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Intraoperative bone and bone marrow sampling: a simple method for accurate measurement of uptake of radiopharmaceuticals in bone and bone marrow.

Accurate estimation of bone marrow uptake of radiopharmaceuticals is of crucial importance for accurate whole body dosimetry. In this study, a method for obtaining normal bone marrow and bone during routine surgery without inconvenience to volunteers is suggested and compared to an indirect method. In five volunteers (group 1), 4 MBq 111In-labelled human polyclonal IgG (111In-IgG) was administered 48 h before placement of a total hip prosthesis. After resection of the femoral head and neck, bone marrow was aspirated from the medullary space with a biopsy needle. In five patients, suspected of having infectious disease (group 2), bone marrow uptake was calculated according to a well-accepted method using regions of interest over the lumbar spine, 48 h after injection of 75 MBq 111In-IgG. Bone marrow uptake in group 1 (4.5 +/- 1.3% D kg-1) was significantly lower than that in group 2 (8.5 +/- 2.1% D kg-1) (P < 0.01). Blood and plasma activity did not differ significantly for both groups. This method provides a system for directly and accurately measuring uptake and retention in normal bone marrow and bone of all radiopharmaceuticals at various time points. It is a safe and simple procedure without any discomfort to the patient. Since small amounts of activity are sufficient, the radiation dose to the patient is low.

Aged↗

Relations of bone mineral content, ash weight and bone mass: implication for correction of bone mineral content for bone size.

Bone mineral content (BMC) was measured by photon absorptiometry in fresh bone specimens: 35 iliac crest and 39 forearms were used. BMC was related to ash weight, fat-free weight and the amount of bone determined by quantitative histologic methods. Ninety-five per cent of the variation in bone mineral concentration (BMC') in the iliac crest could be explained by variation in ash weight, and 88% of the variation in BMC in the forearm could be explained by variation in the amount of bone determined histologically. The total cross-sectional bone area of radius and ulna could be calculated from the width of the structures measured on the photon absorption curves. Using this parameter and the relation between BMC and ash weight, the actual in-vivo bone mineral content in the forearm could be expressed in terms of ash-weight in grams per cm3 of total bone.

Anthropometry↗

The penetration of lincomycin into normal human bone. Determination of penetration into compact bone, spongy bone and bone marrow.

The penetration of lincomycin into normal bone was studied in 10 patients with fracture of the neck of the femur, a separate determination being made of the lincomycin concentration in serum, bone marrow, spongy bone and compact bone. The concentration of lincomycin in bone marrow was found to be at the same level as that in the serum. The concentration in spongy bone amounted in most cases to 50 to 75 percent of the concentration in the serum, whereas the concentration in compact bone varied from 0 to 15 percent of that in the serum.

Aged↗

[Bone and bone related biochemical examinations. Bone and collagen related metabolites. Bone-specific alkaline phosphatase].

Research has long been conducted to elucidate the association between bone and alkaline phosphatase (ALP). ALP-labeled monoclonal antibodies developed in recent years represent a significant advance in the evaluation of bone quality by making bone-specific ALP (BAP) available for standard quantitative measurement. While BAP yields variable results depending on the assay used, the enzyme immunoassay (EIA) or immunoradiometric assay (IRMA), it has enabled the risk of fracture, as well as the efficacy of pharmacological intervention, to be predicted to a certain degree, through evaluation of a parameter other than bone mass. In this regard, BAP is assumed not only to reflect bone strength, but also to serve as an index for bone quality, quite apart from bone mass. Thus, quantitative measurement of BAP currently represents the most effective means available for clinical evaluation of bone quality.

Adult↗

Bone-to-bone, joint-to-bone and joint-to-joint ratios in normal and diseased skeletal states using region-of-interest technique and bone-seeking radiopharmaceuticals.

