Avaliação da cicatrização de feridas em dorso de ratos tratados com mitomicina C tópica e posterior reforço com injetável

Estudo experimental em animais. A mitomicina C vem sendo usada como inibidor de fibroblastos, acarretando, com isso, diminuição do processo cicatricial em feridas cirúrgicas. OBJETIVO: Este trabalho visa avaliar o uso de Mitomicina C para diminuir o processo cicatricial, através de seu uso tópico com reforços posteriores injetáveis. MATERIAL E MÉTODOS: Foi usado um modelo de feridas em dorso de ratos, com retirada circular da pele e cicatrização por segunda intenção. Foram usados 18 ratos, divididos em três grupos: controle; com uso tópico; e com reforço de mitomicina C injetável, mensalmente e por 2 meses. Após 3 meses os animais foram sacrificados e as cicatrizes retiradas cirurgicamente e submetidas a estudo histológico. RESULTADOS: Notou-se sob vários critérios que a cicatrização com o uso tópico é menos intensa, mas ao se usar o reforço injetável os parâmetros voltam a ser comparados ao do grupo controle. DISCUSSÃO: Acreditamos que a administração injetável de mitomicina C nas cicatrizes, pela sua elevada característica tóxica, acarreta destruição tecidual e neoformação cicatricial. CONCLUSÕES: A mitomicina C diminui o processo cicatricial quando usada topicamente, mas acarreta aumento da cicatrização quando nestas feridas são feitos reforços injetáveis.

Natural rubber latex used as drug delivery system in guided bone regeneration (GBR)

In this work, we propose natural rubber latex (NRL) membranes as a protein delivery system. For this purpose Bovine Serum Albumin (BSA) was incorporated into the latex solution for in vitro protein delivery experiments. Different polymerization temperatures were used, from -10 to 27 °C, in order to control the membrane morphology. These membranes were characterized by Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM), as well as the Lowry Method to measure the BSA release. SEM and AFM microscopy analysis showed that the number, size and distribution of pores in NRL membranes can be varied, as well as its overall morphology. We have found that the morphology of the membrane is the predominant factor for higher protein release, compared with pore size and number of pores. Results demonstrated that the best drug-delivery system was the membrane polymerized at RT (27 °C), which does release 66% of its BSA content for up to 18 days. Our results indicate that NRLb could be used in the future as an active membrane that could accelerate bone healing in GBR.

Estudo clínico da aplicação de matriz inorgânica de osso associado a peptídeo sintético de adesão celular (MIO/P-15), PepGen P-15®, em lesões periodontais avançadas de cães

Os mecanismos biológicos desenvolvidos para aumentar a qualidade da regeneração óssea e da reparação tecidual de sítios periodontais específicos continuam a ser um desafio e têm sido complementado pela capacidade de adesão celular do colágeno do tipo I, promovida por um peptídeo sintético de adesão celular (P-15), associado a uma matriz inorgânica de osso (MIO) para formar MIO/P-15. O objetivo deste estudo foi avaliar a perda do nível clínico de inserção e a resposta da bolsa periodontal em dentes após 3 e 6 meses da aplicação de enxerto com MIO/P-15. Vinte e um cães do Hospital Veterinário da Universidade de São Paulo foram anestesiados para realização de tratamento periodontal e 132 faces dentais com perda de nível clínico de inserção foram tratadas, sendo que 36,4% (48 faces) receberam o peptídeo de adesão celular e 63,6% (84 faces) compuseram o grupo controle que recebeu tratamento convencional (retalho muco-gengival e aplainamento radicular). O procedimento foi documentado através de radiografia intra-oral e todas as sondagens de bolsas periodontais foram fotografadas. Depois de 3 e de 6 meses, os animais foram re-anestesiados a fim de se obter novas avaliações, radiografias, fotografias e sondagens periodontais. As 48 faces com perda de nível clínico de inserção que receberam material de enxertia apresentaram taxa de 40% de recuperação do nível clínico de inserção após 6 meses. O grupo controle de faces dentais não apresentou alteração do nível clínico de inserção. A face palatina foi a que apresentou melhor taxa de regeneração (40%) e os dentes caninos e molares mostraram as melhores respostas (57,14% e 65%, respectivamente). Não houve sinais de infecção pós-cirúrgica relacionadas à falta de higienização oral dos animais. Pode-se concluir que o MIO/P-15 auxilia na regeneração e re-aderência das estruturas periodontais, incluindo osso alveolar. Sua aplicação mostrou-se fácil e prática e a incidência de complicações pós-cirúrgicas foi baixa. Ainda assim, mais estudos e pesquisas são necessários para que se avalie a quantidade e a qualidade do osso e do ligamento periodontal formados.

