Development of lamellar gel phase emulsion containing marigold oil (Calendula officinalis) as a potential modern wound dressing.

Appropriate therapeutics for wound treatments can be achieved by studying the pathophysiology of tissue repair. Here we develop formulations of lamellar gel phase (LGP) emulsions containing marigold (Calendula officinalis) oil, evaluating their stability and activity on experimental wound healing in rats. LGP emulsions were developed and evaluated based on a phase ternary diagram to select the best LGP emulsion, having a good amount of anisotropic structure and stability. The selected LGP formulation was analyzed according to the intrinsic and accelerated physical stability at different temperatures. In addition, in vitro and in vivo studies were carried out on wound healing rats as a model. The LGP emulsion (15.0% marigold oil; 10.0% of blend surfactants and 75.0% of purified water [w/w/w]) demonstrated good stability and high viscosity, suggesting longer contact of the formulation with the wound. No cytotoxic activity (50-1000 µg/mL) was observed in marigold oil. In the wound healing rat model, the LGP (15 mg/mL) showed an increase in the leukocyte recruitment to the wound at least on days 2 and 7, but reduced leukocyte recruitment after 14 and 21 days, as compared to the control. Additionally, collagen production was reduced in the LGP emulsion on days 2 and 7 and further accelerated the process of re-epithelialization of the wound itself. The methodology utilized in the present study has produced a potentially useful formulation for a stable LGP emulsion-containing marigold, which was able to improve the wound healing process.

Genistein prevents ultraviolet B radiation-induced nitrosative skin injury and promotes cell proliferation.

Nitric oxide (NO) levels increase considerably after 24h of exposure of skin to ultraviolet B (UVB) radiation, which leads to nitrosative skin injury. In addition, increased NO levels after exposure to UVB radiation are associated with inhibition of cell proliferation. Compared to the UV-control group, UV-genistein at 10 mg/kg (UV-GEN10) group showed tissue protection, decreased lipid peroxide and nitrotyrosine formation, and low CAT activity. Furthermore, NO levels and iNOS labeling remained high. In this group, the reduction in lipid peroxides and nitrotyrosine was accompanied by upregulation of cell proliferation factors (Ki67 and PCNA), which indicated that prevention of nitrosative skin injury promoted cell proliferation and DNA repair. Genistein also prevented nitrosative events, inhibited ONOO(-) formation, which leads to tissue protection and cell proliferation. The UV-GEN15 group did not result in a greater protective effect compared to that with UV-GEN10 group. In the UV-GEN15 group, histological examination of the epidermis showed morphological alterations without efficient protection against lipid peroxide formation, as well as inhibition of Ki67 and PCNA, and VEGF labeling, which suggested inhibition of cell proliferation. These results help to elucidate the mechanisms underlying the photoprotective effect of genistein and reveal the importance of UVB radiation-induced nitrosative damage.

Classification of color images of dermatological ulcers.

We present color image processing methods for the analysis of images of dermatological lesions. The focus of the present work is on the application of feature extraction and selection methods for classification and analysis of the tissue composition of skin lesions or ulcers, in terms of granulation (red), fibrin (yellow), necrotic (black), callous (white), and mixed tissue composition. The images were analyzed and classified by an expert dermatologist into the classes mentioned above. Indexing of the images was performed based on statistical texture features derived from cooccurrence matrices of the RGB (Red, Green, and Blue), HSI (Hue, Saturation, and Intensity), L*a*b*, and L*u*v* color components. Feature selection methods were applied using the Wrapper algorithm with different classifiers. The performance of classification was measured in terms of the percentage of correctly classified images and the area under the receiver operating characteristic curve, with values of up to 73.8% and 0.82, respectively.

