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PubMed1993

Experimental and clinical investigations of human insoluble bone matrix gelatin. A report of 24 cases.

Human bone matrix gelatin (hBMG) was implanted into the quadriceps muscles of mice to determine its osteoinductive activity and immune responses. The host mouse muscle assay reacted positively to the hBMG, providing an experimental basis for clinical application. At two weeks after implantation, cartilage was produced in the mouse muscle, and new bone and bone marrow were formed at three to four weeks after implantation. Human bone matrix gelatin, which was thus shown to have osteoinductive activity, was subsequently implanted in 24 patients. Generally, the bone defects were filled with new bone tissue with increased density within two to four months after operation. Nonunion and delayed unions were healed at two to six months after surgery.

Adult↗
PubMed1993

Demineralized bone matrix in the stabilization of porous-coated implants in bone defects in rabbits.

Three types of grafts were investigated in rabbits to measure fixation strength of bony ingrowth into porous-coated titanium alloy implants. Autogeneic iliac crest bone (ABG), allogeneic demineralized bone matrix (DBM), and DBM augmented with fibrin glue (DBM + FG) grafts were compared with a press-fit implant control group. Initially, the ABG group required eight weeks and the DBM and DBM + FG groups 12 weeks to achieve fixation strength similar to that of the press-fit group at four weeks. Strength increased with time and at 16 weeks reached 83 kg in the ABG group, 71 kg in the DBM group, and 79 kg in the DBM + FG group, compared with 86 kg for the press-fit group. The ratio of the DBM and the DBM + FG group push-out forces to the ABG push-out force improved from 40% at four weeks to more than 80% at 16 weeks. Histologic analysis of bone ingrowth showed that at 12 weeks, bony ingrowth accounted for 21%, 22%, 16%, and 32% of the porous area in the ABG, DBM, DBM + FG, and press-fit groups, respectively. After eight weeks, there was no statistical difference between DBM, DBM + FG, and ABG grafts in either strength or bony ingrowth. The results demonstrate that over long periods, DBM grafts provide fixation stability comparable with that of autogeneic bone graft.

Alloys↗
PubMed2003

Use of demineralized bone matrix in ankle/hindfoot fusion.

Sixty-three patients who underwent complex ankle or hindfoot fusion had demineralized bone matrix placed in their fusion site to stimulate fusion. Thirty-seven patients had Grafton putty, a demineralized bone matrix product, and 26 patients had Orthoblast, a demineralized bone matrix mixed with crushed cancellous allograft bone placed to stimulate their fusion site. All patients were followed clinically and radiographically to fusion or nonunion. Of the 37 patients who had Grafton putty placed to stimulate ankle or hindfoot fusion, five (14%) developed a nonunion. Of the 26 patients who had Orthoblast placed to stimulate fusion, two (8%) developed a nonunion. These differences were not statistically significant. Nonunion rates of approximately 10% continue to be reported for ankle and hindfoot fusion procedures. In an attempt to minimize this complication, various bone graft substitutes have been used. We found no difference in efficacy of the two demineralized bone matrix compounds, and were not able to demonstrate a superior union rate compared to historical controls.

Adult↗
PubMed2006

Homogeneous osteogenesis and bone regeneration by demineralized bone matrix loading with collagen-targeting bone morphogenetic protein-2.

Considerable research has been focused on the development of bone morphogenetic protein-2 (BMP-2) delivery system for homologous and efficient bone regeneration. The aim of the present study was to develop a collagen-based targeting bone repair system. A collagen-binding domain (CBD) was added to the N-terminal of native BMP-2 to allow it bind to collagen specifically. We showed that the collagen-binding bone morphogenetic protein-2 (named bone morphogenetic protein2-h, BMP2-h) had maintained the full biological activity as compared to rhBMP2 lacking the CBD. In vitro functional study also demonstrated that collagen matrix could maintain higher bioactivity of BMP2-h than native BMP-2. When demineralized bone matrix (DBM) impregnated with BMP2-h was implanted subcutaneously in rats, homogeneous bone formation was observed. Moreover, in a rabbit mandible defect model, surgical implantation of collagen matrix loaded with BMP2-h exhibited remarkable osteoinductive properties and excellent homogeneous bone formation. Our studies suggested that this novel collagen-based BMP-2 targeting bone repair system induced better bone formation not only in quantity but also in quality. Similar approaches may also be used for the repair of other tissue injuries.

Animals↗
PubMed1997

Maxillary sinus augmentation using different grafting materials and dental implants in monkeys. Part I. Evaluation of anorganic bovine-derived bone matrix.

