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PubMed1988

Failure of bone induction by bone matrix in adult monkeys.

Extraskeletal bone formation can be induced in rodents by implantation of demineralised bone matrix and such implantation has been used to treat bone defects in man, but it is uncertain if induction or merely conduction occurs. We studied bone induction in primates by excising segments of the fibulae of adult squirrel monkeys, defatting and demineralising them before reimplanting them into the quadriceps of the same animal. As a control experiment, rat matrix was prepared in exactly the same way and implanted in rats. After six weeks the implants were harvested and either ashed and analysed for calcium content or prepared for histology. In the rats, the calcium content indicated that about 20% of the original matrix had been replaced by new bone. In the monkeys the calcium content was about the same as that in normal body fluid and no bone was seen in histological sections. This result casts doubt on the use of demineralised human bone matrix as a bone inductor, although it may function by other mechanisms.

Animals↗
PubMed1973

Studies on bone matrix glycoproteins. Incorporation of (1-14C)glucosamine and plasma (14C)glycoprotein into rabbit cortical bone.

The radioactively labelled constituents present in bone matrix were compared 12 days after injection of either [(14)C]glucosamine or plasma [(14)C]glycoprotein. Both precursors are utilized in the synthesis of organic matrix by bone tissue. Cortical bone from animals injected with [(14)C]glucosamine contains radioactivity derived from glucosamine and plasma glycoproteins and all glycoprotein fractions are labelled. Plasma [(14)C]glycoprotein labels the less acidic glycoproteins to a greater extent than the more acidic components. An antibody has been raised against the less-acidic-glycoprotein fraction of bone. The latter contains a glycoprotein of alpha-mobility that appears to be concentrated specifically in bone tissue and which is present also in plasma. This alpha-glycoprotein accounts for a large proportion of the components labelled and retained in bone matrix after [(14)C]glucosamine injection.

Albumins↗
PubMed2005

Evaluation of demineralized bone matrix paste and putty in periodontal intraosseous defects.

BACKGROUND: Demineralized bone matrix (DBX) paste and putty are particulate demineralized bone matrices in a 2% or 4% hyaluronate carrier, respectively. The purpose of this study was to determine the effectiveness of DBX paste and putty compared to demineralized freeze-dried bone allograft (DFDBA) in the treatment of human intraosseous periodontal defects. METHODS: Sixty systemically healthy individuals between the ages of 31 and 71 years with at least one intraosseous periodontal defect of > or = 3 mm in depth and radiographic evidence of at least 40% to 50% vertical bone loss were accrued. Following initial non-surgical periodontal therapy, sites were randomly selected to receive either DBX paste, DBX putty, or DFDBA (control). Baseline and 6-month reentry soft and hard tissue parameter measurements were made by calibrated examiners. Data were analyzed within and between groups utilizing analysis of variance (ANOVA) and paired and unpaired Student t tests. RESULTS: Probing depth reductions were significantly improved in all treatment groups with DFDBA, DBX paste, and putty patients demonstrating 2.8 mm, 3.6 mm, and 2.3 mm, respectively. Attachment level gains were significantly improved from baseline for all treatment groups with DFDBA, DBX paste, and putty, respectively, demonstrating 2.4 mm, 2.9 mm, and 1.6 mm. Bone fill was similar between all groups with DBX paste, putty, and DFDBA control groups demonstrating 2.0 mm, 2.4 mm, and 2.2 mm, respectively. All groups yielded significant improvements in percent bone fill with DFDBA, DBX paste and putty, respectively, achieving 37%, 42.1%, and 50% with no significant differences between the groups. CONCLUSION: In summary, demineralized bone matrix paste, demineralized bone matrix putty, and demineralized freeze-dried bone allograft all demonstrated similar favorable improvements in soft and hard tissue parameters in the treatment of human intraosseous defects.

Adult↗
PubMed1987

Effects of acetylsalicylic acid on intramuscular bone matrix implants and composite grafts in rats.

Antigen-extracted, autolyzed, demineralized bone matrix and bone matrix combined with isogeneic bone marrow (i.e., a composite graft) were placed in the musculature of young male rats. The influence of acetylsalicylic acid (ASA) on the osteogenic potential of these bone-inducing materials was studied during an 18-day period. ASA (150 mg/kg/12 h) was used to obtain anti-inflammatory serum levels comparable with those in humans. Resorption of the implanted matrix and new formation of bone were quantified using collagen- and mineral-tracing radioisotopes and collagen and calcium analyses. The matrix implants showed a slow tissue turnover and produced only small amounts of new bone, with no significant differences between ASA-treated and control rats. However, during the last 24 h of the study the ASA-treated implants had a significantly hampered rate of new bone formation. The tissue turnover and new bone yield, as well as the rate of new bone formation, increased in the control composite grafts but not in the ASA-treated composite grafts. Thus, the results indicate an inhibitory effect of ASA on the bone-forming capacity of bone marrow.

