Hydrolyzed collagen improves bone metabolism and biomechanical parameters in ovariectomized mice: an in vitro and in vivo study.

Collagen has an important structural function in several organs of the body, especially in bone and cartilage. The aim of this study was to investigate the effect of hydrolyzed collagen on bone metabolism, especially in the perspective of osteoporosis treatment and understanding of its mechanism of action. An in vivo study was carried out in 12-week-old female C3H/HeN mice. These were either ovariectomized (OVX) or sham-operated (SHAM) and fed for 12 weeks with a diet containing 10 or 25 g/kg of hydrolyzed collagen. We measured bone mineral density (BMD) using dual-energy X-ray absorptiometry (DXA). C-terminal telopeptide of type I collagen (CTX), marker of bone resorption, and alkaline phosphatase (ALP), marker of bone formation, were assayed after 4 and 12 weeks. Femur biomechanical properties were studied by a 3-point bending test and bone architecture by microtomography. The BMD for OVX mice fed the diet including 25 g/kg of hydrolyzed collagen was significantly higher as compared to OVX mice. The blood CTX level significantly decreased when mice were fed with either of the diets containing hydrolyzed collagen. Finally, we have shown a significant increase in bone strength correlated to geometrical changes for the OVX mice fed the 25 g/kg hydrolyzed collagen diet. Primary cultures of murine bone cells were established from the tibia and femur marrow of BALB/c mice. The growth and differentiation of osteoclasts and osteoblasts cultured with different concentrations (from 0.2 to 1.0 mg/mL) of bovine, porcine or fish hydrolyzed collagens (2 or 5 kDa) were measured. Hydrolyzed collagens (2 or 5 kDa) in the tissue culture medium did not have any significant effects on cell growth as compared to controls. However, there was a significant and dose-dependent increase in ALP activity, a well-known marker of osteogenesis, and a decrease in octeoclast activity in primary culture of bone cells cultured with hydrolyzed collagens (2 kDa only) as compared to the control. It is concluded that dietary hydrolyzed collagen increases osteoblast activity (as measured in primary tissue culture), which acts on bone remodeling and increases the external diameter of cortical areas of the femurs.

Oxytocin controls differentiation of human mesenchymal stem cells and reverses osteoporosis.

Osteoporosis constitutes a major worldwide public health burden characterized by enhanced skeletal fragility. Bone metabolism is the combination of bone resorption by osteoclasts and bone formation by osteoblasts. Whereas increase in bone resorption is considered as the main contributor of bone loss that may lead to osteoporosis, this loss is accompanied by increased bone marrow adiposity. Osteoblasts and adipocytes share the same precursor cell and an inverse relationship exists between the two lineages. Therefore, identifying signaling pathways that stimulate mesenchymal stem cells osteogenesis at the expense of adipogenesis is of major importance for developing new therapeutic treatments. For this purpose, we identified by transcriptomic analysis the oxytocin receptor pathway as a potential regulator of the osteoblast/adipocyte balance of human multipotent adipose-derived stem (hMADS) cells. Both oxytocin (OT) and carbetocin (a stable OT analogue) negatively modulate adipogenesis while promoting osteogenesis in both hMADS cells and human bone marrow mesenchymal stromal cells. Consistent with these observations, ovariectomized (OVX) mice and rats, which become osteoporotic and exhibit disequilibrium of this balance, have significant decreased OT levels compared to sham-operated controls. Subcutaneous OT injection reverses bone loss in OVX mice and reduces marrow adiposity. Clinically, plasma OT levels are significantly lower in postmenopausal women developing osteoporosis than in their healthy counterparts. Taken together, these results suggest that plasma OT levels represent a novel diagnostic marker for osteoporosis and that OT administration holds promise as a potential therapy for this disease.

Evaluation of macrostructural bone biomechanics.

Bone fragility can be defined as an increased risk of fractures. Advanced age and bone diseases such as osteoporosis increase the fracture risk. Understanding the effects of osteoporosis and its treatments requires a description of the mechanical behavior of bone tissue. To this end, an entire bone can be studied, or the cortical and trabecular components can be investigated separately. We review the biomechanical tests available for measuring the ability of bone to withstand torsional, compressive, tensile, and bending forces.

