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PubMed1989

Osteogenesis in bone defects in rats: the effects of hydroxyapatite and demineralized bone matrix.

Hydroxyapatite (HA) and Demineralized Bone Matrix (DBM) are being investigated as potential osteogenic agents with hopes that these substances can be used to induce bone formation in non-union fractures. This study was done to determine the relative effects of HA and DBM implanted as moldable phospholipid composites in bone defects that result in non-unions. We studied 22 ten-month-old Long-Evans male rats with 5.0 mm unilateral radial defects implanted with HA, DBM, and a combination of both substances. Control defects were left unfilled. Eight weeks after implantation, the histological sections demonstrated a decrease in bone formation with HA relative to controls. The HA crystals were encapsulated by connective tissue stroma made up of collagenous elements, fibroblasts, and blood vessels. There were no indications of bone formation within the fibrous stroma. 45Calcium, alkaline phosphatase, and bone gla protein (BGP) assays demonstrated a 16% increase in bone formation in rats implanted with DBM, an 80% decrease in groups implanted with HA (p = 0.01) and an 80% decrease with DBM plus HA (p = 0.01). Radiologic analysis corresponds well with histological and biochemical results. We conclude that osteogenesis in non-union defects is enhanced by the implantation of DBM, while HA interferes with bone formation in the rat model. In the presence of both substances, HA appears to impede new bone growth, negating any positive effects seen with DBM.

Alkaline Phosphatase↗
PubMed1989

The effects of demineralized bone matrix and direct current on an "in vivo" culture of bone marrow cells.

Bone marrow cells (BMCs) from rabbit femora and tibiae were grown in diffusion chambers implanted in rabbit muscle. At 42 days 80% of the BMC chambers exhibited cartilage formation within them. Demineralized bone matrix added to the marrow cell suspension in the chamber accelerated the appearance and increased the number of chambers with cartilage. Mineralization of the cartilage also occurred earlier in the chambers with bone matrix. In a second experiment, a 5-microA direct current cathode in the bone marrow chamber increased the number of chambers containing cartilage from 50 to 80% at day 25. Mineralization also occurred earlier in the chambers with direct current.

Animals↗
PubMed2005

Stimulation of fracture healing by continuous delivery of demineralized bone matrix proteins and tobramycin.

Demineralized bone matrix (DBM) is an allogenous, bioabsorbable material that has long been used for its osteoinductive and osteoconductive properties. A significant complication experienced by physicians who perform bone defect filling surgery is the risk of subsequent bacterial infections and the inefficiency of oral antibiotics to provide adequate prophylaxis against microorganisms, especially Staphylococcus aureus, Pseudomonas aeruginosa, and Staphylococcus epidermidis. In order to deliver both DBM and an efficient antiobiotic at high local concentrations without deleterious systemic effects, a ceramic sustained delivery system was implanted and monitored over the course of 30 days for bone regeneration, infection, and systemic effects. Twenty-five adult Sprague Dawley albino male rats were used in the experiment. They were randomly divided into five equal groups. Animals in group 1 were used as control, group 2 animals had a created 5mm defect, in group 3 defect induced and implanted with devices loaded with antibiotic (tobramycin) alone, group 4 had a created defect plus DBM, and group 5 had a created defect, plus tobramycin (which is effective against the aforementioned organisms) and DBM. At 30 days post-implantation, the experimental animals showed no significant difference in weight when compared to the control and sham animals. X-rays taken at this time showed the experimental femurs to be totally healed and virtually indistinguishable from control. Initial dissections revealed that the implants were accepted by the hosts as shown by the fibrous, vascularized sheath that surrounded the femurs and capsules. The implants were found to be in close contact with the cancellous bone and none of the sheaths showed signs of infection. Macroscopically, no defect could be detected in the experimental animals, while little regeneration was observed in the femurs of the sham animals.

Animals↗
PubMed1993

Reconstruction of the bone--bone marrow organ by osteogenin, a bone morphogenetic protein, and demineralized bone matrix in calvarial defects of adult primates.

