Hidrogel de quitosana com nanofibras de Policaprolactona (PCL) associado a extratos fitoterápicos no metabolismo celular e ação antimicrobiana: estudo in vitro / Evaluation of the chitosan hydrogel with PCL nanofibers associated with phytotherapeutic extracts in cellular and microbiological activity: in vitro study

O objetivo neste estudo foi produzir hidrogel de quitosana (CH) com PCL e fitoterápicos para uso preventivo de úlcera de pressão. Os hidrogéis de CH foram produzidos com glicerofosfato (GP) e com xantana (X), associados ao PCL e foram caracterizados por estereomicroscopio, intumescimento, molhabilidade e MEV. Posteriormente foram submetidos ao teste de viabilidade (MTT) com fibroblastos HFF-1 e queratinócitos HaCat. O hidrogel que apresentou melhor resultado foi escolhido para continuar na pesquisa. Posteriormente, extratos de Pfaffia panculata K, Juglans regia L, Rosmarinus officinalis L, Zingiber officinale, Própolis e Hamamelis foram colocados em contato com cepas de Staphylococcus aureus (S.a) (ATCC 6538), Streptococcus pyogenes (S.p) (ATCC 19615), Staphylococcus epidermidis (S.e) (ATCC 12228), Pseudomonas aeruginosa (P.a) (ATCC 15442), Escherichia coli (E.c) (ATCC 25922) e Klebsiella Pneumoniae (K.p) (ATCC 4352) na forma planctônica nos testes de CIM e CMM. Os dois melhores extratos fitoterápicos foram avaliados quanto ao sinergismo no teste checkerboard e posteriormente associados ao hidrogel anteriormente eleito. A seguir, o comportamento da HaCat e HFF-1 com os hidrogéis foi analisado por MTT, proteína total, ELISA, genotoxicidade e formação de biofilme monotípico com suspensões padronizadas (107 cel/mL) de S.a, S.e, S.p, P.a, E.c e K.p. Na caracterização e viabilidade o hidrogel CHX PCL apresentou os melhores resultados. Os extratos selecionados após CIM, CMM e checkerboard foram gengibre (G) e própolis (P). O extrato G se destacou na CIM com inibição de K. p e P. a. Os extratos de G e P demonstraram ação microbicida para K. p e P. a e somente o extrato P obteve ação microbicida para S. a na CMM. Houve ação aditiva dos extratos associados no checkerboard para S.p e ação aditiva e sinérgica para S. e. Os grupos de hidrogéis foram compostos por: quitosana xantana (CHX), CHX própolis (CHXP), CHX gengibre (CHXG) e CHX própolis e gengibre associados (CHXPG), todos associados ao PCL. Todos os hidrogéis demonstraram viabilidade celular acima de 70% do grupo controle, permitindo metabolismo celular observado na proteína total. Houve quantificação de IL-6 maior no grupo CHX nas duas linhagens de células enquanto a quantificação de IL-10 não exibiu diferença estatística entre os grupos. Todos os hidrogéis promoveram redução acentuada de biofilme de K.p e E.c. Os grupos CHX, CHXP e CHXG reduziram biofilme de S.e. O grupo CHXG reduziu biofilme de S.p. Para S.a e P.a o grupo CHXPG foi mais eficaz reduzindo biofilme. Concluímos que os hidrogéis apresentaram resultados satisfatórios e promissores, trazendo inovação por associação de biopolímeros e associação de extratos fitoterápicos pouco estudados. Os resultados positivos justificam a continuidade dos estudos com esse biomaterial(AU)

