OBJECTIVETo investigate whether the titrate-resistant acid phosphatase 5 (ACP5) gene polymorphisms were associated with the occurrence or curve severity of adolescent idiopathic scoliosis (AIS).

METHODSThere were 372 AIS patients from January 2006 to December 2008 and 239 normal controls from March 2005 to August 2006 were recruited. The Cobb angles were ≥ 10° in all AIS patients. Using the haplotype data of Han population from the Hapmap Project, two tag SNPs (rs2229531, rs2071484) were defined for ACP5 gene. PCR-restriction fragment length polymorphism was used for the genotyping.

RESULTSNo polymorphism in rs2229531 was found in this study. The genotype and allele frequency distribution in rs2071484 were similar between AIS patients and normal controls (χ(2) = 3.336 and 1.438, P > 0.05). The mean maximum Cobb angles of different genotypes of rs2071484 in ACP5 gene were 38° ± 19° in AA, 34° ± 14° in AG and 38° ± 21° in GG, which were similar with each other among AIS patients who reached skeletal maturity or received surgery treatment (P = 0.157).

CONCLUSIONThe ACP5 gene is neither associated with the occurrence nor the curve severity of AIS.

Acid Phosphatase ; genetics ; Adolescent ; Child ; Female ; Humans ; Isoenzymes ; genetics ; Male ; Polymorphism, Genetic ; Scoliosis ; genetics ; Tartrate-Resistant Acid Phosphatase

OBJECTIVESTo construct the cancellous bone explant model and a method of culturing these bone tissues in vitro, and to investigate the effect of mechanical load on growth of cancellous bone tissue in vitro.

METHODSCancellous bone were extracted from rabbit femoral head and cut into 1-mm-thick and 8-mm-diameter slices under sterile conditions. HE staining and scanning electron microscopy were employed to identify the histomorphology of the model after being cultured with a new dynamic load and circulating perfusion bioreactor system for 0, 3, 5, and 7 days, respectively. We built a three-dimensional model using microCT and analyzed the loading effects using finite element analysis. The model was subjected to mechanical load of 1000, 2000, 3000, and 4000 με respectively for 30 minutes per day. After 5 days of continuous stimuli, the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) were detected. Apoptosis was analyzed by DNA ladder detection and caspase-3/8/9 activity detection.

RESULTSAfter being cultured for 3, 5, and 7 days, the bone explant model grew well. HE staining showed the apparent nucleus in cells at the each indicated time, and electron microscope revealed the living cells in the bone tissue. The activities of AKP and TRAP in the bone explant model under mechanical load of 3000 and 4000 με were significantly lower than those in the unstressed bone tissues (all P<0.05). DNA ladders were seen in the bone tissue under 3000 and 4000 με mechanical load. Moreover, there was significant enhancement in the activities of caspase-3/8/9 in the mechanical stress group of 3000 and 4000 με(all P<0.05).

CONCLUSIONSThe cancellous bone explant model extracted from the rabbit femoral head could be alive at least for 7 days in the dynamic load and circulating perfusion bioreactor system, however, pathological mechanical load could affect the bone tissue growth by apoptosis in vitro. The differentiation of osteoblasts and osteoclasts might be inhibited after the model is stimulated by mechanical load of 3000 and 4000 με.

Acid Phosphatase ; metabolism ; Alkaline Phosphatase ; metabolism ; Animals ; Apoptosis ; Bone Development ; Caspases ; metabolism ; Finite Element Analysis ; Isoenzymes ; metabolism ; Male ; Rabbits ; Stress, Mechanical ; Tartrate-Resistant Acid Phosphatase ; X-Ray Microtomography

