• Journal of Periodontal and Implant Science
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• 제35권4호
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• pp.823-837
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• 2005

The purpose of this study was to study the histopathological correlation between the use of platelet-rich plasma and enamel matrix protein used in conjunction with xenograft. compared to a control group with regards to bone regeneration at the grade III furcation area in beagle dogs. Control group was treated with bovine derived bone $powder(Biocera^{(R)})$, and experimental I group was treated with bovine derived bone powder and Platelet-rich plasma and experimental II group was treated with bovine derived bone powder and Enamel matrix $protein(Emdogain^{(R)})$. The regeneration rate of bone formation was observed and compared histopathologically at 2. 4, and 8 weeks after surgery. The results were as follows: 1. In control group and both experimental groups. inflammatory cells were observed but, new bone formation wasn't. 2. In control group, new cementum on the notch was found in 4 weeks, less mature periodontal ligament when compared to that of experimental group was found and cementum formation was great but, regeneration couldn't be seen in 8 weeks. 3. Experimental I group. new bone formation in the area adjacent to alveolar bone and graft material surrounded by more dense connective tissue were found in 4 weeks. New bone formation up to crown portion was found and periodontal ligament was aligned functionally and cementum more mature. 4. Experimental II group, new bone formation was found under the defect area in 4 weeks and new bone formation around graft material in 8 weeks, too, and there were a number of fibroblasts, blood vessels, acellular cementum, which was less mature when compared to that of experimental I group, and dense collagen fiber like which normal periodontal ligament has in periodontal ligament of experimental II group in 8 weeks. 5. As a result of histologic finding, bone formation rate were 18.0${\pm}$7.87%(control group), 34. 05${pm}$7.25%(experimental I group), 19.33 ${pm}$5.15%(experimental II group) in 4 weeks and 21.89${pm}$1.58%(control group), 38.82${pm}$3.2(experimental I group), 37.65${pm}$9.22%(experimental II group) in 8 weeks. 6. Statistically significant ratio of bone formation was observed in experimental I group in 4 weeks and in experimental II group in 8 weeks. When experimental I group was compared to experimental II group, the ratio of bone formation in experimental I group was higher than that in experimental II group in 4 weeks(p<0.05). This results suggest that platelet-rich plasma showed more new bone formation than enamel matrix protein within 4 weeks. And use of enamel matrix protein in the treatment of periodontal bone defects starts to enhance regeneration after 8 weeks in beagle dogs.

• Journal of Periodontal and Implant Science
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• 제31권3호
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• pp.611-623
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• 2001

Cyclosporin A is a cyclic polypeptide produced by the metabolism of fungi. It is widely used at present as immunosuppressive treatment following organ transplants. It is also used to deal with autoimmune diseases such as rheumatoid arthritis or type II diabetes. Gingival hyperplasia is one of the most frequent side-effects associated with the prescription of Cyclosporin A. The mechanisms involved in Cyclosporin A induced gingival hyperplasia are not yet clear. In vitro Cyclosporin A promotes proliferation of gingival fibroblasts, that Cyclosporin A act as a mitogen. Its action is based on mitosis of gingival fibroblasts regulated by cell cycle regulatory proteins. It was the purpose of the present study to examine the effects of Cyclosporin A on human gingival fibroblasts by means of biological and biochemical criteria. In this present study, we examined change of cell proliferation, cell activity, cell viability and cell cycle progression after application of Cyclosporin A. We also examined expression of cell cycle regulatory proteins by western blot analysis. Human gingival fibroblasts were cultured for 48 hours with application of Cyclosporin A at concentrations of 0.01, 0.1, 1, and 10 ng/ml. Cyclosporin A(1 ng/ml) significantly increased the cell activity of gingival fibroblast. Proliferation and viability of gingival fibroblasts were also increased in group treated with 1 ng/ml of Cyclosporin A compared to control group. In the cell cycle analysis, S phase was increased and G1 phase was decreased in the group treated with 1 ng/ml of Cyclosporin A. Cyclosporin A increased the expression of cdk4 and inhibited the expression of pRB and p21. These results suggest that 1 ng/ml of Cyclosporin A may increase the cell cycle progression of human gingival fibroblasts, and its mechanisms may increase the expression of cdk4 and decrease the expression of pRB and p21.

