Shuzhu Gu

Effects of cadmium on osteoblasts and osteoclasts in vitro

Cadmium (Cd) may have direct effects on bone metabolism and the mechanism is not fully understood. To investigate the effects of Cd on bone metabolism, effects of Cd on osteoblasts and osteoclasts in vitro were observed at cellular and molecular levels. Osteoblasts were cultured by sequential enzyme digestion from Sprague-Dawley rats calvarial bone and osteoclasts were isolated from long bones of new-born male and female Sprague-Dawley rats, and then cells were exposed to different concentrations of Cd (0-2.0 μ mol/L for osteoblasts; 0.03 μmol/L for osteoclasts). As for osteoblasts, cell viability, alkaline phosphatase (ALP) activity, and mineralization were determined. Osteoprotegerin (OPG) and receptor activator of NF-kB ligand (RANKL) were studied via reverse transcription-polymerase chain reaction (RT-PCR). For osteoclasts, after exposure to Cd (0.03 μmol/L) for 72 h and 120 h, number of osteoclasts and pits formation was observed. Cd inhibited the viability, ALP activity, mineralization and up-regulated RANKL mRNA expression in osteoblasts. But Cd had no obvious effect on OPG mRNA expression. For osteoclasts, cadmium (0.03 μmol/L) could increase the numbers of osteoclasts (p<0.05) and enhance pits formation (p<0.05). These results suggested that Cd could inhibit bone formation at high concentrations and enhance bone resorption at low level. OPG/RANKL may constitute an important pathway of Cd effects on bone.

Effects of cadmium on forearm bone density after reduction of exposure for 10 years in a Chinese population.

The main focus of this study was to evaluate long term effects of cadmium on forearm bone mineral density after stopping ingestion of cadmium-polluted rice for 10 years in a Chinese population. A total of 532 persons (338 women and 194 men), living in control, moderately and heavily polluted areas, were included in this study. The residents living in the polluted area ceased ingesting cadmium-polluted rice in 1996. All participants were require to answer a questionnaire and the bone mineral density (BMD) was measured by dual energy X-ray absorptiometry (DXA) at the proximal radius and ulna. Samples of urine and blood were collected for determination of cadmium in urine (UCd) and blood (BCd).The BMD of subjects living in the heavily polluted area was significantly lower than that of those living in control area in both men and women (p<0.01). For the people living in the moderately polluted area, only the womens BMD was greatly lower compared to that in the control area (p<0.05). The BMD declined with the increasing BCd and UCd in both sexes, especially in the highest level (BCd >5 microg/L, UCd >10 microg/g crea) groups (p<0.01). It was found that there were significant differences in the prevalence of osteoporosis among the different areas (chi(2)=13.046, p=0.0003) and different UCd groups (chi(2)=4.511, p=0.0337) in women, but not in men (chi(2)=0.962, p=0.3268; chi(2)=1.906, p=0.1675). But a significant difference exists in the prevalence of osteoporosis among different BCd groups in both genders (chi(2)=9.304, p=0.00229, in women; chi(2)=4603, p=0.0319, in men). This study suggested that cadmium could play a long-term role on bone and more attention should be paid to cadmium effects on bone metabolism after reduction of exposure.

Effects of cadmium on bone microstructure and serum tartrate-resistant acid phosphatase 5b in male rats

This study investigated the effects of cadmium on bone microstructure and serum tartrate-resistant acid phosphatase 5b (Tracp 5b) in male rats. Sprague-Dawley male rats were divided into three groups that were given CdCl2 by subcutaneous injection at doses of 0, 0.1 and 0.5 mg/kg body weight (bw) for 12 weeks, respectively. Before killing at the 12th week, microcomputed tomography scanning was performed on the proximal tibia, and urine samples were collected from all of the rats. All rats were then killed, and their blood was collected for biomarkers assay. Bone tissues were dissected for mineral density determinations and histology. The concentration of cadmium in the blood, urine and bone of rats treated with cadmium were significantly higher than in the control group. The bone mineral density, bone mineral concentrations and bone microstructure index of rats treated with cadmium at 0.5 mg/kg bw were clearly lower than in the control rats. Histological investigation also revealed damage to the bone microstructure caused by cadmium. Tracp 5b concentrations in rats treated with cadmium were dose dependently higher than the control. The concentration of cadmium in blood, urine and bone was significantly correlated with Tracp 5b and bone microstructure parameters. Cadmium was shown to induce bone microstructure damage, especially to trabecular bone. The elevated concentrations of serum Tracp 5b suggest that bone resorption mediated via osteoclasts is an important mechanism for the toxic effects of cadmium on bone.

Cadmium induces differentiation of RAW264.7 cells into osteoclasts in the presence of RANKL.

