Effects of the basic fibroblast growth factor and its anti-factor in the healing and collagen maturation of infected skin wound

PURPOSE: The infection is one of the main factors that affect the physiological evolution of the surgical wounds. The aim of this work is to evaluate the effects of fibroblast growth factor (FGFâ) and anti-FGFâ in the healing, synthesis and maturation of collagen when topically used on infected skin wounds of rats. METHODS: An experimental study was perfomed in 60 male Wistar rats. All animals were divided in two groups (A and B). Each group was divided in three subgroups A1, B1; A2, B2 and A3, B3. After anesthesia with pentobarbital, two open squared wounds (1cm2), 4cm distant to each other, were done in the dorsal skin of all the rats. In group A (n=30) the wounds were contaminated with multibacterial standard solution, and in group B(n=30) the wounds were maintained sterile. These wounds were named F1 (for inflammation analysis) and F2 (for collagen study). The open wounds of A1 and B1 rats were topically treated with saline solution, A2 and B2 were treated with FGFâ and subgroups A3 and B3 were treated with FGFâ and anti-FGFâ. The rats were observed until complete epitelization of F2 wounds for determination of healing time and the expression of types I and III collagen, using Picro Sirius Red staining. Inflammatory reaction in F1 wounds was studied using hematoxilineosin staining. The three variable was measured by the Image Pro-Plus Média Cybernetics software. The statistical analysis was performed by ANOVA and Tukey test, considering p<0.05 as significant. RESULTS: It was observed that infection retarded significantly (p<0.05) the time of wound scarring and the topical application of FCFb reverted the inhibition of healing caused by bacteria. The inflammatory reaction was greater in the subgroup B2 than in B1 and A3, and the difference was significant (p<0.05). It was observed greater expression of type I collagen in all the subgroups treated with FCFb, when compared with the untreated subgroups. Type III collagen was significantly decreased in wounds of B3 rats, comparing to the other subgroups. CONCLUSIONS: The FCFb accelerated the healing of open infected wounds and contributed with maturation of collagen, enhancing the type I collagen density. The anti-FCFb antibody was able to attenuate the production of both type I and III collagen

Pharmacokinetic And Pharmacodynamic Evaluation Of A Nanotechnological Topical Formulation Of Lidocaine/prilocaine (nanorap) In Healthy Volunteers.

Nanorap is a new nanotechnological formulation for topical anesthesia composed of lidocaine (2.5%) and prilocaine (2.5%). The present study evaluated the pharmacokinetics (PK) of Nanorap. For the determination of lidocaine and prilocaine in human plasma a new method using high-performance liquid-chromatography coupled to tandem mass spectrometry was developed. Nanorap pharmacodynamic (PD) and its physical proprieties were also evaluated. Nanorap was administered by topical application of 2g to healthy volunteers and blood samples were collected for the PK analysis. The drugs were extracted from plasma by liquid-liquid extraction with ether/hexane (80/20, v/v). The chromatography separation was performed on a Genesis C18 analytical column 4 µm (100 x 2.1 mm i.d.) with a mobile phase of methanol/acetonitrile/water (40/30/30, for lidocaine, and 50/30/20, for prilocaine, v/v/v) + 2 mM of ammonium acetate and ropivacaine as internal standard. The drugs were quantified using a mass spectrometer with an electrospray source in the ESI positive mode (ES+) configured for multiple reaction monitoring. The PD of Nanorap was evaluated with the use of a visual analogue scale. Nanorap was characterized by cryofracture. The chromatography run time was 5.5 min for lidocaine and 3.3 min for prilocaine and the lower limit of quantification was 0.05 ng/mL for both drugs. Mean Cmax was 6.62 and 1.72 ng/mL for lidocaine and prilocaine, respectively. Median Tmax was 6.5 hours for both drugs. Nanocapsules had a mean size of 88nm and mean drug association of 92.5% and 89% for lidocaine and prilocaine, respectively. The PD study showed that Nanorap has a sufficient analgesic effect (>30% reduction in pain) after 10 minutes of application. A new simple, selective and sensitive method for determination of lidocaine and prilocaine in human plasma was developed. Nanorap generated safe plasma levels of the drugs and satisfactory analgesic effect.