• Journal of Periodontal and Implant Science
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• 제31권3호
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• pp.597-610
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• 2001

Normal gingival fibroblasts functioning is fundamental for the maintenance of periodontal connective tissue as well as wound healing. Nicotine have been found to affect DNA synthesis and cell proliferation, which appear to depend on the type of cells. This in vitro study was done to determine the effects of nicotine, a major component of tobacco, on cell proliferation, viability, activity, cell cycle distribution, and expression of cell cycle regulatory proteins in human gingival fibroblasts. Nicotine has been tested for 2 days or 4 days in 5 different concentrations; $0.1{\mu}g/ml$; $1{\mu}g/ml$; $10{\mu}g/ml$; $100{\mu}g/ml$; $1000{\mu}g/ml$. To assess cell proliferation and viability, viable and non-viable cells were counted by hemocytometer; to evaluate cellular activity, MTT assay was employed; to analyze cell cycle distribution, fluorescent propidium iodide-DNA complex were measured using fluorocytometer; to determine the expression of cell cycle regulatory proteins, western blot analysis was performed. After 2 days and 4 days incubation respectively, at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$, nicotine significantly inhibited proliferation comparing to non-supplemented controls. The cell viability was significantly decreased after 2 days and 4 days at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$ and at $10{\mu}g/ml$ - $1000{\mu}g/ml$ respectively. After 2 days and 4 days, the cellular activity was significantly decreased at concentrations of $10{\mu}g/ml$ - $1000{\mu}g/ml$. Treatment with $100{\mu}g/ml$ nicotine for 48 hours caused an increase in the proportion of G1-phase cells (from 46.41% to 53.46%) and a decrease in the proportion of S-phase cells (from 17.80% to 14.27%). The levels of cyclin $D_1$ and CDK 4 proteins in nicotine-treated fibroblasts were lower than that of controls, whereas the levels of p16 and pRB were higher than that of controls. These results suggest that the decrease of cell proliferation and lengthened Gap phases (G1) by nicotine may due to the increased expression of p16 and pRB as well as decreased expression of cyclin $D_1$ and CDK 4 in human gingival fibroblasts.

• Journal of Periodontal and Implant Science
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• 제28권2호
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• pp.291-310
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• 1998

This study was performed to evaluate the effects of extracts of Drynariae Rhizoma on the characteristics of rat calvaria cells(RCV) and bone marrow cells(RBM) which have the important role on the bone formation in vitro. Drynariae Rhizoma has been known as the useful herbal medicament for treatment of the wound healing including regeneration of bone fracture, and also has been used to treat the periodontal lesions, tooth mobility, gingival bleeding and pus discharge via sulcus in Oriental Medicine. In control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 10% fetal bovine serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, extracts of Drynariae Rhizoma(0.1, 1, 5, 10, $50{\mu}g/ml$) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 3, 7, 14, 21, 30th day, the amount of total protein synthesis and alkaline phosphatase activity of RCV at 2,4th day and those of RBM at 3, 6th day. And also, the calcified nodule of RCV was examed at 3, 5th day in three goup, control, experimental, culture with the PDGF group. The results were as follow ; 1. Both RCV and RBM cells in Drynariae Rhizoma-treated experimental group proliferated more rapidly than nontreated control group. The experimental group below $5{\mu}g/ml$ Drynariae Rhizoma-treated showed more prominent cell proliferation from the 7th day to the 21st day than the control group and above $10\;{\mu}g/ml$ treated group in RCV. 2. Amount of total protein synthesis was more increased in Drynariae Rhizomatreated group than in control group. In $5{\mu}g/ml$ Drynariae Rhizoma-treated group showed most prominent protein synthesis of the any other exrperimental group and control group. 3. Alkaline phosphatase activity also more increased in Drynariae Rhizomatreated group than control group. 4. Mineralized nodules in Drynariae Rhizoma-treated group were more than not in control group but also in PDGF-treated group. From the above results, Drynariae Rhizoma appeared to enhanced the proliferation, protein synthesis, alkaline phosphatase activity and cellular ability of mineralized nodule formation than PDGF. So that, we conclude that Drynariae Rhizoma enhances the activities of bone cells which have the important role on the periodontal regeneration and optimal application of Drynariae Rhizoma was thought to be useful as the means in bone regeneration.