Bone-to-bone, iliosacral joint-to-os sacrum and joint-to-joint ratios were computed using the region-of-interest technique 2 to 3 hrs. after injection of 99mTc Sn-methylene-diphosphonate or 99mTc Sn-pyrophosphate in 139 patients with skeletal diseases (bone tumours, degenerative changes of the spine and joints, inflammatory changes of joints) as well as in 123 patients with normal skeletal states. In the latter group, iliosacral joint-to-os sacrum ratios decreased with increasing age of the patients. In patients with osseous metastases of the spine ratios of 0.80 to 4.0 occurred ( reference area second vertebra below or above the affected vertebra). In degenerative changes of the spine values of 0.80 to 1.69 were computed. These results show, that 74% of the spine metastases could not be differentiated from benign changes of the spine by determining their relative amounts of bone uptake. In bone tumours of the extremities and in rheumatoid or gouty arthritis of the small joints (hands and feet) the highest ratios, i.e. contrasts, occurred referring to a contralateral reference area. Osteoarthritic and inflammatory alterations of the big joints could not be differentiated because of percentual distribution of the increased joint-to-joint ratios turned out to be nearly identical.

Adult↗

A comparative study of bone to bone repair and bone to tendon healing in patella-patellar tendon complex in rabbits.

OBJECTIVES: To study the healing quality of bone to bone and bone to tendon repair in a patella-patellar tendon complex. DESIGN: In vivo animal experiment in 60 mature 32-week-old female rabbits. BACKGROUND: Injuries of patella-patellar tendon complex are not uncommon. However, no studies are available to compare the healing quality between bone to bone and bone to tendon surgical reconstructions used for repair of patella-patellar tendon complex. METHODS: A standard transverse osteotomy was performed at the distal one-third of patella of one hindlimb. Both patellar fragments were reattached for bone to bone group while the patellar tendon was reattached to the remaining patella after removing the distal one-third of patella for bone to tendon group. Patella-patellar tendon complex was harvested at 8, 12 and 24 weeks postoperatively for biomechanical and histological evaluations, with n=8 and n=2 at each healing time points in both groups. The contralateral knee served as a control. RESULTS: No significant differences in the failure loads were found between two groups. However, greater ultimate stress was found in bone to bone group as compared with bone to tendon group at week 8 and 24 (both P<0.05). On average, the ultimate stress of the bone to tendon and bone to bone group only reached 20.6(4.2)% and 28.6(6.7)% of the control values at week 24, respectively. The discrepancy between findings in failure load and failure stress might be explained by an overall larger cross-sectional area of the healing interface in bone to tendon group as compared with bone to bone group. Histology revealed that the bone to bone healing was via endochondral ossification at the healing interface. In bone to tendon group, extensive scar tissue was formed to overbridge the healing interface and remodeled with healing over time. The structural integration at the tendon and bony healing interface was poor and no typical intermitted fibrocartilage zone as seen in normal bone to tendon junction was formed. CONCLUSION: Failure load of bone to bone and bone to tendon healing interface did not differ during repair of patella-patellar tendon complex. However, the healing interface of bone to bone repair in terms of material properties as reflected by failure stress was superior to that of the bone to tendon healing. RELEVANCE: The findings of this experimental study may suggest that the anatomical reconstruction of patella-patellar tendon complex injury may be the primary concern in decision making for selecting either bone to bone or bone to tendon repair. However whenever possible, to initial fracture (bone-to-bone) fixation for ensuring better and predictable repair at the healing interface.

Analysis of Variance↗

Regulation of osteocalcin production and bone resorption by 1,25-dihydroxyvitamin D3 in mouse long bones: interaction with the bone-derived growth factors TGF-beta and IGF-I.

