Novel fibrin-fibronectin matrix accelerates mice skin wound healing

Plasma fibrinogen (F1) and fibronectin (pFN) polymerize to form a fibrin clot that is both a hemostatic and provisional matrix for wound healing. About 90% of plasma F1 has a homodimeric pair of γ chains (γγF1), and 10% has a heterodimeric pair of γ and more acidic γ′ chains (γγ′F1). We have synthesized a novel fibrin matrix exclusively from a 1:1 (molar ratio) complex of γγ′F1 and pFN in the presence of highly active thrombin and recombinant Factor XIII (rFXIIIa). In this matrix, the fibrin nanofibers were decorated with pFN nanoclusters (termed γγ′F1:pFN fibrin). In contrast, fibrin made from 1:1 mixture of γγF1 and pFN formed a sporadic dis- tribution of “pFN droplets” (termed γγF1+pFN fibrin). The γγ′F1:pFN fibrin enhanced the adhesion of primary human umbilical vein endothelium cells (HUVECs) relative to the γγF1+FN fibrin. Three dimensional (3D) culturing showed that the γγ′F1:pFN complex fibrin matrix enhanced the proliferation of both HUVECs and primary human fibroblasts. HUVECs in the 3D γγ′F1:pFN fibrin exhibited a starkly enhanced vascular mor- phogenesis while an apoptotic growth profile was observed in the γγF1+pFN fibrin. Relative to γγF1+pFN fibrin, mouse dermal wounds that were sealed by γγ′F1:pFN fibrin exhibited accelerated and enhanced healing. This study suggests that a 3D pFN presentation on a fibrin matrix promotes wound healing

Evaluation of effect of agonist of receptors GPR120 e GPR40 na wound healing of diabetic wound