Information concerning the efficacy of osteogenin, a bone morphogenetic protein, and demineralized bone matrix in orthotopic sites in nonhuman primates is a prerequisite for potential clinical application in humans. After exposure of the calvaria, 84 cranial defects, 25 mm in diameter, were prepared in 26 adult male baboons (Papio ursinus). Defects were implanted with insoluble collagenous bone matrix (ICBM, the inactive collagenous residue after dissociative extraction of bone matrix with 4 M guanidine hydrochloride) reconstituted with osteogenin fractions isolated from baboon bone matrix by chromatography on heparin-Sepharose and hydroxyapatite-Ultrogel (Og Hep-HA) or osteogenin further purified using Sephacryl S-200 gel filtration chromatography (Og S-200). Baboon osteogenin with the highest biologic activity in a rodent bioassay, as determined by alkaline phosphatase activity, calcium content, and histologic analysis, was used for orthotopic implantation in baboons. Additional defects were implanted with baboon demineralized bone matrix (DBM) or ICBM without osteogenin as control. Defects also were grafted with corticocancellous bone harvested from the iliac crest or left ungrafted to monitor the spontaneous regeneration potential of the adult baboon calvaria. Undecalcified bone sections at 7 microns were prepared from the harvested specimens 30 and 90 days after surgery. Histomorphometry demonstrated that Og S-200 induced copious amounts of bone and osteoid as early as day 30 (P < 0.01 versus ICBM, autogenous grafts and untreated defects). At day 90, in implants of Og S-200, Og Hep-HA, and DBM, bone and marrow formation was extensive, culminating in complete regeneration of the craniotomies. In implants of DBM, bone formed with an intervening phase of cartilage development. This provides the phenotypic evidence of endochondral bone differentiation by induction in defects of membranous calvarial bone in adult primates. These results establish the potential therapeutic application of osteogenin and demineralized bone matrix for the architectural reconstruction of the bone-bone marrow organ in humans.

Animals↗
PubMed1996

Integration of endochondral bone grafts in the presence of demineralized bone matrix.

The use of endochondral bone grafts (EC) and demineralized bone matrix (DBM), which contains a potent osteoinductive matrix, may promote the repair of nonregenerative defects. The purpose of the current work is to assess qualitatively and quantitatively the effect of DBM on the healing of EC bone grafts and to compare it to the healing of EC bone grafts alone. Twenty-four defects in rabbit skulls were filled with EC bone grafts alone, DBM alone, or combined EC and DBM. Histologic and immunohistologic changes were examined in 2 weeks. The amount of new bone formation was quantified by image analysis. Healing of all the groups was characterized by the presence of a cartilage intermediate stage. In the EC bone grafts alone, healing was localized to the host bone/graft interface. In the composite group, amalgamation of the new bone, DBM, and bone graft progressed throughout the whole width and depth of the defect, uniting the graft to the recipient bed. The amount of new bone formed was significantly greater (47%) in the composite group than the EC group. In conclusion, DBM powder augments the bone-induction capacity of the recipient bed as well as the bone graft. The composite EC bone grafts and added DBM possess properties required for an effective graft material and merit further clinical evaluation.

Animals↗
PubMed2006

Effect of xenogenic immobilized bone matrix on the course of wound process.

The effect of xenogenic demineralized bone matrix as a component of film coating on healing of a full-thickness skin wound was studied. Regenerative processes were more intensive in wounds dressed with a complex film containing demineralized bone matrix than in uncovered wounds and wounds covered with films without bone matrix. Film coating with antibiotics suppressed the development of wound microflora. In vitro experiments demonstrated different effects of demineralized bone matrix on some enzymes in rat skin and liver homogenates. Biological activity of xenogenic demineralized bone matrix as a component of wound coating is shown.

Animals↗
PubMed2004

Alteration of noncollagenous bone matrix proteins in distal renal tubular acidosis.

