OBJECTIVE: To investigate the safety and efficacy of demineralized bone matrix (DBM) as a bone graft substitute for anterior cervical discectomy and fusion (ACDF) surgery. METHODS: Twenty consecutive patients treated with ACDF using stand-alone polyestheretherketone (PEEK) cages (Zero-P) with DBM(CGDBM100) were prospectively evaluated with a minimum of 6 months of follow-up. Radiologic efficacy was evaluated with a 6-point scoring method for osseous fusion using plain radiograph and computed tomogrpahy scans. Clinical efficacy was evaluated using the visual analogue scale (VAS), Owestry disability index (ODI), and short-form health questionnaire-36. The safety of the bone graft substitute was assessed with vital sign monitoring and a survey measuring complications at each follow-up visit. RESULTS: There were significant improvements in VAS and ODI scores at a mean 6-month follow-up. Six months after surgery, solid fusion was achieved in all patients. Mean score on the 6-point scoring system was 5.1, and bony formation was found to score at least 4 points in all patients. There was no case with implant-related complications such as cage failure or migration, and no complications associated with the use of CGDBM100. CONCLUSION: ACDF using CGDBM100 demonstrated good clinical and radiologic outcomes. The fusion rate was comparable with the published results of traditional ACDF. Therefore, the results of this study suggest that the use of a PEEK cage packed with DBM for ACDF is a safe and effective alternative to the gold standard of autologous iliac bone graft.
Arm* ; Bone Matrix* ; Bone Transplantation ; Diskectomy ; Follow-Up Studies ; Humans ; Pilot Projects* ; Prospective Studies ; Research Design ; Transplants* ; Treatment Outcome ; Vital Signs

We compared the bone healing capacity of three different demineralized bone matrix (DBM) products applied using different carrier molecules (hyaluronic acid [HA] vs. carboxymethylcellulose [CMC]) or bone compositions (cortical bone vs. cortical bone and cancellous bone) in a rabbit segmental defect model. Overall, 15-mm segmental defects in the left and right radiuses were created in 36 New Zealand White rabbits and filled with HA-based demineralized cortical bone matrix (DBX), CMC-based demineralized cortical bone matrix (DB) or CMC-based demineralized cortical bone with cancellous bone (NDDB), and the wound area was evaluated at 4, 8, and 12 weeks post-implantation. DBX showed significantly lower radiopacity, bone volume fraction, and bone mineral density than DB and NDDB before implantation. However, bone healing score, bone volume fraction, bone mineral density, and residual bone area at 4, 8, and 12 weeks post-implantation revealed no significant differences in bone healing capacity. Overall, three DBM products with different carrier molecules or bone compositions showed similar bone healing capacity.
Animals ; Bone Matrix/*physiology ; Bone Transplantation ; Carboxymethylcellulose Sodium/*pharmacology ; Histology ; Hyaluronic Acid/*pharmacology ; Rabbits ; *Wound Healing ; X-Ray Microtomography ; X-Rays

STUDY DESIGN: Prospective study. PURPOSE: We investigated normative temporal levels of white blood cell (WBC) and absolute neutrophil count (ANC) in uncomplicated anterior cervical discectomy and fusion (ACDF) using allograft and demineralized bone matrix (DBM). OVERVIEW OF LITERATURE: No study has investigated the diagnostic usefulness of WBC and ANC for postoperative infection following ACDF using allograft and DBM. METHODS: Blood samples of 85 patients, who underwent one or two-level ACDF, were obtained and evaluated before surgery and on the first, third, fifth, seventh, fourteenth, thirtieth, and ninetieth postoperative days. No infection was found in all patients for at least one year follow-up period. RESULTS: Mean WBC and ANC values increased significantly and reached peak levels on the first postoperative day. The peaked levels rapidly decreased but still remained elevated above the preoperative levels on the third postoperative day. The levels returned close to the preoperative levels on the fifth postoperative day. The mean WBC and ANC values did not get out of their normal reference ranges throughout the follow-up periods. One-level and two-level ACDF exhibited a similar course of postoperative changes in WBC and ANC values and no significant difference in mean levels of WBC and ANC throughout the follow-up periods. CONCLUSIONS: Uncomplicated ACDF using allograft and DBM showed normal values of WBC and ANC during the early postoperative period. Therefore, significant abnormal values of WBC and ANC at an early postoperative period suggest the possibility of the development of acute postoperative infection after ACDF using allograft and DBM.
Bone Matrix ; Diskectomy ; Follow-Up Studies ; Humans ; Leukocytes ; Neutrophils ; Postoperative Period ; Prospective Studies ; Transplantation, Homologous

OBJECTIVETo observe the effects of microporous porcine acellular dermal matrix (ADM) combined with bone marrow mesenchymal cells (BMMCs) population containing bone mesenchymal stem cells (BMSCs) of rats on the regeneration of cutaneous appendages cells in nude mice.

