The assessment of alveolar bone changes on dental radiographs to indicate progression of periodontal diseases or healing response to therapy is routine procedure. However, the diagnostic accuracy in detecting small alveolar bone changes is very limited. Recently, guided bone regeneration therapy is popular, but the quantification of new bone is somewhat difficult with conventional evaluation method. To quantificate the amount of new bone, various evaluating methods have been introduced including histomorphometry, radiomorphometry, biochemical analysis, X-ray probe microanalysis, scanning electron microscope backscatter method. In this study, guided bone regeneration using resorbable membrane with & without PDGF-BB is quatificated through histomorphmetry to evaluate the efficacy of histomorphometric analysis. 4 beagle dogs and 8 Sprague-Dawley rats were selected as experimental animals. In beagle dog experiment, $4{\times}4mm$ Class II defects were created in maxillary both second premolars, and biodegradable membrane containing PDGF-BB(experimental group) were covered over one defect, and same membrane without PDGF-BB(control group) were covered over the other defect. At 2 weeks, 5 weeks after surgery, each beagle dogs were sacrificed, and the tissues were treated by undecalcified fixation. In Sprague-Dawley rat experiment, 5mm round defect were created in temporal bone, the same membranes were covered on the defects. At 1 week, 2 weeks after surgery, each rats were sacrificed, and undecalcified fixation were taken. After grinding tissue specimen, we analyse them histomorphometrically using image analysis system. In beagle dog 2 weeks specimens, new bone formation area were $0.03123mm^2$ in experimental group,and $0.03012mm^2$ in control group. At 5 weeks specimens, $0.15324mm^2$ in experimental group, and $0.09123mm^2$ in control group. In Sprague-Dawley rat specimens, new bone fomation area were $0.20448mm^2$ in 1 week experimental group, $0.03604mm^2$ in 1 week control group. At 2 weeks specimens, $0.46349mm^2$ in experimental group, $0.17741mm^2$ in control group. The results indicated that histomorphometric analysis of new bone formation using image analysis system is very effective quantification method to evaluate the efficacy of treatment modalities.
Purpose: This study was performed to evaluate the influence of local application of thymoquinone (TQ) on bone healing in experimental bone defects infected with Porphyromonas gingivalis (PG). Methods: Forty-two female rats were randomly divided into 6 groups. A bone defect was created on the right tibia of all animals. The PG, PG/collagen membrane (COL) and PG/TQ/COL groups were infected with PG. In the COL and PG/COL groups, the defects were covered with a COL; in the TQ/COL and PG/TQ/COL groups, the defects were covered with a TQ-containing COL. After 28 days, all animals were sacrificed. Quantitative measurements of new bone formation and osteoblast lining, as well as semiquantitative measurements of capillary density and tissue response, were analyzed. Furthermore, the presence of bacterial infections in defect areas was evaluated. Results: The new bone formation, osteoblast number, and capillary density were significantly higher in the TQ groups than in the control groups (P<0.001, P<0.001, and P<0.01, respectively). In a comparison between the TQ/COL group, with a TQ-containing COL (TQ/COL), and the PG-infected TQ-containing COL (PG/TQ/COL) group, the newly formed bone and capillary density were higher in the TQ/COL group (P<0.01). When the control group was compared to the PG, PG/COL, and PG/TQ/COL groups in terms of tissue response, the differences were statistically significant (P<0.001, P=0.02, and P=0.041, respectively). The intensity of the inflammatory cell reaction was higher in the PG, PG/COL, and PG/TQ/COL groups (P<0.05). Conclusions: Within the limitations of this study, the local application of a TQ-containing COL positively affected bone healing even if the bone defects were infected. The results suggest that TQ increased angiogenesis and showed promise for accelerating bone defect healing. Further research is warranted to support these findings and reach more definitive conclusions.
Several experimental studies showed that the application of small amounts of electric current to bone stimulated osteogenesis at the site of the cathode and suggests that the application of electrical currents to periodontal defects could promote bone and cementum formation. The purpose of this study was to determine the effect of direct microcurrent to the periodontal regeneration of class III furcation defects in dogs. Class III furcation defects were surgically created on the third and the fourth premolars bilaterally in the mandibles of nine mongrel dogs. Experimental periodontitis were induced by placing small cotton pellets into the created defects for 3 weeks. The experimental sites were divided into three groups according to the treatment modalities: Group I-surgical debridement only; Group II-allogenic demineralized freeze dried bone grafting; Group III-allogenic demineralized freeze dried bone grafting and electrical stimulation. For fluorescence microscopic evaluation, calcein, oxytetracycline HCI and alizarin red were injected 2, 4 and 8 weeksfS days prior to sacrifice) after surgery. The animals were sacrificed in the 1st, 2nd, 4th and 8th week after periodontal surgery and the decalcified and undecalcified specimens were prepared for histological and histometrical examination. After the first and the second weeks, gingival recession was more severe in group I than groups II and III. After the fourth and the eighth weeks, there was no difference in the width of junctional epithelium and connective tissue attachment among the three groups, but the width of connective tissue attachment increased in group II at the eighth week, compared to the fourth week. The amount of bone repair in new attachment was significantly greater in group III, compared to groups I and II. New attachment formation was significantly greater in group III, compared to groups I and group II. These results suggest that electrical stimulation using microcurrent generator could be a useful tool for periodontal regenerative therapy in class III furcation defect.