Antimicrobial activity of copaíba (Copaifera langsdorffii) oleoresin on bacteria of clinical significance in cutaneous wounds / Atividade antimicrobiana do óleo-resina de copaíba (Copaifera langsdorffii) em bactérias de significância clínica em úlceras cutâneas

The present study was designed to evaluate the in vitro antimicrobial activity of Copaifera langsdorffii oleoresin, which has been used in folk medicine as an anti-inflammatory, antibacterial, healing among others. The oleoresin was tested against Gram-positive (Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis) and Gram-negative (Pseudomonas aeruginosa and Escherichia coli) bacteria related to infections in cutaneous wounds. Antimicrobial activity was determined by the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Copaiba oleoresin showed antimicrobial activity only against the Gram-positive bacteria with MIC of 200 µg/mL, 400 µg/mL and 1100 µg/mL for S. aureus, S. pyogenes and E. faecalis, respectively. MBC values were the same as MIC for S. aureus and S. pyogenes and for E. faecalis it was 1200 µg/mL. Considering that infection significantly impairs the wound healing process, we believe that the use of copaiba oleoresin as a component of a topical formulation could be a valuable adjunct in the treatment of infected wounds, mainly in the case of wounds infected by Gram-positive microorganisms.

Este trabalho avaliou a atividade antimicrobiana in vitro do óleo-resina da Copaifera langsdorffii, o qual vem sendo utilizado há muitos anos na medicina tradicional popular, principalmente devido às suas propriedades antiinflamatórias, antibacterianas, cicatrizante entre outras. O óleo-resina foi testado em bactérias Gram-positivas (Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis) e Gram-negativas (Pseudomonas aeruginosa e Escherichia coli) relacionadas com infecções de úlceras cutâneas. A atividade antimicrobiana foi determinada pelos testes da Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM). O óleo-resina apresentou atividade antimicrobiana in vitro apenas para as bactérias Gram-positivas, com valores de CIM de 200 µg/mL, 400 µg/mL e 1100 µg/mL para S. aureus, S. pyogenes e E. faecalis, respectivamente. Os valores de CBM foram os mesmos que os valores de MIC para S. aureus e S. pyogenes. O valor de CBM para o microrganismo E. faecalis foi de 1200 µg/mL. Considerando que a presença de infecção significativamente impede o processo normal de cicatrização de úlceras cutâneas, acreditamos que o óleo-resina de copaíba, utilizado como componente de formulações tópicas, poderia ser um adjunto importante no tratamento de úlceras cutâneas infectadas, principalmente nos casos de infecção por microrganismos Gram-positivos.

The inflammatory stimulus of a natural latex biomembrane improves healing in mice

The aim of the present study was to compare healing obtained with biomembranes with the natural healing process (sham) using biochemical and immunohistological assays. C57BL/6 mice were divided into 4 groups of 15 mice each and received different subcutaneous implants: natural latex biomembrane (NLB), denatured latex (DL), expanded polytetrafluorethylene (ePTFE), or sham. On the 2nd, 7th, and 14th days post-treatment, 5 mice per group were sacrificed and biopsied for the following measurements: oxidative stress based on malondialdehyde (MDA), myeloperoxidase (MPO) and hydrogen peroxide by the method of ferrous oxidation-xylenol orange (FOX), as well as glutathione and total proteins; histological evaluation to enumerate inflammatory cells, fibroblasts, blood vessels, and collagen, and immunohistochemical staining for inducible nitric oxide synthase, interleukin-1β, vascular endothelial growth factor (VEGF), and transforming growth factor-β1 (TGF-β1). On day 2 post-treatment, NLB stimulated a dense inflammatory infiltrate mainly consisting of polymorphonuclear cells, as indicated by increased MPO (P < 0.05), but oxidative stress due to MDA was not observed until the 7th day (P < 0.05). The number of blood vessels was greater in NLB (P < 0.05) and DL (P < 0.05) mice compared to sham animals on day 14. NLB induced fibroplasia by day 14 (P < 0.05) with low expression of TGF-β1 and collagenesis. Thus, NLB significantly induced the inflammatory phase of healing mediated by oxidative stress, which appeared to influence the subsequent phases such as angiogenesis (with low expression of VEGF) and fibroplasia (independent of TGF-β1) without influencing collagenesis.

The inflammatory stimulus of a natural latex biomembrane improves healing in mice.