METHODSSplit-thickness dermal grafts, 20 cm×10 cm in size and 0.3 mm in thickness, were prepared from a healthy pig which was sacrificed under sanitary condition. Laser microporous porcine ADM (LPADM) was produced by laser punching, hypertonic saline solution acellular method, and crosslinking treatment, and nonporous porcine ADM (NPADM) was produced by the latter two procedures. Then the appearance observation, histological examination and scanning electron microscope observation were conducted. BMMCs were isolated and cultured from tibia and femur after sacrifice of an SD rat. Osteogenic and adipogenic differentiation experiments were conducted among the adherent cells in the third passage. Then they were inoculated to LPADM and NPADM to construct BMMCs-LPADM and BMMCs-NPADM materials. Twenty-one healthy nude mice were divided into BMMCs-LPADM+NPADM group (A, n = 6), LPADM+split-thickness skin graft group (B, n = 6), BMMCs-LPADM+split-thickness skin graft group (C, n = 6), BMMCs-NPADM+split-thickness skin graft group (D, n= 3) according to randomized block. After anesthesia, a 2 cm×2 cm full-thickness skin defect reaching deep fascia was reproduced in the middle of the back of each nude mouse, and a split-thickness skin graft of the same size was obtained, and then prepared skin grafts were transplanted to cover the wounds respectively. On post transplantation day (PTD) 5, 7, and 14, local condition and adverse effects observation was conducted; one nude mouse was sacrificed each time to harvest all the transplant for tissue structure observation with HE staining. On PTD 7 and 14, neonatal skin appendages in corresponding composite materials were observed with transmission electron microscope.

RESULTS(1) LPADM and NPADM appeared to be porcelain white, soft, and flexible. No cellular component was observed in acellular dermal matrix. Scanning electron microscope showed that the collagen fibers were orderly arranged. LPADM had microporous structure. (2) Cells in the third passage were orderly arranged with the shape similar to fibroblasts with high growth speed. (3) Induced differentiation experiments showed that cells could differentiate into osteoblasts and adipocytes. (4) On PTD 5, the NPADM in group A was dry in part; skin grafts in group D were dry and necrotic, and there was no infection and inflammation in groups A and D; skin grafts in groups B and C survived. On PTD 7 and 14, the overlaying material in group A was black, dry, and hard in part; the skin grafts in group D turned to be completely black, dry, and necrotic, and pale yellow clear exudate was found in subcutaneous area; there was no obvious purulent discharge in groups A and D; the appearance of skin grafts in groups B and C was close to the surrounding skin. (5) On PTD 5 and 7, in groups A, B, and C, vascularization was apparent in the pores of dermal matrix, and red blood cells could be found. In group D, skin grafts were dry and necrotic. On PTD 14, in groups A, B, and C, the pore structure of dermal matrix was fully vascularized in which a large number of red blood cells were visible. In group A, the microporous dermal matrix survived, but the overlaying NPADM was not attached closely. In groups B and C, the skin grafts were closely connected to the dermal matrix, and no cutaneous appendages were observed. In group C, special monolayer cells were found at the junction between skin graft and dermal matrix. (6) Skin grafts in group D failed to survive; they were not observed with the electron microscope. On PTD 7, there were no significant differences among groups A, B, and C. On PTD 14, no sebaceous gland-like cell or sweat gland-like cell and no newborn nerve ending were observed in skin grafts in groups A and B, in spite of the immigration of fibroblasts. In group C, a large number of new capillaries were observed at the junction between the skin graft and dermal matrix; rough endoplasmic reticulum of fibroblasts proliferated exuberantly; newborn unmyelinated nerve endings were observed; single free sweat gland-like cells and sebaceous gland-like cells were observed in superficial dermal matrix.

CONCLUSIONSLPADM, which provides a "cell niche-like" micro-environment for the migration and differentiation of the BMMCs population, when combining with the split-thickness skin graft, can induce exogenous differentiation of BMSCs in vivo, thus achieving the reconstruction of skin appendages.

