Influência da fadiga no comportamento neuromuscular do quadríceps femoral, após reconstrução do ligamento cruzado anterior

Muscle fatigue is a phenomenon that promotes physiological and biomechanical disorders and their changes in healthy subjects have been widely studied and have significant importance for care in preventing injuries, but we do not have many information about its effects in patients after ACL reconstruction. Thus, this study is to analyze the effects of fatigue on neuromuscular behavior of quadriceps after ACL reconstruction. To reach this objective, participants were forty men, twenty healthy (26,90 ± 6,29 years) and twenty after ACL reconstruction (29,75 ± 7,01 years) with a graft of semitendinosus and gracilis tendons, between four to six months after surgery. At first, there was an assessment of joint position sense (JPS) at the isokinetic dynamometer at a speed of 5°/s and target angle of 45° to analyze the absolute error of JPS. Next, we applied the a muscle fatigue protocol, running 100 repetitions of isokinetic knee flexion-extension at 90°/s. Concurrently with this protocol, there was the assessment of muscle performance, as the peak torque (PT) and fatigue index, and electromyographic activity (RMS and median frequency). Finally, we repeated the assessment of JPS. The statistical analysis showed that patients after ACL reconstruction have, even under normal conditions, the amended JPS compared with healthy subjects and that after fatigue, both have disturbances in the JPS, but this alteration is significantly exacerbated in patients after ACL reconstruction. About muscle performance, we could notice that these patients have a lower PT, although there are no differences between the dynamometric and EMG fatigue index. These findings show the necessity about the cares of pacients with ACL reconstruction in respect of the risks of articulate instability and overload in ligamentar graft

Adesão de células mesenquimais da medula óssea de camundongos e do ligamento periodontal de ratos a diferentes superfícies de titânio: estudo comparativo in vitro

The present experiment used cell culture to analyze the adhesion capacity of mouse mesenchymal bone marrow cells and rat periodontal ligament to different titanium surfaces. Grade II ASTM F86 titanium discs 15mm in diameter and 1.5mm thick were used and received 2 distinct surface treatments (polished and cathodic cage plasma nitriding). The cells were isolated from the mouse bone marrow and rat periodontal ligament and cultured in α-MEM basic culture medium containing antibiotics and supplemented with 10% FBS and 5% CO2, for 72 hours at 37ºC in a humidified atmosphere. Subculture cells were cultured in a 24-well plate with a density of 1 x 104 cells per well. The titanium discs were distributed in accordance with the groups, including positive controls without titanium discs. After a 24-hour culture, the cells were counted in a Neubauer chamber. The results show that both the mouse mesenchymal bone marrow cells and rat periodontal ligament cells had better adhesion to the control surface. The number of bone marrow cells adhered to the polished Ti surface was not statistically significant when compared to the same type of cell adhered to the Ti surface treated by cathodic cage plasma nitriding. However a significant difference was found between the control and polished Ti groups. In relation to periodontal ligament cell adhesion, a significant difference was only found between the control and plasma-treated Ti surfaces. When comparing equal surfaces with different cells, no statistically significant difference was observed. We can therefore conclude that titanium is a good material for mesenchymal cell adhesion and that different material surface treatments can influence this process

A influência da criopreservação nas células mesenquimais indiferenciadas do ligamento periodontal de humanos análise comparativa in vitro

Cryopreservation is a process where cells or biological tissues are preserved by freezing at very low temperatures and aims to cease reversibly, in a controlled manner, all the biological functions of living tissues, i.e., maintain cell preservation so that it can recover with high degree of viability and functional integrity. This study aimed to evaluate the influence of cryopreservation on the mesenchymal stem cells originating from the periodontal ligament of human third molars by in vitro experiments. Six healthy teeth were removed and the periodontal cells grown in culture medium containing α-MEM supplemented with antibiotics and 15% FBS in a humidified atmosphere with 5% CO2 at 37° C. Cells isolated from each sample were divided into two groups: Group I - immediate cell culture (not fresh cryopreserved cells) and Group II - cell cryopreservation, during a period of 30 days. Analyses of rates of cell adhesion and proliferation in different groups were performed by counting the cells adhered to the wells, in intervals of 24, 48 and 72 hours after the start of cultivation. The number of cells in each well was obtained by counting viable cells with the use of hemocytometer and the method of exclusion of cells stained by trypan blue. The difference between groups for each of the times was analyzed by Wilcoxon test. Regarding the temporal evolution for each group, analysis was done by Friedman's test to verify the existence of differences between times and, when it existed, the Wilcoxon penalty was applied. The results showed no statistically significant difference between the two groups analyzed in this study. Therefore, we conclude that the cryopreservation process, after a period of 30 days, did not influence the cell type studied, and there was no difference in growth capacity in vitro between the groups