Orientador: Eliana Pereira de AraújoDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de EnfermagemResumo: A progressão mundial do diabetes e as complicações associadas à hiperglicemia evidenciam a ocorrência de feridas crônicas, que resultam em amputação, e impulsionam a busca por novas tecnologias no tratamento dessas lesões. Receptores GPR120 e GPR40 são potenciais alvos terapêuticos no tratamento de distúrbios metabólicos, por exercerem efeitos anti-inflamatórios e sobre a estimulação da proliferação celular. O objetivo desse trabalho foi avaliar o efeito do GW9508, um agonista duplo dos receptores GPR120 e GPR40, na cicatrização de feridas em modelo experimental. Foram utilizados camundongos C57/BL6J machos, adultos não diabéticos e diabéticos induzidos por baixas doses de estreptozotocina. Os animais foram submetidos a confecção de duas feridas excisionais de 6mm na região dorsal. A retração da pele foi estabilizada com prótese de silicone. Após serem aleatoriamente divididos em veículo ou GW9508, receberam tratamento tópico com 20µL de gel de carbopol incorporado ao veículo ou GW9508 a 100µM, uma vez ao dia até 5º ou 12º dia pós-lesão, quando o tecido foi coletado. Os grupos foram comparados quanto a retração macroscópica das feridas, morfologia do tecido cicatricial e expressão gênica por RT-PCR. Para as análises estatísticas aplicamos o teste t de Student em amostras independentes e para mais de duas variáveis, análise de variância (ANOVA), considerando p<0,05. O tratamento tópico com o GW9508 em camundongos não diabéticos aumenta a velocidade de cicatrização de feridas nos dias 6 e 8 pós-lesão, enquanto em camundongos induzidos ao diabetes esse efeito não foi observado. Durante a fase proliferativa, o tratamento com GW9508 diminuiu a expressão gênica de f480, il-1ß, mmp9 e vegf quando comparado ao veículo.O tratamento não modificou quantitativamente a deposição de fibras de colágeno, mas foi capaz de alterar a morfologia do tecido conjuntivo, apresentando derme mais entrelaçada pelas fibras de colágeno e formação de papilas dérmicas, o que torna o tecido cicatricial mais resistente e mais próximo à pele íntegra. A reanálise de dados de sequenciamento de RNA de célula única do tecido da ferida com base nos dados publicados de Guerrero-Juarez et al, mostrou 20 aglomerados de células organizadas principalmente em quatro linhagens compostas por fibroblastos, miofibroblastos, células mielóides e células endoteliais. Os genes modulados pelo tratamento são expressos por clusters celulares de fibroblastos e células de origem mieloide importantes no processo de cicatrização. Nosso estudo mostrou que o tratamento tópico com GW9508 aumenta a velocidade de cicatrização da ferida em camundongos saudáveis, mas não em camundongos diabéticos. O tratamento modula negativamente genes inflamatórios na fase proliferativa expressos por fibroblastos e células mieloides resultando em tecido estruturalmente mais semelhante a pele íntegra. Estudos futuros poderão colaborar para melhor entendimento dos mecanismos envolvidos nesse fenômeno, e identificação da melhor forma do emprego destes agonistasAbstract: Worldwide progression of diabetes and the complications associated with hyperglycemia highlight a number of chronic wounds that result in amputation, and drive a search for new technologies in the treatment of these lesions. GPR120 and GPR40 receptors are potential therapeutic targets in the treatment of metabolic disorders, as they exert anti-inflammatory effects and stimulate cell proliferation. The aim of this study was to evaluate the effect of GW9508, a dual agonist of GPR120 and GPR40 receptors, on wound healing in an experimental model. We use male C57 / BL6J mice, non-diabetic and diabetic adults induced by low doses of streptozotocin. Animals underwent two 6mm excision wounds in the dorsal region. Skin retraction was stabilized with silicone prosthesis. After being randomly divided into vehicle or GW9508, they received topical treatment with 20µL of carbopol gel incorporated into the vehicle or GW9508 at 100µ once daily until the 5th or 12th day after injury, when tissue was collected. The groups were compared for macroscopic wound retraction, scar tissue morphology and gene expression by RT-PCR. For statistical analysis we applied the Student t test on independent samples and for more than two variables, analysis of variance (ANOVA), considering p <0.05. Topical treatment with GW9508 in non-diabetic mice increases wound healing rate on days 6 and 8 post-injury, while in diabetes-induced mice this effect was not observed. During the proliferative phase, GW9508 treatment decreased f480, il-1ß, mmp9 and vegf gene expression when compared to vehicle. The treatment did not quantitatively modify collagen fiber deposition but was able to alter connective tissue morphology , presenting dermis more intertwined by collagen fibers and formation of dermal papillae, which makes scar tissue more resistant and closer to intact skin. Reanalysis of wound cell single cell RNA sequencing data based on published data from Guerrero-Juarez et al, showed 20 cell clusters organized mainly into four strains composed of fibroblasts, myofibroblasts, myeloid cells, and endothelial cells. Treatment-modulated genes are expressed by fibroblast cell clusters and cells of myeloid origin important in the healing process. Our study showed that topical treatment with GW9508 increases wound healing speed in healthy mice, but not in diabetic mice. The treatment negatively modulates proliferative inflammatory genes expressed by fibroblasts and myeloid cells resulting in structurally more tissue-like skin. Future studies may contribute to a better understanding of the mechanisms involved in this phenomenon, and identification of highest use of these agonistsMestradoCuidado e Inovação Tecnológica em Saúde e EnfermagemMestra em Ciências da Saúde18505532013/07607-8CAPESFAPES

Topical topiramate improves wound healing in an animal model of hyperglycemia

Wound healing is severely affected in hyperglycemia and other metabolic conditions. Finding new therapeutic approaches that accelerate wound healing and improve the quality of the scar may reduce the morbidity commonly associated with skin lesions in diabetes. This study evaluated the effect of topical topiramate (TPM) on wound healing in C57 mice. Streptozotocin-induced hyperglycemic mice were subjected to a wound on the back and randomly allocated for treatment with either vehicle or topical TPM cream (2%) once a day for 14 days. Polymerase chain reaction, Western blotting, and microscopy were performed for the analysis. TPM improved wound healing (complete resolution at Day 10, 98% +/- 5 for TPM vs. 81% +/- 28 for vehicle), increased organization and deposition of collagen Type I, and enhanced the quality of the scars as determined by microscopy. In addition, TPM modulated the expression of cytokines and proteins of the insulin-signaling pathway: In early wound-healing stages, expression of interleukin-10, an anti-inflammatory marker, increased, whereas at the late phase, the pro-inflammatory markers tumor necrosis factor-alpha and monocyte chemoattractant protein-1 increased and there was increased expression of a vascular endothelial growth factor. Proteins of the insulin-signaling pathway were stimulated in the late wound-healing phase. Topical TPM improves the quality of wound healing in an animal model of hyperglycemia. The effect of TPM is accompanied by modulation of inflammatory and growth factors and proteins of the insulin-signaling pathway. Therefore, topical TPM presents as a potential therapeutic agent in skin wounds in patients with hyperglycemia214420430FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçãoThe author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: Sao Paulo Research Foundation, OCRC and Coimbra Group of Brazilian Universities Education provided the grants for this stud