Procedures for treatment of molar furcation invasion defects range from open flap debridement, apically repositioned flap surgery, hemisection, tunneling or extraction, to regenerative therapies using bone grafting or guided tissue regenerative therapy, or a combination of both. Several clinical evaluations using regenerative techniques have reported the potential for osseous repair of treated furcation invasions. Regenerative treatment of maxillary molars are more difficult due to the multiple root anatomy and multiple furcation entrances therefore, purpose of this study was to evaluated histologically self-curing glass-ionomer cement and light-curing glass-ionomer cement as a barrier in the treatment of a bi-furcated maxillary premolar. Five adult beagle dogs were used in this experiment. With intrasulcular and crestal incision, mucoperiosteal flap was elevated. Following decortication with 1/2 high speed round bur, degree II furcation defect was made on maxillary third(P3), forth(P4) and fifth(P5) premolar. 2 month later experimental group were self-curing glassionomer cement and light-curing glassionomer cement. After 4, 8 weeks, the animals were sacrificed by vascular perfusion. Tissue block was excised including the tooth and prepared for light microscope with Gomori's trichrome staining. Results were as follows. 1. In all experiment group, there were not epithelial down growth and glass ionomer cement were encapsulated connective tissue. 2. In 4 weeks experiment I group slighly infiltrated inflammatory cells but not disturb the new bone or new cementum formation. 3. In 8 weeks, experiment groups I, II were encapsulated fine connective tissue. 4. Therefore glass-ionomer cement filling to the grade III maxillary furcations with multiple root anatomy and multiple furcation entrances were possible clinical methods and this technique is useful method for Maxillary furcation involvement.
Nerve allografts as a bridge of regeneration is useful in the repair of peripheral nerve defect resulting from trauma, and leprosy. But immunological rejection and complicated scar formation is an unavoidable problem in the application of allogeneic nerves. This article is intended to study of the regeneration of allogeneic nerve grafts in rats with histopathologically, scanning electron microscopically. 24 adult male Sprague-Dawley rats were used as the experimental animals. A 2cm skin incision was made on the lateral aspects of limb, parallel to femur. Segments of sciatic nerve trunk taken from rats, 10mm was resected at the middle of the thigh, nerve graft was inserted between the ends of gaps with perineural and epineural suture method with 10-0 prolene. Obsrevation was made simultaneously at 3 day, 1, 2, 3, 4, 5, 6, 8 weeks after surgery. The results were as follows. 1. In light and electronic microscopic studies, marked degenerative change of the graft nerves were observed at 2 weeks after surgery. 2. After surgery, blood clot fromation was observed at 3 day, granualtion tissue formation was observed at 2 week, and fibrous tissue proliferation was observed at 3 week. 3. In change of nerve fiber, there were Wallerian degeneration at early stage, decrease in degeneration at 4 week but degeneration of myeline was continuded at 8 week. 4. At 4 week, schwann cells proliferate at its cut ends to join with the distal and proximal stump of the damaged nerve. 5. Fibrous scar tissues are formed at 2 weeks and increased progressively in 8 weeks, which was interrupted the regeneration of grafted nerve.
The purpose of this stuffy was to assess and compare the osseous responses to implanted particles of porous synthetic HA (Interpore $200^{(R)}$, Interpore International, U.S.A.), resorbable natural bovine derived HA (Bio-$oss^{(R)}$, Gestlich Pharma, Switzerland) and calcium carbonate(Biocoral $450^{(R)}$, Inoteb, France) in bone defects. Four calvarial defects of 2.5mm diameter were created in earth of 16 Sprague-Dawley rats. The experimental materials were subsequently implanted hi three defects, leaving the fourth defect for control purpose. Four animals were earth sacrificed at 3 days, 1week, 2weeks and 4 weeks after surgery. The tissue response was evaluated under light microscope. Overall, histologic responses showed that all the particles were well tolerated and caused no aberrent tissue responses. There were difference in the amount of newly formed bone at the experimental sites and control site. There was more new bone formation associated with calcium carbonate site. In addition, the calcium carbonate site displayed multinucleated giant cells surrounding calcium carbonate particles after the 1st week, and osteoid tissue within the particle after the 2nd week. After 4 weeks, calcium carbonate particles were resorbed and replaced with new bone. The healing of the natural bovine derived HA site was similar to that of porous synthetic HA, except that new bone growth between the two particles have progressed more in the former site after the 2nd week. In the natural bovine derived HA site, the particle was surrounded by newly formed bone after the 4th week. After 4 weeks, the control site showed more mature bone than other sites. In conclusion, the grafted site were better in new bone formation than non-grafted sites. In particular the calcium Carbonate site showed the ability of osteoinduction and natural bovine denver HA showed osteoconduction in rat calvarial defects. This suggest that calcium carbonate and natural bovine derived HA could enhance the regenerative potential in periodontal defects.
