Radosław P. Radzki

Effect of a carbonated HAP/β-glucan composite bone substitute on healing of drilled bone voids in the proximal tibial metaphysis of rabbits.

A novel elastic hydroxyapatite-based composite of high surgical handiness has been developed. Its potential application in orthopedics as a filler of bone defects has been studied. The biomaterial was composed of carbonated hydroxyapatite (CHAP) granules and polysaccharide polymer (β-1,3-glucan). Cylinders of 4mm in diameter and 6mm in length were implanted into bone cavities created in the proximal metaphysis of tibiae of 24 New Zealand white rabbits. 18 sham-operated animals were used as controls. After 1, 3 or 6 months, the rabbits were euthanized, the bones were harvested and subjected to analysis. Radiological images and histological sections revealed integration of implants with bone tissue with no signs of graft rejection. Peripheral quantitative computed tomography (pQCT) indicated the stimulating effect of the biomaterial on bone formation and mineralization. Densitometry (DXA) analysis suggested that biomineralization of bones was preceded by bioresorption and gradual disappearance of porous ceramic granules. The findings suggest that the CHAP-glucan composite material enables regeneration of bone tissue and could serve as a bone defect filler.

Effect of dietary alpha-ketoglutarate on blood lipid profile during hypercholesterolaemia in rats

Objective. The aim of the study was to determine the effect of α‐ketoglutarate on the blood lipid profile using a rat animal model with experimentally induced hypercholesterolaemia. Material and methods. The female and male (30/30) Wistar rats had ad libitum access to a diet containing cholesterol (1 %) and lard (10 %) throughout the entire experimental period (120 days). On day 60 of the study, both the females and the males were divided into three groups, the first receiving a mixture of drinking water adjusted to pH 4.6 using HCl (control), the other two (experimental groups) receiving a solution containing 0.01 M and 0.1 M α‐ketoglutarate (AKG) (pH adjusted to 4.6). Blood samples were taken on days 0, 30, 60 and 120. Results. The concentrations of total cholesterol, triglycerides, HDL and LDL, respectively, in the blood serum were estimated spectrophotometrically. During the entire experimental period the total cholesterol, triglycerides and LDL levels of the control rats increased, whereas that of HDL decreased. The serum concentrations of total cholesterol, LDL and triglycerides in both the experimental groups receiving AKG decreased (days 60 to 120) (p<0.05), while the HDL concentration tended to increase. The body gain in all groups receiving AKG was significantly lower than in the control group. Conclusions. These observations clearly prove that oral treatment with AKG can decrease the risk of hypercholesterolaemia developing and can lower the body weight. The relative concentrations of the plasma LDL and HDL changed to a more favourable ratio promoting good health.

Alteration in bone geometric and mechanical properties, histomorphometrical parameters of trabecular bone, articular cartilage, and growth plate in adolescent rats after chronic co-exposure to cadmium and lead in the case of supplementation with green, black, red and white tea.

Adolescent male Wistar rats were used to check whether regular consumption of black, red, white, or green tea would have a protective effect on femur development during 12-week exposure to Cd and Pb (7mg Cd and 50mg Pb in 1kg of the diet). The animals were randomly divided (n=12) into a positive control (without Cd, Pb and teas), a negative control group (Cd and Pb), and groups supplemented additionally with green (GT), black (BT), red (RT), and white tea (WT). Heavy metals reduced the geometric and densitometric parameters and the total thickness of articular cartilage irrespective of tea administration and influenced mechanical endurance, growth plate thickness, and trabecular histomorphometry depending on the tea type. It is difficult to indicate which tea has the best protective effects on bone and hyaline cartilage against heavy metal action.

Can 2-oxoglutarate prevent changes in bone evoked by omeprazole?

OBJECTIVE Proton-pump inhibitors, such as omeprazole, are widely used in the prevention and treatment of gastroesophageal diseases. However, an association between proton-pump inhibitors and the increased risk of bone fractures has been observed, especially in patients treated for extended periods. Conversely, 2-oxoglutarate, a precursor of hydroxyproline, the most abundant amino acid in bone collagen, counteracts the bone loss. The aim of the present study was to elucidate the influence of omeprazole on bone and investigate whether dietary 2-oxoglutarate supplementation could prevent the effects of omeprazole. METHODS Eighteen male Sprague-Dawley rats were used. Rats received omeprazole in the diet and 2-oxoglutarate in the drinking water. Body and organ weights and serum concentrations of cholecystokinin and gastrin were measured. The femurs, tibias, and calvarias were collected. Histomorphometric analysis of bone and cartilage tissues was conducted. Bone densitometric and peripheral quantitative computed tomographic analyses of the femur and tibia were performed. RESULTS Omeprazole decreased the femur and tibia weights, the mechanical properties of the femur, the volumetric bone density and content, the trabecular and cortical bone mineral content, the total, trabecular, and cortical bone areas, the mean cortical thickness, and the periosteal circumference of the femur. Omeprazole had a minor effect on the examined bone morphology and exerted negligible effects on the cartilage. 2-Oxoglutarate lowered the gastrin concentration. CONCLUSIONS Omeprazole treatment exerts its effects mostly on bone mineralization and cancellous bone, adversely affecting bone properties. This adverse effect of omeprazole was not markedly abolished by 2-oxoglutaric acid, which acted as an anti-hypergastrinemic agent.