Neotropical freshwater fisheries : A dataset of occurrence and abundance of freshwater fishes in the Neotropics

The Neotropical region hosts 4225 freshwater fish species, ranking first among the world's most diverse regions for freshwater fishes. Our NEOTROPICAL FRESHWATER FISHES data set is the first to produce a large-scale Neotropical freshwater fish inventory, covering the entire Neotropical region from Mexico and the Caribbean in the north to the southern limits in Argentina, Paraguay, Chile, and Uruguay. We compiled 185,787 distribution records, with unique georeferenced coordinates, for the 4225 species, represented by occurrence and abundance data. The number of species for the most numerous orders are as follows: Characiformes (1289), Siluriformes (1384), Cichliformes (354), Cyprinodontiformes (245), and Gymnotiformes (135). The most recorded species was the characid Astyanax fasciatus (4696 records). We registered 116,802 distribution records for native species, compared to 1802 distribution records for nonnative species. The main aim of the NEOTROPICAL FRESHWATER FISHES data set was to make these occurrence and abundance data accessible for international researchers to develop ecological and macroecological studies, from local to regional scales, with focal fish species, families, or orders. We anticipate that the NEOTROPICAL FRESHWATER FISHES data set will be valuable for studies on a wide range of ecological processes, such as trophic cascades, fishery pressure, the effects of habitat loss and fragmentation, and the impacts of species invasion and climate change. There are no copyright restrictions on the data, and please cite this data paper when using the data in publications

NEOTROPICAL ALIEN MAMMALS: a data set of occurrence and abundance of alien mammals in the Neotropics

Biological invasion is one of the main threats to native biodiversity. For a species to become invasive, it must be voluntarily or involuntarily introduced by humans into a nonnative habitat. Mammals were among first taxa to be introduced worldwide for game, meat, and labor, yet the number of species introduced in the Neotropics remains unknown. In this data set, we make available occurrence and abundance data on mammal species that (1) transposed a geographical barrier and (2) were voluntarily or involuntarily introduced by humans into the Neotropics. Our data set is composed of 73,738 historical and current georeferenced records on alien mammal species of which around 96% correspond to occurrence data on 77 species belonging to eight orders and 26 families. Data cover 26 continental countries in the Neotropics, ranging from Mexico and its frontier regions (southern Florida and coastal-central Florida in the southeast United States) to Argentina, Paraguay, Chile, and Uruguay, and the 13 countries of Caribbean islands. Our data set also includes neotropical species (e.g., Callithrix sp., Myocastor coypus, Nasua nasua) considered alien in particular areas of Neotropics. The most numerous species in terms of records are from Bos sp. (n = 37,782), Sus scrofa (n = 6,730), and Canis familiaris (n = 10,084); 17 species were represented by only one record (e.g., Syncerus caffer, Cervus timorensis, Cervus unicolor, Canis latrans). Primates have the highest number of species in the data set (n = 20 species), partly because of uncertainties regarding taxonomic identification of the genera Callithrix, which includes the species Callithrix aurita, Callithrix flaviceps, Callithrix geoffroyi, Callithrix jacchus, Callithrix kuhlii, Callithrix penicillata, and their hybrids. This unique data set will be a valuable source of information on invasion risk assessments, biodiversity redistribution and conservation-related research. There are no copyright restrictions. Please cite this data paper when using the data in publications. We also request that researchers and teachers inform us on how they are using the data