Porous matrices of bioactive polymers such as polyglycolic acid (PGA) or polylactic acid (PLA) can be used as scaffolds in bone tissue growth during bone repair process. These polymers are highly porous and serve as a template for the growth and organization of new bone tissues. We evaluated the effect of PGA and PLA polymers on osteoblastic MC3T3-E1 cell extracellular mineralization. MC3T3-E1 cells were cultured in a time-dependent manner -1, 15, 25d as appropriate - for the period of bone formation stages in one of the five culture circumstances, such as normal osteogenic differentiation medium, PGA-plated, fetal bovine serum (FBS)-plated, PGA/FBS-coplated, and PLA-plated For the evaluation of bone formation, minerals (Ca, Mg, Mn) and alkaline phosphatase activity, a marker for osteoblast differentiation, were measured Alizarin Red staining was used for the measurement of extracellular matrix Ca deposit During the culture period, PGA-plated one was reabsorbed into the medium more easily and faster than the PLA-plated one. At day 15, at the middle stage of bone formation, cellular Ca and Mg levels showed higher tendency in PGA- or PLA-plated treatments compared to non-plated control and at day 25, at the early late stage of bone formation, all three cellular Ca, Mg or Mn levels showed higher tendency as in order of PGA-related treatments and PLA-plated treatments, compared to control even without significance. Medium Ca, Mg or Mn levels didn't show any consistent tendency. Cellular ALP activity was higher in the PGA- or PLA-plated treatments compare to normal osteogenic medium treatment PGA-plated and PGA/FBS-plated treatments showed better Ca deposits than other treatments by measurement of Alizarin Red staining, although PLA-plated treatment also showed reasonable Ca deposit. The results of the present study suggest that biodegradable material, PGA and also with less extent for PLA, can be used as a biomaterial for better extracellular matrix mineralization in osteoblastic MC3T3-E1 cells.
Sinus lift procedure is frequently required for the maxillary molar implant placement. Previous studies have demonstrated alveolar ridge preservation (ARP) can maintain the dimensions of ridge height and width. However, there is a lack of studies which evaluated the effect of ARP to avoid sinus lift procedure. Purpose of this study is to describe a method reducing the need of sinus lift surgery by ARP in maxillary molar areas and to assess the feasibility clinically, radiologically and histologically. Ten maxillary molars in ten patients had severe vertical bone resorption with minimal residual bone height. They were considered having the high possibility of the necessity of sinus lift procedure for dental implant after the extraction. After extraction, open healing ARP with deproteinized bovine bone mineral mixed with 10% collagen and resorbable collagen membranes was performed. After sufficient healing, dental implants were placed, and evaluated clinically and radiologically. Histological observation was conducted just before the implantation in one patient. Implants were successfully placed without sinus lift in all ten cases. All the implants were restored with no sign of complications, and patients are now in a close follow-up up to 20 months post-loading. Histological observation showed minimal inflammatory reaction and newly formed bone was substantially noted. The ARP technique has successfully avoided the sinus lift surgeries. It appears that this procedure may improve the simplicity of the clinical process for the clinicians and reduce the discomfort of patients.
Purpose: Rehabilitation of the edentulous posterior maxilla with dental implants often poses difficulty because of insufficient bone volume caused by pneumatization of the maxillary sinus and by crestal bone resorption. Sinus grafting technique was developed to increase the vertical height to overcome this problem. The present study was designed to evaluate the sinus floor augmentation with anorganic bovine bone (Bio-$cera^{TM}$) using histomorphometric and clinical measures. Patients and methods: Thirteen patients were involved in this study and underwent total 14 sinus lift procedures. Residual bone height was ${\geq}2mm$ and ${\leq}6mm$. Lateral window approach was used, with grafting using Bio-$cera^{TM}$ only(n=1) or mixed with autogenous bone from ramus and/or maxillary tuberosity(n=13). After 6 months of healing, implant sites were created with 3mm diameter trephine and biopsies taken for histomorphometric analysis. The parameters assessed were area fraction of new bone, graft material and connective tissue. Immediate and 6 months after grafting surgery, and 6 months after implantation, computed tomography (CT) was taken and the sinus graft was evaluated morphometric analysis. After implant installation at the grafted area, the clinical outcome was checked. Results: Histomorphometry was done in ten patients.Bio-$cera^{TM}$ particles were surrounded by newly formed bone. The graft particles and newly formed bone were surrounded by connective tissue including small capillaries in some fields. Imaging processing revealed $24.86{\pm}7.59%$ of new bone, $38.20{\pm}13.19%$ connective tissue, and $36.92{\pm}14.51%$ of remaining Bio-$cera^{TM}$ particles. All grafted sites received an implant, and in all cases sufficient bone height was achieved to install implants. The increase in ridge height was about $15.9{\pm}1.8mm$ immediately after operation (from 13mm to 19mm). After 6 months operation, ridge height was reduced about $11.5{\pm}13.5%$. After implant installation, average marginal bone loss after 6 months was $0.3{\pm}0.15mm$. Conclusion: Bio-$cera^{TM}$ showed new bone formation similar with Bio-$Oss^{(R)}$ histomorphometrically and appeared to be an effective bone substitute in maxillary sinus augmentation procedure with the residual bone height from 2 to 6mm.