Bone cells produce multiple growth factors that have effects on bone metabolism and can be incorporated into the bone matrix. Interplay between these bone-derived growth factors and calciotropic hormones has been demonstrated in cultured bone cells. The present study was designed to extend these observations by examining the interactions between either transforming growth factor-beta (TGF-beta) or insulin-like growth factor-I (IGF-I) and 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) in a mouse long bone culture model with respect to osteocalcin production and bone resorption. In contrast to the stimulation in rat and human, in the fetal mouse long bone cultures, 1,25(OH)2D3 caused a dose-dependent inhibition of osteocalcin production. Both the osteocalcin content in the culture medium and in the extracts of the long bones was reduced by 1,25(OH)2D3. This effect was not specific for fetal bone because 1,25(OH)2D3 also reduced osteocalcin production by the neonatal mouse osteoblast cell line MC3T3. TGF-beta inhibited whereas IGF-I dose-dependently increased osteocalcin production in mouse long bones. The combination of TGF-beta and 1,25(OH)2D3 did not result in a significantly different effect compared with each of these compounds alone. The IGF-I effect was completely blocked by 1,25(OH)2D3. In the same long bones as used for the osteocalcin measurements, we performed bone resorption analyses. Opposite to its effect on osteocalcin, 1,25(OH)2D3 dose-dependently stimulated bone resorption. TGF-beta reduced and IGF-I did not change basal (i.e., in the absence of hormones) bone resorption. Our results show that 1,25(OH)2D3-enhanced bone resorption is dose-dependently inhibited by TGF-beta and IGF-I. Regression analysis demonstrated a significant negative correlation between 1,25(OH)2D3-induced bone resorption and osteocalcin production. The specificity for their effect on 1,25(OH)2D3-stimulated bone resorption was assessed by testing the effects of TGF-beta and IGF-I in combination with parathyroid hormone (PTH). Like 1,25(OH)2D3, PTH dose-dependently stimulates bone resorption. However, PTH-stimulated bone resorption was not affected by TGF-beta. Like 1,25(OH)2D3-stimulated bone resorption, IGF-I inhibited the PTH effect but at a 10-fold higher concentration compared with 1,25(OH)2D3. In conclusion, the present study demonstrates growth factor-specific interactions with 1,25(OH)2D3 in the control of osteocalcin production and bone. With respect to bone resorption, these interactions are also hormone specific. The present data thereby support and extend the previous observations that interactions between 1,25(OH)2D3 and bone-derived growth factors play an important role in the control of bone metabolism. These data together with the fact that TGF-beta and IGF-I are present in the bone matrix and potentially can be released during bone resorption support the concept that growth factors may control the effects of calciotropic hormones in bone in a localized and possibly temporal manner. Finally, in contrast to human and rat, in mice 1,25(OH)2D3 reduces osteocalcin production and this reduction is paralleled by stimulation of bone resorption by 1,25(OH)2D3. These data thereby show a dissociation between osteocalcin production and bone resorption.

Animals↗

Autoclaved autograft bone combined with vascularized bone and bone marrow.

In this study, the authors investigated enhancing the results of autoclaved autograft bone by combining it with a vascularized bone graft and supplementing it with autogenous bone marrow for replacement of a 2-cm tibial defect in the rabbit model. The study was divided into 4 groups. Group I consisted of autoclaved autograft bone only; Group II, autoclaved autograft bone and vascularized bone graft; Group III, autoclaved autograft bone and autogenous bone marrow; and Group IV, autoclaved bone and autogenous bone marrow and vascularized bone graft. The vascularized bone graft was provided by the ipsilateral fibula, pedicled on the peroneal vessels, and transposed to sit alongside the autoclaved bone. In Group I, all grafts had nonunion at 16 weeks. In Group II, new bone formation was seen at the proximal and distal site of the autoclaved bone, host bone, and vascularized fibular graft junction as early as 2 weeks, and bony union occurred at the 16th week. The autogenous bone marrow-supplemented groups (III and IV) had new bone formation along the entire length of the autoclaved bone. Bony union at the proximal and distal junction was seen as early as 4 weeks, extending to the entire autoclaved bone by the eight week (98% of the cases with 1 case of infection). Histologic examination showed revascularization and capillary ingrowth into the autoclaved bone at 16 weeks, with a significantly improved torsional stiffness when compared with the groups without autogenous bone marrow supplementation. The addition of vascularized bone graft resulted in an improved union rate as compared with the autoclaved bone alone. Autogenous bone marrow supplementation, in addition to the vascularized bone graft, resulted in rapid new bone formation all around the autoclaved bone, early revascularization and incorporation of the autoclaved bone, and a significantly improved torsional stiffness of the reconstruction.

Animals↗

Placement of autogeneic bone chips or bovine bone mineral in guided bone augmentation: a rabbit skull study.

