Expression of Extracellular Matrix Macromolecules Around Demineralized Freeze-Dried Bone Allografts
University of Queensland, Department of Dentistry, Australia.Journal of Periodontology (Impact Factor: 2.71). 12/1996; 67(11):1233-44. DOI: 10.1902/jop.19188.8.131.523
In the present study histochemical techniques were used to identify specific macromolecular components of the extracellular matrix associated with the tissue reaction to demineralized freeze-dried bone allografts (DFDBA) placed under barrier membranes for ridge augmentation. Small biopsies were obtained from tissues underneath the membranes at various times after placement of the DFDBA and processed for routine immunohistochemistry. Sections were stained with antibodies to osteocalcin, collagen type I, collagen type III, decorin, and biglycan. Non-immune serum, irrelevant antibodies, and omission of the primary antibodies served as negative controls. Histologic examination of the biopsies revealed allograft particles surrounded by well-formed fibrous connective tissue with little or no evidence of new bone formation. Vital autogenous bone fragments were present in the peripheral portions of the biopsies and served as positive controls for comparative purposes with the DFDBA particles. Only 7 out of the 20 biopsies studied were found to have any signs of bone formation around the DFDBA particles and in these such bone formation was irregular and inconsistent around the DFDBA particles. Around the periphery of the allograft particles, osteocalcin, collagen type I, collagen type III, decorin, and biglycan all showed relatively strong staining. Osteocalcin staining was also noted within the vital bone matrix but not in the surrounding fibrous connective tissue. Decorin, biglycan, collagen type I, and collagen type III were also found within the vital bone matrix. None of these antibodies stained the DFDBA particles. The unremarkable osteogenic response of the tissues to the DFDBA particles after healing periods of up to 12 months raises questions as to the predictability of these agents in inducing new bone.
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ABSTRACT: The purpose of this study was to compare extraction socket healing in 8 patients after implantation with either xenogenic bovine bone (n=5 sites), demineralized freeze-dried bone (DFDBA) (n=3 sites), autologous bone (n=3 sites), or human bone morphogenetic proteins in an osteocalcein/osteonectin carrier (hBMP/NCP) (n=2 sites). Three of the patients received 6 commercially pure micro screws which were fixed into extraction sockets, after which the sockets were implanted with either bovine bone (n=3 sites), DFDBA (n=2 sites) or intraoral autologous bone (n=1 site). Biopsies of the extraction sockets were taken from 3 to 6 months after treatment (average, 4.6 months). For comparison of healing between the implanted materials, histologic evaluation and bone scores were determined. Bone scores of 0 indicated an absence of new bone, with dead implanted bone particles entrapped within connective tissue, while a score of 3 indicated the entire field consisted of vital bone. Biopsies from bovine bone sockets revealed dead implanted particles surrounded by connective tissue. Isolated sections showed host bone in contact with the bovine bone particles. Bone scores ranged from 0 to 3. Biopsies from DFDBA-implanted sites revealed dead particles entrapped with dense connective tissue. The bone scores ranged from 0 to 1. Biopsies from sites implanted with hBMP/NCP revealed a combination of woven and lamellar bone with bone scores of 3. Five of the 6 micro screws were processed and evaluated. One screw was mobile at the time of removal and was not evaluated. Bone scores were used to compare new bone formation adjacent to the micro screws. Bone scores ranged from 0 to 2. A score of 0 indicated non-vital implant material in contact with host bone and connective tissue in contact with implant; 2 indicated vital bone in contact with the majority of the implant surface. Retrieved sockets with micro screws implanted with bovine bone (n=2) demonstrated a connective tissue interface between the screws and the surrounding tissues (bone score 0). The adjacent tissues showed dead bovine particles entrapped within fibrous tissue. Retrieved screws implanted with DFDBA (n=2) were surrounded by connective tissue, with dead bone particles enmeshed within fibrous tissue (bone score 0). The screw implanted with intra-oral autologous bone was primarily surrounded by vital bone with a connective tissue interface (bone score 1). Three implant threads were in contact with bone. The results of this study indicate that bovine bone, DFDBA, and intraoral autologous bone do not promote extraction socket healing. Sockets implanted with hBMP/NCP contained vital woven and lamellar bone. Xenogenic bovine bone and DFDBA did not contribute to bone to micro screw contacts and are not recommended for enhancement of vital bone to implant contacts. Intraoral autogenous bone also does not appear to significantly contribute to bone to implant contacts. Intraoral autologous bone, xenogenic bone, and DFDBA appear to interfere with normal extraction socket healing.Journal of Periodontology 05/1998; 69(4):414-21. DOI:10.1902/jop.19184.108.40.2064 · 2.71 Impact Factor
- Periodontology 2000 03/1999; 19(1):104-14. DOI:10.1111/j.1600-0757.1999.tb00150.x · 3.63 Impact Factor
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ABSTRACT: Background: Demineralized freeze-dried bone allografts (DFDBAs) have been proposed as a useful adjunct in periodontal therapy to induce periodontal regeneration through the induction of new bone formation. The presence of bone morphogenetic proteins (BMPs) within the demineralized matrix has been proposed as a possible mechanism through which DFDBA may exert its biologic effect. However, in recent years, the predicability of results using DFDBA has been variable and has led to its use being questioned. One reason for the variability in tissue response may be attributed to differences in the processing of DFDBA, which may lead to loss of activity of any bioactive substances within the DFDBA matrix. Therefore, the purpose of this investigation was to determine whether there are detectable levels of bone morphogenetic proteins in commercial DFDBA preparations. Methods: A single preparation of DFDBA was obtained from three commercial sources. Each preparation was studied in triplicate. Proteins within the DFDBA samples were first extracted with 4M guanidinium HCI for seven days at 40 degrees Celsius and the residue was further extracted with 4M guanidinium HCL/EDTA for seven days at 40 degrees celsius. Two anti-human BMP-2 and -4 antibodies were used for the detection of the presence of BMP's in the extracts. Results: Neither BMP-2 nor BMP-4 was detected in any of the extracts. When recombinant human BMP-2 and -4 were added throughout the extraction process of DFDBA extraction, not only were intact proteins detected but smaller molecular weight fragments were also noted in the extract. Conclusions: These results indicate that all of the DFDBA samples tested had no detectable amounts of BMP-2 and -4. In addition, an unknown substance present in the DFDBA may be responsible for degradation of whatever BMPs might be present.Clinical Implant Dentistry and Related Research 02/2000; 2(2). DOI:10.1111/j.1708-8208.2000.tb00113.x · 3.59 Impact Factor
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