Ultra-som pulsado de baixa intensidade em fraturas diafisárias: aplicação clínica em cães

Os efeitos da estimulação ultra-sônica sobre a consolidação óssea têm sido demonstrados por trabalhos experimentais e clínicos. Este estudo teve por objetivo investigar a aplicação clínica do ultra-som pulsado de baixa intensidade como tratamento adjuvante de fraturas diafisárias em cães. Foram utilizados 16 cães de raças variadas, com faixa etária entre sete meses e seis anos, peso corpóreo entre 2,5 e 43kg, portadores de fraturas diafisárias fechadas recentes localizadas no rádio e ulna, fêmur ou tíbia e fíbula, estabilizadas por procedimentos de osteossíntese (fixação esquelética externa, pinos intramedulares ou a associação desses métodos). Os cães foram divididos em dois grupos: fraturas estabilizadas tratadas por ultra-som de baixa intensidade (grupo tratado, n=8); fraturas estabilizadas, não tratadas por estimulação ultra-sônica, (grupo controle, n=8). Os animais foram avaliados por exames clínicos e radiográficos nos períodos pré-operatório, pós-operatório imediato e a cada 30 dias posteriores aos procedimentos cirúrgicos. Realizou-se tratamento com ultra-som pulsado (sinal senoidal com freqüência de 1,5MHz, largura de pulso de 200µs e freqüência de repetição de 1kHz) de baixa intensidade (30mW cm-2), aplicado de modo estacionário no foco de fratura. A terapia ultra-sônica foi realizada 20 minutos por dia, durante 21 dias consecutivos, a partir do período compreendido entre o 1° e o 9° dia pós-operatório. O teste t de Student, empregado na análise estatística, mostrou diferença significante (P<0,001 e alfa=0,05) entre as médias dos parâmetros de tempo para consolidação óssea observadas nos animais dos grupos tratado (média de 67,5 dias) e controle (média de 106 dias). Este protocolo de estimulação ultra-sônica promoveu sinais clínicos e radiográficos acelerados da consolidação óssea nas fraturas tratadas. Os resultados deste estudo sugerem que o ultra-som pulsado de baixa intensidade pode ser indicado como terapia adjuvante de fraturas diafisárias recentes em cães.

Análise colorimétrica e espectroscópica do muco de caracóis terrestres Achatina sp alimentados com ração diferenciada

Foram estudados os efeitos da adição de plantas medicinais de princípios cicatrizantes (Centelha asiática, Papaína e Confrei) na ração controle de caracóis terrestres, para se avaliar a interferência destas plantas na composição do muco glicoprotéico. Foram utilizados 80 caracóis terrestres Achatina sp, baseados em um peso homogêneo (49 e 40 g e idade média de 10 e 19 meses para Achatina fulica e Achatina monochromatica, respectivamente). Os animais foram distribuídos aleatoriamente em oito grupos experimentais: controle Achatina fulica (FC) e Achatina monochromatica (MC), centelha asiática Achatina fulica (FCe) e Achatina monochromatica (MCe), papaína Achatina fulica (FPa) e Achatina monochromatica (MPa) e confrei Achatina fulica (FCo) e Achatina monochromatica (MCo). Água e ração foram fornecidos ad libitum. Ao final de 150 dias de tratamento, os animais foram submetidos à técnica de extração do muco glicoprotéico, por meio do estímulo manual da glândula podal, responsável pela secreção deste muco. Esta metodologia considerou o bem-estar dos animais, uma vez que os mesmos não foram sacrificados e retornaram ao seu sistema de criação. Os mucos foram analisados por meio de testes colorimétricos e espectroscópicos, que constataram alterações semelhantes, porém apresentaram variação significativa em sua composição glicoprotéica.

