Experimental study on the role of intra-articular injection of MSCs on cartilage regeneration in haemophilia.

Mesenchymal stem cells (MSCs) therapy is a field in progress in cartilage repair strategies. We tried to investigate the functional properties of the joint and cartilage in experimental haemarthrosis (EH) after MSCs intra-articular (IA) injection. One millilitre of fresh autologous blood was injected twice a week for three consecutive weeks in three groups including control haemophilia 10 days (n = 8), control haemophilia 38 days (n = 8) and MSCs (n = 8) group. In later, 10 days after the end of IA blood injections, MSCs IA injection was performed. Eight animals received no treatment as the normal control group. Thirty-eight days after the end of IA blood injections, animals were sacrificed. Joint friction and stress-relaxation tests were done, inflammatory cytokines of synovial membrane and scanning electron microscopy of the cartilage assessed. Joint friction decreased in MSCs in comparison to other groups and was significant with normal control group, (P = 0.011). The mechanical properties of cartilage showed no significant differences between groups. Tumour necrosis factor alpha and interleukin 1 beta decreased and IL-4 very slightly increased in MSCs in comparison to the time-matched control group. Scanning electron microscopy enabled acquisition of good structural properties of the surface and layers of the cartilage after MSCs injection. The hole induced in the medial plateau of the tibia bones, after inducing haemarthrosis, were covered with cartilage-like structure. The results showed that MSCs IA injection has some beneficial effects on cartilage structure and function in haemarthrosis model and is promising in patients with haemophilia.

The effect of supplementation of calcium, vitamin D, boron, and increased fluoride intake on bone mechanical properties and metabolic hormones in rat.

Evidence indicates that optimal nutrition plays a role in bone formation and maintenance. Besides major components of mineralization such as calcium, phosphorus, and vitamin D, other nutrients like boron and fluoride have beneficial role, too. In this study, 34 male Wistar rats were divided into five groups: control diet, fluoride, fluoride + boron, fluoride + calcium + vitamin D, and fluoride + boron + calcium + vitamin D. Boron equal to 1.23 mg, calcium and vitamin D equal to 210 mg + 55 IU and fluoride equal to 0.7 mg/rat/day was added to their drinking water for 8 weeks. Plasma blood samples and bones were collected. Findings are evidence that fluoride + boron intake revealed significant positive effects on bone mechanical properties and bone metabolic hormones. These findings suggest that combined intake of these two elements has beneficial effects on bone stiffness and breaking strength comparing to even calcium + vitamin D supplementation. This evidence dealing with health problems related to bone and skeletal system in humans should justify further investigation of the role of boron and fluoride with other elements in relation to bone.

Comparison between pulsed ultrasound and low level laser therapy on experimental haemarthrosis.

The use of pulsed ultrasound (PUS) and low level laser therapy (LLLT) in patients with haemophilia has been recommended for supportive treatment of acute and chronic phases of haemarthrosis but its role has not been supported by experimental evidence. The purpose of the present study was to evaluate the effect of these modalities on joint swelling, friction and biomechanical parameters of articular cartilage. An experimental rabbit knee haemarthrosis model was used to test the hypothesis that LLLT and PUS favourably impacted on the biotribological and biomechanical properties of cartilage after joint bleeding. To test this, 35 male albino rabbits weighing 1.5-2 kg were used. The left knee of 30 rabbits was injected with 1 mL of fresh autologous blood two times per week for four consecutive weeks to simulate recurrent haemarthrosis; five rabbits served as non-bleeding controls. Ten rabbits were treated with PUS and 10 with LLLT and the remaining 10 were not treated. The treatments were started after 2 days and the treatment duration was planned for 5 days (sessions) in ultrasound and laser groups. A low level Ga-Al-As laser was applied with an 810 nm wavelength, 25 mW power, and 1 J cm(-2) dosage for 200 s duration. The PUS treatment was applied with a duty cycle of 1/9, frequency of 1 MHz, and power of 0.4 W cm(-2) for 150 s. Joint perimeter was measured before the procedure at the beginning of therapies and after cessation of the procedure. Friction and biomechanical parameters were measured immediately after the killing of the animals. The results demonstrate that PUS was more effective in reducing knee joint swelling than LLLT. Moreover, PUS had the unique ability of reducing the joint friction below normal values. However, it was not successful in returning the articular cartilage force and stiffness to normal state. LLLT was more effective in increasing equilibrium force of the articular cartilage than PUS, however, neither therapy normalized this parameter. From these data, we conclude that PUS is more effective than LLLT in reducing joint swelling and articular joint friction after experimental haemarthrosis.

Effect of consumption of fatty acids, calcium, vitamin D and boron with regular physical activity on bone mechanical properties and corresponding metabolic hormones in rats.

