por frações dos extratos de distintas partes da planta em diferentes meses de colheita. O material colhido foi separado, secado e triturado, realizando-se a extração etanólica pela técnica de maceração. Após a obtenção do extrato, este foi seco, dissolvido em Dimetilsulfóxido e vertido em meio BDA fundente. Os tratamentos foram feitos em triplicata, sendo avaliado o crescimento radial do fungo após 5 dias em BOD (25°C). Ocorreu a redução de crescimento do fungo, quando comparadas com a testemunha, obtendo os melhores resultados nas frações mais polares e, as partes mais ativas foram as flores. A constância da atividade, observada nas diferentes partes da planta, é um indicativo do potencial do extrato da planta no controle da antracnose da banana.Plant extracts such as yarrow (Achillea millefolium L.), can help control of post-harvest diseases in Banana (Musa sp.), such as anthracnose (Colletotrichum musae B. & M.A.C.). The objective of this study was to evaluate the inhibition of fungal growth by fractions of different parts of the plant extracts in different months of harvest. The material collected was, separated, dried, crushed and held the ethanol extraction by maceration technique. After obtaining the extract, this was dried, dissolved in dimethylsulfoxide and poured on PDA medium flux. Treatments performed were in triplicate and evaluated the radial growth of the fungus after 5 days under growth chamber (25 ° C). There was a growth reduction of the fungus, compared to the control, obtaining the best results in the polar fractions and the most active parts were the flowers. The constancy of the activity observed in different parts of the plant is indicative of the plant extract potential in controlling anthracnose of banana

Does antibiotic use accelerate or retard cutaneous repair? A systematic review in animal models.

BackgroundThe presence of infections is one of the main factors that leads to delays in healing or non-closure of cutaneous wounds. Although the goal of antibiotic use is to treat or prevent infection, there is currently no agreement on the effectiveness of these products.AimThe aim of this study was to evaluate the efficacy of antibiotic use during the healing process of skin wounds in animal models not intentionally infected, as well as to analyze the advances and limitations of the studies carried out in this field.Main methodsThis systematic review was performed according to the PRISMA guidelines, using a structured search on the MedLine (PubMed) and Scopus platforms to retrieve studies published until August 29, 2018, 13:35p.m. The studies included were limited to those that used excision or incision wound models and that were not intentionally infected. The data for the animal models, antibiotic used, and the main results of the studies were extracted, and compared where possible. Bias analysis and methodological quality assessments were examined through the SYRCLE's Risk of Bias tool.Key findingsTwenty-seven studies were selected. Overall, the effects of the antibiotic on the wound decreased inflammatory cell infiltration and promoted an increased number of fibroblasts, extracellular matrix constituents, re-epithelialization and tissue strength. A great deal of important information about the methodology was not presented, such as: the statistical analysis used, the animal model (sex and age), antibiotic dosage, blinding and randomization of the animals chosen.SignificanceBased on the results found, we believe that antibiotic therapy can be considered a viable alternative for the treatment of cutaneous wounds. However, current evidence obtained from the methodological quality analysis points towards a high risk of bias. This is due to the incomplete characterization of the experimental design and treatment protocol, which compromises the reproducibility of the studies

Strychnos pseudoquina modulates the morphological reorganization of the scar tissue of second intention cutaneous wounds in rats.

Natural substances are used in folk medicines to treat injuries. Strychnos pseudoquina has scarring, antipyretic, and antimalarial actions. The present study aimed to analyze the effect of S. pseudoquina on cutaneous wound healing in rats. The S. pseudoquina extract was submitted to phytochemical prospection. The levels of flavonoids and total phenolic compounds in the extract were 50.7 mg/g and 2.59 mg/g, respectively. Thirty Wistar rats were individualized in cages with food and water ad libitum (registration no. 730/2014). After anesthesia, three circular wounds (12mm diameter) were made in the animals, which were randomly separated into five treatments: Sal, saline; VO, ointment vehicles (lanolin and Vaseline); SS, positive control (silver sulfadiazine 1%); LE 5, freeze-dried extract 5%; and LE 10, lyophilized extract 10%. The animals were treated with the ointment daily for 21 days. Every seven days, the area and the rate of wound contraction were evaluated. Tissue samples were removed for histopathological analysis of the number of mast cells, elastic and collagen fibers, and biochemical analyses, quantification of malondialdehyde (MDA), carbonylated proteins (PCN), superoxide dismutase (SOD), catalase (CAT), transforming growth factor β (TGF-β), Interleukin 10 (IL-10) and tumor necrosis factor (TNF). The number of mast cells, collagen and elastic fibers in the rat wounds were higher in the treatments with the plant. The extract also stimulated the activity of antioxidant enzymes, particularly SOD, presenting high levels, and maintained low levels of PCN. The TGF-β and IL-10 concentration was higher in the LE5 and LE10 treatment of the extract than in the Sal, OV and SS treatments on day 7. The ointment based on S. pseudoquina closed the wound faster and accelerated wound healing in animals

Peptides from Animal Origin: A Systematic Review on Biological Sources and Effects on Skin Wounds