Fermentation of Groundnut Shell Enzymatic Hydrolysate for Fuel Ethanol Production by Free and Sorghum Stalks Immobilized Cells of Pichia stipitis NCIM 3498

Groundnut shell (GS), after separation of pod, is readily available as a potential feedstock for production of fermentable sugars. The substrate was delignified with sodium sulfite. The delignified substrate released 670 mg/g of sugars after enzymatic hydrolysis (50 degrees C, 120 rpm, 50 hrs) using commercial cellulases (Dyadic Xylanase PLUS, Dyadic Inc. USA). The groundnut shell enzymatic hydrolysate (45.6 g/L reducing sugars) was fermented for ethanol production with free and sorghum stalks immobilized cells of Pichia stipitis NCIM 3498 under submerged cultivation conditions. Immobilization of yeast cells on sorghum stalks were confirmed by scanning electron microscopy (SEM). A maximum of ethanol production (17.83 g/L, yield 0.44 g/g and 20.45 g/L, yield 0.47 g/g) was observed with free and immobilized cells of P. stipitis respectively in batch fermentation conditions. Recycling of immobilized cells showed a stable ethanol production (20.45 g/L, yield 0.47 g/g) up to 5 batches followed by a gradual downfall in subsequent cycles.

Sliding wear of weld layers with application to cane mill shaft rebuilding

Three welding procedures used to rebuild worn shafts in sugar cane mills were analysed: two submerged arc welding processes and one flux cored arc welding (FCAW) process. Sliding wear tests were in accordance with ASTM G 77 standard, using rings of welding material, blocks of bronze SAE 67, and oil as lubricant. The worn surfaces of rings and blocks were analysed by scanning electron microscopy to determine the wear mechanisms. High contact pressure, high operating temperature, and low relative speed were applied in sliding wear tests to match the conditions in sugar cane mills. Transferred material and evidence of adhesive junctions were detected. Additionally, hardened fragments produced abrasive grooves on the worn surfaces. The welding deposits that presented strong adhesion on the worn surface showed higher mass loss than the materials that presented more abrasive characteristics. Plastic mechanical properties were measured and related to the mass loss. The tested materials presented similar hardness but different yield stress and hardening coefficient. A relationship between wear, strain hardening coefficient, and yield stress was found. The welding deposit that presented the highest hardening coefficient showed the highest mass loss, with evidence of severe adhesion on the worn surface.

Resprouting from roots in four Brazilian tree species

Previous studies pointed out that species richness and high density values within the Leguminosae in Brazilian forest fragments affected by fire could be due, at least partially, to the high incidence of root sprouting in this family. However, there are few Studies of the factors that induce root sprouting in woody plants after disturbance. We investigated the bud formation on root cuttings, and considered a man-made disturbance that isolates the root from the shoot apical dominance of three Leguminosae (Bauhinia forficata Link., Centrolobium tomentosum Guill. ex Benth, and Inga laurina (Sw.) Willd) and one Rutaceae (Esenbeckia febrifuga (St. Hit.) Juss. ex Mart.). All these species resprout frequently after fire. We also attempted to induce bud formation on root systems by removing the main trunk, girdling or sectioning the shallow lateral roots from forest tree species Esenbeckia febrifuga and Hymenaea courbaril L. We identified the origin of shoot primordia and their early development by fixing the samples in Karnovsky solution, dehydrating in ethyl alcohol series and embedding in plastic resin. Serial sections were cut on a rotary microtome and stained with toluidine blue O. Permanent slides were mounted in synthetic resin. We observed different modes of bud origin on root cuttings: close to the vascular cambium (C. tomentosum), from the callus (B. forficata and E febrifuga) and from the phloematic parenchyma proliferation (L laurina). Fragments of B. forficala root bark were also capable of forming reparative buds from healing phellogen formed in callus in the bark's inner side. In the attempt of bud induction on root systems, Hymenaea courbaril did not respond to any of the induction tests, probably because of plant age. However, Esenbeckia febrifuga roots formed suckers when the main trunk was removed or their roots were sectioned and isolated from the original plant. We experimentally demonstrated the ability of four tree species to resprout from roots after disturbance. Our results suggest that the release of apical dominance enables root resprouting in the studied species. Rev. Biol. Trop. 57 (3): 789-800. Epub 2009 September 30.

