Heat shock proteins (HSP): dermatological implications and perspectives

In recent years, several studies have demonstrated the protective effect of Heat Shock Proteins (HSP) on different organs and tissues under stressful conditions. However, most research explores the performance of those molecular chaperones during immune responses or pathological conditions like cancer, whereas the number of studies related to the performance of HSPs in the skin during diverse natural or physiopathological conditions is very low. Therefore, the aim of this article was to summarize the main concepts concerning the expression and performance of HSPs, from analysis of current medicine and cosmetics publications, as well as exploring the importance of these proteins in the dermatological area in physiological events such as cutaneous aging, skin cancer and wound healing and to present final considerations related to biotechnology performance in this area.

Nitric oxide release using natural rubber latex as matrix

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of extracts of Stryphnodendron polyphyllum Mart. and Stryphnodendron obovatum Benth. on the cicatrisation of cutaneous wounds in rats

Stem bark of the two species Stryphnodendron polyphyllum Mart. and Stryphnodendron obovatum Benth., Leguminosae, was investigated for wound healing, antibacterial and antioxidant activity. These plants contain 12 and 19% tannins in their stem bark, respectively, and are widely used in traditional medicine in Brazil. The total content of phenolics of the crude extract (CE) of Stryphnodendron obovatum was 76.95 +/- 2.98% (CV = 3.87%) and of the ethyl-acetate fraction (EAF) was 89.13 +/- 0.34% (CV = 0.38%); whereas in Stryphnodendron polyphyllum the CE phenolics content was 51.62 +/- 1.53% (CV = 2.96%) and the EAF phenolics content was 59.00 +/- 1.91% (CV = 3.24%). The tannin content of CE from Stryphnodendron obovatum [36.58 +/- 0.35% (CV = 0.98%)] was about 11% higher than in CE from Stryphnodendron polyphyllum [25.43 +/- 0.96% (CV = 3.77%)]. The difference between the species was even greater in the EAF: in Stryphnodendron obovatum the EAF phenolics content was 55.01 +/- 0.36% (CV = 0.65%), whereas in Stryphnodendron polyphyllum the content was 36.16 +/- 0.42% (CV = 1.16%). The healing effect of ointments containing 2.5% crude lyophilised extract (PCE) and 2.5% ethyl-acetate lyophilised fraction (PEA) of the stem bark of Stryphnodendron polyphyllum and Stryphnodendron obovatum was studied in cutaneous wounds of Wistar((R)) rats after 4, 7 and 10 days of treatment. Epithelial cell proliferation in the area of re-epithelialisation of the wounds was evaluated by counting the metaphases blocked by vincristine sulfate. With PCE an increase in epidermal growth was observed after 4 and 7 days of treatment with Stryphnodendron polyphyllum, and after 7 and 10 days of treatment with Stryphnodendron obovatum. Wounds treated with PEA of Stryphnodendron obovatum showed increased epidermal growth only 4 days after the treatment, for Stryphnodendron polyphyllum, epidermal growth was observed after 4 and 7 days of treatment. Both the CE and the EAF fractions of Stryphnodendron polyphyllum and Stryphnodendron obovatum showed antibacterial activity against Staphylococcus aureus with MIC values of 125 and 250 mu g/ml, respectively. Gram-negative bacteria tested were not inhibited by extracts and fractions at concentrations > 1000 mu g/ml. The antioxidant activity through reduction of the DPPH radical in TLC, confirmed the anti-radical properties of these extracts in both species. CE and EAF of both species showed a radical scavenging activity (RSA) and protected DPPH from discolouration, already at 0.032 mu g/ml. The extract from Stryphnodendron polyphyllum were more effective than those Stryphnodendron obovatum, although the former had a lower tannin content. (c) 2005 Elsevier B.V.. All rights reserved.

Effect of Glycolic Extract of Dillenia indica L. Combined With Microcurrent Stimulation on Experimental Lesions in Wistar Rats

This study evaluated the wound healing activity of a glycolic extract of Dillenia indica (GED) prepared from the mature fruits of the plant applied alone or in combination with microcurrent stimulation to skin wounds surgically induced on the back of Wistar rats. Methods. The animals were randomly divided into six groups: (A) negative control group; (B) group receiving microcurrent application (MC; [10 mu A/2 mins]); (C) group treated with GED; (D) group treated with an emulsion containing GED; (E) group treated with GED and MC, and (F) group treated with the emulsion containing GED and MC. Tissue samples were obtained 2, 6, and 10 days after injury and underwent structural and morphometric analysis, Results. There were observed differences in wound healing among the various treatments when compared to the control group. The combination of microcurrent plus extract or microcurrent plus emulsion containing GED was advantageous in all of the studied parameters (P < 0.05) when compared to the other groups with positive effects seen regarding newly formed tissue, number of fibroblasts, and number of newly formed blood vessels. The morphometric data confirmed the structural findings. Conclusion. Microcurrent application alone or combined with GED exerted significant effects on wound healing in this experimental model. This was probably due to the efficacy of microcurrent application since the extract alone did not significantly accelerate the healing process. D indica fruit extract most likely participates in the wound healing process as a result of its anti-inflammatory properties.