The effect of dietary administration of 2-oxoglutaric acid on the cartilage and bone of growing rats

2-Oxoglutaric acid (2-Ox), a precursor to hydroxyproline - the most abundant amino acid in bone collagen, exerts protective effects on bone development during different stages of organism development; however, little is known about the action of 2-Ox on cartilage. The aim of the present study was to elucidate the influence of dietary 2-Ox supplementation on the growth plate, articular cartilage and bone of growing rats. A total of twelve male Sprague-Dawley rats were used in the study. Half of the rats received 2-oxoglutarate at a dose of 0·75 g/kg body weight per d in their drinking-water. Body and organ weights were measured. Histomorphometric analyses of the cartilage and bone tissue of the femora and tibiae were conducted, as well as bone densitometry and peripheral quantitative computed tomography (pQCT). Rats receiving 2-Ox had an increased body mass (P<0·001) and absolute liver weight (P=0·031). Femoral length (P=0·045) and bone mineral density (P=0·014), overall thickness of growth plate (femur P=0·036 and tibia P=0·026) and the thickness of femoral articular cartilage (P<0·001) were also increased. 2-Ox administration had no effect on the mechanical properties or on any of the measured pQCT parameters for both bones analysed. There were also no significant differences in histomorphometric parameters of tibial articular cartilage and autofluorescence of femoral and tibial growth plate cartilage. Dietary supplementation with 2-Ox to growing rats exerts its effects mainly on cartilage tissue, having only a slight influence on bone. The effect of 2-Ox administration was selective, depending on the particular bone and type of cartilage analysed.

Erythrocyte antioxidative enzymes in experimentally induced osteopenia in rats

Abstract The study aimed at the determination of dynamic relationship between mineralisation processes and antioxidative/oxidative status during the development of osteopenia. One hundred and two healthy female Wistar rats at the age of 2 months and initial body weight of 200 g were used in the experiment. The rats were divided into control (CON, n=6), sham operated (SHO, n=48), and ovariectomised (OVX, n=48) groups. Animals from SHO (n=6) and OVX (n=6) groups were sacrificed every week during 8 weeks of the experiment in order to detect dynamic changes in examined parameters. The samples were collected weekly from day 7 to day 56. The femora were examined with the use of DXA (bone mineral density) and pQCT (area, mineral content, volumetric density of trabecular and cortical part of distal femora). The pQCT scans were performed 5 mm from distal end of the tibia. The determination of activity of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in haemolysates of erythrocytes were performed spectrophotometrically. Obtained data showed wave like changes in both enzyme activities and bone parameters and indicated the importance of the 2nd-3rd and 5th-6th week after surgery as a key moment for bone metabolism and activity of enzymatic antioxidative defence during the development of osteopenia induced by bilateral ovariectomy. The obtained results proved that alterations in activity of GSH-Px and SOD, and pQCT ahead the changes registered by DXA by 7 d.

Effects of long-term oral administration of methimazole on femur and tibia properties in male Wistar rats

Physiological concentrations of thyroid hormones are crucial for skeletal growth and development, physiological bone turnover and bone homeostasis maintenance. Methimazole (1-methyl-2-mercaptoimidazole) is an antithyroid drug used for the treatment of the hyperthyroidism in humans and animals. The aim of the study was to determine effects of long-term oral methimazole treatment in male Wistar rats on biochemical bone metabolism markers, as well as morphological, geometric, densitometric and mechanical properties of femur and tibia. Experimental rats were subjected to 90-day-long oral treatment with 0.05% water solution of methimazole and were kept under identical environmental conditions and received the same diet ad libitum as the control group. Serum concentration of osteocalcin (OC) and C-terminal telopeptides of type I collagen (CTX-I) was determined. Femur and tibia were evaluated using quantitative computed tomography (QCT), peripheral QCT (pQCT) and three-point bending test. Final body weight of the experimental group was significantly decreased by 30% (P=0.01). Methimazole treatment significantly decreased serum OC concentration by 21% (P=0.02) and increased CTX-I concentration by 17% (P=0.06). Methimazole decreased morphological, geometric and densitometric parameters of femur and tibia in rats. Mechanical evaluation of bones has shown significantly decreased maximum elastic strength and ultimate strength of femur in rats treated with methimazole by 36% and 40% when compared to the control group (P<0.05). In conclusion, this study has shown that long-term treatment with methimazole inhibits bone formation and accelerates bone resorption processes. The observed negative effects of methimazole treatment on body weight gain and skeletal properties may be considered as additional possible side effects in living organisms to those reported in the previous studies. It may be suggested that long-term antithyroid treatment should be combined with prevention of the negative effects of methimazole on bone tissue and whole body metabolism.

