Efeito do tempo de repouso após fadiga induzida no desempenho neuromuscular do quadríceps femoral: ensaio clínico, controlado, randomizado, cego

Objective: To evaluate the effect of different resting periods, after induced muscle fatigue, in the quadriceps femoris neuromuscular performance, in healthy subjects. Methods: Sixty-four volunteers, of both genders, with an average of 21,8 ± 1,7 years and mean body mass index of 24,2 ± 3,7 Kg/m2 were randomly assigned into 4 groups: control group (was not induced fatigue); Experimental Group 1 (Exp. 1 1 minute of rest after fatigue); Experimental Group 3 (Exp. 3 3 minutes of rest after fatigue) and Experimental Group 5 (Exp. 5 5 minutes of rest after fatigue). The subjects were evaluated to the knee´s joint position sense (JPS), followed by 5 flexion-extension knee concentric isokinetic contractions at 60°/s, with concomitant recording of median frequency (Fmed) of rectus femoris (RF), vastus lateralis (VL) and vastus medialis (VM). Then they underwent a muscle fatigue protocol (30 flexion-extension knee concentric contractions at 60°/s) and were reevaluated on the isokinetic performance, Fmed and JPS. Blood lactate levels were measured before initial assessment, immediately after the fatigue protocol and 5 minutes after the end of the rest period. Results: The adoption of 3 minutes of rest was sufficient to restore the initial conditions for the peak torque normalized by body weight and the VL and VM Fmed. The joint position sense returned to its initial values with 1 minute rest. The lactate concentration remained high regardless of the adopted rest period. Conclusion: The use of 3-minute rest period is sufficient for the reestablishment of the neuromuscular parameters to the pre fatigue values.

Equacões preditivas para as pressões respiratórias estáticas máximas de adolescentes brasileiros

Introduction: The reference values and prediction equations for maximal respiratory pressures (MRP) differ significantly between the available studies. This large discrepancy can be attributed to the different methodologies proposed. Although the importance of MRP is widely recognized, there are no Brazilian studies that provide predictive equations and reference values for PRM adolescents. Objectives: The purpose of this study was to provide normal values and propose predictive equations for maximal static respiratory pressures of Brazilian adolescents. Methods: An observational cross-sectional study, which evaluated 182 adolescents of both sexes aged between 12 and 18 years, enrolled in schools of the state and private in the city of Natal / RN. The selection of schools and participants of the study was randomly through a lottery system. The spirometric evaluation was performed through the digital spirometer One Flow FVC prior to the assessment of respiratory muscle strength. The MICs were measured with MVD digital manometer 300. Statistical analysis was performed using the SPSS 17.0 software STATISTICS, assigning the significance level of 5%. The normality of data distribution was verified using the Kolmogorov-Smirnov (KS). The descriptive analysis was expressed as mean and standard deviation. We used one-way ANOVA test to verify the difference of the averages of MRPs between age and gender and comparing the averages of MRPs between levels of physical activity. The test t'Student unpaired compared the averages of MRPs being ages and sexes. The comparison of mean values obtained in this study PRM with the values predicted using the equations mentioned above was relizada by testing paired t'Student. To verify the correlation between the PRM and the independent variables (age, weight, height) was used Pearson correlation test. Levene's test evaluated the homogeneity of variance. To obtain predictive equations analysis was used stepwise multiple linear regression. Results: There was no significant difference in mean age between the PRM. The male adolescents, regardless of age, showed superiority in MRP values when compared to the opposite sex. Weight, height and sex correlated with the PRM. Regression analysis suggested in this study, pointed out that the weight and sex had an influence in MIP and MEP only in relation to sex influenced. The mean for each PRM adolescents classified as very active were superior to those observed in adolescents classified as irregularly active. Conclusion: This study provides reference values and two models of predictive equations for maximal inspiratory and expiratory pressures, and to establish the lower limits of normality that will serve as an indispensable condition for careful evaluation of respiratory muscle strength in Brazilian adolescents