• Journal of Periodontal and Implant Science
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• 제36권1호
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• pp.223-237
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• 2006

The purpose of the present study was to evaluate the effect of bone graft materials including deproteinized bovine bone(DBB), demineralized freeze-dried bone(DFDB), freeze-dried bone(FDB) on bone formation in guided bone regeneration using perforated titanium membrane(TM). 16 adult male rabbits(mean BW 2kg) were used in this study and 4 rabbits allotted to each test group. Intramarrow penetration(diameter 6.5mm) was done with round carbide bur on calvaria to promote blood supply and clot formation in the wound area. The test groups were devided into 4 groups as follows: TM only(test 1), TM +DBB(test 2), TM +DFDB(test 3), TM +FDB(test 4). Perforated titanium membrane was contoured in rectangular parallelepiped shape(0.5mm pore diameter, 10mm in one side, 2mm in inner height), filled the each graft material and placed on the decorticated carvaria. Perforated titanium membrane was fixed with resorbable suture materials. The animals were sacrificed at 2, 8 weeks after the surgery. Non-decalcified preparations were routinely processed for histologic analysis. The results of this study were as follows: 1. Perforated titanium membrane was biocompatible. 2. Perforated titanium membrane had capability of maintaining the space during the healing period but invasion of soft tissue through the perforations of titanium membrane decreased the space available for bone formation. 3. In test 1 group without bone graft material, the amount of bone formation and bone maturation was better than other test groups. 4. Among the graft materials, the effect of freeze-dried bone on bone formation was best. 5. In the test groups using deproteinized bovine bone, demineralized freeze-dried bone, bone formation was a little. The spacemaking capability of the membrane may be crucial for bone formation. The combined treatment with the perforated titanium membrane and deproteinized bovine bone or demineralized freeze-dried bone failed to demonstrate any added effect in the bone formation. Minimization of size and numbers of perforations of titanium membrane or use of occlusive titanium membrane might be effective to acquire predictable results in the vertical bone formation.

• Journal of Periodontal and Implant Science
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• 제23권3호
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• pp.359-373
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• 1993

The ultimate objective of periodontal therapy is not only stopping the progression of periodontal disease, but also promoting the regeneration of lost periodontal tissue. Guided Tissue Regeneration, which is based on the principle that the goal of periodontal regeneration can be achieved by preventing apical migration of gingival epithelium and blocking cells originating from connective tissue, has been developed and used as a clinical procedure, and although it has shown excellent results in connective tissue healing, there have not been many studies showing its effect on the regeneration of alveolar bone loss due to periodontal disease. The objectives of this study are to investigate the result of 12 months-long treatment following guided tissue regeneration using expanded polytetrafluoroehylene membrane, and to observe the presence of regenerated alveolar bone. Forty-one teeth from 28 patients with clinical diagnosis of periodontitis has been selected. In fifteen of those interproximal intrabony defects, only flap operation had been carried out, and designated as the control group. Twenty-six intrabony defects received e-PTFE membrane following flap operation, and designated as the experimental group. Eleven teeth whose membrane had been exposed were excluded from the experiment. Various measurements including probing depth, loss of attachment, probing bone level and gingival recession have been recorded at 6th month and 12th month, and the significance of the changes has been analyzed. The results are as follows: 1. Probing depth at 6th and 12th month has shown a significant decrease in both groups (p<0.01), but significantly higher decrease was found in the experimental group compared to the control at the month(p<0.05). 2. Loss of attachment at 6th and 12th month has shown a significant decrease in both groups, but significantly higher decrease was found in the experimental group compared to the control (p<0.05). 3. Probing bone level at 6th and 12th month has shown a insignificant decrease in the control group and significant decrease in the experimental group (p<0.01). Significantly higher decrease in probing bone level was found in the experimental group (p<0.05). 4. Gingival recession at 6th and 12th month has shown a statistically significant increase (p<0.05), and the control group showed higher increase compared to the experimental group although no statistical significance was found. As these results have shown, the use of e-PTFE membrane in intrabony pockets results in marked decrease in the loss of attachment and probing bone level. This seems to indicate that e-PTFE membrane may play a role in alveolar bone regeneration in intrabony defects.