Efeitos do tratamento com Hidrogel na cicatrização de úlceras venosas de membros inferiores: revisão sistemática

Chronic venous disease (CVD) is evident among the chronic diseases and affects the elderly population and primarily is responsible for leg ulcers in this population. The use of dressings in the care of a venous ulcer is a fundamental part of the treatment for healing, however, evidence to assist in choosing the best dressing is scarce. The main objective of this study was to evaluate the effectiveness of treatment with hydrogel in the healing of venous ulcers using search methods, synthesis of information and statistical research through a systematic review and meta-analysis. Randomized controlled trials were selected in the following databases: CENTRAL; DARE; NHS EED; MEDLINE; EMBASE; CINAHL. Beyond these databases three websites were consulted to identify ongoing studies: ClinicalTrials.gov, OMS ICTRP e ISRCTN. The primary outcomes were analyzed: complete wound healing, incidence of wound infection and the secondary were: changes in ulcer size, time to ulcer healing, recurrence of ulcer, quality of life of participants, pain and costs of treatment. Four studies are currently included in the review with a total of 250 participants. The use of hydrogel appears to be superior to conventional dressing, gauze soaked in saline, for the healing of venous leg ulcers; 16/30 patients showed complete healing of ulcers (RR 5,33, 95%CI [1,73,16,42]). The alginate gel was shown to be more effective when compared to the hydrogel dressing in reduction of the wound area; 61,2% (± 26,2%) with alginate e 19,4% (± 24,3%) with hydrogel at the end of four weeks of treatment. Manuka honey has shown to be similar to the hydrogel dressings in percentage of area reduction. This review demonstrated that there is no evidence available about the effectiveness of the hydrogel compared to other types of dressings on the healing of venous leg ulcers of the lower limbs, thus demonstrating the need of future studies to assist health professionals in choosing the correct dressing.

Avaliação clínica do efeito da irradiação pós-opratória do laser de baixa intensidade na cicatrização de gengivoplastias em humanos

Many studies have been made to understand the process of tissular cicatrization, as well as the possible effects of laser therapy in the wound healing. However, the influence of low frequency laser irradiation in the repairing process is not completely understood. Our study has the purpose to assess clinically the effect of postoperative irradiation of the low frequency laser in humans, and the gingival repairing process postgingivoplasty performed with the extern bevel technique. Twenty-four patients with inflammatory gingival hyperplasia were enrolled in this study, which did not reduce with basic periodontal procedures, and patients with melanin pigmentation, with esthetic indications. After surgery the test group, randomly selected by a drawing, received laser application with energy density of 4 J/cm2, immediately after surgery and each 48 hours, during a week, with a total of 4 sections. The control group did not receive irradiation. The visual clinical analyses were performed by a single blind examiner, in the 2nd, 4th, 6th, 8th, 15th and 21st days post surgery. For statistic analyses of the data was used a Q-square test. Concerning the color, the results showed a better wound healing during days 6 to 8. when assessed the degree of progress of surgical wound, the results showed that the test group had a better cicatrization compared with the control group in the 2nd, 6th, 8th and 15th days post surgery, and at the 21st day both groups had the same results. Our results confirm that the laser had clinical influence in the repairing process after gingivoplasty surgery during days 2 to 15 post surgery

Influência da laserterapia na proliferação de células-tronco do ligamento periodontal humano

