• Journal of Periodontal and Implant Science
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• v.28 no.1
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• pp.17-35
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• 1998

Chitosan, with a chemical structure similar to hyaluronic acid, has been implicated as a wound healing agent. The purpose of this research was to evaluate the effects of chitosan on the characteristics of periodontal ligament cells, calvaria cells and gingival fibroblasts and to define the effects of chitosan on bone formation in vitro. In control group, the cells were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% Fetal bovine serum, 100unit/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, chitosan($40{\mu}g/ml$) is added into the above culture condition. And then each group was characterized by examining the cell proliferation at 1,3,5,7,9,12,15 day, the amount of total protein synthesis, alkaline phosphatase activity at 3, 7 day and the ability to produce mineralized nodules of rat calvaria cell at 11 day. The results were as follows : 1. At early time both periodontal ligament cells and calvaria cells in chitosan-treated group proliferated more rapidly than in non-treated control group, but chitosan-treated group of periodontal ligament cells at 9 days and calvaria cells at 12days showed lower growth rate than control group. Gingival fibroblast in chitosan-treated group had lower growth rate than in control group but the difference was not statistically significant (P< 0.01).2. Both periodontal ligament cells and calvaria cells in chitosan-treated group showed much protein synthesis than in control group at 3 days, but showed fewer than in control group at 7 days. Amount of total protein synthesis of gingival fibroblast didn't have statistically significant difference among the two groups(P< 0.01). 3. At 3 and 7 days, alkaline phosphatase activity of periodontal ligament cells and calvaria cells was increased in chitosan-treated group, but at 7 days there was not statistically significant difference among the two groups of calvaria cells (P< 0.01). Alkaline phosphatase activity of gingival fibroblast didn't have statistically significant difference among the two groups(P<0.01). 4. Mineralized nodules in chitosan-treated group of rat calvaria cells were more than in control group. In summery, chitosan had an effect on the proliferation, protein systhesis, alkaline phosphatase activity of periodontal ligament cells and calvaria cells, and facilitated the formation of bone. It is thought that these effects can be used clinically in periodontal regeneration therapy.

• Journal of Periodontal and Implant Science
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• v.27 no.1
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• pp.1-17
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• 1997

Tetracycline is known to be effective in eliminating periodontopathogens and have collagenolytic activity. This study was performed to observe the desorption kinetics and structural changes of tetracycline-treated barrier membranes for guided tissue regeneration. Four kinds of barrier membranes were tested : $Tefgen^{(R)}$(American Custom Medical, USA) and $Gore-Tex^{(R)}$(W.L. Gore & Associates Inc., USA) as nonresorbable membranes ; Resolut(polyglycolide & polylactide copolymer, W.L. Gore & Associates Inc., USA) and $Biomend^{(R)}$(collagen, Collatec Co., USA) as resorbable membranes. The membranes were cut into discs(diameter : 4mm) and were immersed in 5% tridodecylmethylammonium chloride(TIMAC) ethanol and air-dried. The membrane discs were absorbed with $100{\mu}g/ml tetracycline solution(pH8) for one minute and dried. For desorption kinetics, TC treated discs were immersed in phosphate buffered saline solution (PBS, pH 7.4). PBS was exchanged daily and TC concentration was measured by absorbance at 276nm on UV spectrophotometer. To measure remaining antibacterial activity, discs of 1 day to 4 weeks after desorption were placed on Mueller Hinton agar containing Bacillus cereus and incubated aerobically in $37^{\circ}C$ for twelve hours and the inhibition diameters were measured. To observe the structural change of membranes after TIMAC treatment or immersion in PBS, the membrane discs were examined under SEM. The results were as follows : 1. Total amounts of TC absorbed into membrane discs($0.7536mm^2$) were $2000{\mu}g$, $1800{\mu}g$, $2625{\mu}g$ and $2499{\mu}g$ for $Tefgen^{(R)}$, $Gore-Tex^{(R)}$, $Biomend^{(R)}$ and $Resolut^{(R)}$. 2. The concentration of TC released from barrier membrane discs was maintained over $4{\mu}g/ml$ until the fifth day in nonresorbable membranes and $Resolut^{(R)}$, but until the fourth day in $Biomend^{(R)}$, Until the ninth day in nonresorbable membranes and until the seventh day in resorbable membranes, the TC concentration was maintained over $1{mu}g/ml$. 3. The four membrane discs in the first day showed similar size of inhibition zone. One to four weeks later, the inhibition zone was much smaller in resorbable membrane discs than nonresorbable membrane discs. 4. Any structural change due to treatment of TIMAC was not observed on the nonresorbable membranes. $Resolut^{(R)}$ did not show any structural change except fibrillar loosening during immersion period, but Biomend showed destruction of membrane structure from the first week of immersion. This study indicates that tetracycline treated barrier membranes lead to the sustained release of tetracycline for over 7 days. This slow release pattern of tetracycline may contribute to the favorable clinical outcome of guided tissue regeneration.

