Estudo de fase clínica I utilizando membrana de quitosana dopada com hormônios derivados de plaquetas

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Análise comparativa de diferentes metodologias para cultivo de células-tronco mesenquimais derivadas de tecido muscular

Pós-graduação em Biociências - FCLAS

Uso de células-tronco para o tratamento de tendinite em eqüinos

Brazil has the fourth largest horse herd in the world, this is due the recognition and appreciation that the different equestrian games are having within the country. Injuries of the tendon, especially in the digital flexor tendon, are the main cause of athletic life reduction among horses. The treatment of tendinitis in horses seeks full recovery of the damage tissue reestablishing the function previously lost, however conventional treatments have proven to be ineffective when considered the quality of the scar tissue and the rate of recurrence. Due to this, the use of adult stem cells to the treatment of musculoskeletal injuries of horses has been studied for some time. This method of treatment consists of aspiration of bone marrow or removal of subcutaneous fat tissue and implantation of these cells in the injured tissue. After obtaining the bone marrow the implantation can be performed with total bone marrow, with the mononuclear fraction of MSC or with cells cultured in vitro. From the fat tissue is used the stromal vascular fraction obtained by collagenase digestion, followed or not by cell culture. According to some studies, cell therapy with material obtained from bone marrow or adipose tissue has shown to be viable, given that these materials are abundant in repair components such as mesenchymal stem cells (MSC), growth factors and other components of the collagen matrix. Several studies using both types of cells have shown great potential and promising clinical results. However, knowledge of the biology and characterization of these cells remain largely unknown, and therefore is needed great care and caution when using stem cells for the treatment of musculoskeletal disorders in horses

Chitosan-collagen scaffolds can regulate the biological activities of adipose mesenchymal stem cells for tissue engineering

Scaffolds of chitosan and collagen can offer a biological niche for the growth of adipose derived stem cells (ADSC). The objective of this work was to characterize the physico-chemical properties of the scaffolds and the ADSC, as well as their interactions to direct influences of the scaffolds on the behavior of ADSC. The methodology included an enzymatic treatment of fat obtained by liposuction by collagenase, ASDC immunophenotyping, cell growth kinetics, biocompatibility studies of the scaffolds analyzed by the activity of alkaline phosphatase (AP), nitric oxide (NO) determination by the Griess-Saltzman reaction, and images of both optical and scanning electron microscopy of the matrices. The extent of the crosslinking of genipin and glutaraldehyde was evaluated by ninhydrin assays, solubility tests and degradation of the matrices. The results showed that the matrices are biocompatible, exhibit physical and chemical properties needed to house cells in vivo and are strong stimulators of signaling proteins (AP) and other molecules (NO) which are important in tissue healing. Therefore, the matrices provide a biological niche for ADSC adhesion, proliferation and cells activities.

Isolamento, cultivo, caracterização e criopreservação de células tronco mesenquimais derivadas de membrana amniótica, geléia de Wharton e sangue do cordão umbilical de fetos bovinos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Fatores indutores e supressores da eritropoiese em caninos doentes renais crônicos

Pós-graduação em Cirurgia Veterinária - FCAV

Derivação de células tronco de tecido ovariano na espécie canina

Pós-graduação em Medicina Veterinária - FCAV

Avaliação do conhecimento de enfermeiros da rede de atenção à saúde no município de Bauru (SP) sobre cuidado aos pacientes com feridas: um estudo transversal

Pós-graduação em Enfermagem - FMB

Matrizes de quitosina-colágeno podem regular as atividades biológicas de células tronco mensenquimais adiposas para a engenharia de tecidos

Pós-graduação em Biofísica Molecular - IBILCE

Effect of low-level laser therapy in an experimental model of osteoarthritis in rats evaluated through raman spectroscopy

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

Islet of Langerhans transplantation. A comparative study of two different methods for isolating islet cells from rat pancreas