The aim of this study was to evaluate clinically, histologically and histometrically the use of anorganic bovine bone matrix (i.e. Bio-oss) as a grafting material for maxillary sinus augmentation procedures. In 4 adult male rhesus monkeys (i.e. Macaca mulatta) the 1st, 2nd and 3rd maxillary molars on one side of the jaws were extracted. The remaining bone between the alveolar crest and the bottom of the sinus was then reduced to 3-4 mm. After 3 months, maxillary sinus augmentation procedures were performed on one side of the jaws in each monkey and the sinuses were grafted with the bovine bone matrix. At that time, 2 IMZ pure titanium plasma coated implants were immediately placed into the augmented sinuses (i.e. simultaneous implants-loaded group). After 4 months, 2 additional similar implants were placed into these previously augmented sinuses (i.e. delayed implants-loaded group). Four months later, the abutment connection was performed and all 4 implants were loaded with a gold-alloy bridge for 6 months (i.e. until sacrifice of the animals). The contralateral side of each monkey received the same treatment with the exception that the extractions were performed 7 months after those in the opposite side and that the implants in this side were not loaded. Thus, 2 additional study groups (i.e. simultaneous implants-unloaded group and delayed implants-unloaded group) were obtained. Clinically, all loaded implants were stable at the day of sacrifice. Histologically, the grafted sinuses exhibited significant bone formation with integration of the bovine bone matrix particles to the new bone. Direct mineralized bone-to-implant contact was greater for the delayed implant placement groups than for the implants installed simultaneously with the sinus augmentation. Furthermore, the percentage of direct mineralized bone-to-implant contact was greater in the residual bone than in the augmented area. It was concluded that the anorganic bovine bone matrix facilitated bone formation and implant osseointegration in the augmented sinuses and that the delayed implant placement in combination with the sinus augmentation procedure seemed to be preferable.

Alveolar Process↗
PubMed1988

The role of mineralization in experimental models of osteogenetic induction with decalcified bone matrix.

The role of mineralization was studied in an experimental model of osteoinduction consisting of the implant of decalcified bone matrix in the rat muscle. After vascular connective tissue invasion of the matrix, the first phenomena is the appearance of hydroxyapatite crystals in the area of chondroid metaplasia from where the deposit of calcium-phosphate is extended to the surrounding decalcified matrix. The recalcified areas act as a substratum for the neo-osteogenesis. The appearances observed reproduced those of enchondral ossification, where the first osteoblasts differentiated on the calcified cartilaginous matrix and began to supply new bone matrix. Non-decalcified bone matrix implants were surrounded by a connective tissue capsule with features identical to those of a foreign body reaction and with total absence of vascular invasion. The absence of induction in this type of implant may be related either to masking of the osteoinducing protein in the calcified matrix, or to the absence of cells with osteogenic differentiation potential due to the lack of cellular invasion of the implant.

Animals↗
PubMed1987

Incorporation of autoclaved autogeneic bone supplemented with allogeneic demineralized bone matrix. An experimental study in the rabbit.

Resected, autoclaved, and reimplanted bone supplemented with allogeneic demineralized bone matrix (DABM) was compared with nonsupplemented bone in the reconstruction of bilateral ulnar defects in seven adult rabbits. The reconstructions were studied in vivo with the use of serial radiography, scintigraphy (99mTc-MDP), and bone mineral determination. After sacrifice at 16 weeks, the ulnar specimens were analyzed with the use of high-resolution radiography, autoradiography (45Ca), and histologic study. Scintigraphically, the uptake was significantly higher in the DABM-supplemented implants during the entire investigation. Bone mineral content was significantly higher at four and eight weeks in the DABM-supplemented implants, compared with the nonsupplemented implants, but later it equalized. New bone formation, as reflected by 45Ca incorporation, still occurred at 16 weeks in both types of implants, but it was more intense in the DABM-supplemented ones. Radiography and histologic analysis showed more new bone in the DABM-supplemented reconstructions than in the nonsupplemented ones. High-resolution radiography showed complete incorporation of all seven DABM-supplemented implants, whereas there was nonunion in five of seven nonsupplemented reconstructions. The current study indicates that resected, autoclaved, and reimplanted diaphyseal segments supplemented with demineralized allogeneic bone matrix incorporate better than nonsupplemented implants of autoclaved autologous bone.

Animals↗
PubMed2006

[Antigen measurement and biomechanical characteristics of the inbred-line Banna mini-pig acellular bone matrix].

