• Journal of Periodontal and Implant Science
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• v.36 no.2
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• pp.385-396
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• 2006

For the regeneration of periodontal tissues, the microenvironment for new attachment of connective tissue fibers should be provided, At this point of view, cementum formation in root surface plays a key role for this new attachment. This study was performed to figure out which factor promotes differentiation of cementoblast Considering anatomical structure of tooth, we selected the cells which may affect the differentiation of cementoblast - Ameloblast, OD11&MDPC23 for odontoblasts, NIH3T3 for fibroblsts and MG63 for osteoblasts. And OCCM30 was selected for cementoblast cell line. Then, the cell lines were cultured respectively and transferred the conditioned media to OCCM30. To evaluate the result, Alizarin red S stain was proceeded for evaluation of mineralization. The subjected mRNA genes are bone sialoprotein(BSP), alkaline phosphate(ALP) , osteocalcin(OC), type I collagen(Col I), osteonectin(SPARC ; secreted protein acidic and rich in cysteine). Expression of the gene were analysed by RT-PCR, The results were as follows: 1. For alizarin red S staining, control OCCM30 didn't show any mineralized red nodules until 14 days. But red nodules started to appear from about 4 days in MDPC-OCCM30 & OD11-OCCM30. 2. For results of RT-PCR, ESP mRNAs of control-OCCM30 and others were expressed from 14 days, but in MDPC23-OCCM30 & OD11-OCCM30 from 4 days. Like this, the gene expression of MDPC23-OCCM30 & OD11-OCCM30 were detected much earlier than others. 3. For confirmation of odontoblast effect on cementoblast, conditioned media of osteoblasts(MG63) which is mineralized by producing matrix vesicles didn't affect on the mineralized nodule formation of cementoblasts(OCCM30). This suggest the possibility that cementoblast mineralization is regulated by specific factor in dentin matrix protein rather than matrix vesicles. Therefore, we proved that the dentin/odontoblast promotes differentiation/mineralization of cementoblasts. This new approach might hole promise as diverse possibilities for the regeneration of tissues after periodontal disease.

• Journal of Periodontal and Implant Science
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• v.26 no.3
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• pp.771-778
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• 1996

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.

• The korean journal of orthodontics
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• v.32 no.6 s.95
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• pp.455-466
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• 2002

The objective of this study was to evaluate the changes that occurred over time in the distracted periodontal ligament space following the rapid retraction of a tooth by periodontal distraction after bone undermining surgery had been conducted in the dogs. The upper second premolars were extracted on the left and right side in 4 male beagle dogs. Immediately after extraction, the interseptal bone distal to the upper first premolar was thinned and undermined by grooving to decrease the bone resistance. Activating an individualized distraction appliance at the rate of 0.225mm twice a day, the upper first premolar was retracted rapidly toward the extraction socket. Periodontal distractions were performed for 5, 10, and 20 days, and 20-day-distraction cases were followed by maintenance periods of 0, 14, 28, and 56 days. After 20 days of rapid retraction, the average distal movement of the upper first premolar was 5.02mm, and the average mesial movement of the upper third premolars serving as an anchorage unit was 0.18 mm. On histological examination, the regeneration of bone occurred in a highly organized pattern. Distracted periodontal ligament space was filled with newly formed bone oriented in the direction of the distraction, and this was followed by extensive bone remodeling. This result was similar to those observed in other bones after distraction osteogenesis. In the periodontal ligament, the relationship between collagen fibers and cementum began to be restored 2 weeks after the distraction was completed, and showed almost normal features 8weeks after the completion of the periodontal distraction. However, on the alveolar side, the new bone formation was still in process and collagen fiber bundles and Sharpey's fibers were not present 8 weeks after the completion of the periodontal distraction. Reactions in the periodontal ligament of the anchorage tooth represented bone resorption on the compressed side and new bone deposition on the tension side as occurred in conventional orthodontic tooth movement. In conclusion, the results of this study showed that periodontal structures on the distracted side of the periodontal ligament were regenerated well histologically following rapid tooth movement.

• Journal of Periodontal and Implant Science
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• v.37 no.1
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• pp.65-75
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• 2007

In this study, author examined the effect of the concentration of the inorganic polyphosphate on the process of the bone regeneration by using the 6 weeks old rabbit with the weight of 2.0kg in average. we performed the experiment by using TR-eITFE membrane filled with collagen immersed with 1%, 2%, and 4% of inorganic polyphosphate, respectively, after removing the proper sized cort-ical bones from the calvaria of rabbit. The experimental results were compared with the one of the following four groups: The control group for membrane only, experimental group I for membrane filled with collagen im-mersed with 1% of inorganic polyphosphate, experimental group II for membrane filled with collagen immerse with 2% of inorganic polyphosphate, experimental group III for membrane filled with colla-gen immersed with 4% of inorganic polyphosphate. The fragments of the tissue with membrane were obtained from each group of the sacrificed rab-bits for 4 or 8 weeks sustained after surgery, were then prestained and coated. New bone formation was assessed by histomorphometric and statistical analysis. We may draw the conclusions from these experiments as following: 1. Collagen was an excellent carrier with a minimal inflammatory reaction and sustaining the form. 2. The sample of the 8th week group has shown the best bone regeneration compared with the cases of all groups including the control group. 3. The samples of collagen immersed with 2% and 4% of inorganic polyphosphate have shown more bone regeneration relative to the sample of the 1% inorganic polyphosphate. 4. The new bone regeneration was shown actively in the group for membrane filled with collagen immersed with 4% of inorganic polyphosphate. With above results, it is strongly suggested the use of inorganic polyphosphate with vehicle under TR-eITFE membrane.