Our previous report on bone histomorphometry in patients with distal renal tubular acidosis (dRTA) revealed decreased bone formation rate (BFR) when compared to healthy subjects. The abnormality improved significantly after alkaline therapy. The modest increase in osteoblastic surface, after correction of metabolic acidosis, could not explain the striking improvement in bone formation, suggesting additional influence of metabolic acidosis on osteoblast function and/or bone matrix mineralization. Osteoblasts and, to a lesser extent, osteoclasts synthesize and secrete bone matrix including type I collagen and various noncollagenous proteins (NCPs). Substantial evidence suggested diverse functions of NCPs related to bone formation, resorption, and mineralization. Metabolic acidosis, through its effect on bone cells, may result in an alteration in the production of NCPs. Our study examined bone histomorphometry with detailed analysis on the mineralization parameters and NCPs expression within the bone matrix of patients with dRTA before and after treatment with alkaline. Seven dRTA patients underwent bone biopsy at their initial diagnosis and again 12 months after alkaline therapy. Bone mineral density (BMD) and bone histomorphometry were obtained at baseline and after the treatment. The expression of NCPs was examined by immunohistochemistry, quantitated by digital image analysis, and reported as a percentage of area of positive staining or mineralized trabecular bone area. Alkaline therapy normalized the low serum phosphate and PTH during acidosis. The reduction in BMD at baseline improved significantly by the treatment. Bone histomorphometry demonstrated the increase in osteoid surface and volume without significant alteration after acidosis correction. In comparison to the normal subjects, osteoid thickness was slightly but insignificantly elevated. Osteoblast and osteoclast populations and their activities were suppressed. The reduction in mineral apposition rate and adjusted apposition rate were observed in conjunction with the prolongation of mineralization lag time. Alkaline therapy improved the mineralization parameters considerably. In addition to the increase in BFR relative osteoblast number after acidosis correction, osteocalcin expression in the bone matrix increased significantly from 16.7% to 22.3%. Six of seven patients had decreased osteopontin expression. In conclusion, the abnormal bone remodeling in dRTA is characterized by low turnover bone disease with some degree of defective mineralization. Alteration of NCPs expression suggested the effect of metabolic acidosis on bone cells. Alkaline therapy increased bone mass through the restoration of bone mineral balance and, perhaps, improved osteoblast function.

Acidosis, Renal Tubular↗
PubMed2003

Evaluation of a cell-permeable barrier for guided tissue regeneration combined with demineralized bone matrix.

AIM: To evaluate whether bone formation by guided tissue regeneration (GTR) and demineralized bone matrix (DBM) can be enhanced by the use of a cell-permeable Teflon barrier allowing the penetration of undifferentiated mesenchymal cells from the surrounding soft tissues. MATERIAL AND METHODS: DBM was produced from the long bones of rats, and its bone-inductive properties were tested in three rats prior to the study by intramuscular implantation. Thirty, 4-month-old, male albino rats of the Wistar strain were used. Following surgical exposure of the mandibular ramus, a cell-permeable Teflon capsule, loosely packed with DBM, was placed with its opening facing the lateral surface of the ramus (test side). At the contralateral side, serving as control, a non-perforated (cell-occlusive) Teflon capsule, loosely packed with the same amount of DBM, was placed. After healing periods of 30, 60, and 120 days, groups of 10 animals were killed, and 40-70 microm thick undecalcified sections of the capsules were produced. RESULTS: Computer-assisted planimetric measurements on the histological sections disclosed similar amounts of newly formed bone in both test and control capsules. After 4 months, the new bone in the control capsules occupied 45.0% of the cross-sectional area of the capsule, while it was 50.5% in the test capsules. This difference was not statistically significant (P<0.05). CONCLUSION: Similar amounts of bone formed in cell-permeable and cell-occlusive capsules grafted with DBM, suggesting that invasion of undifferentiated mesenchymal cells from the surrounding soft tissues into the barrier-protected area is unnecessary for bone formation with GTR.

Animals↗
PubMed1985

Human bone matrix gelatin as a clinical alloimplant. A retrospective review of 160 cases.