In this study, we prepared the acellular bone matrix of the inbred-line Banna mini-pig by using tissue engineering method and evaluated its possible application in bone tissue engineering. Histological analysis, xenoantigen expression and biomechanical measurement were performed on the matrix. HE staining and scanning electron microscopy showed the cellular components were almost removed. Immunohischemical result demonstrated that the xenoantigen, alpha-gal,was also eliminated. There was no statistically significant difference between the acellular bone matrix group and control group. The acellular bone matrix can provide appropriate space structure and strength for grafts. In conclusion, our data suggest that acellular bone matrix is a new kind of ideal bone scaffold material.

Animals↗
PubMed1998

[The role of fetus decalcified bone matrix (FDBM) in inducing pure titanium-bone implant integration].

Because of its high biological compatibility, titanium has been a good biomaterial. The implanted artificial bone made from titanium can contact with the vital and mature osseous tissue directly within 3-6 months, the so-called osteointergration. In order to promote the process of osteointergration, FDBM of rabbit was prepared and was combined with pure titanium so as to speed up osteointergration. The study focused on bone density, bone intergration rate, new bone growth rate around the pure titanium, and the Ca2+ and PO(4)3- density of titanium-bone interface. A control group of pure titanium inplant without FDBM was set up. The results showed FDBM had no antigenicity. It could induce and speed up the new bone formation at titanium-bone interface. The titanium-bone intergration time was within 2 months. It was suggested that there were more bone morphogenesis protein (BMP) or other bone induction and bone formation factors in brephobone than that in child and adult bone. As a kind of bone induction material, FDBM was easy prepared, cheap in price, easy to storage, no antigenicity and obvious bone-inductive function.

Animals↗
PubMed2004

The use of demineralized bone matrix for mastoid cavity obliteration.

OBJECTIVE: To evaluate the use of demineralized bone matrix as a graft material for mastoid cavity obliteration in the treatment and prevention of problematic mastoid cavities. STUDY DESIGN: The study is a retrospective review of patients identified using a computerized otology database. SETTING: Tertiary care referral center. PATIENTS: Patients were included in this study if they underwent mastoid obliteration using demineralized bone matrix. INTERVENTION: Mastoid obliteration was performed for revision of a problematic mastoid cavity (n = 8) or primarily after canal wall down mastoidectomy for recurrent cholesteatoma (n = 3). MAIN OUTCOME MEASURES: Data were collected to evaluate the ability to achieve a dry ear canal. Postoperative healing time and hearing results were also assessed. RESULTS: A dry ear canal was achieved in all patients with a follow-up of 6 to 20 months (average, 14.5 mo). Eight patients (73%) had a well-healed, dry ear canal by their first postoperative visit (9 wk). One patient required 12.5 weeks to heal. Two patients (18%) had more prolonged granulation at the ear canal incision, which resolved in 17 and 28 weeks, respectively. The average preoperative pure-tone average air-bone gap was 47 +/- 14.9 dB (mean +/- SD), compared with postoperative values of 27.6 +/- 12.8 dB (p = 0.0033; paired t test). This represents an average pure-tone average air-bone gap closure of 20 dB. CONCLUSION: The use of demineralized bone matrix as a graft extender in mastoid obliteration allowed creation of a dry, smooth-contoured canal in all patients studied. A significant improvement in hearing was also obtained. Demineralized bone matrix is an acceptable graft alternative for mastoid obliteration.

Adolescent↗
PubMed1988

Bone induction in intramuscular implants by demineralized bone matrix: sequential changes of collagen synthesis.

Implantation of rat demineralized bone matrix into intramuscular pouches has been shown to cause a complex cellular transition of mesenchymal-type cells into well developed mature bone. Demineralized bone matrix was surgically implanted into rat muscle pouches and removed at various intervals between 7 and 28 days. Histological sections of the implants revealed bone formation by endochondral ossification and appositional bone growth. Biochemical analysis of collagen synthesis demonstrated the following: (1) synthesis of type X collagen, a collagen produced by hypertrophic chondrocytes in the growth plate and in fracture callus. (2) Synthesis of a collagenase-sensitive 17k protein which seems to increase in the early stages of bone induction. Pulse chase analysis indicates that 17k is not a degradation product of another protein and appears to be synthesized without a large Mr precursor. The 17k component contains one or more collagenous domains that are partially resistant to proteolysis with pepsin. Our results confirm the appearance of a cartilage intermediate during demineralized bone matrix induced ossification and implicate the existence of proteins which may be useful markers in future studies on matrix mineralization and ossification.