Animals↗
PubMed2001

Effect of gas-plasma sterilization on the osteoinductive capacity of demineralized bone matrix.

The current study evaluated the effect of low-temperature hydrogen peroxide gas plasma sterilization on the osteoinductive capability of human demineralized bone matrix using a rat model. Twelve athymic rats received three separate implants consisting of steam-sterilized demineralized bone matrix (negative control), sterile-harvest demineralized bone matrix (positive control), and gas-plasma-sterilized demineralized bone matrix. A demineralized bone matrix pellet from each sterilization group was placed individually into one of three separate soft tissue pockets created in the epaxial musculature of each rat. All 12 rats were euthanized 9 weeks after implantation. Each implantation site was removed along with 0.5-cm normal tissue around the implant. Histologic examination was done on each implant site to determine the presence or absence of new bone, cartilage, or bone marrow elements. All 12 sterile harvest demineralized bone matrix sites histologically contained new bone elements, whereas none of the negative control or gas plasma sterilized demineralized bone matrix sites contained any of these same elements. The results of this study indicate that demineralized bone matrix sterilized with low-temperature, gas-plasma sterilization loses its osteoinductive capacity in a manner similar to that of steam-sterilized demineralized bone matrix, making low-temperature, gas- plasma sterilization unsuitable as a method of secondary sterilization of demineralized bone matrix.

Animals↗
PubMed1982

Bone formation of transplanted autologous bone matrix in rabbit evaluated by technetium radionuclide bone imaging.

In adult rabbits a 12 mm piece of the radius was bilaterally resected and subsequently decalcified. One piece was placed in one of the radial defects and the other in a skin tube on the back. The other radial defect served as a control. The bone formation process was evaluated by roentgenographic examination and quantified radionuclide bone imaging by technetium diphosphonate scintigraphy at 3 and 6 months postoperatively. Reliability of the scintigraphy method was assessed by means of testing interexamination variability where the same sample was measured twice and where no significant difference was found. In the defect restituted with decalcified bone matrix there was a significantly (p less than 0.01) higher bone formation rate at 3 months postoperatively, with a mean increase of 47% compared to the control side. In the skin tube the implanted matrix showed mineral formation both at 3 and 6 months postoperatively and increased radionuclide uptake compared to the adjacent tissues. Technitium radionuclide imaging was found to be a suitable method for evaluation of the bone formation in these small defects.

Animals↗
PubMed1974

Cyclic electrochemical inactivation and restoration of competence of bone matrix to transform fibroblasts.

Brief exposure of rat bone matrix to highly charged electrochemicals had profound but reversible effects on its competence to transform fibroblasts into chondroblasts and osteoblasts. Suppression and subsequent reactivation of this function were influenced critically by the charge of the electrochemical reagent and the pH of the reaction mixture. In dilute acids, Evans Blue, a long electronegative molecule, suppressed transforming competence, whereas hexadimethrine, a polycationic quaternary ammonium base, failed to do so. A cycle of inactivation-restoration of transforming competence was achieved by sequential treatment of bone matrix with Evans Blue followed by hexadimethrine; an electrochemical complex was demonstrated by histochemistry. In dilute alkalies, hexadimethrine inhibited, whereas, Evans Blue did not suppress the transforming function of the bone matrix. Electric charge characteristics on the surface of the bone matrix are of crucial significance for the fibroblast-transforming attribute.

Alkaline Phosphatase↗
PubMed1994

Effects of pollen from Typha angustata on the osteoinductive potential of demineralized bone matrix in rat calvarial defects.

Typha angustata Bony et Chaub. is a traditional Chinese medicine, commonly used in China for a variety of clinical disorders, including atherosclerosis, cardiovascular diseases, uterus contraction, and wound healing. The effect of the pollen of Typha angustata on the bone inductive capacity of demineralized bone matrix is studied here. Demineralized bone matrix soaked with saline solution was implanted in 8-mm defects in rat calvaria. After surgery all rats received a 0.2-ml injection in the defect sites of Typha angustata extract, plasma, or saline 3 times weekly for 2 to 4 weeks. The repair of bone defects was evaluated by radiography and by histology at 2 and 4 weeks after surgery. Results indicated that the 3% Typha angustata extract and demineralized bone matrix combination produced substantially more bone than demineralized bone matrix alone, while plasma plus demineralized bone matrix induced the same amount of bone formation as Typha angustata extract plus demineralized bone matrix. The osteoinductive potential increased in a dose dependent manner, 3% Typha angustata extract plus demineralized bone matrix produced more bone than the 0.6% Typha angustata extract plus demineralized bone matrix at 2 and 4 weeks. This study demonstrates that an extract of the pollen of Typha angustata is capable of enhancing the osteoinductive potential of demineralized bone matrix.