Função pulmonar de crianças com leucemia aguda

Introduction: The leukemias are the most common malignancy in children and adolescents. With the improvement in outcomes, there is a need to consider the morbidity to generate the protocols used in children under treatment. Aim: To evaluate pulmonary function in children with acute leukemia. Method: This study is an observational cross sectional. We evaluated 34 children distributed in groups A and B. Group A comprised 17 children with acute leukemia in the maintenance phase of chemotherapy treatment and group B with 17 healthy students from the public in the city of Natal / RN, matched for gender, age and height. The thoracic mobility was evaluated by thoracic expansion in the axillary and xiphoid levels. Spirometry was measured using a spirometer Microloop Viasys ® following the rules of the ATS and ERS. Maximal respiratory pressures were measured with digital manometer MVD300 (Globalmed ®). The maximal inspiratory pressure (MIP) and maximal expiratory pressure (MEP) were measured from residual volume and total lung capacity, respectively. The data were analyzed using the SPSS 17.0 software assigning the significance level of 5%. Descriptive analysis was expressed as mean and standard deviation. T'student test was used to compare unpaired values found in group A with group B values, as well as with the reference values used. To compare the respiratory coefficients in the axillary level with the xiphoid in each group, we used paired testing t student. Results: Group A was significantly decreased thoracic mobility and MIP compared to group B, and MIP compared to baseline. There was no significant difference between spirometric data from both groups and the values of group A with the reference values Mallozi (1995). There was no significant difference between the MIP and MEP values and lower limits of reference proposed by Borja (2011). Conclusion: Children with acute leukemia, myeloid or lymphoid, during maintenance phase of chemotherapy treatment have reduced thoracic mobility and MIP. However, to date, completion of clinical treatment, the spirometric variables and the strength of the expiratory muscles appear to remain preserved in children between five and ten years

Efeito do método pilates no desempenho muscular e no equilíbrio corporal de mulheres idosas: ensaio clínico controlado randomizado

Background: The Pilates Method is a modality of exercise that has been growing in recent decades, but few researches has been conducted with elderly and little is known about its benefits in this population. Objective: To evaluate the effect of a program of Mat Pilates exercises in muscle performance and postural balance in elderly women. Materials and Method: This is a randomized controlled trial that evaluated the muscle performance (isokinetic dynamometer Biodex System 3 Pro®) and postural balance (Balance Master System®) of 33 women aged 65-80 years. The experimental group (EG) participated of a 12-week program of Mat Pilates exercises with two weekly sessions. Data normality was verified by the Shapiro - Wilk test and were adopted p value < 0.05 as significance level. Results: There were no differences between groups after training. However, the EG showed an increase in the values of extension and flexion average power to 60 ° / s after training (32.19 W to 37.04 W and 14.48 W to 17.56 W, respectively). Conclusion: The proposed exercise program was not effective in the total work and average power of flexor and extensor of the knee, as well as static and dynamic balance of participants

Efeitos da respiração em freno-labial sobre os volumes pulmonares e o padrão de hiperinsuflação dinâmica em pacientes com asma