The aim of this study was to produce chitosan hydrogel (CH) with PCL and herbal medicines for preventive use of pressure ulcers. The CH hydrogels were produced with glycerophosphate (GP) and xanthan (X), associated with PCL and were characterized by stereomicroscope, swelling, wettability and SEM. Subsequently, they were submitted to a viability test (MTT) with HFF-1 fibroblasts and HaCat keratinocytes. The hydrogel that presented the best result was chosen to continue the research. Subsequently, extracts of Pfaffia panculata K, Juglans regia L, Rosmarinus officinalis L, Zingiber officinale, Propolis and Hamamelis were placed in contact with strains of Staphylococcus aureus (Sa) (ATCC 6538), Streptococcus pyogenes (Sp) (ATCC 19615), epidermidis (Se) (ATCC 12228), Pseudomonas aeruginosa (Pa) (ATCC 15442), Escherichia coli (Ec) (ATCC 25922) and Klebsiella Pneumoniae (Kp) (ATCC 4352) in planktonic form in CIM and CMM tests. The two best herbal extracts were evaluated for synergism in the checkerboard test and subsequently associated with the previously elected hydrogel. Next, the behavior of HaCat and HFF-1 with hydrogels was analyzed by MTT, total protein, ELISA, genotoxicity and monotypic biofilm formation with standardized suspensions (107 cel / mL) of Sa, Se, Sp, Pa, Ec and Kp In the characterization and viability the CHX PCL hydrogel presented the best results. The extracts selected after MIC, CMM and checkerboard were ginger (G) and propolis (P). The G extract stood out in the MIC with inhibition of K. p and P. a. The extracts of G and P showed microbicidal action for K. p and P. a and only the extract P obtained microbicidal action for S. a in CMM. There was an additive action of the associated extracts on the checkerboard for S.p and an additive and synergistic action for S. e. The hydrogel groups were composed of: xanthan chitosan (CHX), CHX propolis (CHXP), CHX ginger (CHXG) and CHX propolis and ginger associated (CHXPG), all associated with PCL. All hydrogels demonstrated cell viability above 70% of the control group, allowing cellular metabolism observed in the total protein. There was a greater quantification of IL-6 in the CHX group in the two cell lines while the quantification of IL-10 did not show statistical difference between the groups. All hydrogels promoted a marked reduction in the biofilm of K.p and E.c. The CHX, CHXP and CHXG groups reduced S.e biofilm. The CHXG group reduced S.p. For S.a and P.a, the CHXPG group was more effective in reducing biofilm. We conclude that the hydrogels presented satisfactory and promising results, bringing innovation through association of biopolymers and association of phytotherapic extracts little studied. The positive results justify the continuity of studies with this biomaterial(AU)

Desenvolvimento de formulações probióticas para a prevenção da candidose bucal: estudo in vitro e in vivo / Development of probiotic formulations for prevention of oral candidiasis: in vitro and in vivo study

Os probióticos são considerados uma alternativa potencial para o controle da candidose, no entanto, existe uma falta de produtos probióticos direcionados para a cavidade bucal. Neste estudo, desenvolvemos formulações probióticas usando gellan gum, um biopolímero natural usado como aditivo alimentar, e investigamos os efeitos dessas formulações em Candida albicans. Para isso, Lactobacillus paracasei 28.4, uma cepa recentemente isolada da cavidade bucal, foi incorporada em várias concentrações de gellan gum (1 a 0,6%). Todas as concentrações testadas foram capazes de incorporar as células de L. paracasei, mantendo a viabilidade bacteriana. As formulações probióticas permaneceram estáveis por 7 dias quando armazenadas à temperatura ambiente ou a 4°C. Entretanto, o armazenamento a longo prazo das formulações probióticas foi conseguido apenas quando L. paracasei 28.4 foi liofilizado. As formulações probióticas proporcionaram uma liberação de células de L. paracasei por 24 horas, o que foi suficiente para inibir o crescimento de C. albicans com efeitos dependentes das concentrações celulares incorporadas no gellan gum. As formulações probióticas também tiveram atividade inibitória contra os biofilmes de Candida, reduzindo o número de células de Candida (p<0,0001), diminuindo a biomassa total (p=0,0003) e prejudicando a formação de hifas (p=0,0002) em relação ao grupo controle não tratado. Contudo, a formulação probiótica de gellan gum a 1% proporcionou uma colonização oral de L. paracasei em camundongos com aproximadamente 6 log de UFC/mL após 10 dias. Essa formulação inibiu o crescimento de C. albicans (p<0,0001), impediu o desenvolvimento de lesões de candidose (p=0,0013) e suprimiu a inflamação (p = 0,0006) quando comparada aos camundongos não tratados. Estes resultados indicam que o gellan gum pode ser um biomaterial promissor como sistema transportador de probióticos para prevenir a candidose oral(AU)

Probiotics are considered a potential alternative for the control of candidiasis, however there is a lack of probiotic products targeted for the oral cavity. In this study, we developed probiotic formulations using gellan gum, a natural biopolymer used as a food-additive, and investigated the effects of this delivery method on Candida albicans. Lactobacillus paracasei 28.4, a strain recently isolated from the oral cavity, was incorporated in several concentrations of gellan gum (1 to 0.6%). All tested concentrations could incorporate L. paracasei cells while maintaining bacterial viability. Probiotic-gellan formulations were stable for 7 days when stored at room temperature or 4°C. Long-term storage of bacterial impregnated gellan gum could be achieved when L. paracasei 28.4 was lyophilized. The probiotic-gellan formulations provided a release of L. paracasei cells over 24 hours that was sufficient to inhibit the growth of C. albicans with effects dependent on the cell concentrations incorporated into gellan gum. The probiotic-gellan formulations also had inhibitory activity against Candida biofilms by reducing the number of Candida cells (p < 0.0001), decreasing the total biomass (p = 0.0003) and impairing hyphae formation (p = 0.0002) in relation to the control group, not treated. However, only probiotic formulation of gellan gum 1% provided an oral colonization of L. paracasei in mice with approximately 6 log of CFU/mL after 10 days. This formulation inhibited the C. albicans growth (p < 0.0001), prevented the development of candidiasis lesions (p = 0.0013), and suppressed the inflammation (p = 0.0006) when compared to the mice not treated. These results indicate that gellan gum is a promising biomaterial as a carrier system of probiotics to prevent oral candidiasis(AU)

