Clinical and Microbiologic Evaluation of Chronic Leg Ulcers: A Cross-sectional Study

OBJECTIVE: This study aimed at evaluating the flora and bacterial load of chronic leg ulcers (CLUs) according to the clinical judgment of colonization or infection.DESIGN: This was an analytical and cross-sectional study.SETTING: This study was conducted in an outpatient wound care unit in the Dermatology Department of the Botucatu School of Medicine-UNESP, Brazil.PARTICIPANTS: The participants were patients with CLUs who did not use systemic antibiotics.METHODS: The ulcers were clinically divided into 3 groups: ulcers with good granulation tissue (GGT), critical colonization (CC), and infection. Secretion was collected from a 1-cm(2) area using a swab and seeded by the semiquantitative method.OUTCOME MEASURES: The main outcome measures were genus and species of the bacteria found in the cultures and result of the semiquantitative culture correlating with the clinical diagnosis of GGT, CC, and infection.MAIN RESULTS: Seventy-seven ulcers were evaluated: 27 with GGT, 29 with CC, and 21 with infection. Gram-negative bacteria were most often found in all groups (81%): Pseudomonas aeruginosa, in granulation and colonized ulcers, and Proteus mirabilis, in infected ulcers. Ulcers from the infected group showed higher bacterial load.CONCLUSIONS: The flora of CLUs was predominantly constituted by gram-negative bacteria, and P aeruginosa was the most prevalent. The bacterial load of infected ulcers was higher as compared with the others, although some ulcers with GGT also presented a high load. The interpretation of microbiologic tests based on the swab techniques and even on semiquantitative analysis requires close clinical correlation.

Serology, isolation, and molecular detection of Leptospira spp. from the tissues and blood of rats captured in a wild animal preservation centre in Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Quantification of Biological Tissue and Construction of Patient Equivalent Phantom (Skull and Chest) for Infants (1-5 years old)

Our main purpose in this study was to quantify biological tissue in computed tomography (CT) examinations with the aim of developing a skull and a chest patient equivalent phantom (PEP), both specific to infants, aged between 1 and 5 years old. This type of phantom is widely used in the development of optimization procedures for radiographic techniques, especially in computed radiography (CR) systems. In order to classify and quantify the biological tissue, we used a computational algorithm developed in Matlab (R). The algorithm performed a histogram of each CT slice followed by a Gaussian fitting of each tissue type. The algorithm determined the mean thickness for the biological tissues (bone, soft, fat, and lung) and also converted them into the corresponding thicknesses of the simulator material (aluminum, PMMA, and air). We retrospectively analyzed 148 CT examinations of infant patients, 56 for skull exams and 92 were for chest. The results provided sufficient data to construct a phantom to simulate the infant chest and skull in the posterior anterior or anterior posterior (PA/AP) view. Both patient equivalent phantoms developed in this study can be used to assess physical variables such as noise power spectrum (NPS) and signal to noise ratio (SNR) or perform dosimetric control specific to pediatric protocols.

Novel Antimicrobial Peptides Bacterial Cellulose Obtained by Symbioses Culture Between Polyhexanide Biguanide (PHMB) and Green Tea

Bacterial cellulose is a highly hydrated pellicle made up of a random assembly of ribbon shaped fibers less than 5 nm wide. The unique properties provided by the nanometric structure have led to a number of diagnostic biological probes, display devices due to their unique size-dependent medical applications. Bacterial cellulose matrix extracellular is a novel biotechnology and unique medicine indicated for ultimate chronic wound treatment management, drug delivery, tissue engineering, skin cancer and offers an actual and effective solution to a serious medical and social problem and to promote rapid healing in lesions caused by Diabetic burns, ulcers of the lower limbs or any other circumstance in which there's epidermal or dermal loss. In this work, it is reported novel antimicrobial peptides (AMPs) bacterial cellulose/polyhexanide biguanide (PHMB) which are produced by symbioses culture between polyhexanide biguanide and green tea culture medium resulting in the pure 3-D structure consisting of an ultra-fine network of novel biocellulose/PHMB nanofibres matrix (2-8 nm), highly hydrated (99% in weight), and with higher molecular weight, full biocompatibility.