Effect Of Calendula Officinalis Cream On Achilles Tendon Healing.

In recent years, the scientific community has undertaken research on plant extracts, searching for compounds with pharmacological activities that can be used in diverse fields of medicine. Calendula officinalis L. is known to have antioxidant, anti-inflammatory, antibacterial, and wound healing properties when used to treat skin burns. Therefore, the purpose of this study was to analyze the effects of C. officinalis on the initial phase of Achilles tendon healing. Wistar rats were separated in three groups: Calendula (Cal)-rats with a transected tendon were treated with topical applications of C. officinalis cream and then euthanized 7 days after injury; Control (C)-rats were treated with only vehicle after transection; and Normal (N)-rats without tenotomy. Higher concentrations of hydroxyproline (an indicator of total collagen) and non-collagenous proteins were observed in the Cal group in relation to the C group. Zymography showed no difference in the amount of the isoforms of metalloproteinase-2 and of metalloproteinase-9, between C and Cal groups. Polarization microscopy images analysis showed that the Cal group presented a slightly higher birefringence compared with the C group. In sections of tendons stained with toluidine blue, the transected groups presented higher metachromasy as compared with the N group. Immunocytochemistry analysis for chondroitin-6-sulfate showed no difference between the C and Cal groups. In conclusion, the topical application of C. officinalis after tendon transection increases the concentrations of collagen and non-collagenous proteins, as well as the collagen organization in the initial phase of healing.

Targeting local tissues by transdermal application: Understanding drug physicochemical properties that best exploit protein binding and blood flow effects

The targeting of topically applied drug molecules into tissues below a site of application requires an understanding of the complex interrelationships between the drug, its formulation, the barrier properties of the skin, and the physiological processes occurring below the skin that are responsible for drug clearance from the site, tissue, and/or systemic distribution and eventual elimination. There is still a certain amount of controversy over the ability of topically applied drugs to penetrate into deeper tissues by diffusion or whether this occurs by redistribution in the systemic circulation. The major focus of our work in this area has been in determining how changes in drug structure and physicochemical properties, such as protein binding and lipophilicity, affect drug clearance into the local dermal microcirculation and lymphatics, as well as subsequent distribution into deeper tissues below an application site. The present study outlines our recent thinking on the drug molecule optimal physical attributes, in terms of plasma and tissue partitioning behaviour, that offer the greatest potential for deep tissue targeting. Drug Dev. Res. 46:309-315, 1999. (C) 1999 Wiley-Liss, Inc.

Bacterial and fungal colonization of burn wounds

A prospective study of fungal and bacterial flora of burn wounds was carried out from February 2004 to February 2005 at the Burns Unit of Hospital Regional da Asa Norte, Brasília, Brazil. During the period of the study, 203 patients were treated at the Burns Unit. Wound swab cultures were assessed at weekly intervals for four weeks. Three hundred and fifty four sampling procedures (surface swabs) were performed from the burn wounds. The study revealed that bacterial colonization reached 86.6% within the first week. Although the gram-negative organisms, as a group, were more predominant, Staphylococcus aureus (28.4%) was the most prevalent organism in the first week. It was however surpassed by Pseudomonas aeruginosa form third week onwards. For S. aureus and P. aeruginosa vancomycin and polymyxin were found to be the most effective drugs. Most of the isolates showed high level resistance to antimicrobial agents. Fungi were found to colonize the burn wound late during the second week postburn, with a peak incidence during the third and fourth weeks. Species identification of fungi revealed that Candida tropicalis was the most predominant, followed by Candida parapsilosis. It is crucial for every burn institution to determine the specific pattern of burn wound microbial colonization, the time-related changes in the dominant flora, and the antimicrobial sensitivity profiles. This would enable early treatment of imminent septic episodes with proper empirical systemic antibiotics, without waiting for culture results, thus improving the overall infection-related morbidity and mortality.

Novel micelle carriers for cyclosporin A topical ocular delivery: in vivo cornea penetration, ocular distribution and efficacy studies.