Objectives: To evaluate how to develop dynamic hyperinflation (DH) during exercise, the influence of pursed-lip breathing in (PLB) on breathing pattern and operating volume in patients with asthma. Methods: We studied 12 asthmatic patients in three moments: (1) anthropometry and spirometry, (2) submaximal incremental cycle ergometer test in spontaneous breathing and (3), submaximal incremental test on a cycle ergometer with PLB using the Opto-electronic plethysmography. Results: Evaluating the end-expiratory lung volume (EEV) during submaximal incremental test in spontaneous breathing, patients were divided into euvolume and hyperinflated. The RFL has increased significantly, the variation of the EEV group euvolume (1.4L) and decreased in group hyperinflated (0.272L). In group volume observed a significant increase of 140% in Vt at baseline, before exercise, comparing the RFL and spontaneous breathing. Hyperinflated group was observed that the RFL induced significant increases of Vt at all times of the test incremental baseline, 50%, 100% load and 66% recovery, 250%, 61.5% and 66% respectively. Respiratory rate decreased significantly with PLB at all times of the submaximal incremental test in the group euvolume. The speed of shortening of inspiratory muscles (VtRcp/Ti) in the hyperinflated increased from 1.6 ± 0.8L/s vs. 2.55 ± 0.9L/s, whereas in the RFL euvolume group ranged from 0.72 ± 0.31L/s vs. 0.65 ± 0.2L/s. The velocity of shortening of the expiratory muscles (VtAb/Te) showed similarity in response to RFL. In group hyperinflated varied vs. 0.89 ± 0.47 vs. 0.80 ± 0.36 and ± 1.17 ± 1L vs. 0.78 ± 0.6 for group euvolume. Conclusion: Different behavior in relation to EEV in patients with moderate asthma were observed, the HD and decreased EEV in response to exercise. The breathing pattern was modulated by both RFL performance as at home, making it more efficient

Treinamento muscular inspiratório para asma: revisão sistemática com metanálise

In asthmatic, the lung hyperinflation leaves the inspiratory muscle at a suboptimal position in length-tension relationship, reducing the capacity of to generate tension. The increase in transversal section area of the inspiratory muscles could reverse or delay the deterioration of inspiratory muscle function. Objective: To evaluate the evidence for the efficacy of inspiratory muscle training (IMT) with an external resistive device in patients with asthma. Methods: A systematic review with meta-analysis was carried out. The sources researched were the Cochrane Airways Group Specialised Register of trials, Cochrane Central Register of Controlled Trials (The Cochrane Library Issue 11 of 12, 2012), MEDLINE, EMBASE, PsycINFO, CINAHL, AMED, ClinicalTrials.gov and reference lists of articles. All databases were searched from their inception up to November 2012 and there was no restriction on the language of publication. Randomised controlled trials that involved the use of an external inspiratory muscle training device versus a control (sham or no inspiratory training device) were considered for inclusion. Two reviewers independently selected articles for inclusion, evaluated risk of bias in studies and extracted data. Results: A total of five studies involving 113 asthmatic patients were included. Three clinical trials were produced by the same group. The included studies showed a significant increase in maximal inspiratory pressure (MD 13.34 cmH2O, 95% CI 4.70 to 21.98), although the confidence intervals were wide. There was no statistically significant difference between the IMT group and the control group for maximal expiratory pressure, peak expiratory flow rate, forced expiratory volume in one second, forced vital capacity, sensation of dyspnea and use of beta2-agonist. There were no studies describing exacerbation events that required a course of oral and inhaled corticosteroids or emergency department visits, inspiratory muscle endurance, hospital admissions and days of work or school. Conclusions: There is no conclusive evidence in this review to support or refute inspiratory muscle training for asthma, once the evidence was limited by the small number of studies included, number of participants in them together with the risk of bias. More well conducted randomized controlled trials are needed, such trials should investigate respiratory muscle strength, exacerbation rate, lung function, symptoms, hospital admissions, use of medications and days off work or school. IMT should also be assessed in the context of more severe asthma

Associação entre diferentes marcadores antropométricos com variáveis espirométricas e de força muscular respiratória em obesos mórbidos