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

Pós-graduação em Biociências - FCLAS

Tissue-engineering-based Strategies for Regenerative Endodontics

Stemming from in vitro and in vivo pre-clinical and human models, tissue-engineering-based strategies continue to demonstrate great potential for the regeneration of the pulp-dentin complex, particularly in necrotic, immature permanent teeth. Nanofibrous scaffolds, which closely resemble the native extracellular matrix, have been successfully synthesized by various techniques, including but not limited to electrospinning. A common goal in scaffold synthesis has been the notion of promoting cell guidance through the careful design and use of a collection of biochemical and physical cues capable of governing and stimulating specific events at the cellular and tissue levels. The latest advances in processing technologies allow for the fabrication of scaffolds where selected bioactive molecules can be delivered locally, thus increasing the possibilities for clinical success. Though electrospun scaffolds have not yet been tested in vivo in either human or animal pulpless models in immature permanent teeth, recent studies have highlighted their regenerative potential both from an in vitro and in vivo (i.e., subcutaneous model) standpoint. Possible applications for these bioactive scaffolds continue to evolve, with significant prospects related to the regeneration of both dentin and pulp tissue and, more recently, to root canal disinfection. Nonetheless, no single implantable scaffold can consistently guide the coordinated growth and development of the multiple tissue types involved in the functional regeneration of the pulp-dentin complex. The purpose of this review is to provide a comprehensive perspective on the latest discoveries related to the use of scaffolds and/or stem cells in regenerative endodontics. The authors focused this review on bioactive nanofibrous scaffolds, injectable scaffolds and stem cells, and pre-clinical findings using stem-cell-based strategies. These topics are discussed in detail in an attempt to provide future direction and to shed light on their potential translation to clinical settings.

Marcadores de superfície de Células-tronco: estudo de reação cruzada inter-espécies

Mesenchymal stem cells (MSCs) are adult multipotent cells with fibroblastoid morphology and adherent to plastic. Furthermore, they can be obtained from different sources. Besides bone marrow, these cells are taken from umbilical cord blood, umbilical vein, saphenous vein, peripheral blood, arteries, liver and fetal pancreas, placenta, dental pulp and adipose tissue. MSCs derived from adipose tissue are important because of the abundant number of cells that can be obtained from this tissue, easy access and little discomfort to the patient. This study compared two techniques for obtaining MSCs from adipose tissue: mechanical dissociation (MD) and enzymatic digestion (ED). We also analyzed the inter-species cross-reactions using commercial monoclonal antibodies directed against surface antigens of stem cells from different species: mouse, horse, rabbit, monkey and human. We found that MD technique is favorable in relation to ED within 15 days of culture, and ED is more efficient in the first days of culture. The data also showed that MD causes less damage to cellular DNA. About inter-species cross-reactions, the monoclonal antibody A69 directed against stem cells from rabbits, which can be used in veterinary medicine, particularly in research involving horses

Ethanol production by Zymomonas mobilis during sucrose fermentation: Optimization of culture conditions using factorial design

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

Automatic classification of prostate stromal tissue in histological images using Haralick descriptors and Local Binary Patterns

In this paper we presente a classification system that uses a combination of texture features from stromal regions: Haralick features and Local Binary Patterns (LBP) in wavelet domain. The system has five steps for classification of the tissues. First, the stromal regions were detected and extracted using segmentation techniques based on thresholding and RGB colour space. Second, the Wavelet decomposition was applied in the extracted regions to obtain the Wavelet coefficients. Third, the Haralick and LBP features were extracted from the coefficients. Fourth, relevant features were selected using the ANOVA statistical method. The classication (fifth step) was performed with Radial Basis Function (RBF) networks. The system was tested in 105 prostate images, which were divided into three groups of 35 images: normal, hyperplastic and cancerous. The system performance was evaluated using the area under the ROC curve and resulted in 0.98 for normal versus cancer, 0.95 for hyperplasia versus cancer and 0.96 for normal versus hyperplasia. Our results suggest that texture features can be used as discriminators for stromal tissues prostate images. Furthermore, the system was effective to classify prostate images, specially the hyperplastic class which is the most difficult type in diagnosis and prognosis.