• Journal of Periodontal and Implant Science
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• v.38 no.1
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• pp.59-66
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• 2008

Purpose: It has been shown that the inorganic polyphosphate is effective for the regeneration of bones through the preliminary animal test of rabbits. The most effective concentration of the polyphosphate, however, is not known yet. Moreover, the effectiveness of carriers inside human body is not confirmed.. Materials and Methods: In this study, we examined the effect of the concentration of the inorganic polyphosphate on the process of the bone regeneration using the 6 weeks old rabbits with the weight of 2.0 kg in average. We performed the experiment using TR-ePTFE membrane(membrane) filled with collagen immersed in 4%, 8% of inorganic polyphosphate, respectively, following removal of the proper sized cortical bones from the rabbit calvaria. The experimental results were compared with the one of the following four groups: The negative control group for membrane only, the positive control group for membrane filled with collagen, the first experimental group for membrane filled with collagen immersed in 4% of inorganic polyphosphate, and the second experimental group for membrane filled with collagen immerse in 8% of inorganic polyphosphate. The fragments of the tissue with membrane obtained from each group of the sacrificed rabbits for 8 or 16 weeks sustained after surgery were then prestained by the Hematoxylin-Eosin stain and coated by resin to form non-decalcified specimens for the histologic examination and analysis. New bone formation was assessed by histomorphometric and statistical analysis. Results: 1. All groups have shown better bone regeneration at 16weeks than 8weeks. 2. Negative control group has shown more bone regeneration relative to the other groups at 8 and 16 weeks. 3. All experimental groups have shown better bone regeneration relative to positive control group. 4. At 16 weeks, the first experimental group has shown more bone regeneration compared to the second experimental group. Exophytic bone formation is not good at the first and the second experimental groups compared with negative control group. But, the use of 4% inorganic polyphosphate was more effective to bone formation than the use of 8% inorganic polyphosphate. Conclusion: With above results, it is suggested the use of inorganic polyphosphate with vehicle under TR-ePTFE membrane.

• Journal of Periodontal and Implant Science
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• v.37 no.2
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• pp.237-249
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• 2007

This study was performed to evaluate the effect of membrane exposure on new bone formation when guided bone regeneration with perforated titanium membrane on atrophic alveolar ridge. The present study attempted to establish a GBR model for four adult beagle dog premolar. Intra-marrow penetration defects were created on the alveolar ridge(twelve weeks after extraction) on the mandibular premolar teeth in the beagle dogs. Space providing perforated titanium membrane with various graft material were implanted to provide for GBR. The graft material were demineralized bovine bone(DBB), Irradiated cancellous bone(ICB) and demineralized human bone powder(DFDB). The gingival flap were advanced to cover the membranes and sutured. Seven sites experienced wound failure within 2-3weeks postsurgery resulting in membrane exposure. The animals were euthanized at 4 weeks postsurgery for histologic and histometric analysis. The results of this study were as follows: 1. There was little new bone formation at 4 weeks postsurgery. irrespectively of membrane exposure. 2. There was significant relationship between membrane exposure and bone graft resorption(P<0.05), but no relation between membrane exposure and infiltrated connective tissue. 3. There was much bone graft resorption on DFDB than ICB and DBB. 4. The less exposure was on the perforated titanium membrane, the more dense infiltrated connective tissue was filled under the membrane when grafted with ICB and DBB. but there was no relationship between the rate of membrane exposure and the percentage of infiltrated connective tissue area and no relationship between the percentage of the area in the infiltrated connective tissue and in the residual bone graft. Within the above results, bone formation may be inhibited when membrane was exposed and ICB and DBB were more effective than DFDB as a bone graft material when guided bone regeneration.

• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.483-509
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• 1999

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.

