Through morphological observation, HE staining, TRAP staining and toluidine blue staining of bone resorption pits to identify osteoclasts which obtained by 1α, 25-(OH)2 VitD3 inducing rabbit bone marrow cells. Three indicators-TRAP staining, TRAP enzyme activity detecting and the number and area of bone resorption pits were adapted to detect the effect of Sargentodoxae caulis on the activity of osteoclasts. Culturing MC3T3-E1 Subclong 14 cells and detecting the effect of S. caulis on differentiation and proliferation of them by MTT and detecting the alkaline phosphatase in cells. The results show that all of the low, middle and high doses of water and alcohol extracts of S. caulis have significant inhibition on osteoclast differentiation and bone resorption ability in a dose-dependent manner. The low and middle doses of water and alcohol extracts of S. caulis can stimulate differentiation and proliferation of MC3T3-ElSubclone 14 cells, which indicates S. caulis can prevent osteoporosis and the function could be achieved by inhibiting osteoclast activity and promoting the proliferation and differentiation of osteoblasts.
Animals ; Bone Resorption ; drug therapy ; physiopathology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Mice ; Osteoclasts ; cytology ; drug effects ; Rabbits

We hypothesized that the formation and differentialtion of osteoclasts are accelerated and the potential of bone resorption is increased in the hemiplegic bone marrow in the early stage of stroke. We randomly divided white female Sprague-Dawley (SD) rats (n = 30) into two groups, stroke (n = 15) and sham group (n = 15). On the 7th day after stroke, after cutting away the epiphyses of the femurs and tibias, diaphyseal channels were flushed using alpha-minimum essential medium (alpha-MEM) and bone marrow cells were collected. Bone marrow stem cells, which were extracted from the femur and tibia, were cultured on the 7th day after middle cerebral artery occlusion. We then estimated the ratio of non-adherent cells to total bone marrow cells that included osteoclast precursor cells. After culturing these cells separately, cells that tested positive on the tartrate resistant acid phosphatase (TRAP) were counted and bone resorption was evaluated by using the OAAS(TM) plate. In comparison to the control group, the stroke group showed a higher increase of non-adherent cells in the hemiplegic side bone marrow. In addition, after the primary culture, the stroke group showed an increased number of TRAP positive cells and a higher degree of bone resorption estimated by OAAS(TM) plate. As a result, osteoclastogenesis and osteoclast differentiation are accelerated and the potential of bone resorption is increased in the hemiplegic bone marrow and these changes are detected as early as within the first week after middle cerebral artery occlusion in SD rats.
Animals ; Bone Marrow Cells/cytology/drug effects ; Bone Resorption/*physiopathology ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Female ; Femur/cytology ; Osteoclasts/cytology ; Rats ; Rats, Sprague-Dawley ; Stem Cells/cytology/metabolism ; Stroke/*metabolism/pathology ; Tartrates/pharmacology ; Tibia/cytology

Osteoarthritis (OA) is an age-related joint disease that is characterized by degeneration of articular cartilage and chronic pain. Oxidative stress is considered one of the pathophysiological factors in the progression of OA. We investigated the effects of grape seed proanthocyanidin extract (GSPE), which is an antioxidant, on monosodium iodoacetate (MIA)-induced arthritis of the knee joint of rat, which is an animal model of human OA. GSPE (100 mg/kg or 300 mg/kg) or saline was given orally three times per week for 4 weeks after the MIA injection. Pain was measured using the paw withdrawal latency (PWL), the paw withdrawal threshold (PWT) and the hind limb weight bearing ability. Joint damage was assessed using histological and microscopic analysis and microcomputerized tomography. Matrix metalloproteinase-13 (MMP13) and nitrotyrosine were detected using immunohistochemistry. Administration of GSPE to the MIA-treated rats significantly increased the PWL and PWT and this resulted in recovery of hind paw weight distribution (P < 0.05). GSPE reduced the loss of chondrocytes and proteoglycan, the production of MMP13, nitrotyrosine and IL-1beta and the formation of osteophytes, and it reduced the number of subchondral bone fractures in the MIA-treated rats. These results indicate that GSPE is antinociceptive and it is protective against joint damage in the MIA-treated rat model of OA. GSPE could open up novel avenues for the treatment of OA.
Analgesics/*administration & dosage ; Animals ; Antioxidants/*administration & dosage ; Bone Resorption ; Disease Models, Animal ; Gene Expression Regulation ; Humans ; Interleukin-1beta/genetics/metabolism ; Iodoacetates/administration & dosage ; Knee Joint/*drug effects/metabolism/pathology ; Male ; Matrix Metalloproteinase 13/genetics/metabolism ; Osteoarthritis/chemically induced/*drug therapy/physiopathology ; Pain ; Plant Extracts/administration & dosage ; Proanthocyanidins/*administration & dosage ; Rats ; Rats, Wistar ; Seeds ; Tomography, Emission-Computed ; Tyrosine/analogs & derivatives/metabolism ; Vitis/immunology