Cornea transplantation is one of the most performed graft procedures worldwide with an impressive success rate of 90%. However, for "high-risk" patients with particular ocular diseases in addition to the required surgery, the success rate is drastically reduced to 50%. In these cases, cyclosporin A (CsA) is frequently used to prevent the cornea rejection by a systemic treatment with possible systemic side effects for the patients. To overcome these problems, it is a challenge to prepare well-tolerated topical CsA formulations. Normally high amounts of oils or surfactants are needed for the solubilization of the very hydrophobic CsA. Furthermore, it is in general difficult to obtain ocular therapeutic drug levels with topical instillations due to the corneal barriers that efficiently protect the intraocular structures from foreign substances thus also from drugs. The aim of this study was to investigate in vivo the effects of a novel CsA topical aqueous formulation. This formulation was based on nanosized polymeric micelles as drug carriers. An established rat model for the prevention of cornea graft rejection after a keratoplasty procedure was used. After instillation of the novel formulation with fluorescent labeled micelles, confocal analysis of flat-mounted corneas clearly showed that the nanosized carriers were able to penetrate into all corneal layers. The efficacy of a 0.5% CsA micelle formulation was tested and compared to a physiological saline solution and to a systemic administration of CsA. In our studies, the topical CsA treatment was carried out for 14 days, and the three parameters (a) cornea transparency, (b) edema, and (c) neovascularization were evaluated by clinical observation and scoring. Compared to the control group, the treated group showed a significant higher cornea transparency and significant lower edema after 7 and 13 days of the surgery. At the end point of the study, the neovascularization was reduced by 50% in the CsA-micelle treated animals. The success rate of cornea graft transplantation was 73% in treated animals against 25% for the control group. This result was as good as observed for a systemic CsA treatment in the same animal model. This new formulation has the same efficacy like a systemic treatment but without the serious CsA systemic side effects. Ocular drug levels of transplanted and healthy rat eyes were dosed by UPLC/MS and showed a high CsA value in the cornea (11710 ± 7530 ng(CsA)/g(tissue) and 6470 ± 1730 ng(CsA)/g(tissue), respectively). In conclusion, the applied formulation has the capacity to overcome the ocular surface barriers, the micelles formed a drug reservoir in the cornea from, where a sustained release of CsA can take place. This novel formulation for topical application of CsA is clearly an effective and well-tolerated alternative to the systemic treatment for the prevention of corneal graft rejection.

In vivo distribution and ex vivo permeation of cyclosporine A prodrug aqueous formulations for ocular application.

Cyclosporine A is a poorly water-soluble, immunosuppressive drug used to treat a variety of ocular diseases. Its limited solubility makes challenging the development of a cyclosporine A-based eye drop for ocular topical application. Based on the prodrug strategy, the practically insoluble cyclosporine A was converted into a freely soluble prodrug. Such a water-soluble prodrug made it possible to develop water-based concentrated eye drops. The prodrug formulations were tested for their ex vivo permeation and in vivo distribution at three concentrations (equivalent to 0.05%, 0.50% and 2.00% w/v cyclosporine A). The ex vivo permeation experiments were performed on corneal and conjunctival epithelia. The in vivo distribution evaluated the total cyclosporine A present in the ocular structures as well as in serum, spleen and cervical lymphatic ganglions. Each prodrug formulation was compared to conventionally used cyclosporine A eye drops at an equivalent concentration. The experimental results showed that the tested eye drops behaved differently. The prodrug formulation was characterized by the following: i) preferential conjunctival penetration, ii) an interesting capacity to create large tissue deposits and iii) a lower risk of systemic complications and immunosuppression. The prodrug aqueous eye drop was demonstrated to be a patient-friendly option for the treatment of ocular diseases requiring high ocular levels of cyclosporine A, pushing the boundaries of the current therapeutic arsenal.