A obesidade é uma epidemia global em alarmante ascensão. Caracterizada pelo excesso de gordura corporal subcutânea, de caráter multifatorial, está relacionada ao surgimento de diversas co-morbidades, entre elas, várias alterações respiratórias, estas se tornam mais intensas quanto maior o grau de obesidade. Não há consenso na relação entre os marcadores de adiposidade geral ou específicos e suas repercussões sobre a função ventilatória, especialmente em relação à sobrecarga muscular respiratória. Objetivo: Analisar a relação entre marcadores antropométricos e variáveis espirométricas e de força muscular respiratória em indivíduos com obesidade mórbida. Métodos: Estudo transversal entre setembro de 2007 e outubro de 2012. Participaram da pesquisa 163 obesos mórbidos (37.1±9.8 anos e IMC=49.0±5.88 Kg/m2) sem alterações espirométricas. Foram observadas as associações entre Índice de Massa Corporal-IMC, adiposidade localizada (Circunferências de Pescoço-CP, Cintura-CC e Quadril-CQ), percentual de gordura corporal através do Índice de Adiposidade Corporal-IAC, volumes e capacidades pulmonares (CVF, VEF1 e VRE) e pressões respiratória estática (PIM e PEM) e dinâmica (VVM). Resultados: O VRE foi o volume mais afetado pela obesidade (apenas 41%predito) e mostrou associação negativa nas relações com todos os marcadores de adiposidade (IMC: r=-0.52; IAC: r=-0.21; CC: r=-0.44; CP: r=-0.25 e CQ: r=-0.28). Há relação inversa entre o percentual de gordura corporal (IAC) com a CVF (r=-0.59), o VEF1(r=-0.56) e o VVM (r=-0.43). As pressões respiratórias são justificadas principalmente pela adiposidade ao redor do pescoço e o IAC. Nossos dados de força muscular respiratória foram melhores associados aos valores de referências sugeridos pelas equações de Harik-Klan et al (1998) para PIM (R²=0.72) e com a equação proposta por Neder et al (1999) para PEM (R²=0.52). Em um modelo de regressão linear, as variáveis de adiposidade não justificam a VVM, já o VEF1 explica 62% da variância da VVM em obesos mórbidos. Conclusão: O percentual da adiposidade corporal e a circunferência do pescoço estão associados com a força muscular e capacidade de gerar fluxo respiratório de obesos mórbidos. Sugerimos a equação elaborada por Harik-Klan et al (1998) para obtenção de valores preditos de PIM e a equação proposta por Neder et al (1999) para valores de normalidade da PEM em sujeitos com obesidade mórbida. Foi possível fornecer uma equação de referência específica para VVM em obesos mórbidos

Efeitos agudos do alongamento estático antes e após exercício e as características neuromusculares do membro inferior: estudo controlado, randomizado e cego

Objective: To evaluate the acute effects of static stretching before and after isokinetic exercise, neuromuscular and biomechanical properties of muscles Biceps Femoris (BF) and semitendinosus (ST). Methods: Eighty-nine volunteers of both genders, healthy and physically active, with a mean age of 22.52 ± 2.6 years and mean BMI 23.86 ± 3.2 kg/m² were randomized into 4 groups: Control Group (CG) made only one Protocol Exercise (PE) without performing the stretching, the Experimental Group 1 (EG1) did stretching before PE; EG2 did the stretching after PE and EG3 did stretching before and after PE. The volunteers were evaluated on the following variables: Range of motion (ROM), soreness, dynamometric variables concentric and eccentric, Neuromuscular Latency Time (NLT) and electromyographic. In the data analysis was assigned a significance level of 5%. Results: ADM and TLNM reported significant reduction in CG, but remained unchanged in GE with p<0,05 and p<0,01, respectively. As for the soreness, no differences between the groups. The electromyographic activity of the BF and ST, in the concentric phase, showed a significant decrease in all groups (p<0,01). However, in the eccentric phase, ST revealed reduction in all groups (p <0.01), except for the CG, while the BF remained unchanged in all groups. The PT showed significant reduction in both conditions (concentric and eccentric) for all groups, with no difference between them (p<0,01). Conclusion: The results of this study do not favor the use of static stretching, even of short duration, before physical activity. However, after exercise or at times unrelated to the sport, he should be given with the aim of avoiding muscle shortening