Low level laser irradiation (LLLI) has been used in Dentistry to promote wound healing and tissue regeneration. The literature shows a positive effect of LLLI on cell proliferation, but little is known about their effectiveness in promoting stem cells proliferation. The aim of this study was to evaluate the effect of LLLI on the proliferative rate of human periodontal ligament stem cells. Extracts of periodontal ligament were isolated from two third molars removed by surgical and/or orthodontic indication. After enzymatic digestion, the cells were grown in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. On the third subculture, the cells were irradiated with a InGaAlP-diode laser, using two different energy densities (0,5J/cm 2 - 16 seconds and 1,0J/cm² - 33 seconds), with wavelength of 660nm and output power of 30mW. A new irradiation, using the same parameters, was performed 48h after the first. A control group (non irradiated) was kept under the same experimental culture conditions. The Trypan blue exclusion test and the mitochondrial activity of the cells measured by MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] essay were performed to assess the cell proliferation in the intervals of 0, 24, 48 e 72 h after irradiation. The data of cell counts were submitted to nonparametrical statistical tests (Kruskal-Wallis and Mann-Whitney), considering a confidence interval of 95%. DAPI (4 -6-Diamidino-2-phenylindole) staining of the cells was performed at 72h interval to evaluate possible nuclear morphological changes induced by LLLI. The results of this study show that the energy density of 1,0 J/cm² promoted greater cell proliferation compared to the other groups (control and 0,5 J/cm²) at intervals of 48 and 72h. The mitochondrial activity measured by MTT essay showed similar results to the Trypan blue cell counting test. The group irradiated with 1,0J/cm² exhibited a significantly higher MTT activity in the intervals of 48 and 72h, when compared to the group irradiated with 0,5J/cm². No nuclear morphological change was observed in the cells from the three groups studied. It is concluded that LLLI has stimulatory effects on the proliferation of human periodontal ligament stem cells. Therefore, the use of laser irradiation in this cell type may be important to promote future advances in periodontal regeneration

Efeito do diclofenaco de sódio na cicatrização da parede abdominal de ratos. Estudo histopatológico, da força de ruptura e do colágeno tecidual

Com o objetivo de estudar os efeitos do diclofenaco sódico sobre a cicatrização da parede abdominal de ratos, foram utilizados 80 animais da linhagem Wistar divididos em dois grupos: Grupo 1: formado por 40 animais submetidos à laparotomia mediana e à injeção intramuscular de soro fisiológico durante quatro dias. Grupo 2: formado por 40 animais submetidos à laparotomia mediana e à injeção intramuscular de diclofenaco sódico durante quatro dias. Animais de ambos os grupos foram analisados no 5°, 7°, 14° e 21° dias de pós-operatório, correspondendo, respectivamente à M1, M2, M3 e M4. em cada momento foram estudados 10 animais e os parâmetros analisados foram a evolução clínica, a força de ruptura, estudo histológico e o conteúdo de colágeno tecidual da parede abdominal. Os animais do grupo tratado apresentaram como complicações, deiscência e/ou hérnia incisional, perda ponderal e taxa de mortalidade, complicações estas não evidenciadas no grupo controle. Observamos também neste grupo, diminuição da força de ruptura no 7° e 14° dia de pós-operatório e diminuição da concentração de colágeno tecidual no 5°, 7° e 14° dia de pós-operatório. Ambos os parâmetros retornaram a valores normais no 21° dia de pós-operatório. Quanto ao estudo histológico, concluímos que a cicatriz da parede abdominal dos animais tratados com D.S. apresentam retardo do processo cicatricial em relação aos seus controles, caracterizado por uma menor fibrogênese, menor densidade de fibras colágenas, além de um número maior de complicações, como microabscessos, em torno dos fios de sutura.

Efeito da heparina sódica e da enoxaparina na consolidação de fratura da tíbia no rato: avaliação clínica e anatomopatológica e biomecânica

Foi realizado estudo experimental em ratos para avaliar o efeito do anticoagulante na consolidação óssea, conforme critérios clínicos, anatomopatológicos e biomecânicos. Manualmente, após perfuração do osso, foi produzida fratura aberta, na diáfise da tíbia direita, mantida sem imobilização, em 72 ratos machos da linhagem Wistar, com 60 dias de idade e peso médio de 242 gramas. Doze horas após a fratura, foi iniciado tratamento anticoagulante, mantido por 28 dias. Via subcutânea, um grupo recebeu heparina sódica na dose de 200UI/Kg de 12 em 12 horas, enquanto outro, recebeu enoxaparina na dose de 1mg/Kg de 12 em 12 horas, doses preconizadas para tratamento do tromboembolismo em humanos. O terceiro grupo, controle, recebeu água destilada. Durante o experimento, os animais foram avaliados clinicamente e após 28 dias, sacrificados. Nos animais dos três grupos, a evolução clínica foi semelhante. Mediante análise anatomopatológica efetuada por estudo descritivo e quantitativo, foi observada presença de fibrose, cartilagem e osso igualmente nos três grupos, sempre com predomínio de tecido ósseo. O estudo biomecânico, realizado por intermédio de ensaios de flexão, demonstrou coeficiente de rigidez e carga máxima semelhantes nos três grupos. Nenhuma diferença clínica, anatomopatológica e biomecânica foi encontrada, resultando todas as fraturas em consolidação de acordo com os critérios adotados, concluindo-se, portanto, que a heparina sódica e a enoxaparina nas doses, via e tempo de administração utilizados não interfiriram na consolidação da fratura da tíbia do rato.