• Journal of Periodontal and Implant Science
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• v.36 no.3
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• pp.613-625
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• 2006

Periodontal ligament(PDL) cells and human gingival fibroblasts(HGFs) play important roles in development, regeneration, normal function, and pathologic alteration. PDL cells and HGFs have the similarity related with general characteristics of fibroblast such as spindle shaped morphology, the presence of vimentin intermediate filament and the synthesis of interstitial collagens and fibronectin. There were many studies about the differences between PDL cells and HGFs, but they were not about whole gene level. In this study, we tried to explain the differences of gene expression profiles between PDL cells and HGFs, and the differences among three individuals by screening gene expression patterns of PDL cells and HGFs, using cDNA microarray. Although there were some variants among three experiments, a set of genes were consistentely and differentially expressed in one cell type. Among 3,063 genes, 49 genes were more highly expressed in PDL cells and 12 genes were more highly expressed in HGFs. The genes related with cell structure and motility were expressed more highly in PDL cells. These are cofilin 1, proteoglycan 1 secretory granule, collagen type I(${\alpha}$ 1), adducin gamma subunit, collagen type III(${\alpha}$ 1), fibronectin, lumican(keratan sulfate proteoglycan), and ${\alpha}$ -smooth muscle actin. Tissue inhibitor of metalloproteinase known as the enzyme controlling extracellular matrix with matrix metalloproteinase is more highly expressed in PDL cells, osteoprotegerin known as osteoclastogenesis inhibitory factor is more highly expressed in HGFs. We performed northern blot to verify cDNA microarray results on selected genes such as tissue inhibitor of metalloproteinase, fibronectin, osteoprogeterin. The result of northern blot analysis showed that each cell expressed the genes in similar pattern with cDNA microarray result. This result indicates that cDNA microarray is a reliable method in screening of gene expression profiles.

• Journal of Periodontal and Implant Science
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• v.41 no.4
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• pp.176-184
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• 2011

Purpose: The final goal of regenerative periodontal therapy is to restore the structure and function of the periodontium destroyed or lost due to periodontitis. However, the role of periosteum in periodontal regeneration was relatively neglected while bone repair in the skeleton occurs as a result of a significant contribution from the periosteum. The aim of this study is to understand the histological characteristics of periosteum and compare the native periosteum with the repaired periosteum after elevating flap or after surgical intervention with flap elevation. Methods: Buccal and lingual mucoperiosteal flaps were reflected to surgically create critical-size, "box-type" (4 mm width, 5 mm depth), one-wall, intrabony defects at the distal aspect of the 2nd and the mesial aspect of the 4th mandibular premolars in the right and left jaw quadrants. Animals were sacrificed after 24 weeks. Results: The results from this study are as follows: 1) thickness of periosteum showed difference as follows (P<0.05): control group ($0.45{\pm}0.22$ mm)> flap-elevation group ($0.36{\pm}0.07$ mm)> defect formation group ($0.26{\pm}0.03$ mm), 2) thickness of gingival tissue showed difference as follows (P<0.05): defect formation group ($3.15{\pm}0.40$ mm)> flap-elevation group ($2.02{\pm}0.25$ mm) > control group ($1.88{\pm}0.27$ mm), 3) higher cellular activity was observed in defect formation group and flap-elevation groups than control group, 4) the number of blood vessles was higher in defect formation group than control group. Conclusions: In conclusion, prolonged operation with increased surgical trauma seems to decrease the thickness of repaired periosteum and increase the thickness of gingiva. More blood vessles and high cellular activity were observed in defect formation group.

