Relationship between biomarkers of cartilage in serum and degenerative joint disease in Lusitano horses

Introduction: Cartilage degradation biomarkers are a potential tool for early diagnosis of degenerative joint disease (DJD). In young horses, Coll2-1 and Coll2-1NO2 have been studied in serum and reported to be useful in the assessment of joint disease. Fib3-2 has been described to be higher in serum of humans with osteoarthritis but was never assessed in horses. The aim of the current study was to evaluate biomarkers’ changes with age, sex and exercise and correlate them with DJD. Material and Methods: Blood collection and radiographic examination were performed in 51 Lusitanian horses. Moreover, inertial sensor-based detection of lameness was used to assess pain together with subjective examination. Results: Females presented significantly higher concentrations of Coll2-1 (p = 0.015) and Coll2-1NO2 (p = 0.014) compared to males. We have found significant influence of high level of work in lower concentration of Coll2-1 (p = 0.001) and significant influence of sex in concentration of Coll2-1NO2 (p = 0.030). There was no influence of sex, age and work on Fib3-2. All biomarkers were increased in the DJD group (n= 35) compared to healthy controls (n = 16). This difference was significant for Coll2-1 (p = 0.015). When sorted by sex and age groups, significant difference in Coll2-1 between disease and healthy controls disappeared in old horses and females. Discussion/ Conclusion: Coll2-1 is a good marker of cartilage degradation in horses with DJD, being more specific in young horses and males. Fib3-2 may be further explored to help identify disease in particular cases.

The Crosslinking Degree Controls The Mechanical, Rheological, And Swelling Properties Of Hyaluronic Acid Microparticles.

Viscosupplements, used for treating joint and cartilage diseases, restore the rheological properties of synovial fluid, regulate joint homeostasis and act as scaffolds for cell growth and tissue regeneration. Most viscosupplements are hydrogels composed of hyaluronic acid (HA) microparticles suspended in fluid HA. These microparticles are crosslinked with chemicals to assure their stability against enzyme degradation and to prolong the action of the viscosupplement. However, the crosslinking also modifies the mechanical, swelling and rheological properties of the HA microparticle hydrogels, with consequences on the effectiveness of the application. The aim of this study is to correlate the crosslinking degree (CD) with these properties to achieve modulation of HA/DVS microparticles through CD control. Because divinyl sulfone (DVS) is the usual crosslinker of HA in viscosupplements, we examined the effects of CD by preparing HA microparticles at 1:1, 2:1, 3:1, and 5:1 HA/DVS mass ratios. The CD was calculated from inductively coupled plasma spectrometry data. HA microparticles were previously sized to a mean diameter of 87.5 µm. Higher CD increased the viscoelasticity and the extrusion force and reduced the swelling of the HA microparticle hydrogels, which also showed Newtonian pseudoplastic behavior and were classified as covalent weak. The hydrogels were not cytotoxic to fibroblasts according to an MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2014.

The Influence Of Altered Occlusion On Pro-inflammatory Cytokine Levels In The Tmj Synovial Tissues Of Rats.

The aim of this study was to evaluate whether altered occlusion affects both the condylar cartilage thickness and the cytokine levels of the TMJs of rats. Thirty adult-male rats (n=30) were randomly assigned to three experimental conditions: a control group that underwent sham operations with unaltered occlusion; an FPDM group that underwent functional posterior displacement of the mandible that was induced by an incisor guiding appliance; and an iOVD group in which the increased occlusal vertical dimension was induced in the molars. The rats were subjected to the FPDM or iOVD model for 14 days and then killed. Both the right and left TMJs were removed and randomly assigned to examination with staining or immunoassay techniques. Toluidine blue staining was used to measure the thicknesses of the four layers of the articular cartilage (i.e., the fibrous, proliferating, mature, and hypertrophic layers). ELISA assays were used to assess the concentrations of the pro-inflammatory cytokines IL-1α, IL-1β, IL-6, and tumour necrosis factor (TNF-α). The measurements of the articular cartilage layers and cytokine concentrations were analyzed with ANOVA and Tukey's tests and Kruskal-Wallis and Dunn tests, respectively (α=5%). The thickness of articular cartilage in the FPDM group (0.3±0.03mm) was significantly greater than those of the control (0.2±0.01mm) and iOVD (0.25±0.03mm) groups. No significant difference was observed between the control and iOVD groups. The four articular cartilage layers were thicker in the FPDM group than in the control and iOVD groups, and the latter two groups did not differ one from each other. Both the FPDM and iOVD groups exhibited higher cytokine levels than did the control (p<0.05) group. Compared to the FPDM group, the iOVD group exhibited significantly higher levels of IL-1β and TNF-α. Both models induced inflammation in the TMJ and caused significant structural changes in the TMJ and surrounding tissues.