End-to-end arterial anastomosis with fibrin glue in larger arteries: histology, hydroxyproline concentration and tensile strength study in carotids of rabbits

A anastomose arterial término-terminal é demorada, requer tempo prolongado de oclusão vascular e esta associada a necrose focal, infiltração leucocitária e, conseqüentemente, à fibrose e calcificação da parede arterial. A cola de fibrina é uma alternativa para a anastomose microvascular e pode evitar estas alterações com menor aderência aos tecidos vizinhos e melhor coaptação das bordas arteriais. OBJETIVO: Comparar o processo cicatricial de anastomoses convencionais com anastomoses feitas com cola de fibrina em artérias maiores. MÉTODOS: em 22 coelhos, ambas carótidas foram seccionadas transversalmente e reconstruídas por meio de anastomose término-terminal com 4 pontos simples de reparo e cola de fibrina de um lado (G1), e com 8 pontos separados do outro lado (G2). Após 3 e 15 dias, os animais foram destinados aleatoriamente para estudo de força tênsil concentração de hidroxiprolina (8 animais) e avaliação histológica das anastomoses (3 animais). As lâminas histológicas foram coradas pelo HE Masson e Picrossirius polarização (PSP). RESULTADOS: Após 3 e 15 dias a força tênsil aumenta em ambos os grupos, de 280,0± 32,6g para 432,2± 131,2g no Grupo 1 e de 221,4± 72,4g para 452,2± 132,0g no Grupo 2; sem diferença estatística entre os grupos em cada período. A concentração de hidroxiprolina expressa como razão hidroxiprolina/proteína, variou de 0,0816± 0,0651 para 0,0622± 0,0184 no Grupo 1 e de 0,0734± 0,0577 para 0,0460± 0,0271 no Grupo 2; sem diferença estatística entre os períodos e grupos. Os estudos histológicos mostraram discreto aumento das reações de inflamação e reparação no Grupo 2. A técnica PSP mostrou predomínio do colágeno tipo I em relação do colágeno tipo II nas anastomoses de ambos os grupos, sem diferença expressiva entre esses grupos. CONCLUSÃO: A anastomose com a cola de fibrina foi menos lesiva para a parede arterial do que a anastomose convencional. Mesmo usando menos pontos, as características de força tênsil e de cicatrização da anastomose com cola de fibrina foram similares em ambos os grupos. Os tempos de realização das anastomoses foram significativamente maiores do que na anastomose convencional.

Low-level laser therapy on the treatment of oral and cutaneous pemphigus vulgaris: case report

Pemphigus vulgaris is a chronic autoimmune mucocutaneous disease that initially is manifested by painful intraoral erosions and ulcers which spread to other mucosa and the skin, generally more than 5 months after oral lesion manifestation. The treatment consists of prednisone alone or in combination with an immunosuppressive agent, and the clinical response is perceived within 2 to 4 weeks. Low-level laser therapy has been effective in accelerating the healing of injured tissue, thus inducing cell proliferation and increasing ATP, nucleic acid, and collagen synthesis. We reported two cases of pemphigus vulgaris that received systemic treatment associated with low-level laser therapy for oral and cutaneous lesions. We observed prompt analgesic effect in oral lesions and accelerated healing of oral and cutaneous wounds. Therefore, the present report suggests LLLT as a noninvasive technique that should be considered as an adjuvant therapy in oral and skin disorders in patients with PV.

Low-level laser therapy for the treatment of epidermolysis bullosa: A case report

Epidermolysis bullosa (EB) is a rare group of diseases characterized by skin fragility. There is no specific treatment, short of protection from trauma, currently available for these patients. Low-level laser therapy (LLLT) was effective as an analgesic and in accelerating cutaneous wound healing after six sessions of therapy in a child with dystrophic EB with cutaneous scarring and blisters on the limbs and trunk.