Animals↗
PubMedDate not supplied

Demineralized bone matrix graft: a scientific and clinical case study assessment.

Osteoinductive demineralized bone matrix results from bone demineralization and is attributed to matrix-associated bone morphogenetic proteins. The osteoinductive potential can vary with donor. Many bioassay methods are available to screen donors, each with its own interpretation, so performance of more than one may be of value. Furthermore, little is known about the relationship between bioassay results and clinical outcomes. A study designed to meaningfully explore these issues would require assay of a large number of donors as well as clinical utilization in a large patient population. A preliminary study was undertaken to gain initial perspective. Using demineralized bone matrix derived from one 33-year-old female donor, 2 methods of bioassay and a clinical case study were performed. The levels of bone morphogenetic proteins 2, 4, and 7 in lyophilized demineralized bone matrix powder were measured (19.65 +/- 0.30 ng/g, 2.49 +/- 0.19 ng/g, and 82.03 +/- 6.89 ng/g, respectively). Also, putty (Osteostim DBM Putty), prepared from powder, was intramuscularly implanted in athymic rats and de novo bone formation quantified (6.7% +/- 3.5% new bone formation with 49% +/- 17% of the implant area associated with new bone formation). The putty, in conjunction with internal fixation, was used in the revision of a medial malleolar nonunion of an obese, 76-year-old woman. Radiographic union with excellent graft incorporation was achieved by 12 weeks postoperatively, with maintenance of an acceptable clinical result during the 14-month follow-up period. These results are interpreted in the broader context of demineralized bone grafting, in general, and an outline for further study is presented.

Adult↗
PubMed2006

Demineralized bone matrix and hydroxyapatite/tri-calcium phosphate mixture for bone healing in rats.

PURPOSE: Hydroxyapatite/tri-calcium phosphate (HA/TCP) mixture is an osteoconductive material used as a bone graft substitute, and demineralised bone matrix (DBM) is an osteoinductive material. A combination of DBM and HA/TCP mixture would probably create a composite with both osteoconductive and osteoinductive properties. The purpose of this study was to determine the effect of the combination of DBM and HA/TCP mixture on healing of rat radius segmental defects. METHODS: Twenty-four adult male Wistar rats were used. Bilateral radial defects were created in each animal. Radial defects were implanted with DBM, HA/TCP mixture and a combination of both substances. Control defects were left unfilled. Ten weeks after implantation, the animals were sacrificed, and the radii were evaluated by radiograhic and histopathological studies. RESULTS: The use of DBM alone demonstrated improved healing on radiographic and histological studies compared to other groups and the control group. There were no differences between the other two groups and the control group. CONCLUSION: The DBM group showed the best healing response. Combined use of DBM and HA/TCP mixture did not improve bone healing, and the osteoinductive properties of DBM were inhibited by HA/TCP mixture.

Animals↗
PubMed2005

Comparative efficacy of 2 different demineralized bone matrix allografts in treating long-bone nonunions in heavy tobacco smokers.

Smoking impairs bone healing and increases the risk for complications associated with nonunions. The efficacies of 2 different allografts, Grafton (demineralized bone matrix [DBM] in a gel-like glycerol carrier) and Orthoblast (DBM with a reverse thermal poloxamer carrier) were examined with respect to nonunions in patients who reported heavy tobacco use. The Grafton allograft was used in 25 patients, and the Orthoblast allograft was used in 13 patients. All patients smoked more than half a pack of cigarettes a day and did not use electric stimulators. A successful graft was defined as healing on the first graft attempt without complications or later regraft. The Grafton and Orthoblast success rates were 52% and 85%, respectively (P = .077). The unique thermal properties of the Orthoblast reverse poloxamer, which may enhance DBM osteoinduction, may account for the difference in success rates. Although results failed to reach statistical significance, the large difference and high likelihood ratio (4.2) between the 2 groups suggest that perhaps not all commercially available allografts may necessarily perform with the same efficacy with respect to heavy smokers.

Adult↗
PubMed1982

The use of decalcified granulated homologous cortical bone matrix in the correction of diaphyseal bone defect. An experimental study in rabbits.