PURPOSE: Our primary aim was to use a rabbit guided bone augmentation model to evaluate whether use of autogeneic bone grafts or bovine bone mineral (BBM) combined with a space-making barrier enhances bone augmentation compared with a barrier alone. MATERIALS AND METHODS: Sixteen rabbits were studied. In each rabbit, 2 titanium cylinders, each with 1 titanium lid, were placed subcutaneously in perforated slits made in the cortical bones, with their open ends facing the parietal bones. One cylinder was left empty and the other was filled with either autogeneic skull bone chips or BBM. Bone labels were injected after 4 and 11 weeks. After 12 weeks, the animals were sacrificed to obtain ground sections for histology and histomorphometry. RESULTS: Significantly more tissue was augmented in the 2 test groups than in the control group. Most of the autografts were resorbed, leaving only minute amounts in the upper third of the cylinders. Slender new bone trabeculae were distributed mainly from the contiguous bone plate that had no contact with the remaining graft material. In the BBM group, most of the BBM remained evenly distributed in the cylinder. In the upper third of the cylinder, the BBM was surrounded by soft connective tissue, while in the lower two thirds, mainly mineralized bone enclosed the BBM. Equal amounts of mineralized bone were found in both test groups. Comparisons of contact between bone and BBM on one hand and bone and bone cylinder wall on the other revealed that the greatest bone contact was with the BBM in the lower third of the cylinder. In the middle and upper third of the cylinder, bone-BBM contact and bone-cylinder wall contact were similar. Fluorescent label intensity was higher in the autograft group than in the BBM group. In all 3 groups the intensity of the early label was similar to that of the late label, indicating that the graft materials do not seem to retard mineralization. DISCUSSION: BBM was found to promote as much new bone as did autogeneic bone. In addition, BBM appears to have at least the same osteoconductive properties as titanium, provided BBM is contained in a stable environment. CONCLUSIONS: Placement of autogeneic bone or BBM in conjunction with a stiff space-making barrier generated more tissue than a barrier only. In this model, autogeneic bone chips and BBM augmented similar amounts of new mineralized bone.

Analysis of Variance↗

Systemic zoledronate treatment both prevents resorption of allograft bone and increases the retention of new formed bone during revascularization and remodelling. A bone chamber study in rats.

BACKGROUND: In osteonecrosis the vascular supply of the bone is interrupted and the living cells die. The inorganic mineral network remains intact until ingrowing blood vessels invade the graft. Accompanying osteoclasts start to resorb the bone trabeculae and gradually replace the bone. If the osteonecrosis occurs in mechanically loaded parts, like in the subchondral bone of a loaded joint, the remodelling might lead to a weakening of the bone and, in consequence to a joint collapse. Systemic bisphosphonate treatment can reduce the resorption of necrotic bone. In the present study we investigate if zoledronate, the most potent of the commercially available bisphosphonates, can be used to reduce the amount or speed of bone graft remodeling. METHODS: Bone grafts were harvested and placed in a bone chamber inserted into the tibia of a rat. Host tissue could grow into the graft through openings in the chamber. Weekly injections with 1.05 microg zoledronate or saline were given subcutaneously until the rats were harvested after 6 weeks. The specimens were fixed, cut and stained with haematoxylin/eosin and used for histologic and histomorphometric analyses. RESULTS: By histology, the control specimens were almost totally resorbed in the remodeled area and the graft replaced by bone marrow. In the zoledronate treated specimens, both the old graft and new-formed bone remained and the graft trabeculas were lined with new bone. By histomorphometry, the total amount of bone (graft+ new bone) within the remodelled area was 35 % (SD 13) in the zoledronate treated grafts and 19 % (SD 12) in the controls (p = 0.001). Also the amount of new bone was increased in the treated specimens (22 %, SD 7) compared to the controls (14 %, SD 9, p = 0.032). CONCLUSION: We show that zoledronate can be used to decrease the resorption of both old graft and new-formed bone during bone graft remodelling. This might be useful in bone grafting procedure but also in other orthopedic conditions, both where necrotic bone has to be remodelled i.e. after osteonecrosis of the knee and hip and in Perthes disease, or in high load, high turnover conditions like delayed union, periprosthetic osteolysis or bone lengthening operations. In our model an increased net formation of new bone was found which probably reflects that new bone formed was retained by the action of the bisphosphonates rather than a true anabolic effect.