Obesity Versus Osteoarthritis: Beyond The Mechanical Overload.

Obesity is currently considered a major public health problem in the world, already reaching epidemic characteristics, according to the World Health Organization. Excess weight is the major risk factor associated with various diseases, such as type 2 diabetes mellitus, hypertension, dyslipidemia and osteometabolic diseases, including osteoporosis and osteoarthritis. Osteoarthritis is the most prevalent rheumatic disease and the leading cause of physical disability and reduced quality of life of the population over 65 years. It mainly involves the joints that bear weight - knees and hips. However, along with the cases of obesity, its prevalence is increasing, and even in other joints, such as hands. Thus, it is assumed that the influence of obesity on the development of OA is beyond mechanical overload. The purpose of this review was to correlate the possible mechanisms underlying the genesis and development of these two diseases. Increased fat mass is directly proportional to excessive consumption of saturated fatty acids, responsible for systemic low-grade inflammation condition and insulin and leptin resistance. At high levels, leptin assumes inflammatory characteristics and acts in the articular cartilage, triggering the inflammatory process and changing homeostasis this tissue with consequent degeneration. We conclude that obesity is a risk factor for osteoarthritis and that physical activity and changes in diet composition can reverse the inflammatory and leptin resistance, reducing progression or preventing the onset of osteoarthritis.

Saude fisica, indicadores antropometricos, desempenho fisico e bem-estar subjetivo em idosos atendidos no ambulatorio de geriatria do HC/UNICAMP

Universidade Estadual de Campinas . Faculdade de Educação Física

Morphological description of Dipturus mennii (Chondrichthyes: Elasmobranchii: Rajidae) and its differentiation from Dipturus trachyderma

Squamation patterns and skeletal anatomy (neurocranium, visceral arches, synarcual cartilage, scapulocoracoid, puboischiadic bar, and mixopterigium) of Dipturus mennii Gomes & Paragó, 2001 are described as a contribution to our limited knowledge of the anatomy of species of Dipturus Rafinesque, 1810. The hyoid and branchial arches, as well as the synarcual cartilage, are described for the first time in this species. We provide morphological comparisons of this species with Dipturus trachyderma (Krefft & Stehmann, 1975), a species that may be confused with D. mennii; we further corroborate, through anatomical features, that these species warrant separate taxonomic recognition. The main differences between D. mennii and D. trachyderma were found in squamation of the nuchal and middisc region, neurocranium, pectoral girdle, and principally the clasper skeleton. The morphology of the pelvic girdle is similar in both species. Dipturus is characterized by having the ventral terminal cartilage J-shaped (as opposed to the Z-shaped ventral terminal cartilage in Zearaja, whose species were, until recently, placed in Dipturus). Additional characters that may be derived for Dipturus include the anterior rostral groove and elevated rostral proportions

Utilização dos subprodutos da fresagem do osso subcondral em substituição ao enxerto autólogo esponjoso em artrodeses de carpo de cães

A artrodese rádio-cárpica é procedimento radical usado no tratamento de fraturas e luxações do carpo, com danos à fibrocartilagem palmar e às estruturas ligamentares de sustentação, decorrentes de hiperextensão. Para este estudo foram selecionados 15 cães, submetidos à artrodese rádio-cárpica, entre abril/02 e julho/04. Foram estudados os casos quanto à formação de calo ósseo e às complicações pós-cirúrgicas. A técnica consistiu na ostectomia da superfície articular do rádio, expondo-a e retificando-a. Os subprodutos desta fresagem foram reintroduzidos como enxerto após sua redução em tamanho com rugina. Para a estabilização articular, em todos os casos, foram usadas placas compressivas. Foram realizados exames radiográficos imediatamente à intervenção cirúrgica e em intervalos de 30 dias, até a total fusão articular. Os resultados obtidos neste estudo, com fusão da articulação em até 300 dias de pós-cirúrgico em 80% dos casos estudados, foram semelhantes aos dispostos para as técnicas que utilizam enxertos autólogos esponjosos, sendo que, em 68% dos pacientes, a recuperação se deu sem qualquer complicação significativa. Pequenas complicações, como o aumento de volume, foram observadas em 22% dos casos, não determinando qualquer alteração no resultado final. Tais resultados viabilizam a técnica descrita, demonstrando a exeqüibilidade da utilização dos subprodutos da fresagem do osso subcondral como enxerto em artrodeses de carpo em cães.