Animals↗
PubMed2004

Concentration of insulin-like growth factor (IGF)-I in iliac crest bone matrix in premenopausal women with idiopathic osteoporosis.

Previous studies have shown a link between low serum insulin-like growth factor-I (IGF-I) and decreased bone mass of patients with osteoporosis. However, whether serum levels are representative for the growth factor concentration or activity available in human bone tissue is controversial. In the present study, IGF-I was assessed in serum and bone matrix extracts from the iliac crest in 19 eugonadal women with idiopathic osteoporosis and in 38 age-matched controls. In addition, the relationship between the skeletal levels of IGF-I and bone mineral density (BMD) or the susceptibility to osteoporotic fractures in women with osteoporosis was examined. Bone matrix extraction was performed based on a guanidine-HCL/ethylendiamine-tetraacetic acid (EDTA) method. No significant difference in both serum and bone matrix IGF-I levels between groups was observed. Serum IGF-I concentrations failed to be associated with bone matrix IGF-I levels in osteoporotic patients. However, in premenopausal women with idiopathic osteoporosis, skeletal IGF-I positively correlated with BMD at the lumbar spine (r = + 0.58, p = 0.01). In contrast, neither femoral neck BMD nor Ward's triangle BMD was associated with bone matrix IGF-I concentrations. A tendency towards lower levels of bone matrix IGF-I in subjects with vertebral fractures as compared to those without fractures was observed in age-adjusted analyses, however the difference failed to remain statistically significant after adjustment for bone mineral density. These data provide no clear evidence for low bone matrix IGF-I as a determinant factor of age-unrelated osteoporosis. However, low skeletal IGF-I concentrations may aggravate osteoporosis in these women.

Absorptiometry, Photon↗
PubMed1984

Effects of pregnancy on bone matrix proteins in Wistar rats.

Using an EDTA extraction procedure, bones from pregnant Wistar rats were analyzed for their content of collagen and non-collagenous components (sialoprotein, proteoglycan and carbohydrate). The bone matrix size was found to be smaller in pregnant rats than in normal rats (19.5% vs 17.5% of the dry weight bone). The EDTA extractability of the bone protein from pregnant rats was higher than that from controls (2.6% vs 1.9% dry weight bone). EDTA extracts from pregnant rats contained higher amounts of soluble collagen (1.6% vs 0.5% of dry weight tissue) and lower amounts of non-collagenous components (1.65% vs 2.23% for hexoses, 2.38% vs 3.95% for sialic acid and 1.24% vs 1.73% for uronic acid). In bone matrix, collagen content was lower in the pregnant rats (9.45% vs 10.6%). Similarly, the amounts of non-collagenous components were slightly decreased in the bone matrix from the pregnant rats. The respective values were: 0.91% vs 0.93% for hexoses, 0.45% vs 0.52% for sialic acid and 0.39% vs 0.50% for uronic acid. These results suggest that in pregnancy collagen and non-collagenous protein content in bone is decreased while the total mineral content is increased.

Animals↗
PubMed2005

Demineralized bone matrix: basic science and clinical applications.

Bone grafting with demineralized bone matrix (DBM) is useful in reconstructive surgery to ultimately provide anatomic alignment, restore function, or augment/change the biomechanics of the foot and ankle. DBM should be used in conjunction with a transplant or implant displaying mechanical strength. DBM can augment cortical grafts used for bridging gaps or defects and lengthening procedures by increasing the connectivity of the structural graft with the host bone. Another useful application is providing a biologic boost to patients who have less-than-ideal physiology. Because DBM has higher concentrations of available bone morphogenic proteins, it can aid in the incorporation of other grafts. Other uses include delayed unions, nonunions, packing joints for arthrodesis, filling resected cysts, and filling gaps of debrided infected bone.

Arthrodesis↗
PubMed1983

Ultrastructural cytochemistry of complex carbohydrates in osteoblasts, osteoid, and bone matrix.