Bone matrix gelatin, prepared by sequential chemical treatment including decalcification with 0.6 N hydrochloric acid [9], was used as an alloimplant for the treatment of benign bone tumours, tumorous conditions of bone, acetabular dysplasia, delayed union, traumatic bone defects and other disorders. The bone matrix gelatin implanted into bone defects was incorporated successfully in 98% of implantations, excluding cases of infection, tumour recurrence and recurrence of tumorous conditions. The material was also implanted into ten bone sites as an onlay but in five it was resorbed without new bone formation. The incorporation of the bone matrix gelatin into the recipient bed was completed from 6 to 33 months (average 14.9 months) after implantation. Wound infection complicated 5 of 165 implantations (3%) in previously uninfected sites. Low grade fever persisting after the tenth post-operative day (a probable sign of immunological reaction) occurred in 4 of 160 implantations (2.5%), excluding cases of infection. Alloimplants of bone matrix gelatin are thus effective in the treatment of bone defects. The risk of complication such as rejection or infection is low.

Adolescent↗
PubMedDate not supplied

Chemosterilized autolyzed antigen-extracted allogeneic (AAA) bone matrix gelatin for repair of defects from excision of benign bone tumors: a preliminary report.

In a series of 33 bone graft operations, antigen-extracted, autolyzed, allogeneic (AAA) bone matrix gelatin was substituted for autologous bone. The period of follow-up was 2.0 to 3.5 years. AAA bone gelatin was resorbed more rapidly than whole bone. AAA bone gelatin was replaced by new bone in the same intervals of time as observed with autologous bone. In treatment of bone tumors with AA bone gelatin, the results of the operation depend upon the nature of the pathologic processes in the host bed. In normal host bed, tight contact between implant and recipient bone is essential for success. The overall results of a preliminary study of 33 cases of 91% successful.

Antigens↗
PubMed1997

Regulation of bone matrix protein expression and induction of differentiation of human osteoblasts and human bone marrow stromal cells by bone morphogenetic protein-2.

We have examined the effects of BMP-2 on the expression of bone matrix proteins in both human bone marrow stromal cells (HBMSC) and human osteoblasts (HOB) and their proliferation and mineralization. Both HBMSC and HOB express BMP-2/-4 type I and type II receptors. Treatment of these two cell types with BMP-2 for 4 weeks in the presence of beta-glycerophosphate and ascorbic acid results in mineralization of their matrix. BMP-2 increases the mRNA level and activities of alkaline phosphatase and elevates the mRNA levels and protein synthesis of osteopontin, bone sialoprotein, osteocalcin, and alpha 1(I) collagen in both cell types. Whereas the mRNA level of decorin is increased, the mRNA concentration of biglycan is not altered by BMP-2. No effect on osteonectin is observed. The effect of BMP-2 on bone matrix protein expression is dose dependent from 25 to 100 ng/ml and is evident after 1-7 days treatment. In the presence of BMP-2, proliferation of HBMSC and HOB is decreased under either serum-free condition or in the presence of serum. Thus, BMP-2 has profound effects on the proliferation, expression of most of the bone matrix proteins and the mineralization of both relatively immature human bone marrow stromal preosteoblasts and mature human osteoblasts.

Adult↗
PubMed1987

[Effect of acid metabolites on the osteoinductive activity of the bone matrix].

The osteoinductive activity of bone matrix demineralized in acid metabolites of glycolysis and Krebs cycle have been studied in rats. The highest osteoinductive effect was reached during the demineralization in oxaloacetic acid, the lowest one--in the malic acid. The regulatory role of glycolysis metabolites and Krebs Cycle in posttraumatic osteogenesis is under discussion.

Animals↗
PubMed1984

The healing of segmental bone defects induced by demineralized bone matrix. A radiographic and biomechanical study.

UNLABELLED: We studied the effect of demineralized bone matrix on the repair of large femoral diaphyseal defects in a rat model by clinical, radiographic, and biomechanical methods. A standard procedure was first developed to create segmental defects that did not heal and in which non-union developed consistently. The effect of demineralized bone matrix on repair was then assessed by physical examination, serial radiographs, and biomechanical studies to determine deformation to failure, stiffness, torsional strength, and energy absorption. By twelve weeks, the defects that had been treated with demineralized bone matrix showed satisfactory repair and remodeling in most animals based on clinical and radiographic evaluation. The biomechanical studies demonstrated that the bone induced by demineralized bone matrix had an energy-absorption capacity and stiffness equal to those of intact rat femoral bone. The bone induced by demineralized bone matrix achieved 35 per cent of the torsional strength of normal bone and an increased capacity to deform under load. These biomechanical properties are similar to those observed in the early stages of normal fracture repair. CLINICAL RELEVANCE: An effective, readily available alternative to autologous bone-graft material would have a variety of clinical uses in orthopaedic surgery, such as augmenting fusions, aiding in the repair of high-risk fractures, and filling or bridging bone defects. Demineralized bone matrix may provide an important tool for these purposes by inducing bone that has the mechanical properties of fracture callus. This would reduce the morbidity associated with harvesting autologous bone and have an advantage over allografts or synthetic biomaterials that require incorporation by the host before they can support mechanical loads.