The mechanism of cadmium effects on bone is not fully understood. In this study, we investigated the effects of cadmium on osteoclasts differentiation and the probable mechanism. RAW264.7 cells were exposed to cadmium (0-60 nmol/L) in the presence or absence of receptor-activated nuclear factor κ B ligand (RANKL) for 5 days. Then, the viability, tartrate-resistant acid phosphatase (TRAP) activity and the formation of TRAP positive multinucleated osteoclasts were observed. Receptor activator of nuclear factor κ B (RANK), tumor necrosis factor receptor associated factor 6 (TRAF6), c-src, c-fos, fos-related antigen 1 (Fra1) expression were determined by reverse transcription polymerase chain reaction. Cadmium increased TRAP activity (20-40%) and TRAP positive cell formation in the presence of RANKL, but had no obvious influence on them without RANKL. RANK, TRAF6, Fra1, c-src and c-fos (at 15-30 nmol/L) expression were enhanced (30-70%) by cadmium in the presence of RANKL, but cadmium had little influence on them in the absence of RANKL. This study demonstrated that cadmium could induce differentiation of osteoclasts precursor into osteoclasts in the presence of RANKL. Even though the changes of gene expression were small, RANKL/RANK and downstream genes may play an important role in cadmium effects on osteoclasts.

Genistein inhibits osteolytic bone metastasis and enhances bone mineral in nude mice

In this study, the effective activity of genistein on osteolytic bone metastasis and bone mineral was investigated. Female BALB/c-nu/nu mice were injected with estrogen receptor-negative human breast cancer cells, MDA-MB-231, into left cardiac ventricle to form osteolytic bone metastases, and administered genistein subcutaneously after radiologically small but defined osteolytic metastases had been observed (protocol 1), simultaneously with cancer cells inoculation (protocol 2) and prophylactically 7 days before inoculation of cancer cells (protocol 3). In all protocols, genistein (10mg/kg/day) markedly reduced the number and volume of osteolytic bone metastases assessed by radiography and the number of osteoclasts. Furthermore, histomorphometrical analysis revealed that genistein markedly increased trabecular area (Tb.Ar%), trabecular thickness (Tb.Th) and trabecular number (Tb.N), and decreased trabecular separation (Tb.Sp). These results thus demonstrate that genistein could inhibit osteolytic bone metastases, suppress bone resorption, increase bone mass and improve bone microstructure in bone metastases of breast cancer.

Combined effects of γ-irradiation and cadmium exposures on osteoblasts in vitro

The combined effects of γ-irradiation and cadmium (Cd) exposures on osteoblasts were observed in the present study. Osteoblasts were exposed to γ-irradiation (0.5 Gy) and Cd (0-0.5 μmol/L). Cell viability, alkaline phosphatase (ALP) activity, mineralization ability, cell apoptosis and genes expression of ALP, osteocalcin (OC) and caspase 3 were observed. Low concentrations of Cd exposure had no obvious influence on cell viability, ALP activity and apoptosis. However, low levels of Cd exposure combined with γ-irradiation induced more toxic effects on osteoblasts than those treated with Cd or irradiation alone. High concentrations of Cd combined with irradiation exposure induced more significant inhibition in cell viability, ALP activity and mineralization ability than those exposed to Cd or irradiation alone. Meanwhile, OC and ALP mRNA expression of cells treated with Cd combined with irradiation were down-regulated more significantly than those treated with Cd or irradiation alone. Cd combined with γ-irradiation could obviously enhance osteoblast apoptosis and up-regulated caspase 3 mRNA expression compared with those treated with Cd or irradiation alone. This study indicated that ionizing irradiation can enhance Cd toxic effects on osteoblast viability and differentiation and apoptosis may play an important role in this progress.

Cadmium exposure induced itai-itai-like syndrome in male rats

Fourteen Sprague-Dawley male rats were randomly divided into 2 groups which were given CdCl2 at the doses of 0 and 1.5 mg /kg for 12 weeks. Before sacrifice, microCT scanning were performed on the proximal tibia and urine were collected for cadmium and N-acetyl-beta-D-glucosaminidase assay, then all of rats were sacrificed and blood was collected for biomarkers measurement; bone tissues were collected for bone mass, histology and biomechanical analysis. The cadmium in blood, urine, bone and kidney of rats treated with cadmium was significantly higher than those in the control group. The bone mineral density, and bone mineral ability of rats treated with cadmium were obviously decreased by 20%–50% compared to controls. Bone microstructure index and trabecular separation of rats treated with cadmium were obviously lower (−50%) and significantly higher (+150%) than that in the control group. Bone biomechanical property decreased by 30%–60% in cadmium treated rats compared to control. Tartrate-resistant acid phosphatase 5b and alkaline phosphatase levels of rats treated with cadmium were significantly higher than those in control, but serum osteocalcin level decreased greatly by cadmium. Obvious proximal tubule damage occurred after cadmium exposure. These observations gave clear and comprehensive evidence that cadmium exposure could induce itai-itai-like syndrome in male rats.