Avaliação in vitro da adesão e proliferação de células mesenquimais do ligamento periodontal humano em diferentes superfícies de titânio

In the last years, many scientific researches in implantology have been focused on alternatives that would provide higher speed and quality in the process of osseointegration. Different treatment methods can be used to modify the topographic and chemical properties of titanium surface in order to optimize the tissue-implant reactions by a positive tissue response. This study aimed to evaluate the adhesion and proliferation of mesenchymal cells from human periodontal ligament on two different titanium surfaces, using cell culture techniques. Grade II titanium discs received different surface treatments, forming two distinct groups: polished and cathodic cage plasma nitriding. Human periodontal ligament mesenchymal cells were cultured on titanium discs in 24-well cell culture plates, at a density of 2 x 104 cells per well, including wells with no discs as positive control. Data obtained by counting the cells that adhered to the titanium surfaces (polished group and cathodic cage group) and to the plastic surface (control group), in the 24, 48 and 72-hour periods after plating, were used to analyze cell adhesion and proliferation and to obtain the cell growing curve in the different groups. The data were submitted to nonparametric analysis and the differences between groups were compared by Kruskal-Wallis and Friedman statistical tests. No statistically significant differences were found in the cells counts between the groups (p>0.05). It was concluded that both treatments produced surfaces compatible with the adhesion and proliferation of human periodontal ligament mesenchymal cells

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

Efeito do kinesio taping® no desempenho neuromuscular do quadríceps e no equilíbrio em sujeitos submetidos à reconstrução do ligamento cruzado anterior: ensaio clínico e randomizado

Objetivo: Investigar os efeitos imediatos do Kinesio Taping® no desempenho neuromuscular do Quadríceps Femoral (QF) de indivíduos submetidos à reconstrução do Ligamento Cruzado Anterior (LCA). Metodologia: trata-se de um ensaio clínico e randomizado composto por 45 indivíduos do sexo masculino que se encontravam entre 12ª e 17ª semanas após reconstrução do LCA. Todos foram submetidos a uma avaliação inicial composta pela análise do equilíbrio postural, através da baropodometria; determinação do Senso de Posição Articular (SPA), seguidas das avaliações isocinéticas excêntricas e concêntricas a 600/s, concomitante com a captação do sinal eletromiográfico do músculo Vasto Lateral (VL). Posteriormente foram alocados de forma aleatória em Grupo Controle (GC), Grupo Placebo (GP) e Grupo Experimental (GE). Os indivíduos do GE foram submetidos ao protocolo sugerido (aplicação do Kinesio Taping® no QF do membro acometido), enquanto os do GP utilizaram a aplicação do Kinesio Taping® sem as recomendações propostas pelo método. Já os indivíduos do GC permaneceram em repouso por dez minutos, sendo todos os indivíduos submetidos a uma reavaliação de forma idêntica à primeira. Foram analisadas as seguintes variáveis: pico de torque médio, pico de torque/peso corporal, potência muscular e erro absoluto do SPA para a dinamometria; amplitude ântero-posterior e médio-lateral para a baropodometria; e a amplitude de ativação muscular (Root Means Square - RMS) por meio da eletromiografia de superfície. Resultados: Nenhuma das variáveis analisadas apresentou diferenças intergrupo ou intragrupo. Conclusão: O Kinesio Taping® não altera o desempenho neuromuscular do quadríceps femoral de indivíduos submetidos à reconstrução do LCA para nenhuma das variáveis analisadas.

Correlação entre torque, equlíbrio e função do joelho após reconstrução do LCA

Background: Several studies emphasize the importance of assessing the knee function after anterior cruciate ligament (ACL) reconstruction. The influence of several variables on the function of these patients has been analyzed, but there is no consensus in the science literature. Purpose: To evaluate the correlation between the torque and balance on the knee function after ACL reconstruction. Methods: 23 males patients with ACL reconstruction were tested. The procedures of the study included analysis of concentric peak torque at 60o/s and 180o/s of quadriceps femoris and hamstring muscle with a isokinetic dynamometer. The balance in single-limb stance was measured with stabilometry. The functional performance were performed by two hop tests. To estimate the subjective function of the patients was applied Lysholm Knee Scoring Scale and a Global Rating scale. Results: The analysis of data showed a moderate positive correlation between knee extensor torque and functional performance tests (r= 0,48; p=0,02). A moderate negative correlation was found between the two variables of the stabilometry center of pressure and average speed of centre of pressure and the Global Rating scale (r = -0.4, p = 0,04 and r = -0,49, p = 0 ,02, respectively). No correlation was found between peak torque and balance in single-limb stance. Conclusion: The results of the present study suggest that knee extensor strength and postural balance have some influence on knee function in patients after ACL reconstruction

Análise citogenética de células-tronco derivadas do subendotélio da veia umbilical humana