To investigate the preventive effect and possible mechanism of puerarin(Pur) in rat model of disuse osteoporosis(DOP),thirty healthy Wistar female rats of 2 months old were randomly divided into control group(Control), hindlimb suspension group(HLS), and puerarin group(HLS+Pur) in hindlimb suspension, with 10 rats in each group. A disuse osteoporosis model was established by tail suspension method, and 15.4 mg·kg~(-1) puerarin suspension was administered to HLS+Pur group every day, and the same volume of distilled water was administered to Control group and HLS group respectively. After 28 days, the rats were sacrificed by abdominal aorta blood collection, the main organs of the rats were removed, and the bone tissues of the rats were dissected. The organ index of the rats was calculated and the histopathology of the organs was observed under microscope. Bone mineral density test and bone biomechanical experiment were performed. Bone histomorphometry results were observed after bone tissue sectioning, and serum biochemical markers of bone metabolism were determined. There was no significant difference in organ index between the groups. There was no obvious abnormality in the pathological examination of the organs. The results of bone mineral density showed that puerarin could significantly increase the bone density of the tibia and vertebrae caused by hindlimb suspension. The mechanical parameters experiments showed that puerarin could effectively increase the maximum load and elastic modulus of the tibia and vertebrae. Fluorescence labeling showed that the fluorosis interval increased and the bone formation increased during puerarin treatment. The VG staining results showed that compared with the HLS group, in the puerarin group, the number of trabecular bone increased, the thickness of the trabecular bone became thicker, and the bone separation became smaller, which greatly improved the bone microstructure after hindlinb suspension. In addition, serum biochemical indicators showed that puerarin could promote bone formation index bone calcium. The content of osteocalcin(OC) increased and inhibited the formation of tartrate-resistant acid phosphatase 5 b(TRACP 5 b). Puerarin has a preventive effect in the rat model of disuse osteoporosis and its effect is good, and its mechanism may be related to promoting bone formation and inhibiting bone resorption.
Animals ; Bone Density ; Female ; Isoflavones ; pharmacology ; Osteocalcin ; metabolism ; Osteoporosis ; drug therapy ; Rats ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase ; metabolism

Studying effects of 50 Hz sinusoidal electromagnetic fields (SEMFs) with different intensities on peak bone mass (PBM) of rats may provide a theoretical basis for application of electromagnetic clinical field. 30 female SD rats, 6 weeks of age, were randomly divided into three groups: the control group, 0.1 mT electromagnetic field group (EMFs) and 0.6 mT EMFs. The EMFs groups were treated for 3 h/day. After 8 weeks, we examined their bone mineral densities (BMD) , measured their bone biomechanical properties, and made serum levels of osteocalcin (OC), tartrate-resistant acid phosphatase 5b (TRACP 5b), and histomorphometry. It was found that the BMD (P < 0.01), maximum mechanical load (P < 0.01) in the 0.1 mT group were significantly higher than those in the control group, and Yield strength (P < 0.05), the analyses of serum bone turnover markers and histomorphometric parameters were better than those in the control group (P < 0.05). However, the 0.6 mT group did not have significantly difference comparing with that in the control group. This study proved that 50 Hz 0.1 mT SEMFs can increased BMD, bone strength, and bone tissue microstructure. Therefore, 50 Hz 0.1 mT SEMFs can improve peak bone mass of rats.
Acid Phosphatase ; blood ; Animals ; Bone Density ; Bone and Bones ; physiology ; Electromagnetic Fields ; Female ; Isoenzymes ; blood ; Osteocalcin ; blood ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase

PURPOSETo investigate the influence of the same mechanical loading on osteogenesis and osteoclastogenesis in vitro.

METHODSPrimary osteoblasts, bone marrow-derived mesenchymal stem cells (BMSCs, cultured in osteoinductive medium) and RAW264.7 cells cultured in osteoclast inductive medium were all subjected to a 1000 μstrain (μs) at 1 Hz cyclic mechanical stretch for 30 min (twice a day).

RESULTSAfter mechanical stimulation, the alkaline phosphatase (ALP) activity, osteocalcin protein level of the osteoblasts and BMSCs were all enhanced, and the mRNA levels of ALP and collagen type I increased. Additionally, extracellular-deposited calcium of both osteoblasts and BMSCs increased. At the same time, the activity of secreted tartrate-resistant acid phosphatase, the number of tartrate-resistant acid phosphatase-positive multinucleated cells, matrix metalloproteinase-9 protein levels of RAW264.7 cells and the extracellular calcium solvency all decreased.