In order to investigate an alternative way to correct diaphysial bone defects, granulated decalcified homologous cortical bone was used as a graft. Because of the suitable anatomic arrangement, the forearm of rabbits was chosen as an experimental model of bone defect. A 2 cm long bone cylinder was removed from the diaphysis of both radii preserving the periosteum. The artificial bone defect was filled with granular decalcified bone on the right side. The left side was used as control and kept empty or filled with undecalcified granular bone. The 18 animals were sacrificed in batches, 3,6, and 9 weeks after the operation. New bone formation was followed by X-rays, routine histology and incorporation of calcein blue, xylenol orange and tetracycline. In the decalcified granular bone grafts new bone formation was already detected at the first week and 9 weeks after the graft operation there was a well developed cylindric ossicle, in 89% of the cases. In some cases a medullary canal was present. No bone formation was found neither in the empty defects nor in the ones filled with undecalcified granular bone grafts.

Animals↗
PubMedDate not supplied

[The stimulation of reparative regeneration with demineralized bone matrix in puncture osteoplasty operations for bone cysts in children].

The authors present the data on positive influence of demineralized osteomatrix (DOM) on the osteogenesis in the experimental conditions. On the basis of data received the method of puncture osteoplastic operation with DOM application was developed. Puncture method was applied in 38 children. In 6 months 31 children cyst were liquidated, osteoarchitectonics was reduced, in 7 children rough trabecular picture remains.

Animals↗
PubMed1993

Characterization of radioiodinated recombinant human TGF-beta 1 binding to bone matrix within rabbit skull defects.

Bone healing is regulated in part by the local production of TGF-beta 1 and other growth factors produced by cells at the site of injury. The single application of recombinant human TGF-beta 1 (rhTGF-beta 1) to calvarial defects in rabbits induces an accelerated recruitment and proliferation of osteoblasts within 3 days. This ultimately results in the formation of new bone and the complete closure of the defect within 28 days. The persistence and localization of [125I]rhTGF-beta 1 within an osseous defect was investigated after applying a single dose of [125I]rhTGF-beta 1 formulated in a 3% methylcellulose vehicle. Normal bone encompassing the defect site, the periosteum, and the gel film covering the dura were harvested at 0, 4, 8, and 24 h and 3, 7, and 16 days after [125I]rhTGF-beta 1 application. The defect site-associated radioactivity was quantitated, visualized by autoradiography, and characterized by TCA precipitation and SDS-PAGE. Radioactivity was observed in autoradiographs of gross specimens, histologic sections of the bone matrix, and periosteal tissue surrounding the defect. There was a time-dependent decrease in TCA-precipitable radioactivity; however, radioactivity was still associated with the bone matrix 16 days after application of [125I]rhTGF-beta 1. SDS-PAGE and autoradiography of the radioactivity in homogenized bone and periosteal samples revealed a 25 kD band, suggesting that the radioactivity remaining at the defect site represented intact [125I]rhTGF-beta 1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1997

Bone matrix degradation by the plasminogen activation system. Possible mechanism of bone destruction in arthritis.

The observed increase in urokinase-type plasminogen activator (u-PA) and its receptor (u-PAR) in synovial tissue of patients with rheumatoid arthritis (RA) suggests pathophysiological involvement of the plasminogen activation (PA) system in inflammatory joint disease. In the present study, we investigated the capacity of the PA system to degrade non-mineralized and mineralized bone-like matrix in vitro as a model for bone destruction. Transfected mouse LB6 cell lines, that expressed either human u-PA or u-PAR, were cultured separately and simultaneously on radiolabelled bone matrix in the presence of plasminogen. Osteoblast-like murine calvarial MC3T3-E1 cells were used to produce a well-characterized, highly organized bone-like matrix, that could be mineralized in the presence of beta-glycerol phosphate. Bone matrix degradation was followed by the release of radioactivity in the culture medium. u-PA-producing cells, in contrast to u-PAR-producing cells, degraded both non-mineralized and mineralized bone matrix. This effect could be inhibited by anti-u-PA antibodies, as well as by tranexamic acid and by aprotinin, indicating that the degrading activity is u-PA mediated and plasmin dependent. Co-cultivation of a small portion of u-PA-producing cells with u-PAR-expressing cells resulted in a marked increase in degradation activity. Reduction of this potentiating effect by suramin or the amino-terminal fragment of u-PA, both competitive inhibitors of u-PA receptor binding, shows that this synergistic effect is due to binding of u-PA to u-PAR. u-PAR must be cell associated, as binding of u-PA to a soluble u-PAR prevented this enhancement. The capability of the PA system to degrade bone matrix in vitro, and the previously demonstrated increased expression of u-PA and u-PAR in synovial tissue of patients with RA, further support a role for the PA system in the development of bone erosions.

3T3 Cells↗