Acellular Dermis ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Extracellular Matrix ; transplantation ; Male ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Nude ; Rats ; Rats, Sprague-Dawley ; Regeneration ; Skin ; cytology ; Skin Transplantation ; Skin, Artificial ; Swine ; Wound Healing

PURPOSE: The purpose of this case series was to evaluate the effect of guided bone regeneration using demineralized allogenic bone matrix with calcium sulfate. MATERIALS AND METHODS: Guided bone regeneration using Demineralized Allogenic Bone Matrix with Calcium Sulfate (AlloMatrix(TM), Wright. USA) was performed at the time of implant placement from February 2010 to April 2010. At the time of the second surgery, clinical evaluation of bone healing and histologic evaluation were performed. The study included 10 patients, and 23 implants were placed. The extent of bony defects around implants was determined by measuring the horizontal and vertical bone defects using a periodontal probe from the mesial, distal, buccal, and lingual sides and calculating the mean and standard deviation of these measurements. Wedge-shaped tissue samples were obtained from 3 patients and histologic examination was performed. RESULTS: In clinical evaluation, it was observed that horizontal bone defects were completely healed with new bones, and in the vertical bone defect area, 15.1% of the original defect area remained. In 3 patients, histological tests were performed, and 16.7-41.7% new bone formation was confirmed. Bone graft materials slowly underwent resorption over time. CONCLUSION: AlloMatrix(TM) is an allograft material that can be readily manipulated. It does not require the use of barrier membranes, and good bone regeneration can be achieved with time.
Bone Matrix ; Bone Regeneration ; Calcium ; Calcium Sulfate ; Humans ; Membranes ; Osteogenesis ; Transplantation, Homologous ; Transplants

To evaluate the biological safety of collagen-based bone repairing material, we implanted the sample or reference substance into rats, and observed relative signs, including the specific inspection targets in animals, blood examination, analysis of immune organ, the pathological examination of organs and tissues, NK cell killing activity assay, lymphocyte group analysis, serum IL-1, IL-6, TNF-alpha detection, detection of immune globulin. Meanwhile, we set control group, sham group, and immunosuppression group. The final results showed that there was no abnormal mental state before and after the experiment. Compared with the control group, the tested group indicated no significant difference in blood test, immune organ analysis, the pathological examination of organs and tissues, NK cell killing activity assay, lymphocyte subset analysis serum IL-1, IL-6, TNF-alphadetection, and detection of immune globulin. Collagen-based bone repairing material produced a slight and transient stimulation on the rats, but created no significant inflammatory responses.
Absorbable Implants ; Animals ; Bone Matrix ; physiology ; transplantation ; Bone Regeneration ; Bone Substitutes ; Collagen ; chemistry ; Implants, Experimental ; Materials Testing ; Prostheses and Implants ; Rats

The purpose of this study was to investigate the repair of the osteoarthritis(OA)-induced cartilage injury by transfecting the new TGF-β3 fusion protein (LAP-MMP-mTGF-β3) with targeted therapy function into the bone marrow-derived mesenchymal stem cells (MSCs) in rats. The recombinant of pIRES-EGFP-MMP was constructed by combination of DNA encoding MMP enzyme cutting site and eukaryotic expression vector pIRES-EGFP. LAP and mTGF-β3 fragments were obtained from rat embryos by RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, so as to construct the recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3. pIRES-EGFP-LAP-MMP-mTGF-β3 was transfected into rat MSCs. The genetically modified MSCs were cultured in medium with MMP-1 or not. The transfected MSCs were transplanted in the rat OA models. The OA animal models were surgically induced by anterior cruciate ligament transaction (ACLT). The pathological changes were observed under a microscope by HE staining, Alcian blue, Safranin-fast Green and graded by Mankin's scale. pIRES-EGFP-LAP-MMP-mTGF-β3 was successfully constructed by means of enzyme cutting and sequencing, and the mTGF-β3 fusion protein (39 kD) was certified by Western blotting. Those genetically modified MSCs could differentiate into chondrocytes induced by MMP and secrete the relevant-matrix. The transfected MSCs could promote chondrogenesis and matrix production in rat OA models in vivo. It was concluded that a new fusion protein LAP-MMP-mTGF-β3 was constructed successfully by gene engineering, and could be used to repair the OA-induced cartilage injury.
Animals ; Base Sequence ; Blotting, Western ; Bone Marrow Cells ; metabolism ; Cartilage, Articular ; pathology ; surgery ; Cell Differentiation ; genetics ; Cells, Cultured ; Chondrocytes ; metabolism ; Chondrogenesis ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Matrix Metalloproteinases ; genetics ; metabolism ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; metabolism ; Microscopy, Fluorescence ; Molecular Sequence Data ; Osteoarthritis ; surgery ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Transforming Growth Factor beta3 ; genetics ; metabolism ; Treatment Outcome