Osteoarthritis (OA) is an age-related joint disease that is characterized by degeneration of articular cartilage and chronic pain. Oxidative stress is considered one of the pathophysiological factors in the progression of OA. We investigated the effects of grape seed proanthocyanidin extract (GSPE), which is an antioxidant, on monosodium iodoacetate (MIA)-induced arthritis of the knee joint of rat, which is an animal model of human OA. GSPE (100 mg/kg or 300 mg/kg) or saline was given orally three times per week for 4 weeks after the MIA injection. Pain was measured using the paw withdrawal latency (PWL), the paw withdrawal threshold (PWT) and the hind limb weight bearing ability. Joint damage was assessed using histological and microscopic analysis and microcomputerized tomography. Matrix metalloproteinase-13 (MMP13) and nitrotyrosine were detected using immunohistochemistry. Administration of GSPE to the MIA-treated rats significantly increased the PWL and PWT and this resulted in recovery of hind paw weight distribution (P < 0.05). GSPE reduced the loss of chondrocytes and proteoglycan, the production of MMP13, nitrotyrosine and IL-1beta and the formation of osteophytes, and it reduced the number of subchondral bone fractures in the MIA-treated rats. These results indicate that GSPE is antinociceptive and it is protective against joint damage in the MIA-treated rat model of OA. GSPE could open up novel avenues for the treatment of OA.
Analgesics/*administration & dosage ; Animals ; Antioxidants/*administration & dosage ; Bone Resorption ; Disease Models, Animal ; Gene Expression Regulation ; Humans ; Interleukin-1beta/genetics/metabolism ; Iodoacetates/administration & dosage ; Knee Joint/*drug effects/metabolism/pathology ; Male ; Matrix Metalloproteinase 13/genetics/metabolism ; Osteoarthritis/chemically induced/*drug therapy/physiopathology ; Pain ; Plant Extracts/administration & dosage ; Proanthocyanidins/*administration & dosage ; Rats ; Rats, Wistar ; Seeds ; Tomography, Emission-Computed ; Tyrosine/analogs & derivatives/metabolism ; Vitis/immunology

OBJECTIVETo compare the effects of Wujia Bugu decoction and Alendronate sodium on protecting bone and muscle loss of hindlimb unloaded rats lasting three weeks.

METHODSMarch to May, 2009, 40 male Wistar rats with age of 6-week, were randomized divided to four groups (10 rats in each group): hindlimb unloaded group treated with Chinese medicine (HUC), hindlimb unloaded group treated with alendronate sodium (HUA), control group (CON), as well as hindlimb unloaded group (HU). During the experiment, rats of HUC was given Wujia Bugu decoction (including the Ciwujia, Shudihuang, Huainiuxi, Muli, etc. with the concentration of 0.704 g/ml) 10 ml/kg weight once a day, HUA was given quantitative alendronate sodium slice dissolve suspension (0.9 mg/ml) once a week. CON and HU were given double-distilled water. The experiment lasted 4 weeks,from the second to the forth week, rats in HU, HUC, HUA were hindlimb unloaded. All rats were sacrificed at the fourth weekend, the content of Ca, P and the activation of ALP in serum, Bone mineral density (BMD) of humerus and femurs, Biomechanical property of tibia and humerus, as well as the weight index of biceps and sural muscles were measured.

RESULTSCompared with CON, serum Ca of HU was significantly increased (P < 0.05), BMD, mechanical properties, muscle index of hindlimb were significantly reduce (P < 0.01), the serum Ca of HUA significantly increased (P < 0.05). Serum ALP of HUC was significantly higher than other three groups (P < 0.01). Compared with HU, femoral BMD of HUC and HUA significantly increased, tibial maximum load, maximum deflection and elastic load had increased tendency; calf muscle atrophy of HUC and HUA was alleviate 50% and 12.5% respectively (P > 0.05), humeral BMD had no significant difference, while the maximum deflection (P < 0.01) and elastic deflection (P < 0.05) in humerus of HUA were significantly lower.