A topical microemulsion for the prevention of allergic rhinitis symptoms: results of a randomized, controlled, double-blind, parallel group, multicentre, multinational clinical trial (Nares study)

Background: Since barrier protection measures to avoid contact with allergens are being increasingly developed, we assessed the clinical efficacy and tolerability of a topical nasal microemulsion made of glycerol esters in patients with allergic rhinitis. Methods: Randomized, controlled, double-blind, parallel group, multicentre, multinational clinical trial in which adult patients with allergic rhinitis or rhinoconjunctivitis due to sensitization to birch, grass or olive tree pollens received treatment with topical microemulsion or placebo during the pollen seasons. Efficacy variables included scores in the mini-RQLQ questionnaire, number and severity of nasal, ocular and lung signs and symptoms, need for symptomatic medications and patients" satisfaction with treatment. Adverse events were also recorded. Results: Demographic characteristics were homogeneous between groups and mini-RQLQ scores did not differ significantly at baseline (visit 1). From symptoms recorded in the diary cards, the ME group showed statistically significant better scores for nasal congestion (0.72 vs. 1.01; p = 0.017) and mean total nasal symptoms (0.7 vs. 0.9; p = 0.045). At visit 2 (pollen season), lower values were observed in the mini-RQLQ in the ME group, although there were no statistically significant differences between groups in both full analysis set (FAS) and patients completing treatment (PPS) populations. The results obtained in the nasal symptoms domain of the mini-RQLQ at visit 2 showed the highest difference (−0.43; 95% CI: -0.88 to 0.02) for the ME group in the FAS population. The topical microemulsion was safe and well tolerated and no major discomforts were observed. Satisfaction rating with the treatment was similar between the groups. Conclusions: The topical application of the microemulsion is a feasible and safe therapy in the prevention of allergic symptoms, particularly nasal congestion.

Seguimiento de áreas cruentas cubiertas con queratinocitos autólogos cultivados en suero autólogo: Estudio multicéntrico en Bogotá

En 2009 se presento la estandarización de cultivos de queratinocitos autólogos cultivados en suero autólogo. En este estudio los autores describen la efectividad de estos parches de regeneración de piel, para la cobertura de áreas cruentas con indicación de injerto de piel parcial. El porcentaje de epitelización del área cruenta fue el punto principal. Métodos: 47 pacientes fueron incluidos consecutivamente, equivalentes a 78 áreas cruentas. Las áreas fueron estratificadas según la profundidad: grupo 1:IIA (n=8) grupo 2: IIB (n=39); grupo 3,III (n=24) y grupo 4, etiología diferente: Otras (n=7). Todas las áreas fueron tratadas con injertos de queratinocitos autólogos cultivados en suero autólogo y se realizo registro fotográfico y del porcentaje de epitelización al día 5, 7, 15 y 30. Resultados: La efectividad de los injertos de queratinocitos autólogos es de 53.16% ± 46.46%. El porcentaje de epitelización es mayor para el grupo 1 (100%) y grupo 2 (62.79%) que para el grupo 3 (27.57%) y el grupo 4 (33.86%). Se encontró relación entre la interacción de las medianas del porcentaje de epitelización entre área corporal y grado de quemadura (p<0.001 KW) siendo mayor para el grupo 1 en todas las áreas, grupo 2 en cara, grupo 3 en tronco y grupo 4 en cara; y el menor porcentaje de epitelización en el grupo 3 y grupo 4 de las áreas ubicadas tronco. Conclusión: Los injertos de queratinocitos autólogos cultivados en suero autólogo son un método de cobertura eficaz para áreas cruentas producidas por quemaduras IIA y IIB independientemente del tamaño y la localización , y para las áreas cruentas pequeñas (<9cm2) de etiología diferente o grado III de profundidad. Palabras Clave: Cultivo queratinocitos, cobertura áreas cruentas, efectividad.

Eficacia del plasma rico en plaquetas en procedimientos médicos estéticos faciales . Evaluación en contorno de ojos.

