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Particulated bone grafts--effectiveness of bone cell supply.

OBJECTIVE: The objective of this study was to measure the amount of viable bone cells present in different types of bone graft. MATERIAL AND METHODS: Bone chips were harvested from the trabecular or cortical bone of the mandible or the iliac crest and either milled or not. The average size of unmilled bone particles was 5 x 5 x 5 mm and that of milled was 2 x 2 x 2 mm. Drill sludge was obtained using either a ball reamer, a diamond ball or an implant drill (the latter from mandibular bone and of average dimension 1 x 1 x 1 mm). A measure of 0.5 g of each category was cultured in Dulbecco's modified Eagle's medium with additives for four weeks. Cell counts were performed. An analysis of the osteocalcin synthesis, the alkaline phosphatase (ALP) activity, the collagen types and the concentration of bone-specific collagen cross-links in medium supernatants was performed. RESULTS: Cells stained positively for osteocalcin and ALP in all groups. Bone-specific collagen cross-links could be quantified and collagen of types I and V was present with no difference in all groups. Unmilled spongy bone chips revealed greater cell counts than milled (P<0.05). Spongy bone chips revealed greater cell counts than cortical bone chips (P<0.05). Drill sludge obtained by hard alloy ball reamer showed the least amount of viable cells (P<0.05). CONCLUSIONS: Bone milling reduces the quantity of osteoblasts. Bone obtained by the ball reamer supplies a smaller number of cells than bone obtained by other methods. Unmilled spongy bone chips appear to offer the greatest amount of viable osteoblasts.

Adult↗

Bone formation at titanium porous oxide (TiUnite) oral implants in type IV bone.

BACKGROUND: Several oral implant design advances have been suggested to overcome poor bone quality, an impediment for successful implant treatment. A novel titanium porous oxide (TPO) surface has been shown to offer favorable results in several settings. The objective of this study was to evaluate the local bone formation and osseointegration at TPO-modified implants in type IV bone. METHOD: Three TPO surface-modified implants (TiUnite) were installed into the edentulated posterior maxilla in each of 8 Cynomolgus monkeys. The animals were injected with fluorescent bone labels at 2, 3, 4 and 16 weeks post-surgery and were euthanized at week 16 when block biopsies were collected for histologic analysis. RESULTS: The predominant observation of the TPO implant surface was a thin layer of new bone covering most of the implant threads. Mean (+/-SE) bone-implant contact for the whole study group was 74.1 +/- 4.8%. There was a significant variability in bone-implant contact between animals (P = 0.0003) and between sites of the same animal (P < 0.0001). The variance in bone-implant contact was 30% larger among sites of the same animal than between different animals (187.5 vs. 144.8, respectively). There was a small but significant difference in bone density immediately outside, compared to within the threaded area of the implants (37.1 +/- 3.2% vs. 32.1 +/- 3.2%, P < 0.0001). Bone density outside the implant threads was significantly correlated (beta = 0.682, P < 0.0001) with the bone density within the threaded area. Bone density within the threaded area was significantly correlated (beta = 0.493, P = 0.0002) with bone-implant contact, whereas bone density outside the implant threads did not have a significant effect (beta = 0.232, P = 0.1). CONCLUSIONS: The results suggest that the TPO surface possesses a considerable osteoconductive potential promoting a high level of implant osseointegration in type IV bone in the posterior maxilla.

Analysis of Variance↗

Increased bone volume and bone mineral content in xylitol-fed aged rats.

BACKGROUND: Our previous studies have shown that dietary xylitol supplementation protects against the loss of bone mineral after ovariectomy. The ovariectomy-induced decrease in trabecular bone volume is significantly retarded by dietary xylitol. OBJECTIVE: To study whether dietary xylitol can protect against bone loss also during aging, a long-term experimental study was performed with rats. METHODS: Twenty-four male Sprague-Dawley rats were divided into two groups. The rats in the control group were fed a basal RM1 diet, while the rats in the experimental group were continuously fed the same diet supplemented with 10% (w/w) xylitol. The rats were killed after 20 months. Their tibiae were used for the analyses of bone density and trabecular bone volume, and their femurs were used for the scanning analyses with peripheral quantitative computed tomography (pQCT). RESULTS: The tibial density of the xylitol-fed aged group (1.73 +/- 0.14 g/mm(3)) was significantly greater than that of the aged group without xylitol (1.56 +/- 0.14 g/ mm(3)). The trabecular bone volume of the xylitol-fed rats was 21.2 +/- 4.0%. It was significantly greater than that of the rats not receiving xylitol (9.3 +/- 4.3%). The pQCT-measured cortical bone mineral density and the pQTC-measured cortical bone mineral content of the femoral diaphysis were significantly greater in the xylitol-fed group than in the control group. The trabecular bone mineral density and the trabecular bone mineral content of the femoral distal metaphysis were also significantly greater in the xylitol-fed group than in the non-xylitol group. The total bone mineral density and the total bone mineral content of the femoral neck in the xylitol-fed aged group significantly exceeded those in the aged group without xylitol supplementation. CONCLUSIONS: A continuous moderate dietary xylitol supplementation leads to increased bone volume and increased bone mineral content in the long bones of aged rats. This indicates a xylitol-induced protection against aging-related osteoporotic changes.

Aging↗

Low estrogen and high parathyroid hormone-related peptide levels contribute to accelerated bone resorption and bone loss in lactating mice.

Providing enough calcium for milk production stresses calcium homeostasis in lactating mammals. A universal response to these demands for calcium appears to be the mobilization of maternal skeletal reserves, and bone loss during lactation has been well documented. However, the regulation of calcium and skeletal metabolism during lactation remains enigmatic. Our study was designed to examine mineral and bone metabolism in lactating mice. We found that mice lose bone rapidly at all sites during lactation. Bone mineral density as determined by dual-energy x-ray absorptiometry was 20 to 30% lower at the spine, femur, and total body in lactating compared with either age-matched virgin or pregnant mice. The decrease in bone mineral density was accompanied by dramatic reductions in bone volume and changes in trabecular architecture. Bone loss was also accompanied by increases in bone turnover as determined by biochemical markers and histomorphometry. PTHrP levels were elevated during lactation and correlated positively with markers of bone resorption and negatively with bone mass at all sites. Estrogen levels were low during lactation and correlated negatively with bone resorption markers. Finally, estrogen and pamidronate treatment lowered rates of bone resorption to baseline virgin levels and mitigated, but did not prevent, bone loss. These data suggest that the combination of estrogen deficiency and elevations in circulating PTHrP during lactation act to stimulate bone resorption and promote bone loss.

Animals↗

Programmed administration of parathyroid hormone increases bone formation and reduces bone loss in hindlimb-unloaded ovariectomized rats.

Gonadal insufficiency and reduced mechanical usage are two important risk factors for osteoporosis. The beneficial effects of PTH therapy to reverse the estrogen deficiency-induced bone loss in the laboratory rat are well known, but the influence of mechanical usage in this response has not been established. In this study, the effects of programed administration of PTH on cancellous bone volume and turnover at the proximal tibial metaphysis were determined in hindlimb-unloaded, ovariectomized (OVX), 3-month-old Sprague-Dawley rats. PTH was administered to weight-bearing and hindlimb-unloaded OVX rats with osmotic pumps programed to deliver 20 microg human PTH (approximately 80 microg/kg x day) during a daily 1-h infusion for 7 days. Compared with sham-operated rats, OVX increased longitudinal and radial bone growth, increased indexes of cancellous bone turnover, and resulted in net resorption of cancellous bone. Hindlimb unloading of OVX rats decreased longitudinal and radial bone growth, decreased osteoblast number, increased osteoclast number, and resulted in a further decrease in cancellous bone volume compared with those in weight-bearing OVX rats. Programed administration of PTH had no effect on either radial or longitudinal bone growth in weight-bearing and hindlimb-unloaded OVX rats. PTH treatment had dramatic effects on selected cancellous bone measurements; PTH maintained cancellous bone volume in OVX weight-bearing rats and greatly reduced cancellous bone loss in OVX hindlimb-unloaded rats. In the latter animals, PTH treatment prevented the hindlimb unloading-induced reduction in trabecular thickness, but the hormone was ineffective in preventing either the increase in osteoclast number or the loss of trabecular plates. Importantly, PTH treatment increased the retention of a baseline flurochrome label, osteoblast number, and bone formation in the proximal tibial metaphysis regardless of the level of mechanical usage. These findings demonstrate that programed administration of PTH is effective in increasing osteoblast number and bone formation and has beneficial effects on bone volume in the absence of weight-bearing and gonadal hormones. We conclude that the actions of PTH on cancellous bone are independent of the level of mechanical usage.

Animals↗

Parathyroid hormone deficiency and excess: similar effects on trabecular bone but differing effects on cortical bone.

Parathyroid hormone (PTH) may be anabolic at trabecular bone and catabolic in cortical bone. As many regions of the skeleton contain both types of bone, the effects of PTH deficiency or excess may be difficult to evaluate using bone densitometry, a technique that integrates the cortical and trabecular compartments of bone. We asked the following questions: 1) Is the higher bone mineral density (BMD) in postsurgical hypoparathyroidism due to higher cortical, not trabecular, bone? 2) Is age-related bone loss slowed in patients with postsurgical hypoparathyroidism? 3) Is lower BMD in primary hyperparathyroidism the result of deficits in cortical, not trabecular, bone? BMD of the lumbar spine, proximal femur, distal radius, and femoral midshaft was measured by postero-anterior (PA) scanning, while bone mineral content (BMC) of the third lumbar vertebra was measured by lateral scanning using dual x-ray absorptiometry in 10 women, ages 64.6 +/- 3.2 yr, with postsurgical hypoparathyroidism and in 25 women, ages 68.7 +/- 1.6 yr, with primary hyperparathyroidism. Measurements were repeated 4.7 +/- 0.6 yr later in 8 patients with hypoparathyroidism and 4.0 +/- 0.4 yr later in 20 age-matched controls. Data were expressed as z scores (SD, mean +/- sem) derived from 405 postmenopausal women. In patients with hypoparathyroidism, bone mass z score of the third lumbar vertebra (vertebral body plus posterior processes) was higher than zero by PA scanning (1.26 +/- 0.58 SD, P < 0.05) and lateral scanning (1.04 +/- 0.60 SD, P = 0.1), and higher at the trabecular-rich vertebral body (1.02 +/- 0.47 SD, P = 0.07) and predominantly cortical posterior processes (0.98 +/- 0.66 SD, P = 0.1) determined by lateral scanning. The BMD z scores were higher than zero at the femoral neck (0.89 +/- 0.48 SD, P = 0.09), but not at the femoral midshaft (0.45 +/- 0.60, NS) and distal radius (0.04 +/- 0.51, NS). During follow-up, femoral neck BMD decreased in controls but not in patients with hypoparathyroidism (slope, -0.00818 +/- 0.00496 g/cm2/year vs. 0.00907 +/- 0.00583 g/cm2/year, respectively, P = 0.06). There was no change in lumbar spine BMD in either group. In 25 women with primary hyperparathyroidism, there were no deficits in BMD at the third lumbar vertebra (vertebral body plus posterior processes) by PA or lateral scanning. By lateral scanning, BMC was increased at the vertebral body (0.64 +/- 0.31 SD, P < 0.01) and reduced at the posterior processes (-0.65 +/- 0.26 SD, P < 0.05). BMD was lower at the midshaft of the femur (-0.82 +/- 0.37 SD, P < 0.05) and at the distal radius (-0.68 +/- 0.20 SD, P < 0.01), but not at the femoral neck (-0.08 +/- 0.20 SD, NS). Longitudinal data were unavailable in hyperparathyroid patients. In summary, trabecular bone is increased by both PTH deficiency and excess. Cortical bone loss is slowed by PTH deficiency and accelerated by PTH excess so that suppression of PTH may reduce age-related bone loss and the risk of fracture. Assessment of BMD in PTH deficiency and excess requires the separate study of cortical and trabecular bone.

Absorptiometry, Photon↗

Effects of cyclic versus daily hPTH(1-34) regimens on bone strength in association with BMD, biochemical markers, and bone structure in mice.

UNLABELLED: We developed a cyclic PTH regimen with repeated cycles of 1-week on and off daily PTH injection and explored its effects on bone strength, BMD, bone markers, and bone structure in mice. Cyclic protocols produced 60-85% of the effects achieved by daily protocols with 57% of the total PTH given, indicating more economic use of PTH. The study supports further exploration of cyclic PTH regimens for the treatment of osteoporosis. INTRODUCTION: To minimize the cost and the catabolic action of hPTH(1-34), a cyclic PTH regimen with repeated 3-month cycles of on-and-off daily injection of hPTH(1-34) was developed in humans and shown to be as effective as a daily regimen in increasing vertebral BMD. However, changes in BMD may not adequately predict changes in bone strength. A murine model was developed to explore the efficacy of a cyclic PTH regimen on bone strength in association with other bone variables. MATERIALS AND METHODS: Twenty-week-old, intact, female C57BL/J6 mice (n = 7/group) were treated with (1) daily injection with vehicle for 7 weeks (control); (2) daily injection with hPTH(1-34) (40 microg/kg/day) for 7 weeks (daily PTH); and (3) daily injection with hPTH(1-34) and vehicle alternating weekly for 7 weeks (cyclic PTH). BMD was measured weekly by DXA, and serum bone markers, bone structure, and strength were measured at 7 weeks. RESULTS: Daily and cyclic PTH regimens increased BMD at all sites by 16-17% and 9-12%, respectively (all p < 0.01). The most dramatic effect of cyclic PTH occurred during the second week of treatment when PTH was off, with femoral and tibial BMD continuing to increase to the same extent as that produced by daily PTH. Both daily and cyclic PTH regimens significantly increased osteocalcin (daily, 330%; cyclic, 260%), mTRACP (daily, 145%; cyclic, 70%), femoral cortical width (daily, 23%; cyclic, 13%), periosteal circumference (daily, 5%; cyclic, 3.5%), and bone strength (max load: daily, 48%; cyclic, 28%; energy absorbed: daily, 103%; cyclic, 61%), respectively. Femoral bone strength was positively correlated with BMD, bone markers, and cortical structure. Neither regimen had an effect on vertebral bone strength. Although actual effects of cyclic PTH were 60-85% of those produced by daily PTH, the effects of cyclic PTH per unit amount administered were slightly greater than those of daily PTH for most measures. CONCLUSIONS: PTH-enhanced femoral bone strength is positively correlated with its effects on femoral BMD, bone markers, and bone structure. Cyclic PTH regimens represent a potential economic use of PTH and warrant further study.

Acid Phosphatase↗

Induction of callus formation by implants of bone morphogenetic protein and associated bone matrix noncollagenous proteins.

Callus formation in the periosteal bone interface in response to bone morphogenetic protein (BMP) and associated bone matrix noncollagenous proteins (BMP/NCP) was investigated in mature adult rabbits. For controls byproducts of BMP/NCP purification, bone marrow, eight nonskeletal tissues, purified matrix gamma-carboxyglutamic acid-rich protein (MGP), and a composite of BMP/NCP and polylactic-polyglycolic acid polymer (PLA/PGA) were also implanted in the periosteal bone interface. Quantitative microcomputer image analysis and histologic studies were performed three weeks after the implantation. BMP/NCP and bone marrow or BMP/NCP implanted over a single drill hole into the marrow cavity produced three times more new bone than the bone marrow alone. BMP/NCP alone produced twice as much new bone as bone marrow alone. Control implants of bovine serum albumin or purified MGP produced no new bone. Autogeneic minced muscle and ten nonskeletal tissue controls produced little or no bone formation. Even at one-fifth of the dose of BMP/NCP, a composite of PLA/PGA incorporating BMP/NCP showed almost the same amount of new bone as BMP alone. Histologically, the response to BMP/NCP consisted of an external callus of calcifying cartilage and woven bone. The response to subperiosteal implants of BMP/NCP or BMP/NCP with bone marrow or with minced muscle occurred with the same sequence of developmental events as seen either in embryonic skeletogenesis or in fracture callus.

Animals↗

Characterization of the mechanical and ultrastructural properties of heat-treated cortical bone for use as a bone substitute.

Heat-treated bovine cortical bone has been proposed as an alternative to bone grafts and synthetic bone substitutes because it may combine the advantages of allografts (high stiffness and strength) and synthetic materials (abundant supply, reduced risk of rejection and disease transfer). Its mechanical properties and ultrastructure, however, are not well characterized. To address this, we compared the compressive (n = 20, bovine bone) and tensile (n = 26, bovine bone) mechanical properties and the ultrastructure (n = 12, human bone) of intact versus 350 degrees C heat-treated cortical bone. The 350 degrees C heat-treated bone had a mean +/- SD elastic modulus similar to the intact bone for both compression (16.3 +/- 2.2 GPa, pooled; p = 0.68) and tension (16.3 +/- 3.7 GPa, pooled; p = 0.95). It also maintained 63% of the intact strength in compression but only 9% in tension (p < 0.001). Infrared scans and X-ray diffraction patterns showed no differences between the 350 degrees C heat-treated and intact bone but large differences between ashed (700 degrees C) and intact bone. Similarly, heat-treated bone previously has been shown to be biocompatible and osteoconductive. We conclude, therefore, that 350 degrees C heat-treated cortical bone may be an excellent load-bearing bone substitute provided that it is loaded in compression only in vivo and is shown by future work to have acceptable fatigue properties.

Aged↗

Effect of acute increases in bone matrix degradation on circulating levels of bone-Gla protein.

Serum bone Gla-protein (BGP), also called osteocalcin, is a specific and sensitive measure of bone turnover in a variety of metabolic bone disorders. Although some BGP diffuses into the circulation after synthesis by osteoblasts, most is incorporated into bone matrix where it remains until bone is resorbed. Thus, serum BGP could reflect bone formation, bone resorption, or a combination of both. The relationship of serum BGP to the components of bone turnover was evaluated in 18 normal women (mean age 48 yr; range 30-70) who received a continuous 24-h intravenous infusion of the 1-34 synthetic fragment of bovine parathyroid hormone. Mean +/- SE for urinary hydroxyproline excretion, an index of bone resorption, increased (from 22.7 +/- 2.2 to 38.5 +/- 3.7 micrograms/100 ml glomerular filtrate [GF], p less than .001), whereas levels of serum alkaline phosphatase, an index of bone formation, were unchanged (from 20 +/- 1 to 20 +/- 1 U/liter, NS). Despite the increase in bone resorption, levels of serum BGP decreased (from 8.8 +/- 0.8 to 6.8 ng/dl, p less than .001). The data suggest that circulating levels of BGP are a measure of bone formation but, at least in subjects with normal renal function, not a measure of bone resorption. Presumably BGP in bone matrix is degraded during osteoclastic resorption into fragments that either are not recognized by an antiserum raised against the native molecule or are rapidly cleared from the circulation.

Adult↗

Assessment of cancellous bone structure: comparison of strut analysis, trabecular bone pattern factor, and marrow space star volume.

Cancellous bone architecture is an important determinant of bone strength. Recently, several approaches to the assessment of bone structure in histological sections have been described; however, no comparative studies of these different methods have been reported. We have developed computerized methods for the simultaneous assessment of several indices of bone structure, including trabecular bone pattern factor, marrow space star volume, node-to-terminus ratio, trabecular number, and trabecular separation. The relationships between these variables has been examined in iliac crest cancellous bone obtained from 41 healthy subjects, 17 male and 24 female, aged 20-80 years (mean 47.9 years). All structural indices assessed showed significant correlations with cancellous bone area (p < 0.0001). Values for trabecular bone pattern factor and marrow space volume were highly correlated (r = 0.789; p < 0.0001). A comparison of indices obtained by strut analysis with trabecular bone pattern factor and marrow space star volume also revealed significant relationships, especially for the terminus-to-terminus strut length (r = 0.704 and r = 0.634, respectively; p < 0.0001) and node to terminus ratio (r = -0.947 and r = -0.788, respectively; p < 0.0001). The node-to-terminus ratio and trabecular bone pattern factor showed significant relationships with age which were independent of sex, cancellous bone area and trabecular width (p < 0.01 and p < 0.005, respectively). Our results demonstrate strong correlations between the different two-dimensional indices of bone structure in cancellous bone from healthy subjects. Trabecular penetration is likely to be an important determinant of all these variables, which may therefore reflect connectivity; however, direct comparison of these methods with three-dimensional techniques is required to establish their true relationship to bone structure.

Adult↗

Bone morphogenetic proteins inhibit adipocyte differentiation by bone marrow stromal cells.

The bone morphogenetic proteins were originally identified based on their ability to induce ectopic bone formation in vivo and have since been identified as members of the transforming growth factor-beta gene superfamily. It has been well established that the bone morphogenetic cytokines enhance osteogenic activity in bone marrow stromal cells in vitro. Recent reports have described how bone morphogenetic proteins inhibited myogenic differentiation of bone marrow stromal cells in vitro. In vivo, bone marrow stromal cells differentiate along the related adipogenic pathway with advancing age. The current work reports the inhibitory effects of the bone morphorphogenetic proteins on adipogenesis in a multipotent murine bone marrow stromal cell line, BMS2. When exposed to bone morphogenetic protein-2, the pre-adipocyte BMS2 cells exhibited the expected induction of the osteogenic-related enzyme, alkaline phosphatase. Following induction of the BMS2 cells with adipogenic agonists, adipocyte differentiation was assessed by morphologic, enzymatic, and mRNA markers. Flow cytometric analysis combined with staining by the lipophilic fluorescent dye, Nile red, was used to quantitate the extent of lipid accumulation within the BMS2 cells. By this morphologic criteria, the bone morphogenetic proteins inhibited adipogenesis at concentrations of 50 to 500 ng/ml. This correlated with decreased levels of adipocyte specific enzymes and mRNAs. The BMS2 pre-adipocytes constitutively expressed mRNA encoding bone morphogenetic protein-4 and this was inhibited by adipogenic agonists. Together, these findings demonstrate that bone morphogenetic proteins act as adipogenic antagonists. This supports the hypothesis that adipogenesis and osteogenesis in the bone marrow microenvironment are reciprocally regulated.

Adipose Tissue↗

Activin A increases the bone mass of grafted bone in C3H/HeJ mice.

Activin A, a member of the TGF-b superfamily, is abundant in bone matrix, but little is known about its physiological role in bone metabolism. The present study was undertaken to determine whether topical activin A can increase the bone mass of isografted bone. The tibiae were bilaterally dissected from a donor C3H/HeJ mouse and transplanted subcutaneously in the dorsal region of a recipient mouse. One isografted tibia was topically infused for either 1, 2, 3, or 4 weeks with activin A, using an osmotic minipump at a dose of 0.02, 0.2, or 2 ng/hr. The other tibia was infused with 0.9% NaCl (control). The following results were obtained: (1) Topical activin A (2 ng/hr) stimulated periosteal bone formation after 2 or 3 weeks. The bone area in a standardized transverse section averaged 1.3 fold that in the control. (2) Numerous cuboidal or conical osteoblasts appeared on the surface of newly formed bone after the infusion of activin A for 2 or 3 weeks. Autoradiographic studies using 3H-proline revealed that the surface area of newly formed bone labelled with autoradiographic silver grains was greater in activin A-treated bone than in the control, suggesting an increased synthesis and secretion of collagen by osteoblasts. (3) Topical activin A increased the number of osteoclasts after 2 to 4 weeks. Furthermore, enhanced or increased bone resorption was observed in the projected anterior site of activin A-treated bone after 4 weeks. These results suggest that topical activin A increases the bone mass of isografted bone by increasing bone turnover.

Activins↗

Effect of exercise on tibial and lumbar vertebral bone mass in mature osteopenic rats: bone histomorphometry study.

The effect of moderate running exercise on tibial and lumbar vertebral bone mass was examined in mature osteopenic rats by bone histomorphometry. Ten 37-week-old female Wistar rats, with bone loss resulting from being fed a relatively low-calcium diet for 14 weeks after ovariectomy at the age of 23 weeks, were randomly divided into two groups of five animals each; control and exercise groups. The exercise consisted of treadmill running at 12 m/min for 1 h per day on 5 days per week for 12 weeks. During the exercise period, all animals were fed a standard calcium diet. After 12 weeks of exercise, bone histomorphometry was evaluated for cancellous bone (secondary spongiosa) of the proximal tibia and the fourth lumbar vertebra and for cortical bone of the tibial shaft. The findings suggested that in the mature osteopenic rat, there was a beneficial effect of moderate running exercise with adequate calcium intake on bone mass only in a weight-bearing long bone, the tibia. The mechanism for increased bone mass appeared to be both decreased bone resorption and increased bone formation in cancellous bone and increased bone formation in cortical bone.

Analysis of Variance↗

[Bone collected with a bone collector during dental implant surgery].

BACKGROUND: Simultaneous implantation and augmentation using autogenous bone collected during implant surgery is a well-established procedure in oral implantology. The aims of this study were (1) to identify any bacterial contamination of bone obtained with a bone collector, and (2) to verify the antimicrobial effect of rinsing the bone collector with a 0.1% chlorhexidine solution prior to augmentation. MATERIAL AND METHODS: A total of 39 patients undergoing a simultaneous implantation and augmentation procedure were examined. All patients rinsed their mouths with a 0.1% chlorhexidine solution for 2 min prior to surgery. Bone was collected with the Osseous Coagulum Trap, while saliva was collected with a separate suction tip. Once bone collection was complete a microbiological swab was taken from the bone collector (sample 1); before the bone was taken from it 200 ml of a 0.1% chlorhexidine solution was aspirated into the collector, after which the bone was removed and the collector sieve was (sample 2) sent for microbiological analysis which included aerobic and anaerobic cultivation of microorganisms and their identification and semiquantitative assessment of microbial growth. RESULTS: Before the collector was rinsed with chlorhexidine microbial contamination was found in 34 (82.7%) of the 39 samples, and 37 different microbial species were identified in cultures. When the collector had been rinsed with 200 ml 0.1% chlorhexidine a significantly lower rate of microbial contamination was found: 66.7% of the samples were sterile. CONCLUSIONS: Despite separate suction techniques for bone dust and saliva and preoperative use of a chlorhexidine mouthwash, bacterial contamination of bone obtained from the mouth with a bone collector has to be anticipated. Not only the physiological bacteria of the oral flora, but also the microorganisms frequently associated with implant failure can be found. Rinsing the bone collector with 200 ml of a 0.1% chlorhexidine solution significantly reduces microbial contamination. The effects on bone vitality must be studied before routine rinsing with chlorhexidine can be recommended.

Adult↗

Dichloromethylenebisphosphonate (Cl2MBP) inhibits bone resorption through injury to osteoclasts that resorb Cl2MBP-coated bone.

Dichloromethylenebisphosphonate (Cl2MBP, formerly Cl2MDP) inhibits bone resorption in vivo and in vitro. The mechanism by which it inhibits osteoclastic bone resorption has not been established. To investigate this, osteoclasts were isolated from rat long bone and incubated with Cl2MBP (10(-9)-10(-5) M) on bone slices. Bone resorption was assessed as plan area resorbed after 6 and 24 h by scanning electron microscopy. Although Cl2MBP inhibited bone resorption in the first 6 h of culture, inhibition was more marked in the incubation period between 6 and 24 h. This pattern of accelerating inhibition is unlike the pattern we have observed using other resorption-inhibitors, and suggested resorption-mediated osteoclast injury. In keeping with this, we found reduced numbers of osteoclasts, and morphological features of cell injury and degeneration of osteoclasts, after incubation with Cl2MBP on bone slices. Bone seemed to be an essential component in Cl2MBP-mediated injury, since osteoclast numbers and morphology on plastic coverslips were unaffected by the bisphosphonate. Moreover, bone slices preincubated with Cl2MBP showed similar effects on resorption and morphology to cultures in in which osteoclasts on bone were continuously immersed in Cl2MBP. Neither non-resorptive cells (macrophages, UMR 106 cells), nor osteoclasts prevented from resorption by calcitonin, showed evidence of cytotoxicity after incubation on bone slices with Cl2MBP. These results suggest that even relatively high concentrations of Cl2MBP in the fluid phase do not affect osteoclasts, nor does contact with Cl2MBP-coated bone surfaces, but that injury to osteoclasts, and a consequent reduction in bone resorption, occurs when osteoclasts excavate bone surfaces upon which Cl2MBP is adsorbed.

Animals↗

The anabolic effect of human PTH (1-34) on bone formation is blunted when bone resorption is inhibited by the bisphosphonate tiludronate--is activated resorption a prerequisite for the in vivo effect of PTH on formation in a remodeling system?

Parathyroid hormone (PTH) and its (1-34) fragment are stimulators of bone turnover that have an anabolic effect increasing trabecular bone mass when administered intermittently by daily subcutaneous injections. Its clinical use in osteoporosis, however, has been limited by the concomitant increased bone resorption and deleterious effect on cortical bone. To evaluate if a treatment combining PTH and a potent inhibitor of bone resorption would retain the anabolic effect of PTH without increasing bone resorption, we analyzed the effects of PTH (1-34) (500 IU/d) with or without the bisphosphonate tiludronate (1 mg/kg per day) for 3 months on biochemical and histological indices of bone turnover in old female sheep, an animal model which has a slow bone remodeling activity that resembles the one of elderly women. As expected, PTH (1-34) induced a significant increase of urinary pyridinoline and hydroxyproline (reflecting bone resorption), and of serum osteocalcin and alkaline phosphatase (reflecting bone formation), that were consistent with an increase of resorption and tetracycline-based formation of bone measured on iliac crest biopsy. In contrast, all biochemical and histological indices of bone turnover were decreased in sheep receiving tiludronate, a potent inhibitor of bone resorption. Surprisingly, in the combined therapy group, biochemical and histological indices of both resorption and formation did not differ from the control groups. Thus, the model of old sheep, which closely resembles the situation in old human, shows that the anabolic effect of PTH on bone is not maintained when PTH is coadministered with a bisphosphonate, in marked contrast to results noted in the growing rat.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaline Phosphatase↗

Calcium phosphate scaffold and bone marrow for bone reconstruction in irradiated area: a dog study.

Squamous cell carcinomas (SCC) of the upper aero-digestive tract are characterized by a high incidence of bone invasion; their treatment often requires large and damaging surgical resections and radiotherapy. Surgical and radiotherapeutic procedures generate irreversible effects on normal tissues, involving injuries on their reparation properties, especially on bone. The quality of life of patients undergoing major surgery and radiotherapy in maxillary and mandible areas is often reduced but could be improved by bone reconstructions. Bone reconstructions are rarely performed because surgery is complex and unsafe in irradiated bone. The aim of the study was to evaluate the bone reconstruction possibilities of macroporous biphasic calcium phosphate (MBCP) associated to autologous bone marrow (BM) graft injected after irradiation. MBCP hollowed blocks were specially designed and implanted in tibia and femur bone before irradiation in a dog model. Implants were removed after 18 weeks. This is the first report of experiments performed after radiation delivery using high fractionated doses approximating usual treatment of SCC in human. The quality of the bone adjacent to implanted MBCP and the bone ingrowth's rates were evaluated. The qualitative and quantitative role of BM grafts associated with the MBCP implants was determined, using scanning electron microscopy linked to quantitative image analysis. A direct contact between newly formed bone and MBCP implants associated to BM graft was observed, without fibrous interposition. The new-bone formation was statistically increased inside the MBCP (P=0.0126) by BM grafts. This study demonstrates that BM graft added to MBCP constitute an appropriate material to be considered in case of bone defect occurring in irradiated tissue, and could be foreseen for use after bone removal for oncologic obligations.

Animals↗