Biodegradable barrier membrane has been demonstrated to have guided bone regeneration capacity on the animal study. The purpose of this study is to evaluate the effects of cultured calvarial cell inoculated on the biodegradable barrier membrane for the regeneration of the artificial bone defect. In this experiment 35 Sprague-Dawley male rats(mean BW 150gm) were used. 30 rats were divided into 3 groups. In group I, defects were covered periosteum without membrane. In group II, defects were repaired using biodegradable barrier membrane. In group III, the defects were repaired using biodegradable barrier membrane seeded with cultured calvarial cell. Every surgical procedure were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). After anesthesia, 5 rats were sacrificed by decapitation to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. The membrane were inserted on the artificial bone defect after 3 days of culture. A single 3-mm diameter full-thickness artificial calvarial defect was made in each animal by using with bone trephine drill. After the every surgical intervention of animal, all of the animals were sacrificed at 1, 2, 3 weeks after surgery by using of perfusion technique. For obtaining histological section, tissues were fixed in 2.5% Glutaraldehyde (0.1M cacodylate buffer, pH 7.2) and Karnovsky's fixative solution, and decalcified with 0.1M disodium ethylene diaminetetraacetate for 3 weeks. Tissue embeding was performed in paraffin and cut parallel to the surface of calvaria. Section in 7${\mu}m$ thickness of tissue was done and stained with Hematoxylin-Eosin. All the specimens were observed under the light microscopy. The following results were obtained. 1 . During the whole period of experiment, fibrous connective tissue was revealed at 1week after surgery which meant rapid soft tissue recovery. The healing rate of defected area into new bone formation of the test group was observed more rapid tendency than other two groups. 2 . The sequence of healing rate of bone defected area was as follows ; test group, positive control, negative control group. 3 . During the experiment, an osteoclastic cell around preexisted bone was not found. New bone formation was originated from the periphery of the remaing bone wall, and gradually extended into central portion of the bone defect. 4 . The biodegradable barrier membrane was observed favorable biocompatibility during this experimental period without any other noticeable foreign body reaction. And mineralization in the newly formed osteoid tissue revealed relatively more rapid than other group since early stage of the healing process. Conclusively, the cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of regeneration of artificial bone defects of alveolar bone. This study thus demonstrates a tissue-engineering the approach to the repair of bone defects, which may have clinical applications in clinical fields of the dentistry including periodontics.
Purpose: To compare the diagnostic accuracy of cone-beam computed tomography (CBCT) with that of parallel(PPA) and oblique projected periapical(OPA) radiography for the detection of different types of peri-implant bone defects. Materials and Methods: Forty implants inserted into bovine rib blocks were used. Thirty had standardized bone defects(10 each of angular, fenestration, and dehiscence defects), and 10 were defect-free controls. CBCT, PPA, and OPA images of the samples were acquired. The images were evaluated twice by each of 2 blinded observers regarding the presence or absence and the type of the defects. The area under the receiver operating characteristic curve (AUC), sensitivity, and specificity were determined for each radiographic technique. The 3 modalities were compared using the Fisher exact and chi-square tests, with P<0.05 considered as statistical significance. Results: High inter-examiner reliability was observed for the 3 techniques. Angular defects were detected with high sensitivity and specificity by all 3 modalities. CBCT and OPA showed similar AUC and sensitivity in the detection of fenestration defects. In the identification of dehiscence defects, CBCT showed the highest sensitivity, followed by OPA and PPA, respectively. CBCT and OPA had a significantly greater ability than PPA to detect fenestration and dehiscence defects(P<0.05). Conclusion: The application of OPA radiography in addition to routine PPA imaging as a radiographic follow-up method for dental implantation greatly enhances the visualization of fenestration and dehiscence defects. CBCT properly depicted all defect types studied, but it involves a relatively high dose of radiation and cost.