The efficacy and safety of urethral injection therapy for urinary incontinence in women: a systematic review

To evaluate the efficacy and safety of different bulking agents for treating urinary incontinence in women, a systematic review including only randomized controlled trials was performed. The subjects were women with urinary incontinence. The primary outcomes were clinical and urodynamic parameters. The results were presented as a weighted mean difference for non-continuous variables and as relative risk for continuous variables, both with 95% confidence intervals. Initially, 942 studies were identified. However, only fourteen eligible trials fulfilled the prerequisites. Altogether, the review included 1814 patients in trials of eight different types of bulking agents, and all studies were described and analyzed. The measured outcomes were evaluated using a large variety of instruments. The most common complications of the bulking agents were urinary retention and urinary tract infection. Additionally, there were certain major complications, such as one case of death after use of autologous fat. However, the lack of adequate studies, the heterogeneous populations studied, the wide variety of materials used and the lack of long-term follow-up limit guidance of practice. To determine which substance is the most suitable, there is a need for more randomized clinical trials that compare existing bulking agents based on standardized clinical outcomes.

Safety and efficiency of carboximethylcellulose hydrogel 2% in leg ulcers / Segurança e eficácia do hidrogel de carboximetilcelulose à 2% em úlceras de perna / Evaluación de la seguridad y eficacia del uso del hidrogel à 2% en ulceras de piernas

Objetivo: Avaliar a eficácia e a segurança do uso do hidrogel a 2%, manipulado na farmácia universitária, no tratamento de pacientes com úlceras de perna. Método: Estudo clínico, prospectivo, não controlado, incluindo 20 pacientes no período de janeiro a outubro de 2010. A avaliação da eficácia foi realizada através da redução e da alteração tecidual do leito da ferida. A segurança foi avaliada por observação e relato dos eventos adversos. Resultados: Os dados mostraram que 33,3% das lesões apresentaram cicatrização total. Quanto à segurança, apenas 2 pacientes apresentaram dor, aumento de exsudato e infecção da ferida como reações adversas possíveis, segundo o algoritmo de Naranjo, ao uso do hidrogel 2% como curativo. Conclusão: Os resultados permitem considerar que o hidrogel de CMC a 2% mostrou-se eficaz e seguro para o tratamento desse tipo de lesão.

Objective: The aim of this study is to assess the efficiency and safety on using hydrogel carboxymethylcellulose 2%, handled at university pharmacy in treatment of patients with leg ulcers. Method: A non-controlled prospective clinic study with 20 patients took place from January to October 2010. Results: The efficiency assessment was performed through the wound area reduction and wound bed tissue alteration; the safety was assessed through observation and report of adverse events. The data showed that 33.3% of the lesions presented total healing. Regarding the safety, only 2 patients reported pain, increase of the exudate and wound infection as possible adverse reaction to the use hydrogel as dressing, according to the Naranjo algorithm. Conclusion: The results let us consider that the hydrogel 2% can be considered efficient and safe for this type of lesion.

Objetivo: Es evaluar la eficacia y la seguridad del uso del Hidrogel 2%, manoseado en una farmacia docente, en el tratamiento de enfermos con úlceras de pierna. Método: El estudio clínico, no controlado, incluyó 20 pacientes en el periodo entre enero y octubre de 2010. Resultados: La evaluación de su eficacia fue comprobada por la reducción de la herida y de la alteración de la piel en su interior. La seguridad fue evaluada por observación y el relato de los eventos adversos. Los dados han evidenciados que 33,3% de las lesiones presentaron cicatrización total. Cuanto a la seguridad, solamente dos pacientes presentaron reacciones previsibles al uso del Hidrogel 2% como curativo. Conclusión: Se puede concluir que el Hidrogel 2% se muestra eficaz y seguro para el tratamiento de esto tipo de lesión.