Mesenchymal stem cells (MSCs) are known as a population of multi-potential cells able to proliferate and differentiate into multiple mesodermal tissues including bone, cartilage, muscle, ligament, tendon, fat and stroma. Several applications of the study of EC can be emphasized the therapeutic techniques such as guided bone regeneration by implantation of EC in the affected site, without the need for bone grafts, using titanium as a vehicle. The process of cryopreservation is essential for the maintenance of cell cultures, since the cell line is frozen, it can be maintained in liquid nitrogen for an indefinite period and then thawed for therapeutic or experimental purposes. The aim of this study was to isolate a population of MSCs derived from the subendothelium of the umbilical vein human (MSCs-SUVH) to assess cytogenetic analysis by the possibility of appearance of chromosomal changes in two different situations: MSCs-SUVH regarding the process of cryopreservation and MSCs-SUVH grown on the surface of titanium. Flow cytometry analysis revealed that, this cell population was positive for the markers CD29, CD73 and CD90, but there was no expression of hematopoietic lineage markers, such as CD14, CD34 and CD45 and demonstrated capacity for osteogenic differentiation. The chromosomes obtained from the primary culture of MSCs-SUVH were analyzed by GTW banding technique, and results are described as guidelines to ISCN 2005. There was not the emergence of clonal chromosomal changes in the MSCs-SUVH in different situations analyzed. However one of the strings presented a balanced paracentric inversion, probably a cytogenetic constitutional alterations, which was present before and after the experimental situations that the MSCs-SUVH was submitted

Análise imuno-histoquímica das proteínas RANK, RANKL e OPG em dentes de ratos reimplantados

The RANK / RANKL / OPG sy stem plays an important role in bone formation and resorption . This finding has been regarded as one of the m ost important advances in the understanding of bone biology with respect to osteoclastogenesis. The aim of this study was to investigate the expression of RANKL / RANK / OPG markers in reimplanted t eeth of rats, and to observe the relationship between the expression of these markers and to oth and bone resorption. Thirty male Wistar rats (Rattus norvegicus albinos) had their maxillary right incisors extrac ted , and were divided into 2 groups according to the period that the extracted teeth were kept in dry air before reimplantation : G1 (n = 15) - 5 minutes , and G2 (n = 15) - 60 minutes . After reimplantation, teeth were analyzed at intervals of 1, 3 and 7 da ys. After these experimental periods, the animals were euthanized. Longitudinal sections with 5μm thick were obtained and stained with Hematoxylin and Eosin for histological analysis , while 3μm thick sections were subjected to immunohistochemical analysis of OPG , RANK and RANKL. The results showed that the RANK / RANKL / OPG system actively participates in both the repair process, as well as tooth and bone resorption . Extr a - alveolar time of 60 minutes before replantation caused minor expressions of RANKL a nd OPG, not influencing the expression of RANK; RANKL immunostaining showed higher in both groups when compared to other biomarkers, participating in all phases of bone and tooth resorption; RANKL was associated to both osteoclastogenesis and c ell ular proliferation , and was expressed in both groups.

Efeitos imediatos da plataforma vibratória no desempenho neuromuscular do quadríceps femoral, após reconstrução do LCA: ensaio controlado, randomizado e cego

Objective: to investigate the immediate effect of the vibrating platform on the neuromuscular performance of the quadriceps femoris and on the postural oscillation of subjects submitted to Anterior Cruciate Ligament (ACL) reconstruction. Materials and methods: this study is a randomized and blind clinical trial. Forty-four male volunteers (average age of 27,4 ±6,2 IMC of 26,85± 3,8 Kg/m² and post surgery timeframe of 17± 1,4 weeks) were randomized into two groups: OFF platform (n=22, protocol of exercise over the vibrating platform off) and ON platform (n=22 protocol of exercise over the vibrating platform on, 50Hz frequency and 4mm of amplitude). All volunteers were submitted to assessment the isokinetic evaluation of the quadriceps femoris (isometric and isokinetic at 60°/s) and of the electromyography activity of the muscles Vasto Lateralis (VL) and Vasto Medialis (VM), besides the postural oscillation (baropodometry) in two distinct moments: before and immediately after the intervention protocol. The data was analyzed through the SPSS 20.0 software, with a 5% significance level. To verify the homogeneity of the groups it was used an ANOVA one way, and a ANOVA mixed model to compare the intra and inter groups. Results: it was observed differences between the pre and the post, to latero lateral velocity, isometric torque peak and total work in comparison with intragroup. However, it wasn’t verified any difference in comparing the intergroup in the preevaluation and in the post-evaluation protocol over the vibrating platform. Conclusion: the use of the vibrating platform doesn’t change as an immediate manner the isokinetic performance of the quadriceps femoris, the electromyography activity of the VL and the VM, also doesn’t interfere with the postural oscillation of individuals that were submitted to the ACL reconstruction.