CONCLUSIONHerbal prescription and alendronate sodium can effectively protect the bone and muscle loss of hindlimb unloaded rats, improve its mechanical structure. Herbal prescription has advantages of relieving mechanical properties change. The effects of Wujia Bugu decoction and alendronate sodium are similar in treating space weightlessness bone loss.

Alendronate ; administration & dosage ; Animals ; Bone Density ; drug effects ; Bone Resorption ; drug therapy ; physiopathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Femur ; drug effects ; physiopathology ; Hindlimb Suspension ; Humans ; Humerus ; drug effects ; physiopathology ; Male ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo explore the mechanisms of puncturing Shenshu point in improving osteoporosis.

METHODSSerum levels of testosterone (T) and osteocalcin (BGP) in senescence accelerated mouse prone 6 (SAMP6, test animals) and senescence accelerated mouse resistant 1 (SAMR1, for control) were determined by radioimmunoassay and their femoral biomechanical properties were determined with three-point bending test before and after puncturing to observe the effect of puncturing on the femoral biomechanical properties and bone mineral contents.

RESULTSCompared with the SAMR1 control group, the serum level of T (20.91 +/- 3.41 nmol/L) decreased (11.09 +/- 1.48 nmol/L in SAMP6 mouse), BGP (6.7 +/- 2.07 microg/L) increased (12.29 +/- 2.29 microg/L in SAMP6 mouse), femoral bending strength lowered and fragility increased. These changes were all improved to some extent or normalized, serum T level 15.05 +/- 2.63 nmol/L and BGP 8.88 +/- 1.85 microg/L after needling at Shenshu point showed significant difference when compared with those in SAMP6.

CONCLUSIONPuncturing Shenshu point could effectively prevent the bone loss in SAMP6 mice, increase their bone strength, the therapeutic effect is partly by way of promoting the secretion of sex hormone, improving bone metabolism, suppressing bone transformation rate and increasing bone minerals.

Acupuncture Points ; Aging ; Animals ; Biomechanical Phenomena ; Body Weight ; Bone Resorption ; Calcium ; metabolism ; Femur ; metabolism ; physiology ; physiopathology ; Male ; Mice ; Minerals ; metabolism ; Osteocalcin ; blood ; Osteoporosis ; blood ; metabolism ; physiopathology ; prevention & control ; Punctures ; methods ; Testosterone ; blood

OBJECTIVETo investigate the effects of the decoction of Rhizoma Dioscorea septemlobae (RD) on the bone metabolism in ovariectomized rats.

METHODThirty female, 3-month-old Wistar rats without pregnancy and deliver were randomly divided into 6 groups: sham (sham-operation), ovariectomy (OVX), OVX + diethylstilbestrol, OVX + high dose RD (4 g x kg(-1) x d(-1)), OVX + middle dose RD (2 g x kg(-1) x d(-1)) and OVX + low dose RD (1 g x kg(-1) x d(-1)) (n = 5 in every group). After 12-week period of continuous treatment, the urinary samples and blood samples were collected for the determination of serum estrodiol (E2), calcium (Ca), phosphorus (P), bone glaprotein (BGP), alkaline phosphatase (ALP), urinary calcium/creatinine (Ca/Cr), phosphorus/ creatinine (P/Cr) and deoxypyridioline/creatinine (DPD/Cr). The uteri were removed and weighed. The bone mineral density (BMD) and the biomechanical parameters of the femur of the rats in every group were determined, respectively.

RESULTThe coefficient of uteri in every dose group of OVX + RD was significantly higher than that in the OVX group (P < 0.01). The concentration of serum ALP, BGP and urinary DPD/Cr, Ca/Cr in the OVX group was significantly higher than that in the sham group (P < 0.05), respectively, However, that in the every dose of OVX + RD was lower than that in the OVX group, respectively. There was no significan difference in the concentration of serum Ca, P and urinary P/Cr in every group, respectively. The bone mineral density (BMD) in the OVX group was (0.032 +/- 0.007) g x cm(-2) and was significantly lower than that in the sham group (P < 0.01). However, the value in the group of every dose OVX + RD was significantly higher than that in the OVX group (P < 0.05, P < 0.01), respectively. The maximum loading, deflection and the maximum strain of the femur in the OVX group were (125.78 +/- 15.48) N, (1.87 +/- 0.22) mm, (9.34 +/- 1.10) % and were significantly lower than those in the sham group (P < 0.05, P < 0.01), respectively. The maximum loading and maximum stress were increased in different extent in the every dose group of OVX + RD, respectively.

CONCLUSIONThe decoction of RD can inhibit bone absorption, decline bone turnover and improve the loss of bone in ovariectomized rats.