Animals↗

Concentration of insulin-like growth factor (IGF)-I and -II in iliac crest bone matrix from pre- and postmenopausal women: relationship to age, menopause, bone turnover, bone volume, and circulating IGFs.

Insulin-like growth factor-I (IGF-I) and -II are important local regulators of bone metabolism, but their role as determinants of human bone mass is still unclear. In the present study, we analyzed the concentration of IGF-I and -II in the bone matrix of 533 human biopsies from the iliac crest that were obtained during surgery for early breast cancer. There was an inverse association of bone matrix IGF-I concentration with age that was unaffected by menopause. Bone matrix IGF-I was positively associated with histomorphometric and biochemical parameters of bone formation and bone resorption and with cancellous bone volume. Based on the estimates of the linear regression analysis, women with a bone matrix IGF-I concentration 2 SD above the mean had a 20% higher bone volume than women with a bone matrix IGF-I concentration 2 SD below the mean. In contrast, serum IGF-I was neither correlated with bone turnover nor with bone volume and was only weakly associated with bone matrix IGF-I when adjusted for the serum concentration of IGF binding protein-3. Bone matrix IGF-II was positively associated with the osteoblast surface, but in contrast to IGF-I, tended to be positively associated with age and was unrelated to cancellous bone volume. In summary, our study suggests the following. 1) The concentration of IGF-I in cancellous bone undergoes age-related decreases that are similar to those of circulating IGF-I. 2) Menopause has no effect on this age-related decline. 3) Physiological differences in bone matrix IGF-I are associated with differences in iliac crest cancellous bone volume. 4) Bone matrix IGF-I is a better predictor of cancellous bone volume than circulating IGF-I. 5) The role of IGF-II in human bone tissue is clearly distinct from that of IGF-I.

Adult↗

Aspects of bone healing and bone substitute incorporation. An experimental study in rabbit skull bone defects.

After an initial assessment of the experimental approach this experimental project was undertaken to study the regenerative response and incorporation of various implanted materials in calvarial bone defects in rabbits. Four 5-mm full-thickness defects were trephined in the frontal and parietal bones of the rabbits. After removal of the circular bone plugs the defects were used as recipient sites for inlay bone tissue and bone substitutes. In five separate studies the impact of bone regeneration of autogeneic bone grafts or eight bone substitutes were evaluated mainly by contact radiography, light microscopy and morphometry. The observation periods were four and 15 weeks. The main findings were: Autogeneic bone chips offered only minor advantages over controls in the model used and, also, differences in bone regeneration between diversified amounts of bone chips were negligible. In contrast, after bone paste implantation a cellular and mature bone was rapidly produced. Natural bone mineral (Bio-Oss) and synthetic dense hydroxylapatite ceramic proved to be biocompatible and a definite long term bone regeneration around all implants regardless of granulae size or resorbability was observed. Initial bone regeneration in and around the Bio-Oss particles was more extensive. Demineralized bone matrix of membranous and enchondral origins displayed extensive osteoinductive capacity and early bone production significantly exceeded that of control groups. The embryonic origin implied minor effects on the initial regenerative response only. Lyophilized bone allografts of two embryonic origins showed a low osteoinductive potential and a similar fashion of bone regeneration. No marked difference between these groups were displayed. HTR-polymer alone or combined with membraneous mineralized autogeneic bone chips showed a more rapid early bone regeneration than the groups containing lactomer beads, resorbable gel and controls. Also, the HTR-material proved to be a well-tolerated implant material by the recipient tissue bed.

Animals↗

[Combined use of autologous micro-morselized bone with bone morphogenetic protein and type I collagen graft in repairing rabbit bone defects].