Tratamento de feridas cutâneas extensas usando tecido dérmico acelular porcino com e sem cobertura impermeável

FUNDAMENTOS: O tecido dérmico acelular porcino é alternativa para o tratamento de feridas cutâneas. OBJETIVOS: Avaliar a resposta clínica e inflamatória do implante de tecido dérmico acelular porcino, com e sem cobertura impermeável. MÉTODOS: Estudo pareado, longitudinal, criando-se duas feridas cutâneas no dorso de 16 ratos (quatro animais/grupo), em que foi implantado tecido dérmico acelular coberto ou não por impermeável. Os animais foram avaliados e sacrificados sete, 15, 30 e 60 dias após a cirurgia, sendo removidos os tecidos acelulares e adjacentes para avaliação histológica e morfométrica. RESULTADOS: A cobertura impermeável permaneceu sobre o tecido acelular porcino até cerca de 15 dias. O grupo sem impermeável apresentou maior desidratação, com crosta fibrinoleucocitária, edema e reação inflamatória na derme. Sessenta dias após a cirurgia, animais do grupo sem impermeável ainda apresentavam ulcerações, afinamento do epitélio e ausência de queratina, enquanto nos do grupo com impermeável a pele já se encontrava normal. CONCLUSÃO: O tecido dérmico acelular porcino com cobertura impermeável apresentou resultados clínicos e histológicos melhores do que os do tecido dérmico acelular porcino sem impermeável para tratamento de feridas cutâneas extensas.

Modelos experimentais de deficiência limbar em coelhos: análise clínica

Objetivo: Comparar em coelhos três modelos experimentais de destruição das células germinativas (CG) do limbo corneano quanto a aspectos clínicos. Métodos: Foram utilizados 54 coelhos, 108 olhos, subdivididos em 3 grupos experimentais: (G1), (G2) e (G3), cada um formado pelos OE de 18 coelhos, submetidos às técnicas experimentais (T1), (T2) e (T3), respectivamente, e um grupo controle, formado pelos 54 olhos contralaterais (OD). Nas três técnicas foi utilizado o n-heptanol. Na T1, o n-heptanol foi aplicado por 5 minutos, para remoção do epitélio límbico. Na T2, além da aplicação do n-heptanol, realizou-se peritomia e remoção da conjuntiva perilímbica até 4 mm do limbo, juntamente com a escarificação do tecido episcleral. Na T3, além dos procedimentos da T2, foi realizada dissecção lamelar do limbo abrangendo aproximadamente 1,5 mm na periferia da córnea e 2 mm na superfície escleral. em todas as córneas dos animais foram estudados seis parâmetros clínicos: neovascularização, perda da transparência, irregularidade da superfície, reparação epitelial, erosão ou defeito epitelial, granuloma e outras lesões corneanas. Resultados: A neovascularização corneana iniciou-se mais precocemente com a T1 e T2; ocorreu em 100% dos casos com as três técnicas, de forma não homogênea, variando de leve a intensa; permaneceu estável a partir do 28º dia até o final do experimento (56º dia), foi maior nos quadrantes superior e inferior e menor nos quadrantes nasal e temporal. A perda da transparência e a irregularidade da superfície corneana foram menores com a T1 que com a T2 e T3, que foram similares entre si. Houve, nas três técnicas experimentais, latência de três dias para o início da reepitelização, que se completou com a T1 no 7º dia e com a T2 e T3 no 14º dia. Erosão epitelial corneana e granuloma corneano foram encontradas de forma similar nas três técnicas experimentais. Conclusões: A T2 e T3 mostraram-se adequadas como possíveis modelos de ampla remoção das CG límbicas, levando a resultados similares nos diversos parâmetros estudados. A T1 se mostrou adequada como modelo de remoção parcial do epitélio límbico. Ocorreu conjuntivalização e neovascularização nas três técnicas experimentais.

Evaluation of tissue response to periodontal dressings: Histological study in tooth sockets of rats

Although the use of periodontal dressings is currently limited, there are some indications for their use. Selection of any material that will have direct contact with live tissues, such as periodontal dressings, should be careful in order to allow surgical wound healing. The aim of this study was to evaluate the intensity of inflammatory response and bone formation in tooth sockets of rats after implantation of three periodontal dressings. After removal of the right maxillary incisors of 84 male rats, each tooth socket received implantation of a polyethylene tube, 63 of which were filled with non-eugenol periodontal dressing and the remaining 21 tubes remained empty (control group). Histological evaluation assessed the intensity of inflammatory response and presence and location of bone tissue formation at postoperative periods of 7, 14 and 28 days. Statistical analysis was performed by the Kruskal-Wallis test at 5% significance level. Regarding the inflammatory infiltrate, at 28 days, there was statistically significant difference between one of periodontal dressings and control group (p < 0.05). Analysis of postoperative periods, showed that the control group presented statistically significant reduction in the inflammatory infiltrate comparing the 14- and 28-day periods (p < 0.05). Regarding bone tissue formation, there was difference in control group between the 7- and 28-day periods (p < 0.05). Within the experimental conditions, it may be concluded that no differences were found in the inflammatory response among the groups at 7 and 14 days and that Voco pac (TM) dressing induced a more intensive inflammatory reaction at 28 days.