Glycosaminoglycans (GAGs) and glycoproteins are essential components for osteogenesis. We have examined rat osteoblasts, osteoid, transitional zone, and fully calcified bone matrix, utilizing Spicer's high-iron diaminethiocarbohydrazide-silver protein (HID-TCH-SP) method for sulfated glycoconjugates and Thiéry's periodate-TCH-SP (PA-TCH-SP) method for vicinal glycol-containing glycoconjugates. HID-TCH-SP stained cytoplasmic granules of osteoblasts. Stain deposits in the extracellular matrix were observed in decreasing amounts in osteoid, the transitional zone, and fully calcified bone matrix. Enzyme digestion with testicular hyaluronidase removed most HID-TCH-SP stain deposits. PA-TCH-SP staining was observed with increasing intensity in rough endoplasmic reticulum, Golgi saccules, and cytoplasmic granules. Collagen fibrils in osteoid were weakly stained with PA-TCH-SP, and their staining appeared even weaker in fully calcified bone matrix. In contrast, collagen fibrils in calcified cartilage stained intensely with the PA-TCH-SP method. Focal circular profiles (0.1-0.5 mumol in diameter), which lacked collagen fibrils but reacted moderately with PA-TCH-SP, were frequently seen in the transitional zone and fully calcified bone matrix, but were only occasionally present in osteoid. The presence of testicular hyaluronidase-resistant GAG and acid phosphatase in these focal areas suggests that they represent sites of GAG degradation. The eventual loss of HID-TCH-SP staining in the bone matrix suggests that removal of sulfated glycoconjugates may be a requisite for expansion of initial calcification sites and/or complete calcification.

Acid Phosphatase↗
PubMed1981

Radioisotopic and morphometric evaluation of the effects of beta-aminopropionitrile on chick bone matrix formation and its mineralization.

Bones from 2-week-old chicks, fed a diet containing 0.05% beta-aminopropionitrile (beta-APN), were studied for 3H-tetracycline and 3H-proline incorporation at 7, 14 and 21 days of the experiment. Liquid scintillation counts showed a comparable amount of 3H-proline incorporation in the controls and experimentals. However, the incorporation of 3H-tetracycline was significantly lower in experimentals at all intervals. Fluorescent microscopy after cold tetracycline injections at similar intervals showed a comparable amount of linear bone apposition in controls and beta-APN-fed chicks. The bone ash content in beta-APN-fed chicks was lower at all intervals whereas the serum calcium level was similar to the controls. Microradiography showed that cortical and marrow areas, in cross sections, were not significantly different in the controls and beta-APN-fed chicks. On the basis of these data it is suggested that beta-APN at the dose used does not inhibit the bone matrix synthesis but inhibits its mineralization.

Animals↗
PubMed1978

Ultrastructure of chondrogenetic interactions between bone matrix gelatin and mesenchymal cells.

The ultrastructure of post fetal chondrogenesis was investigated in chondrogenetic implants and explants of bone matrix gelatin: (A) in direct contact with living cells; (B) separated from living cells by cellulose acetate membranes; (C) in serum or in a culture medium with no contact with living cells. Ruthenium red (RR) staining coarse granules free of interconnecting filaments aggregated in interstitial fluid and inside the surface of the bone matrix gelatin within 72 hours in systems with either direct or indirect contact with living connective tissue cells. No RR granules formed when control denatured or autolysed bone matrix gelatin were substituted for undenatured bone matrix gelatin. No RR granules formed in bone matrix gelatin incubated in serums or media in the absence of living cells in the system. These observations suggest the possibility that 4 days before overt transmembrane cartilage cell differentiation, covert (presumptive) chondroblasts synthesize and secrete precursor substance for aggregation of filament-free RR granules. The relationship between chondromorphogenesis and filament-free RR granules requires further investigation.

Animals↗
PubMed2003

Comparative evaluation of the osteoinductivity of two formulations of human demineralized bone matrix.

In the United States, demineralized bone matrix (DBM) is considered a transplantable tissue and therefore is regulated primarily by the American Association of Tissue Banks. Even though DBM is not subjected to the same regulations relative to performance claims as medical devices are, one would expect different processing methods might yield DBM preparations of different osteoinductive potential. The purpose of this study was to use an established athymic rat model to compare the osteoinductive properties of two commercially available human DBMs prepared using different methods but having essentially identical product claims. Sixteen female athymic rats were used to test equivalent volumes of two lots each of Grafton Putty (Osteotech, Inc., Eatontown, NJ), Osteofil (Regeneration Technologies, Inc., Alachua, FL), and rat DBM. At 28 days after implantation, qualitative and semiquantitative microscopy showed no significant differences in bone formation between the two lots from each source, but rat DBM produced significantly more bone than Grafton, which produced significantly more bone than Osteofil. Our results suggest that methods of graft processing may represent a greater source of variability than do differences among individual donors. Whether these differences relate to methods of demineralization, carrier, dose of DBM per volume, or to some other factor remains to be determined.

Animals↗