Two different methods for isolation of islet of Langerhans on control of metabolic abnormalities of alloxan-induced diabetic rat were tested. Sixty rats were randomly assigned to four experimental groups: GI included 10 non-diabetic control rats, GII included 10 diabetic control rats, without treatment, GIII included 20 diabetic rats (10 inbred and 10 outbred rats) that received islet of Langerhans transplantation (ILT) using islet cells prepared by collagenase, and GIV included 20 diabetic rats (10 inbred and 10 outbred rats) submitted to ILT using islet cells prepared by nonenzymatic method. Clinical and laboratory parameters at beginning and 4, 7, 14, 21 and 30 days of follow-up were recorded. Outbred rats were immunosuppressed with cyclosporin A, diabetes was induced by e.v. alloxan administration, and islet cells were isolated from normal donor Lewis rats and injected into the portal vein. ILT corrected the body weight gain, polyuria, polydipsia, polyphagia, and the high levels of blood and urine glucose in 73.7% of rats treated by enzymatic method and in 64.7% of those ones treated by nonenzymatic method. However, there was no significantly difference between the two methods (P > 0.50). We did not also observe significantly difference between the two methods when ILT was performed either in inbred or outbred rats. We concluded that ILT performed by nonenzymatic method may be an alternative treatment for diabetes due to be less expensive and to have possible advantages in the isolation process.

Characterization of biomodified dentin matrices for potential preventive and reparative therapies

Biomodification of existing hard tissue structures, specifically tooth dentin, is an innovative approach proposed to improve the biomechanical and biochemical properties of tissue for potential preventive or reparative therapies. The objectives of the study were to systematically characterize dentin matrices biomodified by proanthocyanidin-rich grape seed extract (GSE) and glutaraldehyde (GD). Changes to the biochemistry and biomechanical properties were assessed by several assays to investigate the degree of interaction, biodegradation rates, proteoglycan interaction, and effect of collagen fibril orientation and environmental conditions on the tensile properties. The highest degree of agent–dentin interaction was observed with GSE, which exhibited the highest denaturation temperature, regardless of the agent concentration. Biodegradation rates decreased remarkably following biomodification of dentin matrices after 24 h collagenase digestion. A significant decrease in the proteoglycan content of GSE-treated samples was observed using a micro-assay for glycosaminoglycans and histological electron microscopy, while no changes were observed for GD and the control. The tensile strength properties of GD-biomodified dentin matrices were affected by dentin tubule orientation, most likely due to the orientation of the collagen fibrils. Higher and/or increased stability of the tensile properties of GD- and GSE-treated samples were observed following exposure to collagenase and 8 months water storage. Biomodification of dentin matrices using chemical agents not only affects the collagen biochemistry, but also involves interaction with proteoglycans. Tissue biomodifiers interact differently with dentin matrices and may provide the tissue with enhanced preventive and restorative/reparative abilities.

Avaliação da eficácia dos biocurativos em ratos wistar com lesões de pele causadas por queimaduras