• Journal of Periodontal and Implant Science
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• 제23권3호
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• pp.411-421
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• 1993

The purpose of this study was to assess the effect of polycaprolactone strip with minocycline on the periodontal pocket in humans and the various clinical parameters in rapidly progressive periodontitis. Nine patients with rapidly progressive periodontitis were selected for the study. They had not taken antibiotics for 6 months and had no history of dental treatment for 6 months before the study. They were in good general health. By the split-mouth method, patients received a supragingival scaling, experimental group (9sites) were subjected to subgingival placement of polycaprolactone strips(1 strip) containg 30% minocyclne and control group (9 site) were subjected to subgingival placement of not polycaprolactone strips(1 strip) containing 30 A Minocycline. Strips were replaced with freshly filled ones at 1 week and 2 week. All strips were removed from pockets at 3 week Clinical examination (plaque index, gingival index, probing pocket depth) and distribution on the bacteria morphology of subgingival plaque were monitored on baseline (0 week), 1, week, 2 week, 4 week and 8 week. The result were as follows : 1. Plaque index in experimental group was not significantly reduced during all weeks(P<0.05), but slightly reduced at 2, 4 and 8 weeks and that in control group was not significantly reduced during monitoring period. 2. Probing pocket depth was significantly reduced at 2, 4 and 8 weeks(P<0.05) in experimental group, but that in control group was not siginificantly changed during monitoring period. 3. Gingival index was significantly improved at 2, 4 and 8 weeks(P<0.05) in experimental group but that in control group was not significantly changed. 4. Percentage of cocci was significantly increased at 2, 4 and 8 weeks in experimental group but that in control group was not significantly changed. 5. Percentage of non-motile rods in both group were not significantly changed when compared with those of baseline(0 week) (P<0.05). 6. Percentage of motile rods was siginificantly reduced at 1, 2 and 4 weeks in experimental group (p<0.05) but that in control group was not significantly changed. 7. Percentage of spirochetes was siginificantly reduced during all weeks(P<0.05) but that in control group was not significantly changed. The result showed that polycaprolactone containing 30% minocycline effect the clinical index and bacterial morphotype.

• Journal of Periodontal and Implant Science
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• 제43권4호
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• pp.147-152
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• 2013

Purpose: An animal periodontitis model is essential for research on the pathogenesis and treatment of periodontal disease. In this study, we have introduced a lipopolysaccharide (LPS) of a periodontal pathogen to the alveolar bone defect of experimental animals and investigated its suitability as a periodontitis model. Methods: Alveolar bone defects were made in both sides of the mandibular third premolar region of nine beagle dogs. Then, the animals were divided into the following groups: silk ligature tied on the cervical region of tooth group, Porphyromonas gingivalis LPS (P.g. LPS)-saturated collagen with silk ligature group, and no ligature or P.g. LPS application group as the control. The plaque index and gingival index were measured at 0 and 4 weeks postoperatively. The animals were then euthanized and prepared for histologic evaluation. Results: The silk ligature group and P.g. LPS with silk ligature group showed a significantly higher plaque index at 4 weeks compared to the control (P<0.05). No significant difference was found in the plaque index between the silk ligature group and P.g. LPS with silk ligature group. The P.g. LPS with silk ligature group showed a significantly higher gingival index compared to the silk ligature group or the control at 4 weeks (P<0.05). Histologic examination presented increased inflammatory cell infiltration in the gingival tissue and alveolar bone of the P.g. LPS with silk ligature group. Conclusions: An additional P.g. LPS-saturated collagen with silk ligature ensured periodontal inflammation at 4 weeks. Therefore, P.g. LPS with silk ligature application to surgically created alveolar bone defects may be a candidate model for experimental periodontitis.