Ossificação endocondral em embriões e fetos de bovinos

Estudou-se a ossificação endocontral de 18 embriões e 12 fetos de até três meses de gestação, os quais foram coletados de úteros gestantes em frigoríficos e abatedouros. Os úteros foram dissecados e, em seguida, realizou-se uma incisão dorsal até o cérvix para avaliações macroscópicas dos embriões e fetos. Para o estudo microscópico foram realizadas técnicas de inclusão, seguidas de marcação dos depósitos de cálcio e fósforo, responsável pela ossificação dos moldes de cartilagem. Foram identificados hipertrofia da cartilagem e morte dos condrócitos e aumento da área de depósito de cálcio e fósforo, por volta da 10ª semana gestacional (74 dias). Durante a 11ª semana de gestação (81 dias), os grupamentos de carbonato de cálcio e fósforo espalharam-se por todo o osso, sendo mais intenso na diáfise.

Effects of different levels of protein intake and physical training on growth and nutritional status of young rats

This study aimed to investigate the effects of physical training, and different levels of protein intake in the diet, on the growth and nutritional status of growing rats. Newly-weaned Wistar rats (n=48) were distributed into six experimental groups: three of them were subjected to physical swim training (1 h per day. 5 d per week, for 4 wk, after 2 wk of familiarization) and the other three were considered as controls (non-trained). Each pair of groups, trained and non-trained, received diets with a different level of protein in their composition: 14%. 21% or 28%. The animals were euthanized at the end of the training period and the following analyses were performed: proteoglycan synthesis as a biomarker of bone and cartilage growth, IGF-I (insulin-like growth factor-I) assay as a biomarker of growth and nutritional status. total RNA and protein concentration and protein synthesis measured in vivo using a large-dose phenylalanine method. As a main finding, increased dietary protein, combined with physical training, was able to improve neither tissue protein synthesis nor muscle growth. In addition, cartilage and bone growth seem to be deteriorated by the lower and the higher levels of protein intake. Our data allow us to conclude that protein enhancement in the diet, combined with physical exercise, does not stimulate tissue protein synthesis or muscle mass growth. Furthermore, physical training, combined with low protein intake, was not favorable to bone development in growing animals

Exercise x BCAA supplementation in young trained rats: what are their effects on body growth?

The purpose of this study was to evaluate whether Branched-chain amino acids (BCAAs) supplementation had any beneficial effects on growth and metabolic parameters of young rats submitted to chronic aerobic exercise. Thirty-two young rats (age: 21-d) were randomly assigned to four experimental groups (n = 8): Supplemented Trained (Sup/Ex), Control Trained (Ctrl/Ex), Supplemented Sedentary (Sup/Sed) and Control Sedentary (Ctrl/Sed). The trained groups underwent a five-week swimming protocol and received supplemented (45 mg BCAA/body weight/day) or control ration. Trained animals presented a lower body length and a higher cartilage weight, regardless of supplementation. Physical activity was responsible for a substantial reduction in proteoglycan synthesis in cartilage tissue, and BCAA supplementation was able to attenuate this reduction and also to improve glycogen stores in the liver, although no major differences were found in body growth associated to this supplementation.

Effects of Laser on the Synovial Fluid in the Inflammatory Process of the Knee Joint of the Rabbit

Objective: The purpose of this study was to evaluate the effects of low-level laser (LLL) energy on the clinical signs of inflammation and the cellular composition of synovial fluid (SF) in the inflamed knee of the rabbit. Background Data: There are few findings related to the effects of LLL on SF in inflammatory processes and there is little knowledge about the optimal parameters for reducing joint inflammation. Materials and Methods: Inflammation in the right knee of 36 rabbits was induced by intracapsular injection (0.2 mL) of Terebinthina commun (Tc). The animals were randomly assigned to three groups: acute experimental group (AEG), chronic experimental group (CEG), and control group (CG), which only received Tc. Each group was divided in two subgroups of six animals each. The AEG and CEG groups began to receive laser treatment 2 and 5 d after the induction of inflammation, respectively. Laser irradiation at a wavelength of 830 nm, power output of 77 mW, and power density of 27.5 W/cm(2) was applied daily for 7 d for either 0.12 sec or 0.32 sec, resulting in doses of 3.4 J/cm(2) and 8 J/cm(2), respectively. Body mass, joint perimeter, joint temperature, and the morphology of the SF were analyzed. Results: There was no statistically significant differences between groups in the body mass, joint perimeter, and SF morphology. Conclusion: Laser irradiation with the selected parameters produced only a few subtle differences in the inflammatory signs and the SF. The lack of effects may have been due to the short irradiation time.