Combined effects of exercise and propranolol on bone tissue in ovariectomized rats.

UNLABELLED: The bone response to physical exercise may be under control of the SNS. Using a running session in rats, we confirmed that exercise improved trabecular and cortical properties. SNS blockade by propranolol did not affect this response on cortical bone but surprisingly inhibited the trabecular response. This suggests that the SNS is involved in the trabecular response to exercise but not in the cortical response. INTRODUCTION: Animal studies have suggested that bone remodeling is under beta-adrenergic control through the sympathetic nervous system (SNS). However, the SNS contribution to bone response under mechanical loading remains unclear. The purpose of this study was to examine the preventive effect of exercise coupled with propranolol on cancellous and cortical bone compartments in ovariectomized rats. MATERIALS AND METHODS: Six-month-old female Wistar rats were ovariectomized (OVX, n = 44) or sham-operated (n = 24). OVX rats received subcutaneous injections of propranolol 0.1 mg/kg/day or vehicle and were submitted or not submitted to treadmill exercise (13 m/minute, 60 minutes/day, 5 days/week) for 10 weeks. Tibial and femoral BMD was analyzed longitudinally by DXA. At death, the left tibial metaphysis and L(4) vertebrae were removed, and microCT was performed to study trabecular and cortical bone structure. Histomorphometric analysis was performed on the right proximal tibia. RESULTS: After 10 weeks, BMD and trabecular strength decreased in OVX rats, whereas bone turnover rate and cortical porosity increased compared with the Sham group (p < 0.001). Either propranolol or exercise allowed preservation of bone architecture by increasing trabecular number (+50.35% versus OVX; p < 0.001) and thickness (+16.8% versus OVX; p < 0.001). An additive effect of propranolol and exercise was observed on cortical porosity but not on trabecular microarchitecture or cortical width. Biomechanical properties indicated a higher ultimate force in the OVX-propranolol-exercise group compared with the OVX group (+9.9%; p < 0.05), whereas propranolol and exercise alone did not have any significant effect on bone strength. CONCLUSIONS: Our data confirm a contribution of the SNS to the determinants of bone mass and quality and show a antagonistic effect of exercise and a beta-antagonist on trabecular bone structure.

The mode of bone conservation does not affect the architecture and the tensile properties of rat femurs.

The bone samples used in clinical and experimental trials must be the less damaged as possible to avoid alterations of their properties. However, the mode of storage might possibly alter the bone properties, particularly microarchitecture and strength. The aim of our study was to analyze the effects of deep-freezing and alcohol conservation techniques on the densitometric, microarchitectural and biomechanical parameters of rat femurs. The left femurs were elongated in uniaxial tension up to breakdown in order to calculate biomechanical parameters. The densitometric and microarchitectural properties of right femurs were evaluated using dual-energy X-ray absorptiometry and microcomputed tomography, respectively. Results showed no significant difference in the parameters investigated between deep-freezing, alcohol storage and fresh femurs when comparing each parameter separately. Therefore, one month storage in alcohol or deep-freezing seemed to induce no harmful effect on densitometric, microarchitectural and biomechanical parameters of rat femurs.

Does training have consequences for the walk-run transition speed?

A number of authors when studying the walk-run transition phenomenon focused either on the mechanical or energy expenditure whilst only a few used both parameters concurrently. Moreover the literature demonstrates that the contribution of these variables changes along with the level and method of training. Consequently the purpose of this study is to find, by analyzing concurrently these two variables, if the walk-run transition speed is linked to the type of training. To this end we calculated two theoretical transition speeds: one based on the metabolic energy expenditure St(1) and the second one based on the internal work St(2). Subjects were divided into three groups (untrained, sprint and endurance-trained men) who were required to walk and run on a treadmill at increasing speeds. Firstly we show that the relationship between St(1) and St(2) differs depending on the groups. Sprinters have a significantly lower St(2) than St(1) whereas the opposite is found for untrained subjects. We also show that the transition speed is linked to the subject's type of training. To conclude it seems that acquiring running techniques through specific training has consequences for the walk-run transition phenomenon.