Animals↗
PubMed1997

Expression of bone matrix proteins in human breast cancer: potential roles in microcalcification formation and in the genesis of bone metastases.

The skeleton is the privileged target of metastatic human breast cancer cells. Bone metastases are indeed found in virtually all advanced breast cancer patients and generate major morbidity. The high osteotropism of breast cancer cells suggests that they exhibit a selective affinity for mineralized tissues. The observation that mammary malignant cells are able to induce hydroxyapatite crystals deposition within the primary tumour suggests that they can generate a microenvironment that favors the crystallization of calcium and phosphate ions into the bone specific hydroxyapatite. Osteonectin (OSN), osteopontin (OPN) and bone sialoprotein (BSP), 3 bone matrix proteins involved in bone matrix mineralization, are expressed in human breast cancers. BSP, an RGD (Arg-Gly-Asp) containing phosphoprotein, initiates hydroxyapatite deposition and mediates attachment of osteoclast to the same crystals prior to their resorption. Detection of BSP at both the protein and the mRNA levels in human breast cancer and in human breast cancer cell lines (MCF-7, T47-D and MDA-MB 231) indicates that mammary malignant cells synthesize directly BSP rather than uptaking it from the serum. Interestingly, the level of BSP expression correlates with the development of bone metastases and with poor survival. These data suggest that the ectopic expression of bone matrix proteins could be involved in conferring osteotropic properties to circulating metastatic breast cancer cells. These observations open new alleys of investigation for the identification of the molecular mechanisms responsible for the genesis of bone metastases.

Biomarkers, Tumor↗
PubMed1995

Comparison between allograft plus demineralized bone matrix versus autograft in anterior cervical fusion. A prospective multicenter study.

STUDY DESIGN: This study analyzed the fusion results of an allograft-demineralized bone matrix composite versus autograft in a prospective series of patients undergoing surgery for cervical disc disease. OBJECTIVES: To determine the fusion rates of allograft-demineralized bone matrix composite in anterior cervical fusion as compared with the gold standard autograft. SUMMARY OF BACKGROUND DATA: For the anterior cervical fusion, the use of freeze-dried allograft is well documented in the literature, citing its effectiveness and inferior fusion rates. The use of demineralized bone matrix in conjunction with freeze-dried allograft in anterior cervical fusion has not been reported. METHODS: This study was done in a prospective fashion in two medical centers. One group received autograft from the anterior iliac crest, whereas others received freeze-dried allograft augmented with demineralized bone matrix (Grafton, Osteotech, Inc., Shrewsbury, New Jersey). For the autograft group, the standard Smith-Robinson grafting technique was used. For the allograft composite group, demineralized bone matrix was pasted onto the freeze-dried allograft and into the disc space before graft insertion. The autograft group consisted of 38 patients with age ranging 26-71 years (mean, 46.1 years) and follow-up periods of 12-33 months (mean, 18.4 months). There were 19 one-level, 17 two-level, and two three-level fusions. Similarly, the allograft group consisted of 39 patients with age ranging 28-80 years (mean, 48.0 years) with follow-up period of 12-31 months (mean, 17.5 months). There were 19 one-level, 16 two-level, and four three-level fusions. Clinical and radiographic follow-up evaluations were completed at 3-month intervals. Radiographs taken 12 months after surgery were analyzed blindly. RESULTS: Pseudarthrosis developed in 46.2% of patients (33.3% of levels) in the allograft-demineralized bone matrix group compared with 26.3% (22% of levels) in the autograft group (P = 0.11 for patients, P = 0.23 for levels). For patients undergoing two-level fusions, 37.5% of allograft-demineralized bone matrix failed compared with 23.5% of autografts. For single-level fusions, 47.4% of allograft patients developed a pseudarthrosis compared with 26.3% in the autograft group. Graft collapse of > or = 3 mm was noted in 11% of the autograft group versus 19% in the allograft-demineralized bone matrix group (P = 0.32). Graft collapse of > or = 2 mm occurred in 24.4% of autograft patients compared with 39.7% of the allograft-demineralized bone matrix group (P = 0.09). Smokers had an increased rate of pseudarthrosis (47.1%) compared with nonsmokers (27.9%, P = 0.13). CONCLUSIONS: The study revealed that the allograft-demineralized bone matrix construct gives a higher rate of graft collapse and pseudarthrosis when compared with autograft in a prospective series, although the differences were not statistically significant. The pseudarthrosis rate in the series may be high because of the large percentage of smokers and radiographic evaluation techniques. For the purpose of solid radiographic fusion, the use of autograft is recommended in anterior cervical surgery until other acceptable osteoinductive materials are developed.