OBJECTIVE: To study the effect of combined use of autologous micro-morselized bone with bone morphogenetic protein(BMP) and type I collagen graft on the treatment of segmental bone defects. METHODS: The bulk bone of rabbit iliac crest was ground into micro-morselized bone, which was combined with BMP and type I collagen. The model of 1.5 cm bone defect was established in the middle shaft of the radius. Fifty-six rabbits were assigned to four repairing methods: autologous micro-morselized bone graft with BMP and type I collagen, autologous micro-morselized bone graft with type I collagen, autologous micro-morselized bone graft alone, and control group. The defect-repairing capability of each group was assessed by radiographic, histological, bone densitometry and biomechanical studies. RESULTS: X-ray manifested that at the end of 8 weeks after operation, the bone defect treated with autologous micro-morselized bone graft with BMP and type I collagen was repaired completely, and at the end of 12 weeks after operation the bone defect treated with autologous micro-morselized bone and type I collagen was cured completely, but the bone defect treated with autologous micro-morselized alone was completely repaired. No healing was found in the control group. In the bone densitometry detection, the material with BMP exhibited the strongest defect-repairing capability in terms of amount increased and quality of the new bone at the end of 8 weeks and 12 weeks. The group with BMP has the best mechanical strength of all groups at the end of 12 weeks. CONCLUSION: Autologous micro-morselized bone graft with BMP/type I collagen and autologous micro-morselized bone graft with type I collagen prove to be effective in repairing segmental bone defects. The autologous micro-morselized bone combined BMP and type I collagen is an excellent bone repairing material considering the satisfactory osteogenesis, osteo-conduction, and osteo-induction seen in this method.

Animals↗

Bone morphogenetic proteins in bone stimulate osteoclasts and osteoblasts during bone development.

UNLABELLED: In this study, overexpression of noggin, a BMP antagonist, in developing bone caused significantly decreased osteoclast number as well as bone formation rate, resulting in increased bone mass with immature bone quality. BMP signaling plays important roles in normal bone development and regulation of bone resorption. INTRODUCTION: Bone morphogenetic proteins (BMPs) act on various types of cells. Although involvement of BMP signals in osteoblast differentiation has been studied extensively, the effects of BMPs on osteoclasts have not been widely researched. Consequently, the net effects of BMPs on bone remain unclear. The purpose of this study was to delineate more fully the role of BMPs in skeletal biology. MATERIALS AND METHODS: We generated transgenic mice that express BMP4 or noggin in bone under the control of the 2.3-kb alpha1(I) collagen chain gene (Col1a1) promoter, and analyzed their bone phenotype. We also analyzed bone of transgenic mice expressing BMP4 specifically in cartilage. RESULTS: Mice overexpressing BMP4 in bone developed severe osteopenia with increased osteoclast number. Mice overexpressing noggin, a BMP antagonist, in bone showed increased bone volume associated with decreased bone formation rate and decreased osteoclast number. The noggin-transgenic tibias exhibited reduced periosteal bone formation and reduced resorption of immature bone in marrow spaces, associated with frequent fractures at the diaphysis. Co-culture of primary osteoblasts prepared from noggin-transgenic calvariae and wildtype spleen cells resulted in poor osteoclast formation, which was rescued by addition of recombinant BMP2, suggesting that noggin inhibits osteoclast formation by attenuating BMP activities in noggin-transgenic mice. The expression levels of Rankl were not decreased in primary osteoblasts from noggin transgenic mice. Immunoblot analysis showed increased phosphorylation of Smad1/5/8 in osteoclast precursor cells after 20-minute treatment with BMPs, suggesting that these cells are stimulated by BMPs. Mice overexpressing BMP4 in cartilage had enlarged bones containing thick trabeculae, possibly because of expansion of cartilage anlagen. CONCLUSIONS: Overexpression of noggin in bone revealed that BMP signals regulate bone development through stimulation of osteoblasts and osteoclasts.

Animals↗

[Experimental study on repair of bone defect in femoral head by enhanced autogenous bone combined with bone morphogenetic protein].

OBJECTIVE: To compare and evaluate the capability of pure autogenous bone and the enhanced autogenous bone combined with bone morphogenetic protein in bone repair of femoral head. METHODS: Eighteen femoral heads of 9 dogs were drilled by trephine, 4 mm in diameter, followed by respective implantations of autogenous bone grafting (group B) and of the enhanced autogenous bone composite, combined with bone morphogenetic protein (group C), with the self-repair of bone defect as the control (group A). Three, six, nine weeks after the operation, radiological examination, computerized tomography, light and electronic microscopes were performed to investigate the bone healing of the defect in the femoral head. RESULTS: In group A, it could be observed that there was hematoma organization and delayed woven bone formation in the 3rd week after operation, and there was little replacement of woven bone by bone trabecula in the 9th week; in group B, the autogenous bone implanted were dead in the 3rd week and maintained in situ in the 9th week; in group C, active new bone formation, either endochondral or intramembranous ossification, was found in the 3rd week and entire repair of the bone defect by bone trabecula in the 9th week after operation. CONCLUSION: The enhanced autogenous bone combined with bone morphogenetic protein could promote reconstruction of the bone defect in femoral head, superior to pure autogenous bone which could provide a framework for the new bone formation.