Introducción: En la actualidad se están implementando nuevas técnicas, para el tratamiento de líneas de expresión facial. El Plasma Rico en Plaquetas (PRP) utiliza factores de crecimiento humano autólogos con fines médicos estéticos. El objetivo de este trabajo fue evaluar el tratamiento con plasma rico en plaquetas en el manejo del rejuvenecimiento periocular. Materiales y métodos: Estudio descriptivo retrospectivo en una cohorte de 27 pacientes entre 30 a 70 años de ambos sexos,tratados con PRP sin tratamientos médicos estéticos previos . Se compararon fotografías del sistema VISIA®, previo y posterior el PRP, para determinar los cambios del área periocular. Con análisis comparativo de medias utilizando pruebas t de student. Resultados: De 27 historias clínicas revisadas 96,3% eran mujeres, la edad promedio fue de 52.67 años. Se observaron cambios clínicos satisfactorios en el manejo del foto envejecimiento periocular con mejoría estadísticamente significativa entre el promedio inicial y el post tratamiento en arrugas, textura y porfirinas (p: 0.000). No se observaron diferencias estadísticamente significativas entre los grupos de edad (p = 0,62). Los pacientes analizados posterior al tratamiento se encuentran en mejor estado que el 63,78% de la población de su mismo sexo, edad y fototipo de piel. Los eventos adversos fueron disminuyendo en su frecuencia en cada una de las sesiones siguientes. Discusión: El PRP proporciona una mejoría global en los parámetros de envejecimiento periocular lo cual se correlaciono con los estudios previos in vitro. Conclusiones: El PRP es seguro y eficaz en contorno de ojos.

Descripción del estado acido base en pacientes con quemaduras térmicas agudas: serie de casos

Introducción: Los pacientes con lesiones térmicas presentan alteraciones fisiológicas complejas que hacen difícil la caracterización del estado ácido-base y así mismo alteraciones electrolíticas e hipoalbuminemia que pudieran estar relacionados con un peor pronóstico. Se ha estudiado la base déficit (BD) y el lactato, encontrando una gran divergencia en los resultados. Por lo anterior, el análisis físico-químico del estado ácido-base podría tener un rendimiento superior a los métodos tradicionales. Metodología: Se realizó el análisis de una serie de casos de 15 pacientes mayores de 15 años, con superficie corporal quemada mayor al 20% que ingresaron a una unidad de cuidado intensivo (UCI) de quemados, dentro de las siguientes 48 horas del trauma. Para el análisis se utilizaron tres métodos distintos: 1) método convencional basado en la teoría de Henderson-Hasselbalch, 2) anión-gap (AG) y anión-gap corregido por albúmina, 3) análisis físico-químico del estado ácido-base según la teoría de Stewart modificado por Fencl y Figge. Resultados: Por el método de Henderson-Hasselbalch, 8 pacientes cursaron con acidosis metabólica, 4 pacientes con una BD leve, 5 pacientes con una BD moderada y 5 pacientes con una BD severa. El AG resultó menor a 16 mmol/dl en 10 pacientes, pero al corregirlo por albumina sólo 2 pacientes cursaron con AG normal. La diferencia de iones fuertes (DIF) se encontraba anormalmente elevada en la totalidad de los pacientes. Conclusión:El análisis del AG corregido por albumina y el análisis físico-químico del estado ácido-base, podrían tener mayor rendimiento al identificar las alteraciones metabólicas de estos pacientes.

Efeitos da aplicação tópica do mel de melipona subnitida em feridas infectadas de ratos

Avaliar efeitos do uso tópico do mel da abelha silvestre Melipona subnitida na evolução de feridas infectadas de pele. Método: Ratos Wistar foram distribuídos aleatoriamente em grupos de 6, anestesiados com tiopental sódico 20mg/Kg IP e cetamina 30mg/Kg IM e submetidos a exérese de segmento de 1 cm2 de pele total do dorso. Os ratos do grupo C (não infectado) foram tratados com solução salina sobre a ferida diariamente e no grupo MEL (não infectado) as feridas foram tratadas com mel uma vez por dia. Nos grupos C/I e MEL/I as feridas foram inoculadas com solução polimicrobiana. Culturas foram feitas 24 horas após. Caracterizada a infecção, as feridas foram tratadas com solução salina e mel, respectivamente. No terceiro dia de tratamento foi feita nova cultura. Após epitelização foi contado o tempo de cicatrização e as feridas foram biopsiadas para histopatologia e dosagem de TNF-a, IL-1b e IL-6 no tecido. Resultados: O tempo médio de cicatrização do grupo MEL/I foi menor que nos demais grupos(P<0,05). Verificou-se que a densidade de colágeno, leucócitos, fibroblastos e dosagem de citocinas (especialmente TNF) foi maior no grupo infectado e tratado com mel que nos demais grupos. Houve significante redução de bactérias Gram-negativas e positivas nas feridas após o tratamento com mel. Conclusão: O uso tópico de mel de Melipona subnitida em feridas infectadas da pele de ratos estimulou a resposta imunológica, reduziu a infecção e o tempo de cicatrização