Fernandez, Maria;Martinez, Adela;Modrono, Sagrario;De La Roza, Begona
Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
/
2001.06a
/
pp.1269-1269
/
2001
The Bovine Spongiform Encephalopathy (BSE) is one of the more important problems that have affected the economy of european cattles and the Public Safety. Their transmission is mainly through digestive system, and the compound feeds made with animal proteins are one source of infection for healthy cows. Nowadays the official method for meat and bone meal (MBM) detection in compound feeds is a microscopy technique. However, this methodology is subjective, and that alter the fact to make one exhaustive quantitative analysis and one differentiation between mammalian and poultry bones. In addition, the separation of the differents fractions in a sample by density before the analysis, requires the use of organochlorates products as $CCl_4$, which produce serious damages in the atmosphere ozone content. NIR methodology is another possible way to confirm and identifying animal ingredients in compound feeds, Its capabilities for quantitative and qualitative analysis of foods and feeds has been enought demonstrated. The objective of this work was to use NIR as a tool to make an qualitative and quantitative analysis and a prediction of the meat and bone meal presence in compound feeds from North Spain cattle farms. Using a global population of compound feeds, on make three different groups depending of MBM percentage presence (0, 0-100, 100), to build and validate one calibration equation to determine MBM content and make one discriminant analysis between these three groups. The preliminary dates obtained with another differents samples of known composition showed promising results.
Purpose: The present study investigated the impact of 2 different suture techniques, the conventional crossed mattress suture (X suture) and the novel hidden X suture, for alveolar ridge preservation (ARP) with an open healing approach. Methods: This study was a prospective randomized controlled clinical trial. Fourteen patients requiring extraction of the maxillary or mandibular posterior teeth were enrolled and allocated into 2 groups. After extraction, demineralized bovine bone matrix mixed with 10% collagen (DBBM-C) was grafted and the socket was covered by porcine collagen membrane in a double-layer fashion. No attempt to obtain primary closure was made. The hidden X suture and conventional X suture techniques were performed in the test and control groups, respectively. Cone-beam computed tomographic (CBCT) images were taken immediately after the graft procedure and before implant surgery 4 months later. Additionally, the change in the mucogingival junction (MGJ) position was measured and was compared after extraction, after suturing, and 4 months after the operation. Results: All sites healed without any complications. Clinical evaluations showed that the MGJ line shifted to the lingual side immediately after the application of the X suture by $1.56{\pm}0.90mm$ in the control group, while the application of the hidden X suture rather pushed the MGJ line slightly to the buccal side by $0.25{\pm}0.66mm$. It was demonstrated that the amount of keratinized tissue (KT) preserved on the buccal side was significantly greater in the hidden X suture group 4 months after the procedure (P<0.05). Radiographic analysis showed that the hidden X suture had a significant effect in preserving horizontal width and minimizing vertical reduction in comparison to X suture (P<0.05). Conclusions: Our study provided clinical and radiographic verification of the efficacy of the hidden X suture in preserving the width of KT and the dimensions of the alveolar ridge after ARP.
Purpose: This study examined the biomechanical stability of four different plating techniques in the experimental model of mandibular subcondyle fracture. Methods: Twenty standardized bovine tibia bone samples ($7{\times}1.5{\times}1.0cm$) were used for this study. Each of the four sets of tibia bone was cut to mimic a perpendicular subcondyle fracture in the center area. The osteotomized tibia bone was fixed using one of four different fixation groups (A,B,C,D). The fixation systems included single 2.0 mm 4 hole mini adaption plate (A), single 2.0 mm 4 hole dynamic compression miniplate (B), double fixation with 2.0 mm 4 hole mini adaption plate (C), double fixation with a 2.0 mm 4 hole mini adaption plate and 2.0 mm 4 hole dynamic compression miniplate (D). A bending force was applied to the experimental model using a pressure machine (858 table top system, $MTS^{(R)}$) until failure occurred. The load for permanent deformation, maximum load of failure were measured in the load displacement curve with the chart recorder. Results: Double fixation with a 2.0 mm 4 hole mini adaption plate and a 2.0 mm 4 hole dynamic compression miniplate (D) applied to the anterior and posterior regions of the subcondyle experimental model showed the highest load to failure. Conclusion: From this study, double fixation with an adaption plate and dynamic compression miniplate fixation technique produced the greatest biomechanical stability. This technique may be considered a useful means of fixation to reduce the postoperative internal maxillary fixation period and achieve early mobility of the jaw.