Animals↗

Effects of estrogen replacement therapy on bone turnover in subchondral bone and epiphyseal metaphyseal cancellous bone of ovariectomized cynomolgus monkeys.

UNLABELLED: ERT decreases the severity of OA in OVX cynomolgus monkeys. We show that bone formation is greater in subchondral bone compared with epiphyseal/metaphyseal cancellous bone of the proximal tibia in these animals and that ERT decreases bone formation in both sites. ERT may decrease the risk of OA by decreasing bone formation in the SC bone. INTRODUCTION: Estrogen replacement therapy (ERT) decreases the risk of osteoporosis and osteoarthritis (OA) in postmenopausal women and has been shown to have direct effects on cells of the bone and cartilage. The effects of ERT have been studied extensively in cancellous bone, but subchondral (SC) bone directly beneath the articular cartilage has not been specifically evaluated. MATERIALS AND METHODS: Adult feral female cynomolgus monkeys were bilaterally ovariectomized (OVX) to simulate menopause; treated with ERT, soy phytoestrogens (SPE), or no hormones (OVX control group) for 3 years; and labeled with calcein before necropsy. At necropsy, the proximal tibias of 20 randomly selected animals from each treatment group were embedded in bioplastic and sectioned. Areas and labels were measured in a carefully defined region of the SC bone and epiphyseal/metaphyseal cancellous (EMC) bone, and derived dynamic and static indices were compared between the SC and EMC bone and among the three treatment groups. Student's t-tests and ANOVA were used to compare the data. RESULTS AND CONCLUSIONS: In both the SC and EMC bone, most of the values for the dynamic indices were highest in the OVX control group, intermediate in the SPE group, and lowest in the ERT group. The mineralizing surface, double-labeled surface, and bone formation rate (surface referent) were significantly higher in the SC bone compared with the EMC bone in the OVX control group. The trabecular bone volume was higher in the SPE-treated group compared with the OVX control group. In conclusion, the bone turnover indices were higher in the SC bone compared with the EMC bone, and ERT decreased these indices in both sites. In addition, SPE was protective against loss of bone volume.

Age Factors↗

Comparison of bone scintigraphy with bone markers in the diagnosis of bone metastasis in lung carcinoma patients.

