Denise Cantarelli Machado

Sources of processed lipoaspirate cells: influence of donor site on cell concentration.

Background: Recently, adipose tissue harvested by liposuction has been identified as a source of processed lipoaspirate cells. The aim of this study was to determine the concentration of processed lipoaspirate cells in adipose tissue obtained by liposuction from different harvest areas in women. Methods: A prospective cross-sectional study was conducted in 25 women in whom liposuction in four or more different zones in the same procedure was indicated. After selective liposuction, the material was sent to the laboratory, where it was processed for extraction of processed lipoaspirate cells, which were separated from the adipose tissue, quantified, and characterized through determination of c-kit expression. The following harvest regions were evaluated: upper abdomen, lower abdomen, trochanteric region, inner thigh, knee, and flank. The cell concentration obtained at each site was compared by analysis of variance for mixed models. Results: A significant difference was found for cell concentration obtained at the different harvest sites. The cell concentration in the lower abdomen was greater than in other areas, but no significant difference was found in relation to the inner thigh. Conclusions: The lower abdomen and the inner thigh may have higher processed lipoaspirate cell concentrations. These sites may turn out to be better sources of adult mesenchymal stem cells.

Differentiation of human adipose-derived adult stem cells into neuronal tissue: Does it work?

Adipose tissue contains many cells and proteins that are of value not only for their potential therapeutic applications, but also for the low cost of their harvest and delivery. Mesenchymal stem cells (MSC) were originally isolated from the bone marrow, although similar populations have been isolated from adipose and other tissues. At one time, neural tissues were not regarded as regenerative populations of cells. Therefore, the identification of cell populations capable of neuronal differentiation has generated immense interest. Adipose tissue may represent an alternative source of cells that are capable of neuronal differentiation, potentially enhancing its use in the treatment of neurological disease. The aim of this review is to cover the current state of knowledge of the differentiation potential of human adipose-derived stem (ADAS) cells, specifically their ability to give rise to neuronal cells in vitro. This review presents and discusses different protocols used for inducing human ADAS cells to differentiate in vitro, and the neuronal markers utilized in each system.

Effects of human aging on periodontal tissues

Loss of teeth is frequently associated with periodontal disease in older adults. The aim of this review was to present the effects of aging on the periodontal tissues. Aging alone does not lead to critical loss of periodontal attachment in healthy elderly persons. The effects of aging on periodontal tissues are based on molecular changes in the periodontal cells, which intensify bone loss in elderly patients with periodontitis. These effects may be associated with (1) alterations in differentiation and proliferation of osteoblasts and osteoclasts; (2) an increase in periodontal cell response to the oral microbiota and mechanical stress leading to the secretion of cytokines involved in osseous resorption; and (3) systemic endocrine alterations in the elderly people.

Prevention of seizures and reorganization of hippocampal functions by transplantation of bone marrow cells in the acute phase of experimental epilepsy

In this study, we investigated the therapeutic potential of bone marrow mononuclear cells (BMCs) in a model of epilepsy induced by pilocarpine in rats. BMCs obtained from green fluorescent protein (GFP) transgenic mice or rats were transplanted intravenously after induction of status epilepticus (SE). Spontaneous recurrent seizures (SRS) were monitored using Racines seizure severity scale. All of the rats in the saline-treated epileptic control group developed SRS, whereas none of the BMC-treated epileptic animals had seizures in the short term (15 days after transplantation), regardless of the BMC source. Over the long-term chronic phase (120 days after transplantation), only 25% of BMC-treated epileptic animals had seizures, but with a lower frequency and duration compared to the epileptic control group. The density of hippocampal neurons in the brains of animals treated with BMCs was markedly preserved. At hippocampal Schaeffer collateral-CA1 synapses, long-term potentiation was preserved in BMC-transplanted rats compared to epileptic controls. The donor-derived GFP(+) cells were rarely found in the brains of transplanted epileptic rats. In conclusion, treatment with BMCs can prevent the development of chronic seizures, reduce neuronal loss, and influence the reorganization of the hippocampal neuronal network.