The ultimate aim of periodontal treatment is periodontal regeneration, which necessiates the regeneration of bone tissues. This paper investigated the effect of growth factor on bone cells. Platelet-derived growth factor(PDGF) is the one of the polypeptide growth factor that has been reported as a biological mediator which regulates activities of the cell proliferation, migration and metabolism of undifferentiated mesenchymal cells. The purpose of this study is to evaluate the effects of PDGF on bone nodule formation and ALP activity of MC3T3-El cells. Cells were seeded at $1{\times}10^5cells/well$ in alpha-modified eagle medium containing 10% fetal bovine serum, lOml beta-glycerophosphate and $50{\mu}g/ml$ of ascorbic acid. PDGF 0, 0.1, 1, 10 ng/ml were added to the cells at a confluent state and cultured for 3, 7, 14, 21, 28 days. We examined bone nodule formation and alkaline phosphatase activity. The results were as follows : There were bone nodule formation at day 21 both in control and all the experimental groups, and at day 28, all the experimental groups showed much more bone nodules than control groups. Compared to control-l group, ALP activity was increased in PDGF O.1ng/ml group and was decreased in 1,10ng/ml PDGF treated groups.{P< 0.05, P< 0.01) Compared to control-2, ALP activity was decreased in all the experimental groups except PDGF 0.1ng/ml in 21 day group. In the time-response effect, ALP activity was increased by the day 14 in all the experimental groups and thereafter ALP activity was decreased.(P<0.05, P< 0.01) In the dose-response effect, ALP activity was decreased as the dose of PDGF was increased, and after 21 day ALP activity was lowest in 1 ng/ml group, ALP activity was highest in the day 7 in control group and 0.1 ng/ml, 14 day experimental group. In conclusion, PDGF is considered more effective in the proliferation than differentiation of osteoblast-like cells, and it may be useful to study the combined effect of PDGF and other growth factors on osteoblast-like cells.
Bone morphogenetic protein-2/4 (BMP-2/4) are members of Transforming Growth $Factor-{\beta}\;(TGF-{\beta})$ superfamily and they may differentiate the osteoprogenitor cell and induce formation of cartilage and bone in vivo. This study was performed to investigate the effects of bone morphogenetic protein-2/4 on the characteristics of rat periodontal ligament cells(RPDL) and rat calvaria cells(RCV). In the control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 20% fetal bovine serum, $100{\mu}/ml$ penicillin, $100{\mu}/ml$ streptomycin. In the experimental groups, recombinant human bone morphogenetic protein-2/4 (25ng, 100ng, 250ng/ml) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 2, 3, 5, 7th day, the amount of total protein synthesis and alkaline phosphatase activity at 2, 5, 7th day. And also, the calcified nodule was examed. The results were as follows ; 1 . Both RCV and RPDL cells in both control and experimental groups proliferated during the entire experimental period, but there is no stastically significant difference according to the BMP-2/4 concentration. 2 . Amount of total protein synthesis of both cells in both groups was steadily increased until 5th day, but all experimental groups were significantly different from the control group at 7th day. 3. Alkaline phosphatase activity of both cells in both groups was increased during the entire experiment period. In RCV cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 7th day. In RPDL cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 5th day, and all experimental groups were significantly different from the control group at 7th day. 4. In the both of the cultured Rat Periodontal ligament and calvaria cell treated with BMP-2/4 to compared with control group, it revealed more rapid cell polarization, cell aggregation and hyperchromatic stained on HE agent, and even though only 1 day treated with BMP-2/4 both RPDL and RCV showed more rapid cell reaction than control group. More sensivitve cell reaction of RCV were observed than RPDL in this experiment. From the above results, we could conclude that BMP-2/4 influenced the induction, proliferation and differentiation of bone forming cells
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