Estudio comparativo de la cicatrización en quemaduras con tratamiento en base a miel de ulmo (eucryphia cordifolia) y vitamina C oral versus hidrogel en cobayos (cavia porcellus) / Comparative study of healing in burns treatment based on ulmo honey (eucryphia cordifolia) and oral vitamin C in guinea pig (cavia porcellus) versus hydrogel

La quemadura es una herida traumática que resulta en un daño local y sistémico, con cambios oxidativos. La miel posee actividad antioxidante, antiinflamatoria y antibacteriana. A su vez, la vitamina C mejora la barrera epidérmica y reduce la contracción de heridas, entre otros. El objetivo de este estudio fue evaluar morfológicamente el efecto del tratamiento con miel de ulmo + vitamina C en cobayo y comparar su capacidad cicatrizante y debridante con hydrogel+tull en heridas causadas por quemaduras tipo B al día 10 post-injuria. Se utilizaron 15 cobayos (Cavia porcellus) distribuidos en los grupos A (miel de ulmo tópica y vitamina C oral), B (hidrogel-tull), y Control (suero fisiológico al 0,9 por ciento). Se observó el debridamiento de las heridas, y se realizaron curaciones diarias hasta la obtención de biopsias al día 10 post-injuria. Secciones de 4 um fueron teñidas con HE, Tricrómico de Masson y PAS. En el grupo A se produjo un rápido debridamiento respecto al grupo B y Control. El tratamiento en estudio presentó una fase proliferativa avanzada al día 10 post-injuria, con tejido vascularizado, presencia de fibras colágenas y fibroblastos. La epidermis se regeneró por completo, presentando una lámina basal visible mediante la tinción PAS. El grupo B presentó una etapa fibroblástica proliferativa inicial, observándose elementos agudos a nivel dérmico. La fase de epitelización se completó solo en el 50 por ciento de las muestras. La miel ulmo + vitamina C mostraron ser sustanciasefectivas como material de tratamiento en la cicatrización de quemaduras tipo B de piel en cobayo.

A burn is a traumatic injury resulting in local and systemic injury with oxidative changes. Honey has antioxidant, anti-inflammatory and antibacterial properties. At the same time, among other benefits vitamin C improves the epidermal barrier and reduces wound contraction. The aim of this study was to evaluate morphologically the effect of regimen with Ulmo honey + vitamin C in guinea pigs and compare its healing and debriding potential with Hydrogel + tull in injuries caused by type B burns at day 10 post-injury. We used 15 guinea pigs (Cavia porcellus) distributed in groups A (Ulmo honey and vitamin C topical, oral), B (hydrogel-tull), and control (0.9 percent saline). Wound debridement was observed daily and wound treatment was performed daily until biopsies were obtained at day 10 post-injury. Four 4 um sections were stained with HE, Masson's trichrome and PAS. In group A there was rapid debridement when compared with groups B and Control. Treatment study presented advanced proliferative phase at day 10 post-injury with vascularized tissue, and presence of collagen fibers and fibroblasts. The epidermis was completely regenerated, presenting a visible basal lamina by PAS staining. Group B presented an initial proliferative fibroblastic phase, showing acute dermal elements. Epithelialization phase was completed in only 50 percent of the samples. Ulmo honey + vitamin C substances have been shown to be effective as treatment material in healing of type B burns in guinea pig skin.

Hydrogel of polysaccharide of sugarcane molasses as carrier of bone morphogenetic protein in the reconstruction of critical bone defects in rats

PURPOSE: To evaluate the benefit of using carriers such as the biopolymer gel (hidrogel of polysaccharide of sugarcane molasses) associated with the bone morphogenetic proteins (BMP's) in the repair of critical bone defects in calvaria of Wistar rats. METHODS: Forty-two rats were submitted to a surgical calvaria bone defects. These animals were divided into two experimental groups, positive control group and negative control group. The Group I the calvaria defect was filled up with biopolymer gel, biological membrane, BMP and lyophilized graft. The Group II was treated with biopolymer gel, BMP and lyophilized graft. And the group III (positive control group) was treated with BMP, lyophilized graft and biological membrane. In the negative control group (Group IV) a defect was made in the rat calvaria and the animals were sacrificed immediately after the surgery. The animals of experimental groups and positive control group were slaughtered after subsequent periods of 90 and 180 days. In these periods, the histological analysis and image assessment by cone bean tomographic imaging were obtained. RESULTS: There was highest bone tissue formation with statistically significant results in the groups that associated biopolymer gel and membrane (Group I), followed by the group III (BMP, lyophilized graft and biological membrane). The lower bone formation occurred in the group not using the sugarcane biopolymer gel (Group II). The radiolucent areas of the analyzes of 180 days among the groups studied were respectively, 14.98 mm², 26.65 mm² and 35.81 mm². CONCLUSION: The biopolymer gel showed to be an excellent bone morphogenetic protein carrier, probably by facilitating the controlled release of these proteins in the process of bone repair.