Use of Cassava Peel as Carbon Source for Production of Amylolytic Enzymes by Aspergillus niveus

Aspergillus niveus produced high levels of alpha-amylase and glucoamylase in submerged fermentation using the agricultural residue cassava peel as a carbon source. In static conditions, the amylase production was substantially greater than in the agitated condition. The optimized culture conditions were initially at pH 5.0, 35 degrees C during 48 hours. Amylolytic activity was still improved (50%) with a mixture of cassava peel and soluble starch in the proportion 1:1 (w/w). The crude extract exhibited temperature and pH optima approximately 70 degrees C and 4.5, respectively. Amylase activity was stable for 1 h at 60 degrees C, and at pH values between 3.0 and 7.0. The enzyme hydrolysed preferentially maltose, starch, penetrose, amylose, isomaltose, maltotriose, glycogen and amylopectin, and not hydrolysed cyclodextrin (alpha and beta), trehalose and sucrose. In the first hour of reaction on soluble starch, the hydrolysis products were glucose and maltose, but after two hours of hydrolysis, glucose was the unique product formed, confirming the presence in the crude extract of an alpha-amylase and a glucoamylase.

Photodynamic Therapy Can Be Effective as a Treatment for Herpes Simplex Labialis

Background Data and Objective: Herpes is a common infectious disease that is caused by human herpesviruses. Several treatments have been proposed, but none of them prevent reactivation of the virus. This article describes the use of photodynamic therapy (PDT) as a treatment for herpes lesions, and reports on four cases. Materials and Methods: PDT was used as an adjuvant therapy for the treatment of herpes labialis in four patients. A special type of 0.01% (m/V) of methylene blue solution was applied to the vesicular stage of herpesviral disease and the lesions were irradiated with laser energy (wavelength 660 nm, energy density 120 J/cm(2), output power of 40 mW, 2 min per point, 4.8 J of energy/point, at four points). After 24 h the patients returned and phototherapy was repeated with the same equipment, this time with 3.8 J/cm(2) and 15 mW, for a total dose of 0.6 J. The same procedure was repeated 72 h and 1 wk later. Results: Treatment with low-level laser therapy can be considered as an option in the treatment of herpes labialis, and decreases the frequency of vesicle recurrence and provides comfort for patients. No significant acute side effects were noted and the lesions healed rapidly. Conclusion: Treatment of herpes labialis with PDT was effective, had no side effects, and when associated with laser phototherapy, accelerated the healing process.

Effect of Laser Phototherapy on the Release of TNF-alpha and MMP-1 by Endodontic Sealer-Stimulated Macrophages

Objective: Our aim was to analyze the effect of laser phototherapy on the secretory activity of macrophages activated by interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS), and stimulated by substances leached from an epoxy resin-based sealer (AH-Plus) and a calcium hydroxide-based sealer (Sealapex). Background Data: Laser phototherapy can modulate the inflammatory process, improving wound healing. This type of therapy could be useful for modulating postoperative symptoms seen after endodontic treatment. Materials and Methods: Cytotoxicity was indirectly assessed by measuring mitochondrial activity. Macrophages were stimulated by the leached substances or not (controls), and the groups were then irradiated or not. The secretion of pro-inflammatory cytokines (TNF-alpha and MMP-1) was analyzed using ELISA. Two irradiations at 6-h intervals were done with an As-Ga-Al diode laser (780 nm, 70 mW, spot size 4.0 mm(2), 3 J/cm(2), for 1.5 sec) in contact mode. Results: The sealers were non-cytotoxic to macrophages. The production of TNF-alpha was significantly decreased by laser phototherapy, regardless of experimental group. The level of secretion of MMP-1 was similar in all groups. Conclusion: Based on the conditions of this study we concluded that in activated macrophages, laser phototherapy impairs the secretion of the pro-inflammatory cytokine TNF-alpha, but has no influence on MMP-1 secretion.

High-Intensity Laser and Photodynamic Therapy as a Treatment for Recurrent Herpes Labialis

Objective: The aim of this study was to report the treatment of recurrent herpes labialis (RHL) using a high-intensity laser or methylene blue (MB)-mediated photodynamic therapy (PDT) in combination with low-level laser therapy (LLLT). Materials and Methods: Four clinical cases of patients diagnosed with RHL are described in this report. Two patients were subjected to high-intensity laser therapy (HILT) followed by LLLT, and two patients received MB-mediated PDT, again followed by LLLT. LLLT was conducted at 24, 48, 72 h, and 7 d after HILT or PDT. Patients were followed up after 6 mo. Results: Throughout the follow-up period, all patients reported pain relief and did not show any signs or symptoms of RHL. A favorable healing process was observed in all cases. None of the patients reported pain as a consequence of the treatment. Conclusion: These results suggest that HILT and MB-mediated PDT, in combination with LLLT, may constitute a benefit when treating vesicles in RHL.