• Journal of Periodontal and Implant Science
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• v.29 no.2
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• pp.355-373
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• 1999

Many researches have been reported that collagen as cellular stroma, matrix of grafting materials, mediator of agents for the purpose of promoting healing process invivo, but the responses in vivo were seen various. The goal of this experiment is to assess the effect of collagen on bony healing, through histological evaluation of implanted collagen on the calvarial defect in rats. 2-month-old Sprague-Dawley, 24 rats were used and 12 rats assigned to each group of control and test. Defect of 5mm in diameter was made on the calvarial bone with trephine bur. Following thorough saline rinse, defect of control group was left in empty and that of experimental group was filled with fibrillar collagen($COLLATAPE^{(R)}$, COLLA-TEC. INC. U.S.A.) soaked in saline. 3 rats in each group were sacrificed at 3, 7, 14, 21 days after operation respectively, and the tissue blocks were prepared for light microscope with H-E for evaluation of overall healing, with TRAP(tartrate resistant acid phosphatase) for evaluation of osteoclastic activity and with immunohistochemical staining for macrophages. The results were as follows : 1. In the control group, inflammatory responses were disappeared at day 14, but, in the experimental group inflammatory infiltrates were reduced at day 21. Thus, the experimental group showed more severe soft tissue inflammation than control group. 2. Both control and experimental group showed slight appositional growth at day 7 and gradual bony growth to 21th day. But, complete bony healing of the defect was not shown. There was no significant difference in bony healing between control and experimental group 3. Specific response of macrophages for implanted collagen was observed at day 14 in the experimental group. In conclusion, although fibrillar collagen caused inflammation of soft tissue during initial healing period, inflammatory responses by fibrillar collagen didn't inhibit bony regeneration and implanted collagen was biodegradaded by macrophages. Thus, we expect that fibrillar collagen can be used for useful mediator of graft materials or growth factors.

• Journal of Periodontal and Implant Science
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• v.47 no.4
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• pp.194-210
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• 2017

Purpose: The aim of the present study was to evaluate the healing of post-extraction sockets following alveolar ridge preservation clinically, radiologically, and histologically. Methods: Overall, 7 extraction sockets in 7 patients were grafted with demineralised bovine bone mineral and covered with a porcine-derived non-crosslinked collagen matrix (CM). Soft tissue healing was clinically evaluated on the basis of a specific healing index. Horizontal and vertical ridge dimensional changes were assessed clinically and radiographically at baseline and 6 months after implant placement. For histological and histomorphometric analysis, bone biopsies were harvested from the augmented sites during implant surgery 6 months after the socket preservation procedure. Results: Clinically, healing proceeded uneventfully in all the sockets. A trend towards reduced horizontal and vertical socket dimensions was observed from baseline to the final examination. The mean width and height of resorption were 1.21 mm (P=0.005) and 0.46 mm (P=0.004), respectively. Histologically, residual xenograft particles ($31.97%{\pm}3.52%$) were surrounded by either newly formed bone ($16.02%{\pm}7.06%$) or connective tissue ($50.67%{\pm}8.42%$) without fibrous encapsulation. The CM underwent a physiological substitution process in favour of well-vascularised collagen-rich connective tissue. Conclusions: Socket preservation using demineralised bovine bone mineral in combination with CM provided stable dimensional changes of the alveolar ridge associated with good reepithelialisation of the soft tissues during a 6-month healing period.

• Journal of Periodontal and Implant Science
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• v.37 no.4
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• pp.655-669
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• 2007

Reactive oxygen species (ROS) have been implicated in the pathogenesis of various diseases. And vitamin C has shown a protective effect for the tissues. The aim of this study was to evaluate the effects of $H_2O_2$ and ascorbic acid on matrix metalloproteinase-1 (MMP-1), tissue inhibitor of metalloproteinase (TIMP: TIMP-1, TIMP-2), Type 1 collagen, fibronectin, and PDLs22 level in human periodontal ligament fibroblasts (hPDLF) via reverse transcription-polymerase chain reaction (RT-PCR). hPDLF was obtained from a healthy periodontium and cultured in Dulbecco's modified Eagles's medium plus 10% fetal bone serum. The concentration of ascorbic acid in hPDLF was $50{\mu}g/ml$, and that of $H_2O_2$ in hPDLF was 0.03% and 0.00003%. Ascorbic acid only, $H_2O_2$ only and mixture of ascorbic acid and $H_2O_2$ were applied with hPDLF for 1-, 3-, and 30-min. respectively. The gene expression of MMP-1-, TIMP-1-, TIMP-2-, Type 1 collagen-, fibronectin-, and PDLs22-mRNA in hPDLF was analysed via RT-PCR. The results were as follows; 1. hPDLF in response to 30-min. incubation with 0.03% $H_2O_2$ did not show any gene expression. 2. In all the experimental groups, the gene expression of fibronectin mRNA showed the decreased tendency compared to control. 3. In all the experimental groups, the gene expression of TIMP-1 mRNA showed the tendency similar to control. 4. hPDLF in response to 30-min. incubation with 0.03% $H_2O_2$ and ascorbic acid increased mRNA induction for MMP-1. 5. In all the experimental groups, hPDLF increased mRNA induction for PDLs22, collagen type 1, and TIMP-2 compared to control. Within the limited experiments, $H_2O_2$ and ascorbic acid increased mRNA induction for PDLs22, collagen type 1, TIMP-2 in hPDLF. More research will be needed in order to confirm the relative importance of the different roles of ROS and antioxidants in hPDLF from a periodontal regeneration or repair standpoint.