• Journal of Periodontal and Implant Science
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• v.36 no.1
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• pp.27-37
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• 2006

Although the main purpose of periodontal treatment to regenerate is the complete regeneration of periodontal tissue due to periodontal disease, most of the treatment cannot meet such purpose because healing by long epithelial junction. Therefore, diverse materials of resorbable and non-resorbable have been used to regenerate the periodontal tissue. Due to high risk of exposure and necessity of secondary surgical procedure when using non-resorbable membrane, guided tissue regeneration using the resorbable membrane has gain popularity, recently. However, present resorbable membrane has the disadvantage of not having sufficient time to regenerate date to the difference of resorption rate according to surgical site. Meanwhile, other than the structure stability and facile manipulation, acellular dermal matrix has been reported to be a possible scaffold for cellular proliferation due to rapid revascularization and favorable physical properties for cellular attachment and proliferation. The purpose of this study is to estimate the influence of acellular dermal matrix on periodontal ligament, cementum and alveolar bone when acellular dermal matrix is implanted to 1-wall alveolar bone defect. 4 dogs of 12 to 16 month old irrelevant to sex , which below 15Kg of body weight, has been used in this study. ADM has been used for the material of guided tissue regeneration. The 3rd premolar of the lower jaw was extracted bilaterally and awaited for self-healing. subsequently buccal and lingual flap was elevated to form one wall intrabony defect with the depth and width of 4mm on the distal surface of 2nd premolar and the mesial surface of 4th premolar. After the removal of periodontal ligament by root planing. notch was formed on the basal position. Following the root surface treatment, while the control group had the flap sutured without any treatment on surgically induced intrabony defect. Following the root surface treatment, the flap of intrabony defect was sutured with the ADM inserted while the control group sutured without any insertion. The histologic specimen was observed after 4 and 8 weeks of treatment. The control group was partially regenerated by periodontal ligament, new cementum and new alveolar bone. the level of regeneration is not reached on the previous formed notch. but, experimental group was fully regenerated by functionally oriented periodontal ligament fiber. new cementum and new alveolar bone. In conclusion, we think that ADM seems to be used by scaffold for periodontal ligament cells and the matrix is expected to use on guided tissue regeneration.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.89-112
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• 2002

The earliest reports of the use of electrical energy to directly stimulate bone healing seem to be in 1853 from England, the techniques involved the introduction of direct current into the non-united fracture site percutaneously via metallic needles, with subsequent healing of the defect. One endpoint of the periodontal therapy is to generate structure lost by periodontal diseases. Several procedural advances may support regeneration of attachment, however, regeneration of alveolar bone does not occur consistently. Therefore, factors which stimulate bone repair are areas for research in periodontal reconstructive therapy. Effects of cytokines or growth factors on bone repair are examples of such areas. Another one is electrical current which occurs in bone naturally, so that such bone may be particularly susceptible to electrical therapy. The purposes of this study were to observe the effects of electrical stimulation on the normal periodontium, to determine whether the electricity is the useful means for periodontal regeneration or not. Forty rats weighted about 100 gram were used and divided into 4 groups, the first group, there was no electrical stimulation with the connection of electrodes only. In the second group, there was stimulated by the 10 mA during 10 minutes per a day, in the third group was stimulated by the 25 mA , and the fourth by the 50 mA. At 3, 5, 10 and 15 days post-appliance , two rats in each group were serially sacrificed. and the maxillae and the mandible processed to paraffin, and the specimens were prepared with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows : 1. There was the distinct reversal line on the lingual alveolar crest, whereas a little changes in the labial alveolarcrest to the duration and amount of currents. 2. In 50 mA group, the cells were highly concentrated at the apex of anterior teeth, and was observed the necrotic tissue. In posterior root apex, the hypercementosis was appeared, and newly formed cementum layer has been increased continuously with the time. 3. The periodontal ligament fiber and Sharpey's fiber were arranged in order, and the bone trabeculae were increased as the experiment proceeded by, relatively the bone marrows were decreased. 4. In the pulp tissue, the blood vessels were increased with blood congestion in the experimetal specimens remarkably, and the dentinal tubules were obstructed . 5. The osteoblasts in alveolar bone proper had been showed highly activity, and also observed the formation of bone trabeculea. In the conclusion, it was suggested that the electrical stimulation has influence on the periodontium and the pulp tissue. However, there might be the injurious effects.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.129-142
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• 2002

Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.

• Journal of Periodontal and Implant Science
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• v.36 no.4
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• pp.797-808
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• 2006

Objective : The purpose of this study was to evaluate the physicochemical properties and cytocompatibility of microporous, spherical biphasic calcium phosphate(BCP) ceramics with a 60/40 $hydroxyapatite/{\beta}$ -tricalcium phosphate weight ratio for application as a bone graft substitute. Materials and Methods : Microporous, spherical BCP granules(MGSB) were prepared and their basic characteristics were compared with commercially available BCP(MBCP; Biomatlante, France) and deproteinized bovine bone mineral(Bio-Oss; GBistlich-Pharma, Switzerland, BBP; Oscotec. Korea), Their physicochemical properties were evaluated by scanning electron microscopy, X-ray diffractometry, Fourier-transform infrared spectroscopy, inductively coupled plasma atomic emission spectrometer, and Brunauer-Emmett-Teller method. Cell viability and proliferation of MC3T3-El cells on different graft materials were evaluated. Results : MGSB granules showed a chemical composition and crystallinity similar with those in MBCP, they showed surface structure characteristic of three dimensionally, well-interconnected micropores. The results of MTT assay showed increases in cell viablity with increasing incubation times. At 4d of incubation, MGSB, MBCP and BBP showed similar values in optical density, but Bio-Oss exhibited significantly lower optical density compared to other bone substitutes(p <0,05). MGSB showed significantly greater cell number compared to other bone substitutes at 3, 5, and 7d of incubation(p <0,05), which were similar with those in polystyrene culture plates. Conclusion: These results indicated the suitable physicochemical properties of MGSB granules for application as an effective bone graft substitute. which provided compatible environment for osteoblast cell growth. However, further detailed studies are needed to confirm its biological effects on bone formation in vivo.

• Journal of Periodontal and Implant Science
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• v.34 no.3
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• pp.543-549
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• 2004

Chitosan has been widely researched as bone substitution materials and membranes in orthopedic/periodontal applications. Chitosan nanofiber membrane was fabricated by chitosan nanofiber using electrospinning technique. The structure of the membrane is nonwoven, three-dimensional, porous, and nanoscale fiber-based matrix. The aim of this study was to evaluate the biocompatibility of chitosan nanofiber membrane and to evaluate its capacity of bone regeneration in rabbit calvarial defect. Ten mm diameter round cranial defects were made and covered by 2 kinds of membranes (Gore-Tex membrane, chitosan nanofiber membrane) in rabbits. Animals were sacrificed at 4 weeks after surgery. Decalcified specimens were prepared and observed by microscope. Chitosan nanofiber membrane maintained its shape and space at 4 weeks. No inflammatory cells were seen on the surface of the membrane. In calvarial defects, new bone bridges were formed at all defect areas and fused to original old bone. No distortion and resorption was observed in the grafted chitosan nanofiber membrane. However bone bridge formation and new bone formation at the center of the defect could not be seen in Gore-Tex membranes. It is concluded that the novel membrane made of chitosan nanofiber by electrospinning technique may be used as a possible tool for guided bone regeneration.

• Journal of Periodontal and Implant Science
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• v.38 no.1
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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.