Protective immune response against methicillin resistant Staphylococcus aureus in a murine model using a DNA vaccine approach.

Methicillin resistant Staphylococcus aureus (MRSA) are a major pathogen responsible for serious hospital infections worldwide. These bacteria are resistant to all beta-lactam antibiotics due to the production of an additional penicillin binding protein, the PBP2a, encoded by the mecA gene, which shows low affinity for this class of antibiotics. In this study, we cloned an internal region from the transpeptidase domain from the PBP2a into a mammalian expression vector, to be used as DNA vaccine in a Murine model. After three sets of DNA vaccination, the immune response represented by antibodies against a fragment of PBP2a was evaluated by enzyme linked immunosorbent assay (ELISA), showing a significant antibody response. The antibacterial effect of the DNA vaccine was evaluated by intraperitoneal immunization and challenge with a sublethal dose of MRSA for 7 days in mice. After the challenge, the number of bacteria from kidneys from immunized and non-immunized mice were determined. Kidneys from immunized mice had 1000 times less on bacteria than the positive controls (non-immunized mice). The response specificity indicates no effects against the normal PBPs from staphylococci and no effects against Gram positive rods from normal intestinal flora. Our results indicate that the immunization against the PBP2a from MRSA using a DNA vaccine approach could be used as a new strategy to efficiently fight these multiresistant bacteria.

Bone marrow mononuclear cells reduce seizure frequency and improve cognitive outcome in chronic epileptic rats.

AIMS Epilepsy affects 0.5-1% of the worlds population, and approximately a third of these patients are refractory to current medication. Given their ability to proliferate, differentiate and regenerate tissues, stem cells could restore neural circuits lost during the course of the disease and reestablish the physiological excitability of neurons. This study verified the therapeutic potential of bone marrow mononuclear cells (BMMCs) on seizure control and cognitive impairment caused by experimentally induced epilepsy. MAIN METHODS Status epilepticus (SE) was induced by lithium-pilocarpine injection and controlled with diazepam 90 min after SE onset. Lithium-pilocarpine-treated rats were intravenously transplanted 22 days after SE with BMMCs obtained from enhanced green fluorescent protein (eGFP) transgenic C57BL/6 mice. Control epileptic animals were given an equivalent volume of saline or fibroblast injections. Animals were video-monitored for the presence of spontaneous recurrent seizures prior to and following the cell administration procedure. In addition, rats underwent cognitive evaluation using a Morris water maze. KEY FINDINGS Our data show that BMMCs reduced the frequency of seizures and improved the learning and long-term spatial memory impairments of epileptic rats. EGFP-positive cells were detected in the brains of transplanted animals by PCR analysis. SIGNIFICANCE The positive behavioral effects observed in our study indicate that BMMCs could represent a promising therapeutic option in the management of chronic temporal lobe epilepsy.

Effect of titanium surface roughness on human bone marrow cell proliferation and differentiation: an experimental study

PURPOSE To assess the proliferation and differentiation of human bone marrow-derived cells cultured on titanium surfaces with different roughness characteristics. METHODS Cells obtained from the iliac crest of an adult human donor were routinely processed and cultured on titanium surfaces of varying roughness, according to their preparation method: polishing only (smooth surface) and polishing followed by etching with HF/HNO3 for 15 and 30 minutes (rough surfaces). Surfaces were assessed using scanning electronic microscopy and profilometry. RESULTS Titanium disks etched with acid for 15 minutes allowed greater cell proliferation in all culture periods. The level of osteopontin and osteocalcin expression was increased in both acid-etched groups, which indicates an advanced stage of differentiation of cells into osteoblasts. CONCLUSIONS Increased surface roughness accelerates the differentiation of undifferentiated mesenchymal cells into osteogenic lineage cells, but does not necessarily favor cell proliferation. An intermediate surface roughness of 0.5microm (acid etching for 15 minutes) favors both initial and final cell responses.