• Journal of Periodontal and Implant Science
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• 제38권1호
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• pp.97-102
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• 2008

Purpose: Periodontal intrabony defects have great deal of importance since they contribute to the development of periodontal disease. Current treatment regimens for intrabony defects involve grafting of numerous bony materials, GTR using biocompatible barriers, and biomodification of root surface that will encourage the attachment of connective tissue. Xenograft using deproteinized bovine bone particles seems to be very convenient to adjust because it doesn't require any donor sites or imply the danger of cross infections. These particles are similar to human cancellous bone in structure and turned out to be effective in bone regeneration in vivo. We here represent the effectiveness of grafting deproteinized bovine bone particles in intrabony defect and furcation involvements that have various numbers of bony walls. Materials and methods: Open flap debridement was done to remove all root accretions and granulation tissue from the defects within persisting intrabony lesions demonstrating attachment loss of over 6mm even 3 months after nonsurgical periodontal therapy have been completed. Deproteinized bovine bone particles($BBP^{(R)}$, Oscotec, Seoul) was grafted in intrabony defects to encourage bone regeneration. Patients were instructed of mouthrinses with chlorohexidine-digluconate twice a day and to take antibiotics 2-3 times a day for 2 weeks. They were check-up regularly for oral hygiene performance and further development of disease. Probing depth, level of attachment and mobility were measured at baseline and 6 months after the surgery. The radiographic evidence of bone regenerations were also monitored at least for 6 months. Conclusion: In most cases, radio-opacities increased after 6 months. 2- and 3-wall defects showed greater improvements in pocket depth reduction when compared to 1-wall defects. Class I & II furcation involvements in mandibular molars demonstrated the similar results with acceptable pocket depth both horizontally and vertically comparable to other intrabony defects. Exact amount of bone gain could not be measured as the re-entry procedure has not been available. With in the limited data based on our clinical parameter to measure pocket depth reduction following $BBP^{(R)}$ grafts, it was comparable to the results observed following other regeneration techniques such as GTR.

• Journal of Periodontal and Implant Science
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• 제38권2호
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• pp.125-134
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• 2008

Purpose: Bone morphogenetic protein (BMP) is a potent differentiating agent for cells of the osteoblastic lineage. It has been used in the oral cavity under a variety of indications and with different carriers. However, the optimal carrier for each indication is not known. This study evaluated the bone regenerative effect of rhBMP-2 delivered with different carrier systems. Materials and Methods: 8 mm critical-sized rat calvarial defects were used in 60 male Sprague-Dawley rats. The animals were divided into 6 groups containing 10 animals each. Two groups were controls that had no treatment and absorbable collagen membrane only. 4 groups were experimentals that contained rhBMP-2 only and applied with absorbable collagen sponge($Collatape^{(R)}$), $MBCP^{(R)}$, Bio-$Oss^{(R)}$ each. The histological and histometric parameters were used to evaluate the defects after 2- or 8-week healing period. The shape and total augmented area were stable in all groups over the healing time. Results: New bone formation was significantly greater in the rhBMP-2 with carrier group than control group. rhBMP-2/ACS was the highest in bone density but gained less new bone area than rhBMP-2/$MBCP^{(R)}$ and rhBMP-2/Bio-$Oss^{(R)}$. The bone density after 8 weeks was greater than that after 2 weeks in all groups. However, rhBMP-2 alone failed to show the statistically significant difference in new bone area and bone density compared to control group. Also $MBCP^{(R)}$ and Bio-$Oss^{(R)}$ particles remained after 8 weeks healing period. Conclusion: These results suggest that rhBMP-2 with carrier system is an excellent inductive agent for bone formation and we can use it as the predictable bone tissue engieering technique. Future study will likely focus on the kinetics of BMP release and development of carriers that is ideal for it.