Green tea polyphenol epigallocatechin-3-gallate and cranberry proanthocyanidins act in synergy with cathelicidin (LL-37) to reduce the LPS-induced inflammatory response in a three-dimensional co-culture model of gingival epithelial cells and fibroblasts

Objectives: The human antimicrobial peptide cathelicidin (LL-37) possesses anti-inflammatory properties that may contribute to attenuating the inflammatory process associated with chronic periodontitis. Plant polyphenols, including those from cranberry and green tea, have been reported to reduce inflammatory cytokine secretion by host cells. In the present study, we hypothesized that A-type cranberry proanthocyanidins (AC-PACs) and green tea epigallocatechin-3-gallate (EGCG) act in synergy with LL-37 to reduce the secretion of inflammatory mediators by oral mucosal cells. Methods: A three-dimensional (3D) co-culture model of gingival epithelial cells and fibroblasts treated with non-cytotoxic concentrations of AC-PACs (25 and 50 mg/ml), EGCG (1 and 5 mg/ml), and LL-37 (0.1 and 0.2 mM) individually and in combination (AC-PACs + LL-37 and EGCG + LL-37) were stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS). Multiplex ELISA assays were used to quantify the secretion of 54 host factors, including chemokines, cytokines, growth factors, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs). Results: LL-37, AC-PACs, and EGCG, individually or in combination, had no effect on the regulation of MMP and TIMP secretion but inhibited the secretion of several cytokines. ACPACs and LL-37 acted in synergy to reduce the secretion of CXC-chemokine ligand 1 (GRO-a), granulocyte colony-stimulating factor (G-CSF), and interleukin-6 (IL-6), and had an additive effect on reducing the secretion of interleukin-8 (IL-8), interferon-g inducible protein 10 (IP-10), and monocyte chemoattractant protein-1 (MCP-1) in response to LPS stimulation. EGCG and LL-37 acted in synergy to reduce the secretion of GRO-a, G-CSF, IL-6, IL-8, and IP-10, and had an additive effect on MCP-1 secretion. Conclusion: The combination of LL-37 and natural polyphenols from cranberry and green tea acted in synergy to reduce the secretion of several cytokines by an LPS-stimulated 3D coculture model of oral mucosal cells. Such combinations show promising results as potential adjunctive therapies for treating inflammatory periodontitis.

Response of human pulps to different in-office bleaching techniques: preliminary findings

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Bacterial cellulose biocomposites for guided tissue regeneration

Bacterial cellulose (BC) has become established as a remarkably versatile biomaterial and can be used in a wide variety of scientific applications, especially for medical devices. In this work, the bacterial cellulose fermentation process is modified by the addition of chondroitin sulfate and hyaluronic acid (1% w/w) to the culture medium before the bacteria is inoculated. Besides, biomimetic precipitation of calcium phosphate of biological interest from simulated body fluid on bacterial cellulose was studied. Chondroitin sulfate and hyaluronic acid influences in bacterial cellulose were analyzed using transmission infrared spectroscopy (FTIR), XRD (X-ray diffraction) and scanning electron microscopy (SEM). FTIR analysis showed interaction between bacterial cellulose nanobiocomposites and calcium phosphate and XRD demonstrated amorphous calcium phosphate and calcium chloride on bacterial cellulose nanobiocomposites. SEM images confirmed incorporation of calcium phosphate in bacterial cellulose nanobiocomposites surface with different calcium phosphate particles morphology.

Determination of the mercury fraction linked to protein of muscle and liver tissue of Tucunaré (Cichla spp.) from the Amazon Region of Brazil

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

Effect of GaAIAs low-level laser therapy on the healing of human palate mucosa after connective tissue graft harvesting: randomized clinical trial

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

Combined therapy with mineral trioxide aggregate, and guided tissue regeneration for a large radicular cyst: a 13-year follow-u

Biomaterials such as membrane barriers and/or bone grafts are often used to enhance periapical new bone formation. A combination of apical surgery and these biomaterials is one of the latest treatment options for avoiding tooth extraction. In case of periapical lesions, guided tissue regeneration (GTR) is attempted to improve the self-regenerative healing process by excluding undesired proliferation of the gingival connective tissue or migration of the oral epithelial cells into osseous defects. In many cases, GTR is necessary for achieving periodontal tissue healing. This report describes the healing process after surgery in a challenging case with a long-term followup. In this case report, endodontic surgery was followed by retrograde sealing with mineral trioxide aggregate (MTA) in the maxillary right central incisor and left lateral incisor. Apicectomy was performed in the maxillary left central incisor and a 1-mm filling was removed. The bone defect was filled with an anorganic bone graft and covered with a decalcified cortical osseous membrane. No intraoperative or postoperative complications were observed. After 13 years of follow-up, the patient showed no clinical signs or symptoms associated with the lesion and radiographic examination showed progressive resolution of radiolucency. In conclusion, the combination of apical surgery and regenerative techniques can successfully help the treatment of periapical lesions of endodontic origin and is suitable for the management of challenging cases