This study was designed to evaluate the utility of the bone markers total alkaline phosphatase (TAP), bone-specific alkaline phosphatase (BAP), aminoterminal propeptide of type I collagen (PINP), carboxyterminal propeptide of type I collagen (PICP), pyridinoline crosslinks (PYD), deoxypyridinoline crosslinks (DPD), cross-linked carboxyterminal telopeptide of type I collagen (ICTP), cross-linked carboxyterminal telopeptide of type I collagen (CTx, beta-CrossLaps) and tartrate-resistant acid phosphatase 5b (TRAP 5b) in comparison with bone scintigraphy for the diagnosis of bone metastasis in lung carcinoma patients. The study population consisted of 49 patients with bone metastasis confirmed by plain radiography and/or computed tomography, 89 patients without bone metastasis, 12 patients with benign lung diseases and 18 healthy persons. All patients were of male gender. The bone markers were measured using commercially available tests. Serum and urine were collected from fasting patients at the time of bone scan between 7.00 and 8.00 a.m. The sensitivity of bone scintigraphy was 100%, its specificity 76.4%, resulting in a diagnostic efficiency of 84.8%. The positive predictive value was calculated to be 70% and the negative one to be 100%. The concentrations of the bone markers TAP, BAP, PINP, PYD, DPD and ICTP were significantly higher in patients with bone metastasis than in those without bone metastasis (p<0.01). The levels of PICP and CTx only tended to be higher in the patients with bone metastasis compared to those without bone metastasis. There was no significant difference in the TRAP 5b levels between the two groups. There was also no difference in the marker levels between osteoblastic, osteolytic and mixed osteoblastic-osteolytic lesions. Contrary to BAP, PICP, CTx and TRAP 5b, the markers TAP, PINP, PYD, DPD and ICTP were found to be higher (p<0.01-0.05) in patients with bone metastasis than in patients with benign lung diseases. In addition, PYD, DPD and ICTP differentiated patients with benign lung diseases from the healthy controls. Based on cut-off values that correspond to 95% specificity in the group of healthy persons, the sensitivity of the marker assays were as follows (specificity in brackets): TAP 33.3% (97.5%), BAP 22% (100%), PINP 18.4% (97.5%), PICP 2.1% (95.2%), PYD 91.8% (24.1%), DPD 83.7% (34.5%), ICTP 75.5% (44.6%), CTx 45.8% (77.5%) and TRAP 5b 14% (84%). The corresponding data for the diagnostic efficiency were as follows: TAP 73.6%, BAP 77.1%, PINP 67.7%, PICP 61.1%, PYD 48.5%, DPD 55.2%, ICTP 56.1%, CTx 65.6% and TRAP 5b 58.7%, respectively. The positive predictive values ranged from 20% (PICP) to 100% (BAP) and the negative values from 62.7% (PICP) to 84% (PYD). In the ROC analysis, TAP, followed by RAP, PINP and PYD, showed the best performance. The levels of TAP, BAP, PINP, PYD, DPD and ICTP were found to be higher in the patients with bone metastasis compared to those with metastastic lesions in other sites (p<0.01, except for ICTP having a p value of < 0.05). The levels of TAP, BAP, PYD, DPD and ICTP increased significantly with the number of metastases. There was also a steady increase in T scores of the markers PINP, PYD, DPD and ICTP with the extent of the metastatic bone disease. It is concluded that the currently available bone markers cannot replace bone scintigraphy, either for screening or in the diagnosis of bone metastasis, in lung carcinoma patients. However, a panel consisting of TAP, BAP, PINP, PYD, DPD and ICTP may be of some value as an adjunct tool to bone scintigraphy for this purpose.

Acid Phosphatase↗

Trabecular bone turnover, bone marrow cell development, and gene expression of bone matrix proteins after low calcium feeding in rats.

Low-calcium-fed animals have been accepted as one of the experimental models showing a reduction in bone mass. However, the effects of short-term low-calcium feeding on bone turnover, the development of osteoprogenitor cells, and gene expression of bone matrix proteins have not been reported. In this study, we examined the effect of a low-calcium diet on rat tibia and analyzed the changes in the bone by histomorphometry, bone marrow cell culture, and in situ and Northern hybridization of the bone matrix proteins. Rats were fed either a low-calcium diet (0.05% Ca) or a normal calcium diet (0.5% Ca) using the pair feeding technique. They were killed at day 0, 12 h, and days 1, 2, and 3. In the low-calcium group, the serum parathyroid hormone (PTH) level was temporarily increased in 12 h after feeding the low-calcium diet. Bone mineral density in the trabecular bone was significantly decreased from 1 day after the low-calcium feeding, but cortical bone did not show any changes during the experimental period. The bone volume per tissue volume in the proximal tibia also decreased from day 1 in the low-calcium group. The number of osteoclasts and osteoblasts on the trabecular bone surface was increased in the low-calcium group compared with the normal-calcium group. An ex vivo study showed that the number of progenitors of osteoclasts and osteoblasts in bone marrow was also increased in the low-calcium group of rats. The localization of type I collagen mRNA was observed in osteoblasts in the low-calcium group. The Northern hybridization study showed that the gene expression of type I collagen, osteopontin, and osteocalcin was increased at day 3 in the low-calcium group. These results indicated that the trabecular bone surface quickly responded to the low-calcium feeding and that bone remodeling activity was activated probably by PTH. The changes in bone marrow cell populations and the gene expression of bone matrix proteins are closely associated with increased bone turnover induced by the low-calcium diet, resulting in rapid bone loss of the trabecular bone.

Animals↗