CONCLUSIONThe results demonstrated that 1000 μs cyclic mechanical loading enhanced osteoblasts activity, promoted osteoblastic differentiation of BMSCs and restrained osteoclastogenesis of RAW264.7 cells in vitro.

Animals ; Biomechanical Phenomena ; Cell Differentiation ; Cells, Cultured ; Mice ; Mice, Inbred C57BL ; Osteoblasts ; cytology ; Osteoclasts ; physiology ; Osteogenesis ; physiology ; Tartrate-Resistant Acid Phosphatase ; metabolism

OBJECTIVE: To analyze the level of serum N-terminal propeptide of type I precollagen (PINP) and tartrate resistant acid phosphatase 5b(TRACP-5b) in patients with femoral neck fracture(FNF) after minimally invasive anterior lateral approach with total hip arthroplasty and the effects of hip function. METHODS: From September 2016 to May 2017, 98 cases of femoral neck fracture were divided into control group and observation group, 49 cases in each group. There were 49 patients in control group, including 30 males and 19 females, ranging in age from 63 to 72 years old, who underwent minimally invasive anterolateral total hip arthroplasty. There were 49 patients in observation group, including 29 males and 20 females, ranging in age from 62 to 73 years old, who underwent minimally invasive anterolateral total hip arthroplasty. The perioperative conditions(operation time, bleeding volume, incision length, hospitalization time), bone metabolism indexes including PINP, TRACP-5b, fibroblast growth factor(FGF), bone gla-protein(BGP), propetide carboxy-terminal procollagen (PICP), bone-specific alkaline phosphatase(BAP), and pain mediators such as prostaglandin E2 (PGE2) and 5-hydroxytrytamine (5-HT) levels were compared between the two groups. The hip joint function and complications were evaluated. RESULTS: The operation time of the observation group was longer than that of the control group(<0.05); there was no significant difference in bleeding volume, incision length and hospitalization time between two groups(>0.05). PINP, fibroblast growth factor, BGP, PICP and BAP in observation group were higher than those in control group 1 month after operation, and TRACP-5b was lower than those in control group(<0.05); PGE2 and 5-HT in observation group 1 month after operation were lower than those before operation, and lower than those in control group(<0.05). The excellent and good rate of hip function in observation group was higher than that in control group (<0.05), and the incidence of complications was not significantly different from that in the control group (>0.05). CONCLUSIONS Minimally invasive anterolateral approach total hip arthroplasty is safe and reliable, and can improve hip function, improve bone metabolism, promote fracture healing, alleviate pain in patients with femoral neck fracture, which is worthy of promotion.
Aged ; Arthroplasty, Replacement, Hip ; Collagen Type I ; Female ; Femoral Neck Fractures ; surgery ; Hip Joint ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Tartrate-Resistant Acid Phosphatase ; Treatment Outcome

OBJECTIVETo study the change and relationship among bone mineral density (BMD), collagen composition and biomechanical properties of the callus in the healing process of osteoporotic fracture.

METHODSThe osteoporotic rat model and fracture model were established through bilateral ovariectomy (OVX) and osteotomy of the middle shaft of the right hind tibiae, respectively. Ninety female SD rats were randomly divided into OVX group and sham group. With the samples of blood and callus, roentgenographic and histological observation were performed for the assessment of the healing progress of the fracture, and the serum concentration of TRAP-5b, proportion of type I collagen, BMD and biomechanical properties of the callus were measured.

RESULTSThe OVX group experienced a significant delay of fracture healing. The mean serum concentration of TRAP-5b of rats in the OVX group was much higher than that in the sham group after the operation (P less than 0.05), but the difference at the same time point after fracture was smaller than that before fracture (P less than 0.05). The BMD of the callus in both groups reached the peak value at the 6 th week after fracture while the proportion of the type I collagen and the biomechanical strength reached the peak at the 8th week.