Background. Skin wounds are closely correlated with opportunistic infections and sepsis risk. Due to the need of more efficient healing drugs, animal peptides are emerging as new molecular platforms to accelerate skin wound closure and to prevent and control bacterial infection. Aim. The aim of this study was to evaluate the preclinical evidence on the impact of animal peptides on skin wound healing. In addition, we carried out a critical analysis of the studies’ methodological quality. Main Methods. This systematic review was performed according to the PRISMA guidelines, using a structured search on the PubMed-Medline, Scopus, and Web of Science platforms to retrieve studies published until August 25, 2020 at 3 : 00 pm. The studies included were limited to those that used animal models, investigated the effect of animal peptides with no association with other compounds on wound healing, and that were published in English. Bias analysis and methodological quality assessments were examined through the SYRCLE’s RoB tool. Results. Thirty studies were identified using the PRISMA workflow. In general, animal peptides were effective in accelerating skin wound healing, especially by increasing cellular proliferation, neoangiogenesis, colagenogenesis, and reepithelialization. Considering standardized methodological quality indicators, we identified a marked heterogeneity in research protocols and a high risk of bias associated with limited characterization of the experimental designs. Conclusion. Animal peptides show a remarkable healing potential with biotechnological relevance for regenerative medicine. However, rigorous experimental approaches are still required to clearly delimit the mechanisms underlying the healing effects and the risk-benefit ratio attributed to peptide-based treatments

Plant Extracts in the Bone Repair Process: A Systematic Review

Bone lesions are an important public health problem, with high socioeconomic costs. Bone tissue repair is coordinated by an inflammatory dynamic process mediated by osteoprogenitor cells of the periosteum and endosteum, responsible for the formation of a new bone matrix. Studies using antioxidant products from plants for bone lesion treatment have been growing worldwide. We developed a systematic review to compile the results of works with animal models investigating the anti-inflammatory activity of plant extracts in the treatment of bone lesions and analyze the methodological quality of the studies on this subject. Studies were selected in the PubMed/MEDLINE, Scopus, and Web of Science databases according to the PRISMA statement. The research filters were constructed using three parameters: animal model, bone repair, and plant extracts. 31 full-text articles were recovered from 10 countries. Phytochemical prospecting was reported in 15 studies (48.39%). The most common secondary metabolites were flavonoids, cited in 32.26% studies (n=10). Essential criteria to in vivo animal studies were frequently underreported, suggesting publication bias. The animals treated with plant extracts presented positive results in the osteoblastic proliferation, and consequently, this treatment accelerated osteogenic differentiation and bone callus formation, as well as bone fracture repair. Possibly, these results are associated with antioxidant, regenerative, and anti-inflammatory power of the extracts. The absence or incomplete characterization of the animal models, treatment protocols, and phytochemical and toxicity analyses impairs the internal validity of the evidence, making it difficult to determine the effectiveness and safety of plant-derived products in bone repair

Antioxidant and Anti-Inflammatory Potential of <i>Brassica oleracea</i> Accelerates Third-Degree Burn Healing in Rats

Burns account for more than 265,000 deaths per year in the world, mainly associated with infections. Therefore, the shorter the healing time, the better the prognosis. Based on this, the objective of the present study was to investigate the effect of an ointment based on Brassica oleracea var. capitata extract in the third-degree healing process. Twenty-five male Wistar rats (335 ± 16 g, three months of life) were individualized in cages with food and water ad libitum. After anesthesia, two circular third-degree burn wounds (12 mm in diameter) were made on the animals, which were randomly separated into five treatments (n = 5/group), i.e., SAL: saline solution 0.9%; OV: ointment vehicle; SS: silver sulfadiazine 1%; PB1: 10% B. oleracea extract; and PB2: 20% B. oleracea extract. The animals were treated with the ointment daily for eight days. Every four days, the area and the wound contraction index were evaluated. Tissue samples were taken for histopathological analysis (cellularity, blood vessels, and extracellular matrix components) and analysis of oxidative/nitrosative status (antioxidant enzymes, lipid, and protein oxidation markers, as well as nitric oxide (NO) and hydrogen peroxide (H2O2)). The ointment based on B. oleracea var. capitata at 10 and 20% concentrations increased the number of cells, blood vessels, and fibrous components of the extracellular matrix and the activity of antioxidant enzymes, promoting a fast and efficient cutaneous repair in third-degree burn wounds

Levels of transforming growth factor beta (TGF-β), Interleukin 10 (IL-10) and tumor necrosis factor (TNF-α) in scar tissue of rats treated with <i>Strychnos pseudoquina</i> extract.

<p>F1: treated tissue after seven days, F2: treated tissue, after 14 days. Sal: saline solution, OV: vehicle, SS: silver sulfadiazine, LE 5: <i>S</i>. <i>pseudoquina</i> 5%; LE 10: <i>S</i>. <i>pseudoquina</i> 10%. Data represented as mean ± SD. *^† p<0.05, statistical difference between treatments: * vs. Sal and OV, *^†vs. Sal, OV and SS (Kruskal-Wallis test).</p

Doxycycline hyclate may damage testicular germinal epithelium in Wistar rats

Broad-spectrum antimicrobial doxycycline acts as an inhibitor of protein synthesis and it is widely used in the clinical treatment of various infections by microorganisms that are sensitive to the drug, as well as in animal feed. Its liposolubility guarantees its high tissue bioavailability, being associated with several biochemical changes in the organism and potentially adverse effects on reproduction. This study aims to evaluate the effects of the action of doxycycline on spermatogenesis to provide a complete analysis of the tubular and interstitial compartments and to identify possible changes in the testicular parenchyma. Adult male Wistar rats were divided into three groups: one control (water), and two treated with doxycycline at the doses of 10mg/kg and 30mg/kg, for 30 days. After euthanasia and sample processing, the following parameters were evaluated: a) tubular diameter and height of the seminiferous epithelium; b) volumetric proportions (%) and volumes (mL) of the components of the testicular parenchyma; c) counting testicular germ cell populations; d) evaluation of cell viability. The results of the comparative evaluation between the experimental groups demonstrated a significant increase in the diameter and area of the tubular lumen and a reduction in the count of spermatogonia in the experimental group that received doxycycline hyclate at a dose of 30mg/kg. In the same experimental group, an increase in the overall yield of spermatogenesis was found as a consequence of the increase in the mitotic index