Efeitos agudos de diferentes intensidades de pressão expiratória positiva sobre os volumes pulmonares em pacientes com acidente vascular encefálico

observar os efeitos agudos de diferentes intensidades de Pressão Expiratória Positiva (PEP) sobre a cinemática do complexo toracoabdominal de pacientes acometidos por Acidente Vascular Encefálico (AVE). Métodos: Foram selecionados 21 indivíduos com AVE e 16 indivíduos saudáveis pareados por idade sexo e IMC para grupo controle. Avaliamos função pulmonar, pressões dos músculos respiratórios e os volumes pulmonares por meio da Pletismografia Optoeletrônica durante três diferentes intensidades de PEP 10, 15 e 20 cmH2O. Resultados: o efeito da PEP no volume corrente (VC) do grupo AVE em relação ao grupo controle foi diferente. Enquanto o grupo controle aumentou o VC em relação a respiração tranquila em 343%, 395,2% e 431,8% nas PEP10, PEP15 e PEP20 cmH2O o grupo AVE aumento 186%, 218.8% e 209.5% (p < 0.0001). A PEP também influenciou de forma diferente em relação ao Tempo inspiratório com intensidades diferentes no grupo controle e AVE (p < 0.0001). No ciclo de trabalho foi observado um aumento no grupo controle nas PEP10 (p < 0.001) e PEP15 (p < 0.05) e no grupo AVE foi observada uma redução PEP20 (p < 0.01) quando comparada com a respiração tranquila. Os volumes operacionais do grupo AVE foi observado aumento do volume inspiratório final da parede torácica (Vifpt) e do Volume expiratório final da parede torácica (Vefpt) diferente do grupo controle que gerou aumento do Vifpt acompanhado de diminuição do Vefpt durante as três intensidades de PEP. Conclusão: A hiperinsuflação observada no grupo AVE demonstra que essa terapêutica deve ser utilizada com cautela especialmente nas intensidades maiores que 10 cmH2O para essa população

Efeito do tratamento com as drogas antipsicóticas haloperidol e clozapina sobre a infecção de toxoplasma gondii em cultura de células retinianas embrionárias

T. gondii is an obligate intracellular protozoan and the main cause of retinochoroiditis in humans. The aim of this study was to evaluate the effect of the antipsychotic drugs haloperidol and clozapine on the course of infection by T. gondii of cultured embryonic retinal cells. Embryo retinas of Gallus gallus domesticus (E12) were used for the preparation of mixed monolayer cultures of retinal cells. Cultures were maintained on plates of 96 and 24 wells by 37°C in DMEM medium supplemented with 5% fetal bovine serum for 2 days. After this period, cultures were simultaneously infected with tachyzoites of T. gondii and treated with the antipsychotics haloperidol and clozapine for 48 hours. Treatment effects were determined by both assessing cell viability with the MTT method and evaluating infection outcomes in slides stained with Giemsa. The treatment with haloperidol and clozapine cells infected with T. gondii resulted in higher viability of these cells, suggesting a possible prevention of neuronal degeneration induced by T. gondii. Additionally, intracellular replication of this protozoan in cells treated with haloperidol and clozapine were significantly reduced, possibly by modulation of the parasite s intracellular calcium concentration

Eletromiografia de superfície da musculatura supra-hióidea durante a deglutição de idosos sem doenças neurológicas e idosos com Parkinson

OBJETIVO: investigar as diferenças entre os achados eletromiográficos dos músculos supra-hióideos direito e esquerdo durante a deglutição de um líquido e um pastoso fino, em sujeitos idosos com doença de Parkinson e idosos sem doença neurológica. MÉTODOS: 30 sujeitos de ambos os sexos participaram deste estudo, 15 idosos sem doença neurológica e os demais com doença de Parkinson. Todos foram submetidos a um protocolo de avaliação clínica e avaliação eletromiográfica de superfície. RESULTADOS: foram constatadas diferenças significantes nos valores entre os sujeitos (f=14.4 e p= 0.000) e entre os lados (f=4.4 e p=0.0037). em relação ao tempo de duração da contração muscular durante a deglutição as diferenças foram significantes apenas entre as consistências (f=8.2 e p=0.005). CONCLUSÃO: a amplitude eletromiográfica foi menor nos sujeitos com doença de Parkinson e maior nos músculos supra-hiódeos do lado direito em ambos os grupos. A duração da contração muscular não diferiu significativamente entre os sujeitos com doença de Parkinson e idosos sem doença neurológica e foi menor para a consistência líquida.