CONCLUSIONSThe deficiency of estrogen after the ovariectomy could induce the up-regulation of the osteoclasts activities, whereas the potency of further activation after fracture was depressed. Although the synthesis of collagen together with its mineralization determines the biomechanical properties of new bone, the accumulation of collagen could be assessed as an index in the prediction of biomechanical strength of bones independent of the bone mineral deposition.

Acid Phosphatase ; blood ; Animals ; Biomechanical Phenomena ; Bone Density ; Bony Callus ; physiology ; Collagen ; chemistry ; Collagen Type I ; blood ; Fracture Healing ; physiology ; Isoenzymes ; blood ; Male ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase

OBJECTIVETo investigate the effect of pulsed electromagnetic fields (PEMs) on inducing osteoclastic like cell (OLC) formation changes and apoptosis in ovariectomized (OVX) rats bone marrow culture in vitro.

METHODSThirty healthy three-month-old female Wistar rats were either sham-operated (Sham) or ovariectomized (OVX) and randomly divided into three groups: group A (OVX + PEMs, 18 rats), group B (OVX, 6 rats) and group C (Sham, 6 rats); group A was again randomly divided into three groups: A1, A2, A3. The frequencies adopted were 1.5, 2, 75 Hz and 30 minutes for once a day. All rats were fed with normal diet for 3 months, then the bone marrow of all rats were cultured, 2 days later, group A cells (including group A1, A2, A3) were collected and exposed to different frequencies PEMs for 2 weeks (30 min/day). In order to observe the changes of osteoclasts and count their numbers, cells were taken for Wright Giemsa staining, tartrate-resistance acid phosphatase (TRAP) staining and Hoechst 33258 staining.

RESULTSTRAP staining results indicated the number of OLC in group C was the least, then was group A2, A3, A1, B. The number of OLC in group B was remarkably increased (P < 0.01; vs group C, A2). The number of OLC in group B was significantly increased (P < 0.05; vs group A1, A3). Hoechst 33258 staining results indicated the number of apoptosis of OLC in group C was more than other groups, which of group C, A2 was significantly increased (P < 0.05; vs group B).

CONCLUSIONPEMs had decreased the formation of OLC and increased the number of apoptosis of OLC in ovariectomized (OVX) rats bone marrow culture in vitro, the effects of 2 Hz was the best. PEMs would be a new way of osteoporosis therapy.

Acid Phosphatase ; metabolism ; Animals ; Apoptosis ; radiation effects ; Bone Marrow Cells ; cytology ; enzymology ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Female ; Isoenzymes ; metabolism ; Osteoclasts ; cytology ; enzymology ; radiation effects ; Ovariectomy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase

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

OBJECTIVETo investigate the effects of fluoride on activities of tartrate-resistant acid phosphate (TRAP) and matrix metalloproteinase-9 (MMP-9) in rat osteoclasts cultured in vitro.

METHODSOsteoclast was isolated mechanically from long bones of neonatal rats and cultured in vitro. Histochemical stain was applied to detect the effects of fluoride on activities of TRAP and in-situ hybridization was used to study the expression of MMP-9 mRNA in rat osteoclasts in vitro.

RESULTSNumber of TRAP positive cells was 154.2, 160.0, 170.6, 179.0 and 180.0 per cm(2), respectively for the rats with varied doses of fluoride, in a dose-response pattern but without statistical significance. The expression of MMP-9 mRNA increased with elevating dose of fluoride, especially in the rats with 1.00, 2.00 and 4.00 mg/L of fluoride, to 94.50, 94.64 and 104.97, respectively, significantly different from those in control group.

CONCLUSIONSFluoride can enhance the MMP-9 mRNA expression in cultured osteoclasts of rats.

Acid Phosphatase ; metabolism ; Animals ; Animals, Newborn ; Biomarkers ; Cells, Cultured ; Dose-Response Relationship, Drug ; Fluorides ; pharmacology ; Isoenzymes ; metabolism ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Osteoclasts ; cytology ; drug effects ; enzymology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase