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PubMed2003

Reconstruction of cartilage, bone, and hematopoietic microenvironment with demineralized bone matrix and bone marrow cells.

Highly specialized hard tissues, such as cartilage, bone, and stromal microenvironment supporting hematopoiesis, originate from a common type of mesenchymal progenitor cell (MPC). We hypothesized that MPCs present in bone marrow cell suspension and demineralized bone matrix (DBM) that possess natural conductive and inductive features might constitute a unit containing all the essential elements for purposive bone and cartilage induction. Using a rodent preclinical model, we found that implantation of a composite comprising DBM and MPCs into A) a damaged area of a joint; B) an ablated bone marrow cavity, and C) a calvarial defect resulted in the generation of A) a new osteochondral complex comprising articular cartilage and subchondral bone; B) trabecular bone and stromal microenvironment supporting hematopoiesis, and C) flat bone, respectively. The new tissue formation followed differentiation pathways controlled by site-specific physiological conditions, thus developing tissues that precisely met local demands.

Animals↗

PubMed2003

Does quantitative ultrasound of bone reflect more bone mineral density than bone microarchitecture?

Relationships among quantitative ultrasound of bone (QUS), bone mineral density (BMD) and bone microarchitecture have been poorly investigated in human calcaneus.Twenty-four specimens, from 12 men and 12 women (mean age 78 +/- 10 years; range 53-93), removed from cadavers were studied. The feet were axially sectioned above the ankle. Two variables were measured for QUS (Achilles, Lunar): broadband ultrasound attenuation (BUA) and speed of sound (SOS). A third variable, the stiffness index (SI), which is a combination of both BUA and SOS, was also calculated. BMD (a lateral view) was measured on a QDR 2000 densitometer (Hologic). Bone microarchitecture was assessed by computed tomography (CT) using a conventional CT-system. Fifteen sagittal sections (1 mm in width and 2 mm apart) were selected for CT. Methods used for characterizing bone microarchitecture consisted in structural (trabecular network characterization) and a fractal analyses. The relationships between QUS and bone microarchitecture were assessed by simple linear regression analysis with and without adjustment for BMD (partial correlation) and by backward stepwise regression analysis. Strong relationships were found between BMD and QUS. Adjusted r(2) values were 0.545 for SOS and 0.717 for SI. Two microarchitectural variables were also significantly correlated with both SOS and SI: apparent trabecular separation (App Tr Sp) and trabecular bone pattern factor (App TBPF). After adjustment for BMD few correlations between QUS and microarchitectural variables were always significant. Adjusted squared semipartial coefficients of correlation (rsp2) values between SOS and bone microarchitecture were 6%, 6.8%, 13.2% and 4.6% for App BV/TV, App Tr Sp, App TBPF and fractal dimension (FD), respectively. For SI, corresponding figures were 3.7%, 4.1%, 5.2% and 3.2%. Backward stepwise regression analysis using BMD and microarchitecture showed a slight increase of r(2) values that varied from 8.4% for SI to 17.8% for SOS, compared with BMD alone. The current study suggests that although BMD is a major determinant of acoustic properties of human calcaneus, significant density independent relationships with bone microarchitecture should also be taken into account.

Aged↗

PubMed2006

Evaluation of tibial bone-tunnel changes with X-ray and computed tomography after ACL reconstruction using a bone-patella tendon-bone autograft.

Thirteen patients who underwent anterior cruciate ligament (ACL) reconstruction with bone-patella tendon-bone autografts (BTB) using interference screws were the subjects of this study. We analysed the relationship between bone-tunnel changes and clinical results 2 years after ACL reconstruction. To investigate changes, X-ray images were used to evaluate bone-tunnel enlargement, and computed tomography (CT) was used to evaluate the sclerotic area around the bone tunnel. The KT-2000 was used to measure the discrepancy of tibial anterior displacement between the affected and nonaffected sides (DTAD). There was no correlation between bone-tunnel enlargement and DTAD. On the other hand, in the CT evaluation, there was a significant correlation between the sclerotic area and DTAD. Our results suggest that it is more significant to analyse the area of sclerotic change than bone-tunnel enlargement for clinical evaluation. We cannot evaluate bone-tunnel changes correctly with two-dimensional X-rays and cannot analyse the sclerotic area using X-rays. Therefore, we recommend that CT, with which it is possible to analyse the sclerotic area, be used to evaluate bone-tunnel changes and clinical results.

Adult↗

PubMed2001

Influence of bone environment on ceramic osteointegration in spinal fusion: comparison of bone-poor and bone-rich sites.

Quantitative experimental data showed differences in bone quality and ceramic incorporation between bone-rich and bone-poor implantation sites. Bone in-growth was significantly lower for ceramic implanted at a lumbar intertransverse than a laminar site. Bone-marrow enrichment of the lumbar intertransverse site (regarded as bone-poor) greatly facilitated ceramic osteointegration. The vertebral interbody site, despite theoretical richness in osteogenic precursor cells, might be bone-poor at the time of grafting as compared to the reference iliac crest site. These data have important clinical implications concerning the potential benefit of enriching both bone-poor and bone-rich sites.

Bone Marrow Transplantation↗

PubMed1996

Bone marrow capacity for bone cells and trabecular bone turnover in immobilized tibia after sciatic neurectomy in mice.

Trabecular bone turnover and bone marrow capacity for the development of bone cells in the tibia were assessed after sciatic neurectomy (NX) in mice. The right hindlimbs of 6-week-old DDY mice were neurectomized and left hindlimbs were sham-operated and served as NX controls. Histomorphometrical analyses of the trabecular bone of the proximal tibia demonstrated the initial decrease in bone formation rate for the first 14 days and the subsequent increase in osteoclast surface for the next 14 days. The number of adherent stromal cells per tibia obtained for the NX limbs was reduced on days 7 and 10 postsurgically, and then recovered on day 12. However, the alkaline phosphatase activity of the cells was persistently depressed. The formation of osteoclast-like multinucleated cells in the marrow cultures obtained from NX limbs at days 10, 12, and 14 showed a significant increase in the medium containing parathyroid hormone (PTH). The number of colonies cultured for colony forming units-fibroblastic (CFU-f) that developed from the marrow cells did not differ in the NX and the contralateral limbs at any time during the period. On the other hand, the number of colonies cultured of colony forming units for granulocytes and macrophages (CFU-GM) was markedly increased for both the NX and the contralateral tibiae at days 12 and 14. This study clearly demonstrates that there are two stages in the development of osteopenia after NX. During the first 14 days, trabecular bone formation and number of marrow stromal cells are reduced. In the second 14 day period, the trabecular osteoclast number is increased and osteoclast formation from the bone marrow cells is enhanced in the presence of PTH. However, neither the CFU-f nor the CFU-GM assay could identify the changes in osteogenic or osteoclastogenic potential of the bone marrow. These in vitro assays provide limited information on the shifts in bone marrow cell lineages and the local environment producing osteopenia in the immobilized limb in vivo.

Alkaline Phosphatase↗

PubMed2006

Bone regeneration after enucleation of mandibular cysts: comparing autogenous grafts from tissue-engineered bone and iliac bone.

OBJECTIVE: The aim of this study was to compare bone regeneration after grafting enucleated mandibular cyst cavities using either autogenous osteoblasts cultured on a biomaterial or autogenous spongiose iliac bone. STUDY DESIGN: Twenty patients with 22 mandibular cysts were assessed. Eleven cysts were filled in with tissue-engineered bone (autogenous osteblasts cultured on demineralized bone matrix Osteovit) and 11 with spongiose iliac bone as controls. Panoramic radiographs were taken preoperatively, immediately postoperatively, and 3, 6, and 12 months after surgery. Radiolucency was computer analyzed using gray-level histograms. RESULTS: In both groups bone regeneration took place in a similar fashion. After 3 and 6 months there were few differences in bone density between the groups. However, in radiographic controls after 12 months ossification was considerably stronger in cysts grafted with tissue-engineered bone. CONCLUSION: These results advocate for the clinical application of tissue-engineered bone as an alternative viable filling material for cysts.

Adolescent↗

PubMed1999

Osteogenic effect of a gastric pentadecapeptide, BPC-157, on the healing of segmental bone defect in rabbits: a comparison with bone marrow and autologous cortical bone implantation.

Gastrectomy often results in increased likelihood of osteoporosis, metabolic aberration, and risk of fracture, and there is a need for a gastric peptide with osteogenic activity. A novel stomach pentadecapeptide, BPC-157, improves wound and fracture healing in rats in addition to having an angiogenic effect. Therefore, in the present study, using a segmental osteoperiosteal bone defect (0.8 cm, in the middle of the left radius) that remained incompletely healed in all control rabbits for 6 weeks (assessed in 2 week intervals), pentadecapeptide BPC-157 was further studied (either percutaneously given locally [10 microg/kg body weight] into the bone defect, or applied intramuscularly [intermittently, at postoperative days 7, 9, 14, and 16 at 10 microg/kg body weight] or continuously [once per day, postoperative days 7-21 at 10 microg or 10 ng/kg body weight]). For comparison, rabbits percutaneously received locally autologous bone marrow (2 mL, postoperative day 7). As standard treatment, immediately after its formation, the bone defect was filled with an autologous cortical graft. Saline-treated (2 mL intramuscularly [i.m.] and 2 mL locally into the bone defect), injured animals were used as controls. Pentadecapeptide BPC-157 significantly improved the healing of segmental bone defects. For instance, upon radiographic assessment, the callus surface, microphotodensitometry, quantitative histomorphometry (10 microg/kg body weight i.m. for 14 days), or quantitative histomorphometry (10 ng/kg body weight i.m. for 14 days) the effect of pentadecapeptide BPC-157 was shown to correspond to improvement after local application of bone marrow or autologous cortical graft. Moreover, a comparison of the number of animals with unhealed defects (all controls) or healed defects (complete bony continuity across the defect site) showed that besides pentadecapeptide intramuscular application for 14 days (i.e., local application of bone marrow or autologous cortical graft), also following other pentadecapeptide BPC-157 regimens (local application, or intermittent intramuscular administration), the number of animals with healed defect was increased. Hopefully, in the light of the suggested stomach significance for bone homeostasis, the possible relevance of this pentadecapeptide BPC-157 effect (local or intramuscular effectiveness, lack of unwanted effects) could be a basis for methods of choice in the future management of healing impairment in humans, and requires further investigation.

Animals↗

PubMed1992

Sex differences in cancellous and cortical bone strength, bone mineral content and bone density.

The purpose of this study was to examine sex differences in cancellous and cortical bone strength, bone mineral content (BMC) and bone density of excised cadaver vertebral and phalangeal bones. The samples were age-matched. Bone strength was measured as the mechanical force required to crush or break the bones. Two parameters of bone strength were used on the vertebrae; the force at the first deviation from linearity and the mean force during the consolidation before final failure. The force at first deviation from linearity was not significantly different between the sexes, but there was a significant difference in the consolidation force. The mean men's phalangeal strength was twice that of the women's. BMC and BMC/BW of both types of bone were statistically different between the sexes. Radiographic photodensity measures on the vertebrae showed no sex differences. Cortical diameters of the finger bones were significantly greater in males.

Aged↗

PubMed1984

Effect of propranolol treatment on bone mass, bone mineral content, bone remodelling, parathyroid function and vitamin D metabolism in hyperthyroidism.

The effect of propranolol 160-640 mg/day for 3 months on the accelerated loss of bone matrix and mineral in hyperthyroidism was studied in seventeen patients. A rise in serum thyroxine (P less than 0.01) during the first 3 weeks was followed by a fall (P less than 0.02). Serum triiodothyronine declined during the study (P less than 0.02). The enhanced bone mineral mobilization and collagen turnover continued during treatment and the bone mineral content decreased 3.2% (P less than 0.01). The secondary adaptive changes in serum parathyroid hormone and vitamin-D metabolites and in renal phosphate handling stayed unchanged. Iliac crest bone biopsies after tetracycline double-labelling showed initially a high bone turnover (P less than 0.01) with a reduced amount of cortical and trabecular bone (P less than 0.05). Following treatment bone formation rate decreased at both cellular and tissue level (P less than 0.01). No significant changes were observed in the amount of cortical and trabecular bone. The investigation shows that propranolol, in contrast to antithyroid medication, lacks any curative effect on the accelerated bone loss in hyperthyroidism.

Adult↗

PubMed1994

[Bone mineral density of lumbar spine and its relations to biological and lifestyle factors in middle-aged and aged Japanese women (Part 2). Effects of age and menopause on bone mineral density evaluated by biochemical markers of bone metabolism].

Bone mineral density of the lumbar spine (BMD) and biochemical markers for bone turnover were examined to study the mechanisms of age-related and menopause-related bone loss. We measured BMD of the lumbar spine and serum bone alkaline phosphatase (B-AIP) and bone gla-protein (BGP) as markers of bone formation and fasting urinary creatinine-adjusted hydroxyproline (Hyp/Cr) and calcium (Ca/Cr) as those of bone resorption in 166 community-dwelling Japanese women. A highly significant positive correlation between age and each of the biochemical markers, except for Ca/Cr, was observed. This relationship was not linear. Marked elevation in the levels of the markers was found in women in their sixth decade women compared with those in their fifth. All the markers correlated inversely with the BMD and these relationships remained significant after elimination of the effect of age by partialization. When analyzing the subjects in each five-year age group, the positive correlation of Hyp/Cr with Ca/Cr was significant in the subjects aged 45 to 49 and the negative correlation of Hyp/Cr with BMD was significant in those aged 50 to 54. B-AIP correlated positively with BGP in the subjects aged between 50 and 54 and inversely with BMD in those aged between 55 and 59. These correlations were significant. Thus, intercorrelations between the markers were observed five years earlier than were correlations between the markers and BMD. Such associations appeared earlier in terms of the markers for bone resorption than in terms of the markers for bone formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1997

Biochemical bone markers, bone mineral content, and bone mineral density in rats with experimental nephrotic syndrome.

The human nephrotic syndrome (NS) is accompanied by important alterations of mineral and bone metabolism. The purpose of the present study was to examine bone metabolism in rats with experimental NS and normal creatinine clearance, and to evaluate the reversibility of this alteration. NS was induced by three injections of puromycin aminonucleoside (PAN) on days 0, 21, and 35 (10, 5, and 5 mg/100 g body weight, respectively). The biochemical markers of bone formation (osteocalcin and alkaline phosphatase) and bone resorption (hydroxyproline and pyridinoline), bone mineral content (BMC), and bone mineral density (BMD), determined by dual-energy x-ray absorptiometry (DEXA), were studied on days 0, 7, 14, 28, 42, 56, 84, and 112. Proteinuria was present throughout the study. Hypoproteinemia was seen on days 7, 28, 42, and 56, returning to control values on days 84 and 112. In serum, osteocalcin (OC) concentration increased (p < 0.001), and alkaline phosphatase (ALP) decreased (p = 0.002). In urine, hydroxyproline increased (p < 0.001), but urinary pyridinoline was not different from the control group throughout the study. Increased serum parathyroid hormone concentration and decreased levels of 25-hydroxy and 1,25-dihydroxyvitamin D were found from day 7. During the intense proteinuria, bone resorption predominates and decreased BMC and BMD ensues in PAN-nephrotic rats. PAN-nephrotic rats showed low BMC and BMD compared to control group (p < 0.001). At the end of the study, when proteinuria persisted but total serum protein returned to control values, the biochemical bone markers, BMC, and BMD returned to normal. In conclusion, PAN-nephrotic rats had reversible bone alterations that were related to the magnitude of proteinuria and the concentration of total serum protein.

Alkaline Phosphatase↗

PubMed1997

[A biomechanical study on the bone repairing ability of recombinant human bone morphogenetic protein-2-coral composited artificial bone].

To improve bone repairing ability of coral, recombinant human bone morphogenetic protein-2 (rhBMP-2)-coral composited artificial bone was obtained by combining coral with rhBMP-2 in this experiment. The composite was implanted into rabbit calvarial critical-size defects and coral alone was implanted as control. The bone repairing ability was assessed by radiography, histology and biomechanics. The results showed that implants were absorbed gradually after they was implanted into the defects. In the meantime, the new bone was formed and the mechanical strength was reinforced uninterruptedly in bone defect regions. At 12 weeks, the implants were replaced completely by bone, and the mechanical strength was entirely restored. These suggest that the composite possess a superior bone repairing ability and may be one of the most perfect bone graft substitutes currently available.

Animals↗

PubMed1987

New bone formation and bone marrow differentiation induced in rats by extracellular bone matrix implantation: effect of local preirradiation on the process.

Subcutaneous implantation of demineralized diaphyseal rat bone matrix in ACI rats initiates a developmental cascade that results in the formation of new endochondral bone and an associated hematopoietic bone marrow differentiation. Irradiation (1500 rad, 60Co) of the implantation site 24 h prior to implantation suppresses the formation of endochondral bone and bone marrow. All phases of the developmental cascade, including chemotaxis, proliferation, and differentiation, are arrested by the irradiation. Simultaneous implantation with the extracellular matrix of bone marrow, bone, pieces of a four-day-old implant or of thoracic muscle--but not of brain, liver, or spleen tissue--results in the development of endochondral bone and bone marrow at the irradiated site. Concurrent implantation with the extracellular matrix of in vitro growing fibroblasts of marrow or ossicle origin does not restore the developmental cascade.

Alkaline Phosphatase↗

PubMed1982

Bone scintigraphy in evaluating the viability of composite bone grafts revascularized by microvascular anastomoses, conventional autogenous bone grafts, and free non-revascularized periosteal grafts.

We studied the value of bone scintigraphy in the assessment of anastomotic patency and bone-cell viability in free bone grafts revascularized by microvascular anastomoses in twenty-seven dogs. The dogs were divided into three different groups, and scintigraphy was carried out using technetium-labeled methylene diphosphonate in composite bone grafts revascularized by microvascular anastomoses, conventional autogenous bone grafts, and periosteal grafts placed in different recipient beds. The viability of the grafts was evaluated by histological examination and fluorescence microscopy after triple labeling with oxytetracycline on the first postoperative day, alizarin complexone on the fourth postoperative day, and DCAF on the eleventh postoperative day. A positive scintiscan within the first week following surgery indicated patent microvascular anastomoses, and histological study and fluorescence microscopy confirmed that bone throughout the graft was viable. A positive scintiscan one week after surgery or later does not necessarily indicate microvascular patency or bone-cell survival, because new bone formed by creeping substitution on the surface of a dead bone graft can result in this finding.

Animals↗

PubMed2006

Enhanced bone regeneration at a segmental bone defect by controlled release of bone morphogenetic protein-2 from a biodegradable hydrogel.

The objective of this study is to investigate the feasibility of a biodegradable hydrogel of gelatin as the controlled release carrier of bone morphogenetic protein-2 (BMP-2) suitable for enhancement of bone regeneration at a segmental bone defect. Hydrogels with three different water contents were prepared through glutaraldehyde crosslinking of gelatin with an isoelectric point of 9.0 under varied reaction conditions. Segmental critical-sized defects (20 mm) were created at the ulnar bone of skeletally mature New Zealand white rabbits, and gelatin hydrogels incorporating BMP-2 (17 microg/hydrogel) were implanted into the defects. When bone regeneration was evaluated by soft x-ray observation and bone mineral density (BMD) measurement, the gelatin hydrogels incorporating BMP- 2 exhibited significantly high osteoinduction activity compared with that of free BMP-2, although the activity depended on the water content of the hydrogels. Significantly higher BMD enhancement was observed in the gelatin hydrogel with a water content of 97.8 wt% than that with the lower or higher water content. We concluded that the biodegradable gelatin hydrogel is a promising controlled release carrier of BMP-2 for bone regeneration at the segmental bone defect.

Absorbable Implants↗

PubMed1992

Holding power of metallic screws in cancellous bone and metallic plates in cortical bone. An experimental study on rabbits using polymethylmethacrylate and bone graft as supporting material.

The holding power of metallic screws in cancellous bone and metallic plates in cortical bone was tested in sixty rabbits (n = 60). These metallic devices were fixed either without support or with the support of plastic material or autogenous bone graft. The fixation firmness was studied radiographically and by a test of the bonding strength. After six weeks, the mean value of the holding power of screws in cancellous bone without support was 440 N, and that of screws supported by polymethylmethacrylate 674 N and by bone graft 540 N. Corresponding values after twelve weeks were 436 N, 664 N, and 616 N. Fixation with polymethylmethacrylate was better than either fixation without support (P < 0.001, t-test) or fixation with bone graft (P < 0.01), whereas fixation with bone graft was better than fixation without support (P < 0.07). The best fixation was achieved by polymethylmethacrylate according to group mean values. Our investigation of the holding power of plates in cortical bone did not reveal any statistically significant differences between the methods used.

Animals↗

PubMed2002

Stress and strain distribution behavior in the bone due to the effect of cancellous bone, dental implant material and the bone height.

The stress and strain distribution in the bone surrounding a dental implant have been analyzed using the finite element and optimization techniques. The effect of removing cancellous bone completely or not was investigated. Two models were used, the first model without cancellous bone and the second with it. The elastic modulus of the implant material and the length of the implant neck or the height of bone surrounding the implant were used as design variables in the two models. In the first model a higher level of stress in the cortical bone surrounding the neck of the implant was found. While in the second model, it was found surrounding the tip of the implant. The result indicates that the stress concentration factor in the bone of the first model is reduced by 30% compared to the initial design. However, when the implant was surrounded by sleeve of cancellous bone (second model) the stress concentration is reduced by 16% for cortical bone and 15% for cancellous bone. This reduction help to reduce fatigue failure and bone resorption.

Coated Materials, Biocompatible↗

PubMed1992

Bone blood flow and in vitro proliferation of bone marrow and trabecular bone osteoblast-like cells in ovariectomized rats.

Ovariectomy in the rat induces a rapid osteopenia associated with an elevated bone turnover. One hundred and twenty-day-old rats were ovariectomized (OVX) or sham-operated (n = 6-8 per group and per time period studied). 45Ca accretion rate and bone blood flow (microspheres trapping technique) in the femurs were determined at 28, 42, 84, and 119 days after ovariectomy. Both parameters were markedly increased by 84 days and subsided thereafter. At the 42nd day, when bone turnover was maximal, bone marrow and trabecular bone cultures were obtained from sham-operated and ovariectomized animals (n = 10/group). Proliferation rate of bone marrow cells and trabecular osteoblast-like cells estimated by fibroblast colony-forming units (FCFU) efficiency and cell counting was markedly increased in primary and secondary cultures in ovariectomy. These data fitted well with the enhanced number of osteoblasts observed in situ in the long bone metaphyses of estrogen-depleted animals. As estrogens were shown in the literature to inhibit proliferation of the red cell line and of other hemopoietic lines, it is possible that estrogens, through a general mechanism, inhibit hemopoietic and stromal lines and also the proliferation of bone marrow-derived trabecular bone cells.

Animals↗