The consumption of fatty acids, nutrients, and regular physical activity, individually influence bone mechanical properties in rats. To investigate their effects in combination, male rats were divided into the seven groups: G1: regular food and drinking water; G2: same as Gr.1 + physical activity (Whole body vibration; WBV); G3: same as Gr.2 + Calcium, Vit. D, Boron; G4: same as Gr.3 + canola oil; G5: same as Gr.3 + sunflower oil; G6: same as Gr.3 + mix of sunflower oil and canola oil; and G7: same as Gr.3 + coconut oil; and treated for 8 weeks. Analysis between the control with the groups 2 and 3 revealed that vibration in the G2 increased the body weight (P = 0.04), with no other major difference in plasma and bone indices. Comparison between the control with the G4-G7 (the oil groups) revealed that the rats in the G5 had a lower body weight (15 % less) and a significant increase in plasma levels of Estradiol in the G7 was noted. In addition, levels of Testosterone in the G4 and G7, and Free Testosterone in the G7 had a remarkable increase. Similar trend was observed for plasma levels of Vit. D in the G4 and G5. The stiffness and the breaking strength of the femur in the G7, and the breaking strength of the lumbar in the G7 compared to the control and the G4 and G5 was significantly higher and tended to increase in comparison to the G6. Better and stronger measurements observed for coconut oil is warranted to further study its effect on biomechanical properties of bones.

Biotribological and biomechanical changes after experimental haemarthrosis in the rabbit knee.

Changes in articular cartilage after haemarthrosis have not been completely elucidated in haemophilic arthropathy. Insights into the pathophysiological mechanisms of blood-induced joint damage mainly derived from histological, inflammatory and biochemical investigations. A structure-function relationship is another reasonable way to determine the joint overall health status. Cartilage, a viscoelastic connective tissue, is at least a biphasic material that should also work under minimal friction. Pendulum friction tester measures the mechanical aspects of joint lubrication and quantifies the biotribological properties of the joint. Indentation test is an in situ method characterizing the biomechanical properties of the cartilage. Gross, biotribological and biomechanical properties were determined in a rabbit model of experimental haemarthrosis. A sample of 1 mL of fresh autologous blood was injected in the left knee of rabbit's joint twice weekly for four consecutive weeks. The right knee and animals in the control group were left untreated. After 8 days, joint perimeter, biotribological and biomechanical tests were performed. In a consistent manner, all data showed detrimental effects of the blood on the overall cartilage function under loading. Non-weight bearing and early blood aspiration seem wise to be considered after haemarthrosis.

Effects of tripolar TENS on slow and fast motoneurons: a preliminary study using H-reflex recovery curve method.

This study aimed at examining the effect of tripolar TENS of vertebral column on the activity of slow and fast motoneurons on 10 healthy non-athlete women aged 22.7 +/- 2.21 yrs. H-reflex recovery curve of soleus (slow) and gastrocnemius (fast) muscles were recorded before and after applying tripolar TENS. For recording of this curve, rectangular paired stimuli were applied on tibial nerve (with 40-520 ISI, frequency of 0.2 Hz and pulse width of 600 micros). Our findings showed that maximum H-reflex recovery in gastrocnemius muscle appeared in the shorter ISI, while in soleus muscle, it appeared in the longer ISI and its amplitude slightly decreased after applying tripolar TENS. It is suggested that tripolar TENS excites not only the skin but also Ia and Ib afferents in the dorsal column. A Synaptic interaction of these afferents in spinal cord causes the inhibition of type I MNs and facilitation of type II MNs. This effect can be used in muscle tone modulation.

Whole body vibration is a safe exercise training method and induces no impaired alterations on rat plasma parameters.

UNLABELLED: Whole body vibration (WBV) has been regarded as an exercise training method and as a non-pharmacological supportive treatment option appearing to be efficient for chronic disease conditions, such as bone disorders and cardio-respiratory fitness. Since, data on the safety and efficacy of vibration on plasma parameters are lacking, therefore, it was decided to assess the effects of WBV on the plasma parameters in adult male Wistar rat model. METHODS: Male Wistar rats, weighing 140­180 g, were divided into control and the vibration group. Vibration training consisted of vertical sinusoidal whole body vibration for 8 weeks, followed by blood collection. RESULTS: The vibrated rats weighed more than the control group (approximately 14% more). Plasma CK, E2 and IL-6 levels were significantly higher in the vibration group compared with the controls. The mean of Vit. D level was 15% higher; hsCRP level was 11% lower and IL-6 level was 32% higher in the vibration group. No difference was observed for other selected plasma parameters. DISCUSSION: The potential effects of physiological responses of WBV on several physiological systems are without deteriorations in plasma parameters.