Mesenchymal progenitor cells from canine fetal tissues: yolk sac, liver, and bone marrow

During fetal development, mesenchymal progenitor (MP) cells are co-localized in major hematopoietic territories, such as yolk sac (YS), bone marrow (BM), liver (LV), and others. Studies using mouse and human MP cells isolated from fetus have shown that these cells are very similar but not identical to adult mesenchymal stem cells (MSC). Their differentiation potential is usually restricted to production of highly committed osteogenic and chondrogenic precursors. Such properties of fetal MP cells can be very useful for tissue regeneration, when a great number of committed precursors are required. The objectives of this study were to isolate and characterize MP cells from canine YS, BM, and LV in early and late stages of fetal development. Gestational stage was identified, and cell culture conditions were evaluated for efficient isolation of canine MP cells. All canine fetal MP cells expressed vimentin, nestin, and CD44 proteins. Cytokeratin 18 expression was observed in BM-and LV-MP cells, and vascular endothelial (VE)-cadherin expression was observed only in YS-MP cells. A small number of MP cells (5%) from LV and YS expressed Oct3/4 protein. The differentiation potential of canine fetal MP cells varied significantly: YS- and BM-MP cells differentiated into bone and cartilage, whereas LV-MP cells differentiation was limited to osteogenic fate. None of the canine fetal MP cells were able to differentiate into adipose cells. Our data suggest that canine fetal MP cells are an appropriate in vitro model to study MP biology from hematopoietic territories and they are a source of committed osteogenic and chondrogenic precursors for regenerative medicine.

Human fallopian tube: a new source of multipotent adult mesenchymal stem cells discarded in surgical procedures

Background: The possibility of using stem cells for regenerative medicine has opened a new field of investigation. The search for sources to obtain multipotent stem cells from discarded tissues or through non-invasive procedures is of great interest. It has been shown that mesenchymal stem cells (MSCs) obtained from umbilical cords, dental pulp and adipose tissue, which are all biological discards, are able to differentiate into muscle, fat, bone and cartilage cell lineages. The aim of this study was to isolate, expand, characterize and assess the differentiation potential of MSCs from human fallopian tubes (hFTs). Methods: Lineages of hFTs were expanded, had their karyotype analyzed, were characterized by flow cytometry and underwent in vitro adipogenic, chondrogenic, osteogenic, and myogenic differentiation. Results: Here we show for the first time that hFTs, which are discarded after some gynecological procedures, are a rich additional source of MSCs, which we designated as human tube MSCs (htMSCs). Conclusion: Human tube MSCs can be easily isolated, expanded in vitro, present a mesenchymal profile and are able to differentiate into muscle, fat, cartilage and bone in vitro.

Preclinical Studies with Umbilical Cord Mesenchymal Stromal Cells in Different Animal Models for Muscular Dystrophy

Umbilical cord mesenchymal stromal cells (MSC) have been widely investigated for cell-based therapy studies as an alternative source to bone marrow transplantation. Umbilical cord tissue is a rich source of MSCs with potential to derivate at least muscle, cartilage, fat, and bone cells in vitro. The possibility to replace the defective muscle cells using cell therapy is a promising approach for the treatment of progressive muscular dystrophies (PMDs), independently of the specific gene mutation. Therefore, preclinical studies in different models of muscular dystrophies are of utmost importance. The main objective of the present study is to evaluate if umbilical cord MSCs have the potential to reach and differentiate into muscle cells in vivo in two animal models of PMDs. In order to address this question we injected (1) human umbilical cord tissue (hUCT) MSCs into the caudal vein of SJL mice; (2) hUCT and canine umbilical cord vein (cUCV) MSCs intra-arterially in GRMD dogs. Our results here reported support the safety of the procedure and indicate that the injected cells could engraft in the host muscle in both animal models but could not differentiate into muscle cells. These observations may provide important information aiming future therapy for muscular dystrophies.

The relative effects of severe burn injury and pre- and post-natal protein deprivation on mandibular condyle morphology

The mandible has a mixed embryological origin, and its growth is associated with the secondary cartilage of the condyle process (CP). In this area, growth depends on an array of intrinsic and extrinsic factors that influence protein metabolism. In the present study, we used an adolescent rat model to evaluate the growth and development of the CP under conditions of pre- and postnatal protein deficiency, combined with or without the stress of severe burn injury (BI). We found that protein deficiency severely undermined the growth of the CP, by altering the thickness of its constituent layers. BI is also capable of affecting CP growth, although the effect is less severe than protein deficiency. Interestingly, the summed effect of protein deficiency and BI on the CP is less severe than protein deficiency alone. A possible explanation is that the increased carbohydrates in a hypoproteic diet stimulate the production of endogenous insulin and protein synthesis, which partially compensates for the loss of lean body mass caused by BI.