Parâmetros reprodutivo e metabólico de vacas da raça Guzerá suplementadas no pré e pós-parto

Foram utilizadas 30 vacas da raça Guzerá (10 por tratamento), suplementadas no pré-parto (PRE), pós-parto (POS) e sem suplementação (SS) com os objetivos de avaliar no pós-parto as variações do peso corporal (PC), do escore de condição corporal (ECC) (1 = muito magra a 9 = muito gorda) e dos níveis plasmáticos de colesterol total (NPCT), o índice de prenhez (IP) até 112 dias pós-parto e a viabilidade econômica dos tratamentos PRE e POS, que constou de 1 kg/dia/animal de concentrado (16% PB; 3000 kcal ED/kg). O PC e o ECC foram obtidos a cada 28 dias, e colheitas de sangue semanalmente para análise dos NPCT até 112 dias pós-parto. O PC e o ECC tiveram interação dias pós-parto x tratamento. O tratamento PRE aumentou o PC e o ECC das vacas ao parto: 477,80 kg e 6,20 respectivamente, em comparação com os tratamentos POS e SS de 453,37 kg e 5,5; 447,57 kg e 5,28, respectivamente. O PC e o ECC do tratamento PRE foi superior de 28 a 84 dias comparado ao SS, enquanto o tratamento POS foi superior apenas de 84 a 112 dias pós-parto. O tratamento POS não diferiu do PRE no PC de 56 a 112 dias pós-parto e no ECC de 28 a 112 dias pós-parto. O tratamento POS proporcionou aumento do PC e ECC das vacas, já os tratamentos PRE e SS perderam PC e ECC até 112 dias pós-parto. O PC e ECC dos tratamentos PRE, POS e SS aos 112 dias pós-parto foram de 457,79 kg e 5,38; 461,87 kg e 5,66; 436,57 kg e 4,78, respectivamente. Os NPCT tiveram efeito dos dias pós-parto, mas não de tratamento, aumentando com os dias pós-parto. Os IP não foram influenciados pelos tratamentos PRE, POS e SS com 50,00; 62,20 e 57,14%, respectivamente. Economicamente a suplementação não foi viável.

Chemical attributes of a savannah Typic Hapludox soil under management systems

O trabalho teve por objetivo estudar as modificações de atributos químicos de um Latossolo Vermelho após dois anos de manejo com adubação orgânica e/ou mineral com diferentes sistemas de cultivo. O experimento foi realizado na área experimental da Universidade Estadual Paulista - UNESP, localizada no município de Selvíria, Estado do Mato Grosso do Sul, no ano agrícola 2004/2005 e 2005/2006. Os tratamentos foram: cultivo convencional; cultivo mínimo e semeadura direta. As adubações foram: testemunha (sem adubação); adubação mineral (300 kg ha-1 da fórmula 20-00-20); adubação orgânica (esterco bovino - 20 Mg ha-1); adubação orgânica (esterco bovino) + ½ adubação mineral recomendada para a cultura utilizada; 20 e 30 Mg ha-1 de lodo de esgoto. em um ano foi utilizada a soja como cultura e no seguinte o sorgo. Avaliaram-se os atributos químicos do solo em quatro camadas. Os atributos químicos do Latossolo Vermelho foram modificados no primeiro ano após as adubações; a adubação com esterco, lodo de esgoto e a combinação do esterco+adubação mineral foram eficazes em modificar os atributos químicos do solo estudado; o lodo de esgoto foi mais eficaz na recuperação do P do solo e, a semeadura direta contribuiu para o aumento de K no solo