Alveolar Process ; physiology ; Animals ; Biocompatible Materials ; therapeutic use ; Bone Matrix ; transplantation ; Bone Regeneration ; physiology ; Bone Transplantation ; methods ; Dental Implantation ; methods ; Esthetics, Dental ; Guided Tissue Regeneration, Periodontal ; methods ; Humans ; Prosthodontics ; methods ; Tooth Extraction ; methods

OBJECTIVETo evaluate the effect of maxillary sinus elevation with gene-enhanced tissue engineering bone in dogs.

METHODSbone marrow stromal cells (BMSC) derived from dog marrow were cultured, and transduced with the adenovirus carrying bone morphogenetic protein-2 (BMP-2) gene (AdBMP-2), the adenovirus carrying green fluorescent protein (GFP) gene (AdGFP) in vitro. The bone formation ability of gene modified BMSC with scaffold was examined in nude mice and in elevated maxillary sinus of dog. Student's t-test was used for statistical analysis with SPSS 11.0 software package.

RESULTSGene transfection efficiency reached up to (83.95 ± 2.43)% as demonstrated by GFP expression. Ectopic bone formation was detected in nude mice. As for maxillary sinus floor elevation in a dog model, new bone formation area in the AdBMP-2 gene transduced BMSC with Bio-Oss group was significantly higher than in BMSC with Bio-Oss group at 120 d (P < 0.05).

CONCLUSIONSAdBMP-2 gene transduced BMSC can stimulate ectopic bone formation in nude mice, and promote bone formation and maturation in the dog maxillary sinus.

Adenoviridae ; genetics ; Animals ; Bone Matrix ; transplantation ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Bone Transplantation ; methods ; Cells, Cultured ; Dogs ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Nude ; Minerals ; Osteogenesis ; Recombinant Proteins ; genetics ; metabolism ; Sinus Floor Augmentation ; methods ; Tissue Engineering ; methods ; Transfection

BACKGROUND: Injectable calcium sulfate is a clinically proven osteoconductive biomaterial, and it is an injectable, resorbable and semi-structural bone graft material. The purpose of this study was to validate the clinical outcomes of injectable calcium sulfate (ICS) grafts as compared with those of a demineralized bone matrix (DBM)-based graft for filling in contained bony defects created by tumor surgery. METHODS: Fifty-six patients (41 males and 15 females) with various bone tumors and who were surgically treated between September 2003 and October 2007 were included for this study. The patients were randomly allocated into two groups, and either an ICS graft (28 patients) or a DBM-based graft (28 patients) was implanted into each contained defect that was developed by the surgery. The radiographic outcomes were compared between the two groups and various clinical factors were included for the statistical analysis. RESULTS: When one case with early postoperative pathologic fracture in the DBM group was excluded, the overall success rates of the ICS and DBM grafting were 85.7% (24/28) and 88.9% (24/27) (p > 0.05), respectively. The average time to complete healing was 17.3 weeks in the ICS group and 14.9 weeks in the DBM group (p > 0.05). Additionally, the ICS was completely resorbed within 3 months, except for one case. CONCLUSIONS: Although the rate of resorption of ICS is a concern, the injectable calcium sulfate appears to be a comparable bone graft substitute for a DBM-based graft, with a lower cost, for the treatment of the bone defects created during surgery for various bone tumors.
Absorbable Implants ; Adolescent ; Adult ; Biocompatible Materials/*administration & dosage ; Bone Demineralization Technique ; Bone Matrix/*transplantation ; Bone Neoplasms/radiography/surgery/*therapy ; Bone Substitutes/*administration & dosage ; Calcium Sulfate/*administration & dosage ; Child ; Child, Preschool ; Curettage ; Female ; Humans ; Infant ; Injections ; Male ; Middle Aged ; Wound Healing ; Young Adult