Alkaline Phosphatase ; blood ; Animals ; Bone Density ; drug effects ; Bone Remodeling ; drug effects ; Bone Resorption ; blood ; physiopathology ; urine ; Calcium ; urine ; Creatinine ; urine ; Dioscorea ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Estradiol ; blood ; Female ; Femur ; drug effects ; metabolism ; physiopathology ; Osteocalcin ; blood ; Osteoporosis ; blood ; physiopathology ; urine ; Ovariectomy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Weight-Bearing

This study was aimed to assess the effects of fluid shear stress on the bone resorption in rat osteoclasts. The osteoclasts were derived from the lumbar vertebrae marrow cells which were isolated from the 6-month-old female Sprague Dawley rats, cultured on the slide at 1 x 10(6) cell/ml, and induced with 1, 25 (OH)2 Dihydroxyvitamin D3. The slide containing osteoclasts was taken out on day 7 after culture and then put into the flow chamber. The loads of fluid shear stress applied to the osteoclasts were 5.97, 11.36, 16.08 and 20.54 dyne/cm2, respectively, for 30 minutes. The osteoclasts unloading fluid shear stress were used as control. The bone resorptive pits were studied by light microscopy and scanning electron microscopy. The tartrate-resistant acid phosphatase (TRAP) secreted by osteoclasts was detected with ultraviolet spectrophotometry. The results showed that fluid shear stress can increase the activity of TRAP and significantly increase the number and area of bone resorptive pits made by osteoclasts,and the effect of fluid shear stress on the bone resorption of osteoclasts is the same as that on the activity of TRAP. The reaction of the osteoclasts to the fluid shear stress in this study also suggested that the bone resorption of osteoclasts be increased in a magnitude of fluid shear stress-dependent manner, and that the changes of TRAP activity be closely related to the changes of the number and area of resorptive pits of the osteoclasts.
Acid Phosphatase ; metabolism ; Animals ; Bone Resorption ; physiopathology ; Calcitriol ; pharmacology ; Cells, Cultured ; Female ; Isoenzymes ; metabolism ; Lumbar Vertebrae ; cytology ; Osteoclasts ; cytology ; enzymology ; physiology ; Rats ; Rats, Sprague-Dawley ; Shear Strength ; Tartrate-Resistant Acid Phosphatase

Gushukang is a compound Chinese herbal preparation. Pharmacological studies indicated that Gushukang can increase bone density, inhibit bone resorption, promote bone formation, and restore bone microstructure, as well as improve bone biomechanical parameters and promote healing of bone fracture. Clinical observations demonstrated that it has favorable efficacy in preventing and treating osteoporosis.
Bone Density ; drug effects ; Bone Resorption ; prevention & control ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Osteogenesis ; drug effects ; Osteoporosis ; drug therapy ; physiopathology ; Phytotherapy

This study was designed to investigate the effects of some Traditional Chinese Medicine (TCM) agents on bone resorption and morphometric features of osteoclasts as well as their relationships. TCM ShengGuZaiZaoSan and XianLingGuBao, were used to treat the experimental fracture. Thirty 6-month-old Chinchilla rabbits were used for the establishment of animal models each with a 3 mm bone defect in the middle of left radius as well as of right radius. These models were divided randomly into 3 groups : ShengGuZaiZaoSan Group (Group A), XianLingGuBao groups (Group B) and control-group (Group C). Every group was further divided into 2 subgroups: a former sacrificed group (14 days after operation) and a latter sacrificed group (31 days after operation). After the rabbits being killed, the samples of their undecalcified calli were subjected to the morphometry study of bone resorption and osteoclasts. Group A had more bone resorption, compared with Group B and C. Both Groups A and B exhibited some changed morphometric features of osteoclasts as compared with Group C (P < 0.05). Simple correlation analysis indicated that bone resorption is mainly correlated with osteoclast numbers, and that in individual group, bone resorption is correlated with osteoclast form factor, area and mean photodensity (P < 0.05). These allow us to conclude that ShengGuZaiZaoSan can increase bone resorption and accelerate bone remodeling by increasing osteoclast numbers at the former stage and can enhance osteoclast function at the latter stage. These changes are beneficial to fracture healing.
Animals ; Bone Remodeling ; drug effects ; Bone Resorption ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Fracture Healing ; drug effects ; physiology ; Male ; Osteoclasts ; drug effects ; pathology ; Phytotherapy ; Rabbits ; Radius Fractures ; drug therapy ; pathology ; physiopathology ; Random Allocation