Influência da laserterapia na proliferação de células-tronco do ligamento periodontal humano

Low level laser irradiation (LLLI) has been used in Dentistry to promote wound healing and tissue regeneration. The literature shows a positive effect of LLLI on cell proliferation, but little is known about their effectiveness in promoting stem cells proliferation. The aim of this study was to evaluate the effect of LLLI on the proliferative rate of human periodontal ligament stem cells. Extracts of periodontal ligament were isolated from two third molars removed by surgical and/or orthodontic indication. After enzymatic digestion, the cells were grown in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. On the third subculture, the cells were irradiated with a InGaAlP-diode laser, using two different energy densities (0,5J/cm 2 - 16 seconds and 1,0J/cm² - 33 seconds), with wavelength of 660nm and output power of 30mW. A new irradiation, using the same parameters, was performed 48h after the first. A control group (non irradiated) was kept under the same experimental culture conditions. The Trypan blue exclusion test and the mitochondrial activity of the cells measured by MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] essay were performed to assess the cell proliferation in the intervals of 0, 24, 48 e 72 h after irradiation. The data of cell counts were submitted to nonparametrical statistical tests (Kruskal-Wallis and Mann-Whitney), considering a confidence interval of 95%. DAPI (4 -6-Diamidino-2-phenylindole) staining of the cells was performed at 72h interval to evaluate possible nuclear morphological changes induced by LLLI. The results of this study show that the energy density of 1,0 J/cm² promoted greater cell proliferation compared to the other groups (control and 0,5 J/cm²) at intervals of 48 and 72h. The mitochondrial activity measured by MTT essay showed similar results to the Trypan blue cell counting test. The group irradiated with 1,0J/cm² exhibited a significantly higher MTT activity in the intervals of 48 and 72h, when compared to the group irradiated with 0,5J/cm². No nuclear morphological change was observed in the cells from the three groups studied. It is concluded that LLLI has stimulatory effects on the proliferation of human periodontal ligament stem cells. Therefore, the use of laser irradiation in this cell type may be important to promote future advances in periodontal regeneration

Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação

Dental pulp stem cells have been widely investigated because of their ability to differentiate into both dental and non-dental cells, with potential use in therapies involving tissue engineering. The technique of cell cryopreservation represents a viable alternative for the conservation of these cells, since it stops reversibly, in a controlled manner, all of cell biological functions in an ultra low temperature. The present study aimed to evaluate, using in vitro experiments, the influence of a cryopreservation protocol on the biologic acti vity of stem cells from human exfoliated deciduous teeth (SHED). Cells obtained from the pulp of three deciduous teeth on end-stage exfoliation or with indicated extraction were expanded in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. At second subculture (P2), a group of cells were submitted to cryopreservation for 30 days in 10% DMSO diluted in fetal bovine serum, at -80º C, while the remind cells continued under normal conditions of cell culture. Cell proliferation was evaluated in both groups (not cryopreserved or cryopreserved) by Trypan blue stain essay at intervals of 24, 48 and 72h after plating. Cell cycle analysis of SHEDs submitted or not to the cryopreservation protocol was performed in the same intervals. Events related to cell death were studied by Annexyn V and PI expression under flow cytometry at the intervals of 24 and 72h. The presence of nuclear morphological changes was evaluated by DAPI staining at 72h interval. It was observed that both groups exhibited an upward cell proliferation curve, without considerable changes in cell viability throughout the experiment. The distribution of cell in the cell cycle phasis was consistent with cell proliferation in both groups. There were no nuclear morphological damages in the end range of the experiment. therefore, it is concluded that the proposed cryopreservation protocol is efficient for storing the studied cell type, allowing its use in future experimental studies