Associação entre manifestações estomatológicas e características socioeconômicas e culturais de crianças brasileiras e norte-americanas infectadas pelo HIV

OBJETIVO: Investigar as caracteristicas socioeconomicas e culturais de criancas infectadas pelo HIV e de suas familias, relacionando-as com a presenca de manifestacoes estomatologicas da AIDS, e identificar as manifestacoes estomatologicas mais frequentes na amostra estudada. METODO: A amostra foi composta por 184 criancas de ambos os sexos, com idade entre 0 e 13 anos, atendidas no Hospital de Clinicas de Porto Alegre e Hospital Sao Lucas da Pontificia Universidade Catolica, em Porto Alegre, Brasil, e no hospital universitario da Universidade do Estado de Nova Iorque em Stony Brook, Estado de Nova Iorque, Estados Unidos. Os dados foram analisados pelos testes do qui-quadrado, t de Student e analise de variância. RESULTADOS: Dos 184 pacientes, 63,59% eram do Hospital de Clinicas de Porto Alegre; 22,28% do hospital de Stony Brook; e 14,13% do Hospital Sao Lucas. Na amostra brasileira, 42,66% das criancas residiam com os pais naturais (principalmente com a mae), enquanto que nos Estados Unidos 56,10% das criancas moravam com pais adotivos. Em relacao a renda, 39,86% das familias no Brasil ganhavam entre 180 e 450 dolares mensais; 33,57% nao tinham renda ou nao forneceram seus dados. Nos Estados Unidos, a pergunta sobre renda nao foi autorizada, mas todas as familias foram classificadas pelos assistentes sociais do hospital como tendo renda abaixo de 1 000 dolares. O HIV foi adquirido por transmissao vertical em 97,20% dos casos no Brasil e 97,56% nos Estados Unidos. A frequencia das manifestacoes estomatologicas foi maior nas criancas brasileiras (72,73%) do que nas norte-americanas (53,66%) (P = 0,01). CONCLUSOES: As condicoes socioeconomicas e culturais, estrutura e renda familiar, acesso a informacao sobre AIDS e a adesao aos esquemas terapeuticos influenciaram a frequencia das manifestacoes estomatologicas nas duas amostras.

Cytotoxicity of Silver Solder Employed in Orthodontics

OBJECTIVE To test the null hypothesis that the silver soldering employed in orthodontics is not cytotoxic for fibroblasts. MATERIALS AND METHODS This in vitro study was performed using a culture of mice fibroblasts (lineage NIH/3T3), divided into four groups (n = 10 each): control, negative control (stainless steel archwire), positive control (amalgam disks), and test group (silver soldering). After cell culture in complete Dulbecco modified eagle medium and achievement of confluence in 80%, the suspension was added to the plates of 24 wells containing the specimens and incubated in an oven at 37 degrees C for 24 hours. The plates were analyzed on an inverted light microscope, photomicrographs were obtained, and the results were recorded as response rates based on modifications of the parameters of Stanford according to the size of the diffusion halo of the toxic substance and quantity of cell lysis. RESULTS The results revealed a maximum response rate for the silver soldering group, as well as severe inhibition of cell proliferation and growth, more round cells with mostly darkened and granular aspects, suggesting lysis with cell death. A similar response was seen in the positive control group. CONCLUSION The hypothesis is rejected. The silver soldering used in orthodontics represents a highly cytotoxic material for the cells analyzed.

Can Local Supply of Bone Marrow Mononuclear Cells Improve the Outcome from Late Tubular Repair of Human Median and Ulnar Nerves

The purpose of this non-randomised retrospective study was to compare nerve regeneration after reconnection with silicone tubes with two different strategies. A total of 44 patients with injured median or ulnar nerves in the forearm were surgically treated. In one group of patients, a silicone tube alone was placed in the nerve gap. In a second group, the silicone tube was filled with autologous bone marrow mononuclear cells obtained by aspiration from the iliac crest. Motor function, sensation and the effect of pain on function were assessed 1 year after surgery. The tubes filled with bone marrow cells showed better recovery than the empty tubes. The use of bone marrow mononuclear cells in addition to tube re-connection may promote better nerve regeneration than conventional tubular repair.