Venous Lake of the Lips Treated Using Photocoagulation with High-Intensity Diode Laser

Objective: To evaluate the effectiveness of photocoagulation with high-intensity diode laser in the treatment of venous lake (VL) lesions. Background Data: VL is a common vascular lesion characterized by elevated, usually dome-shaped papules, ranging in color from dark blue to dark purple, seen more frequently in elderly patients. They often occur as single lesions on the ears, face, lips, or neck. Once formed, lesions persist throughout life. Although these lesions are usually asymptomatic, they can bleed if injured. Methods: Seventeen patients (7 men and 10 women) with VL on the lip were treated using a noncontact diode laser (wavelength 808 nm, power output 2-3 W in continuous wave). Results: After only one irradiation exposure, all lesions were successfully treated. Healing was completed in approximately 2 to 3 weeks, and none of the patients experienced complications. Postoperative discomfort and scarring were not present or were minimal. Conclusion: Photocoagulation with high-intensity diode laser is an effective, bloodless procedure for the treatment of VL.

Low- and High-Intensity Lasers in the Treatment of Herpes Simplex Virus 1 Infection

Herpes simplex virus (HSV) is one of the most common viral infections of the human being. Although most of the seropositive persons do not manifest symptoms, infected individuals may present recurrent infections, characterized by cold sores. HSV-1 infection can result in potentially harmful complications in some patients, especially in those with compromised immunity. We report a clinical case of a patient with severe oral HSV-1 infection in the lower lip. The treatment of the lesions with the association of high-intensity (erbium-doped yttrium aluminum garnet, 2.94 mu m, 80 mJ/pulse, 2-4 Hz) and low-intensity (indium gallium aluminum phosphide, 660 nm, 3.8 J/cm(2), 10mW) lasers has not been reported in the literature. During treatment, no systemic or topical medication was used. Pain sensitivity was completely gone after the first irradiation with the low-intensity laser. During the healing process, lesions were traumatized twice, on the days 4 and 7. Even though the lesions were completely healed within 10 days.

Influence of Laser Photobiomodulation on Collagen IV During Skeletal Muscle Tissue Remodeling After Injury in Rats

Objective: The aim of the present study was to determine the effect of GaAlAs low-level laser therapy (LLLT) on collagen IV remodeling of the tibialis anterior (TA) muscle in rats after cryolesion. Background: Considerable interest exists in skeletal muscle regeneration in situations such as repair after exercise-induced muscle injury, after muscle transplantation, in muscular dystrophy, exercise-induced muscle injury, and the recovery of strength after atrophy due to disuse. A number of studies have demonstrated the potential of LLLT in facilitating the muscle-healing process; however, no consensus is found in the literature regarding the best laser-irradiation parameters. Methods: Adult male Wistar rats (n = 45) were used and randomly divided into three groups: control (n = 5); nontreated cryolesioned group (n = 20), and LLLT-cryolesioned group (n = 20). The cryolesioned groups were analyzed at 1, 7, 14, and 21 days after the injury procedure. Laser irradiation was performed 3 times per week on the injured region by using the GaAlAs laser (660 nm; beam spot of 0.04 cm(2), output power of 20 mW, power density of 500 mW/cm(2), and energy density of 5 J/cm(2), for 10 sec). The muscles were removed, frozen, cryosectioned, and then stained with hematoxylin-eosin for the visualization of general morphology or used for immunohistochemical analysis of collagen IV. Results: It was demonstrated that LLLT promotes an increase in collagen IV immunolabeling in skeletal muscle in the first 7 days after acute trauma caused by cryoinjury, but does not modify the duration of the tissue-repair process. Even with LLLT, the injured muscle tissue needs similar to 21 days to achieve the same state of organization as that in the noninjured muscle. Conclusion: The collagen IV content is modulated in regenerating skeletal muscle under LLLT, which might be associated with better tissue outcome, although the histologic analysis did not detect tissue improvement in the LLLT group.