Purpose: The aim of this study was to determine the clinical feasibility of using dehydrothermally cross-linked collagen membrane (DCM) for bone regeneration around peri-implant dehiscence defects, and compare it with non-cross-linked native collagen membrane (NCM). Methods: Dehiscence defects were investigated in twenty-eight patients. Defect width and height were measured by periodontal probe immediately following implant placement (baseline) and 16 weeks afterward. Membrane manipulation and maintenance were clinically assessed by means of the visual analogue scale score at baseline. Changes in horizontal thickness at 1 mm, 2 mm, and 3 mm below the top of the implant platform and the average bone density were assessed by cone-beam computed tomography at 16 weeks. Degradation of membrane was histologically observed in the soft tissue around the implant prior to re-entry surgery. Results: Five defect sites (two sites in the NCM group and three sites in the DCM group) showed soft-tissue dehiscence defects and membrane exposure during the early healing period, but there were no symptoms or signs of severe complications during the experimental postoperative period. Significant clinical and radiological improvements were found in all parameters with both types of collagen membrane. Partially resorbed membrane leaflets were only observed histologically in the DCM group. Conclusions: These findings suggest that, compared with NCM, DCM has a similar clinical expediency and possesses more stable maintenance properties. Therefore, it could be used effectively in guided bone regeneration around dehiscence-type defects.
Background Many topical hemostatics are widely applied for bleeding control. They can be classified into two categories according to their mechanism of action on the clotting cascade in a biologically active or passive manner. Passive hemostatics include cellulose and gelatin. We performed an experimental study to compare the effect of passive hemostatics in wound healing by applying them to a rectus abdominis muscle defect of white mice. Methods Surgicel is a sterile absorbable knitted fabric prepared by the controlled oxidation of regenerated cellulose. Spongostan is an absorbable hemostatic gelatin sponge. In 30 mice, a $1{\times}1$ cm defect was created on the rectus abdominis muscle and the materials were applied in three ways: control group, cellulose (Surgicel) group, gelatin (Spongostan) group. For the histologic analysis, biopsies were performed at 3 and 28 days. Results After 3 days, the cellulose group showed limited granulation formation with acute inflammatory reactions similar to the control group. At the 28th day, moderate amounts of granulation tissue formation was observed with milder inflammatory reactions than the control group. In the gelatin group, after 3 days, gelatin remnants were observed surrounded by severe inflammatory changes. After 28 days, the same quantity of gelatin remnants could be still observed. Conclusions This study suggests that cellulose is associated with minimal morbidity in wound healing, while the use of gelatin shows severe adverse tissue reactions with delayed wound healing. Consequently, cellulose is better than gelatin when considering wound healing.
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: This study was aimed to evaluate the effect of the deproteinated bovine bone powder (DBBP) coated with calcium phosphate (Ca-P) on osseous regeneration in the calvarial bone defect of rat. Materials and Methods : The DBBP (Control group, n=6) and the Ca-P coated DBBP (Experimental group, n=6) were grafted in the critical sized calvarial bone defect (8 mm) of rat weighing 250 g. The animals were sacrificed at 1, 4 week. The biopsy specimens were decalcified with 5%formaldehyde and embedded in paraffin. The rats were sacrificed at 8 week received tetracycline (1 week), calcein blue (4 week), and alizarin red (7 week), and the biopsy specimens were taken. The specimens were embedded in methylmethacrylate and ground to 10 ${\mu}m$ thin sections were made. All of the specimens were stained with H & E and Masson's trichrome and examined under light microscope. The specimens at 8 week were examined under fluorescent microscope. Results : In the Control group, the grafted DBBP was surrounded with connective tissue, and osteoblasts were observed partially around the grafted particles at 1 week. At 4 week, some osteoid was observed and, new bone formation was observed at the periphery of grafted materials at 8 week, In the Experimental group, some osteoid was seen at the periphery of the grafted Ca-P coated DBBP at 1 week, and osteoblast and newly formed bone were observed around the grafted materials. At 8 week, newly formed bone was observed at the periphery of the grafted materials. Conclusion: These results suggest that Ca-P coated DBBP group was more and faster than DBBP group in new bone formation and Ca-P could contribute to enhance bone formation in the critical sized calvarial bone defect of rat.
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