Adult↗
PubMedDate not supplied

Treatment of unicameral bone cyst with demineralized bone matrix.

Eleven patients with active unicameral bone cysts were treated primarily with placement of demineralized bone matrix in the cyst by using a two-needle technique and a custom large-bore needle. Cyst healing was rated according to the Neer classification, and the average time of healing was 4.5 months. The demineralized bone matrix demonstrated an ability to obliterate the cyst in nine of 11 patients by using a single injection within 4-5 months, and at 2 years' follow-up, no cysts were deemed active or recurrent.

Bone Cysts↗
PubMed2007

Quantification of various growth factors in different demineralized bone matrix preparations.

Besides autografts, allografts, and synthetic materials, demineralized bone matrix (DBM) is used for bone defect filling and treatment of non-unions. Different DBM formulations are introduced in clinic since years. However, little is known about the presents and quantities of growth factors in DBM. Aim of the present study was the quantification of eight growth factors important for bone healing in three different "off the shelf" DBM formulations, which are already in human use: DBX putty, Grafton DBM putty, and AlloMatrix putty. All three DBM formulations are produced from human donor tissue but they differ in the substitutes added. From each of the three products 10 different lots were analyzed. Protein was extracted from the samples with Guanidine HCL/EDTA method and human ELISA kits were used for growth factor quantification. Differences between the three different products were seen in total protein contend and the absolute growth factor values but also a large variability between the different lots was found. The order of the growth factors, however, is almost comparable between the materials. In the three investigated materials FGF basic and BMP-4 were not detectable in any analyzed sample. BMP-2 revealed the highest concentration extractable from the samples with approximately 3.6 microg/g tissue without a significant difference between the three DBM formulations. In DBX putty significantly more TGF-beta1 and FGFa were measurable compared to the two other DBMs. IGF-I revealed the significantly highest value in the AlloMatrix and PDGF in Grafton. No differences were accessed for VEGF. Due to the differences in the growth factor concentration between the individual samples, independently from the product formulation, further analyzes are required to optimize the clinical outcome of the used demineralized bone matrix.

Bone Matrix↗
PubMed1985

The osteo-inductive properties of bone matrix from rats pretreated with indomethacin.

Decalcified bone matrix pieces were prepared from growing rats treated for 3 weeks with subcutaneous injections of indomethacin 2 mg/kg/day or saline, and implanted into growing rats. Of the 16 recipients with 'saline implants', 8 received saline and another 8 indomethacin 2 mg/kg/day in subcutaneous injections. In another 16 recipients with 'indomethacin implants', 8 received saline and 8 indomethacin. As in previous studies, it was found that indomethacin exerted a mild inhibition of new bone formation expressed as decreased amount of ash weight in implants. The amount of ash weight and 45Ca specific activity of implants in recipients with implants from indomethacin-treated donor animals was not altered. These findings indicate that indomethacin is unable to modify the osteo-inductive properties of bone matrix.

Animals↗