Accidents caused by thermal, chemical, electrical or radioactive agents cause skin lesions causing burns of varying degrees. The therapeutic approach aims to restore damaged tissues and involves a wide range of products on the market. This study aims to evaluate the use of biological dressing, biotech product developed at the Blood Center of Botucatu / UNESP obtained from fresh frozen plasma or platelet concentrate with in vitro addition of thrombin and calcium gluconate. This addition in the platelet concentrate, intended to release the active growth factors of the platelets granules on the healing process. The study of the effectiveness of Platelet Gel home made in Wistar rats was established, in agreement with scald burns, comparing efficacy and cost of Platelet Gel with usual hospital -based treatment collagenase + chloramphenicol plus cost analysis through pharmacoeconomics. We used 25 Wistar rats were divided into 3 treatment groups: Group A, Collagenase + Chloramphenicol; Group B, Platelet Gel and C, control. The products were applied every other day for 30 days in animals. In group A, there was the presence of erythema and crust in all animals. The exudates was indentified 2/10 animals. For the Group B, we observed the presence of erythema and crust at all and no presence of exudates. In group C all the animals showed erythema with no presence of exudates and scab occurred in 1/10. Statistical analysis showed significant difference ( p < 0.0 ) for crust formation between Groups B and C. In the histological analysis, group A showed a slight amount of blood vessels and collagen fibers, moderate amounts of macrophages and fibroblasts was observed while B and C groups showed moderate amounts of blood vessels, macrophages and fibroblasts and discreet presence of collagen fibers. The re-epithelialization occurred in most animals of all groups without significant statistical differences. For the aspects of pharmacoeconomics, the platelet gel presented a better cost - effectiveness in relation to treatment based on collagenase / chloramphenicol. In light of the ethical aspects of the raw material is the result of spontaneous blood donation, the proposal should have biological dressings productions the responsibility of public blood transfusion centers for free distribution. This may point to the production chain of Brazilian blood banks like special blood components for use no intravenous.

Expression of soluble and functional full-length human matrix metalloproteinase-2 in Escherichia coli

Characterization of the matrix metalloproteinase-2 (MMP-2) substrates and understanding of its function remain difficult because up to date preparations containing minor amounts of other eukaryotic proteins that are co-purified with MMP-2 are still used. In this work, the expression of a soluble and functional full-length recombinant human MMP-2 (rhMMP-2) in the cytoplasm of Escherichia coli is reported, and the purification of this metalloproteinase is described. Culture of this bacterium at 18 degrees C culminated in maintenance of the soluble and functional rhMMP-2 in the soluble fraction of the E. coli lysate and its purification by affinity with gelatin-sepharose yielded approximately 0.12 mg/L of medium. Western Blotting and zymographic analysis revealed that the most abundant form was the 72-kDa MMP-2, but some gelatinolytic bands corresponding to proteins with lower molecular weight were also detected. The obtained rhMMP-2 was demonstrated to be functional in a gelatinolytic fluorimetric assay, suggesting that the purified rhMMP-2 was correctly folded. The method described here involves fewer steps, is less expensive, and is less prone to contamination with other proteinases and MMP inhibitors as compared to expression of rhMMP-2 in eukaryotic tissue culture. This protocol will facilitate the use of the full-length rhMMP-2 expressed in bacteria and will certainly help researchers to acquire new knowledge about the substrates and biological activities of this important proteinase. (C) 2011 Elsevier B.V. All rights reserved.

Maternal Moderate Physical Training during Pregnancy Attenuates the Effects of a Low-Protein Diet on the Impaired Secretion of Insulin in Rats: Potential Role for Compensation of Insulin Resistance and Preventing Gestational Diabetes Mellitus

The effects of pregestational and gestational low-to-moderate physical training on insulin secretion in undernourished mothers were evaluated. Virgin female Wistar rats were divided into four groups as follows: control (C, n = 5); trained (T, n = 5); low-protein diet (LP, n = 5); trained with a low-protein diet (T + LP, n = 5). Trained rats ran on a treadmill over a period of 4 weeks before mate (5 days week(-1) and 60 min day(-1), at 65% of VO2max). At pregnancy, the intensity and duration of the exercise were reduced. Low-protein groups were provided with an 8% casein diet, and controls were provided with a 17% casein diet. At third day after delivery, mothers and pups were killed and islets were isolated by collagenase digestion of pancreas and incubated for a further 1 h with medium containing 5.6 or 16.7 mM glucose. T mothers showed increased insulin secretion by isolated islets incubated with 16.7 mM glucose, whereas LP group showed reduced secretion of insulin by isolated islets when compared with both C and LP + T groups. Physical training before and during pregnancy attenuated the effects of a low-protein diet on the secretion of insulin, suggesting a potential role for compensation of insulin resistance and preventing gestational diabetes mellitus.