Simvastatin improves the healing of infected skin wounds of rats

This study explores the potential of the simvastatin to ameliorate inflammation and infection in open infected skin wounds of rats. Methods: Fourteen Wistar rats weighing 285±12g were used. The study was done in a group whose open infected skin wounds were treated with topical application of sinvastatina microemulsion (SIM, n=7) and a second group with wounds treated with saline 0.9 % (SAL, n=7). A bacteriological exam of the wounds fluid for gram positive and gram negative bacteria, the tecidual expression of TNFá and IL-1â by imunohistochemical technique, and histological analysis by HE stain were performed. Results: The expression of TNFa could be clearly demonstrated in lower degree in skin wounds treated with simvastatin (668.6 ± 74.7 ìm2) than in saline (2120.0 ± 327.1 ìm2). In comparison, wound tissue from SIM group displayed leukocyte infiltration significantly lower than that observed in SAL group (p<0.05). Culture results of the samples taken from wound fluid on fourth post treatment day revealed wound infection in only one rat of group simvastatin (SIM), where Proteus mirabilis, Escherchia coli and Enterobacter sp were isolated. In the rats whose wounds were treated with saline (SAL), polymicrobial infection with more than 100,000 CFU/g was detected in all the wounds. Conclusion: In addition to its antiinflammatory properties, the protective effects of simvastatin in infected open skin wounds is able to reduce infection and probably has antibacterial action. The potential to treat these wounds with statins to ameliorate inflammation and infection is promising

Risk factors for acquisition of Methicillin-resistant Staphylococcus aureus among patients from a burn unit in Brazil

Methicillin-resistant Staphylococcus aureus (MRSA) is an important agent of colonization and infection in burn units. in order to identify risk factors for MRSA acquisition in a Brazilian burn unit, we performed two retrospective studies. In the first ("cohort" study), 175 patients who were not colonized with MRSA on admission were followed to assess risk factors for MRSA acquisition. in the second ("case-case-control" study), 143 individuals from the previous study who were negative for both MRSA and Methicillin-susceptible S. aureus (MSSA) on admission were followed. Case-control studies were performed to investigate risk factors for MRSA and MSSA acquisition. MRSA and MSSA were recovered from 75 and 23 patients, respectively. In the "cohort" study, only the number of wound excisions (Odds Ratio [OR] = 1.55, 95% Confidence Interval [CI] = 1.21-1.98, P = 0.001) was associated with MRSA acquisition. in the "case-case-control" study, burns involving head (OR=3.43, 95%CI = 1.50-7.81, P = 0.003) and the number of wound excisions (OR = 1.83, 95%CI = 1.27-2.63, P = 0.001) were significant risk factors for MRSA. Burns involving perineum were negatively associated with MSSA acquisition (OR = 0.16, 95%CI = 0.03-0.75, P = 0.02). In conclusion, the acquisition of MRSA was related to the site of the burn and to the surgical manipulation of tissues, but not to the use of antimicrobials. (C) 2009 Elsevier Ltd and ISBI. All rights reserved.

Morphomettic changes on honeybee Apis mellifera L. workers fat body cells after juvenile hormone topic application at emergence

The effect of topical application of juvenile hormone (JH) over the lifetime of worker bees was evaluated in Apis mellifera, by measuring the area of the two cell types, trophocytes and oenocytes, found in the fat body. Topical application of 1 mu l of a 1 mu g/mu l solution of JH in acetone to the abdomens of newly emerged workers produced an increase in cell size, in both types of cell of 5-day-old treated workers in relation to the untreated control. The treatment was more effective on the oenocytes, since there were significant differences compared to the averages of the treatments and the interaction of the treatments with the age of the workers. The developmental pattern seemed to differ from the treated group. However, subsequent effects were probably dependent on different, natural variations in hormonal levels. (c) 2007 Elsevier Ltd. All rights reserved.