The peripheral quantitative computed tomographic and densitometric analysis of skeletal tissue in male Wistar rats after chromium sulfate treatment

INTRODUCTION This study evaluates the effects of three different doses of chromium sulphate on bone density and the tomographic parameters of skeletal tissue of rats. MATERIAL AND METHODS The experiment was performed on 40 male Wistar rats which received, by gavage, during 90 days, a chromium sulphate in either a daily dose of 400, 600 or 800 µg/kg BW. At the end of experiment, the rats were scanned using the densitometry method (DXA) to determine the bone mineral density, bone mineral content of total skeleton and vertebral column (L2-L4) and parameters of body composition (Lean Mass and Fat Mass). The isolated femora were scanned using peripheral a quantitative computed tomography method (pQCT) for a separate analysis of the trabecular and cortical bone tissue. The ultimate strength, work to ultimate and the Young modulus of femora was also investigated by the three-point bending test. RESULTS The negative impact of chromium was observed in relation to bone tissue. All doses significantly decreased total skeleton density and mineral content, and also had impact upon the isolated femora and vertebral column. Trabecular volumetric bone mineral density and trabecular bone mineral content measured by pQCT in distal femur metaphysis were significantly lower in the experimental groups than in the control. Higher doses of chromium also significantly decreased values of ultimate strength and Young modulus in the investigated femora. CONCLUSIONS The results of the experiment demonstrate that chromium sulphate is dose dependent, and exerts a disadvantageous effect on the skeleton, as it decreases bone density and resistance.

Effect of sodium butyrate and Yucca schidigera extract on bone characteristics in growing pigs

Abstract Introduction: The aim of this study was to investigate the influence of diet supplementation with sodium butyrate and Yucca schidigera extract (0.2% and 0.3%) on femur quality of growing pigs (n = 45). Material and Methods: At the age of 28, 35 and 56 d, five piglets from each group fed a different diet were euthanised and the femora were collected for further analyses. The bone characteristics were assessed based on weight, length, densitometric analysis of BMC and BMD, pQCT analysis (area, mineral content, volumetric density of trabecular and cortical part of metaphysis and diaphysis, respectively), ultimate strength, and geometrical parameters (cross-sectional area and second moment of inertia). Results: There were no significant differences in femur bone parameters among experimental groups on the 28th d of life. On the 35th d of life, piglets with 0.2% supplementation of sodium butyrate and Yucca schidigera extract had significantly lower values of weight and second moment of inertia, and significantly higher trabecular BMD and BMC compared to other experimental groups. In 56-day-old pigs, the higher values were observed in both experimental groups regarding BMC, ultimate strength, geometrical parameters, cortical BMC, diaphyseal total area, and endosteal circumference (P < 0.05). Significant differences between experimental groups were observed only in bone weight and cortical thickness. Conclusion: This study proved that simultaneous supplementation with sodium butyrate and Yucca schidigera extract positively influences bone quality in pigs in the post-weaning period. However, there were no differences in bone characteristics between the addition of 0.2% and 0.3% preparations.

The protective and therapeutic effect of exclusive and combined treatment with alpha-ketoglutarate sodium salt and ipriflavone on bone loss in orchidectomized rats

ObjectiveThis study investigated the effect of alpha-ketoglutarate sodium salt (AKG) and ipriflavone (IP) treatment on the mineralization of the tibia in male rats during the development and after the establishment of osteopenia.DesignOne hundred and twenty eight male rats were randomly selected and submitted to either sham-operation (SHO) or orchidectomy (ORX), after which each group were then randomly divided between the two experiments. In Experiment-1, treatment with AKG or/and IP started after a 7-day recovery period, whereas in Experiment-2, the experimental protocol proceeded after a 60-day period of osteopenia establishment. AKG was then administered as an experimental drinking, at a concentration of 1.0 mol/l. As a control, a placebo solution was administered. IP at 50 mg/kg b.w., and physiological saline–PhS (as a control for IP) were applied daily via gavage.MeasurementsAfter 60 days of experimental treatment, in both experiments, the rats were sacrificed, their body weight recorded, while blood serum (Osteocalcin, CTX) and isolated tibia (weight, length, pQCT, DXA, 3-point bending test) were stored for further analysis.Results and conclusionsOur results show that during the development of osteopenia, AKG and IP when applied exclusively, counteracts osteopenia development, whereas their usage after the establishment of osteopenia, significantly limits the development of bone disorders. Furthermore, combined treatment of AKG and IP exceeded the effects of their sole usage. In addition, during the development of osteopenia, AKG and IP not only inhibited bone resorption, but markedly stimulated the formation of bone tissue. Finally, after the development of osteopenia, combined treatment with AKG and IP protected the bone tissue against orchidectomy-induced bone loss.