The aim of the present study was to compare healing obtained with biomembranes with the natural healing process (sham) using biochemical and immunohistological assays. C57BL/6 mice were divided into 4 groups of 15 mice each and received different subcutaneous implants: natural latex biomembrane (NLB), denatured latex (DL), expanded polytetrafluorethylene (ePTFE), or sham. On the 2nd, 7th, and 14th days post-treatment, 5 mice per group were sacrificed and biopsied for the following measurements: oxidative stress based on malondialdehyde (MDA), myeloperoxidase (MPO) and hydrogen peroxide by the method of ferrous oxidation-xylenol orange (FOX), as well as glutathione and total proteins; histological evaluation to enumerate inflammatory cells, fibroblasts, blood vessels, and collagen, and immunohistochemical staining for inducible nitric oxide synthase, interleukin-1ß, vascular endothelial growth factor (VEGF), and transforming growth factor-ß1 (TGF-ß1). On day 2 post-treatment, NLB stimulated a dense inflammatory infiltrate mainly consisting of polymorphonuclear cells, as indicated by increased MPO (P < 0.05), but oxidative stress due to MDA was not observed until the 7th day (P < 0.05). The number of blood vessels was greater in NLB (P < 0.05) and DL (P < 0.05) mice compared to sham animals on day 14. NLB induced fibroplasia by day 14 (P < 0.05) with low expression of TGF-ß1 and collagenesis. Thus, NLB significantly induced the inflammatory phase of healing mediated by oxidative stress, which appeared to influence the subsequent phases such as angiogenesis (with low expression of VEGF) and fibroplasia (independent of TGF-ß1) without influencing collagenesis.

Retrospective analysis of leg ulcers casesat the university hospital, Faculty ofMedicine of Ribeirão Preto, Universityof São Paulo (1991-2001)

Introduction: Leg ulcers are a syndrome characterized by loss of tegument that affect the lower extremities usually caused by vascular systemdysfunction, representing a high morbidity chronic disease especially among the aged. This study determined the epidemiological characteristics ofleg ulcer patients seen at Clinical Hospital Ribeirão Preto from 1991 to 2001, and their main etiologies and the resolving nature of the treatmentsapplied were investigated.Method: This was a descriptive retrospective study (case series) analyzing 199 medical records of leg ulcer patients, coded by the Medical Recordsfrom 1991 to 2001.Results: 51% of the patients were women and 49% men, with respective median ages of 65 and 66 years, and 81% were white. Etiologically, 49%were classified as venous ulcers, 15% arterial, 9% neuropathic, 2% anemic, and 20% were not classified. Approximately 40% of the patients hadbeen wounded for more than 1 year, 13% died, 40% discharged, and 45% evolved with chronic disease. Among the discharged patients, 64% ofthose with arterial ulcers and 80% of those with neurotrophic ulcers underwent amputation of the segment involved. Regarding venous ulcers, only37% of the patients were discharged, after local dressing in 56% of cases and after skin graft in only one.Comment: Leg ulcer disease was found to be an important chronic disease among the patients seen at the Clinical Hospital Ribeirão Preto from 1991to 2001, with severe consequences regarding the arterial and neurotrophic ulcers, although the etiologic diagnosis was unknown or neglected in20.1% of the cases (AU)

Introdução: Úlcera de perna é uma síndrome caracterizada por perda circunscrita ou irregular do tegumento, que acomete extremidades dos membrosinferiores cuja causa geralmente está na disfunção do sistema vascular. Doença crônica de alta morbidade principalmente na população idosa. Oestudo busca características epidemiológicas dos pacientes com úlceras de perna atendidos no Hospital das Clínicas de Ribeirão Preto, de 1991 a2001, conhecer suas principais etiologias e a resolubilidade dos tratamentos realizados.Metodologia: Estudo descritivo e retrospectivo (série de casos), da análise de 199 prontuários médicos dos pacientes com úlceras de perna, codificadospelo Arquivo Médico, de 1991 a 2001.Leg ulcers are a syndrome characterized by the circumscribedor irregular loss of tegument (dermis or epidermis)that may reach subcutaneous tissue and underlying tissues.The disease involves the lower extremities and thecause usually is related to some dysfunction of the arterialor venous vascular system.[1] The classification may bebased on etiology and therefore leg ulcers are divided intothose due to venous insufficiency (venous ulcers), to arterialinsufficiency (arterial ulcers), to neuropathy (neuropathiculcer/diabetic foot), to red blood cell abnormalities(anemic ulcers), to obliterating thromboangiitis, as well asthose due to causes such as trauma, neoplasia, infections,panniculitis, and pyoderma gangrenosum.[1-4] Among themain predisposing factors are age (elderly individuals), sex(females), obesity, profession (prolonged standing up), vulnerabilityof the legs to traumas and infections, (..) (AU)