• Journal of Periodontal and Implant Science
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• 제27권1호
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• pp.217-234
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• 1997

The purpose of present study is to compare the effect of treatment using $Guidor^{(R)}$ as a barrier membrane in conjunction with platelet-derived growth factor and insulin like growth factors on experimental dehiscence defects. Following the resection of premolar crowns, roots were submerged. After 12 weeks of healing period, experimental dehiscence defects of 4mm in height and 4mm in width were surgically created on the mid-facial aspect of the lower premolar roots in each of 4 adult dogs. After root planning and demineralization of the root surface with citric acid, the control groups received 4% methylcellulose gel only, the test group I received 4% methylcellulose gel and were covered by $Guidor^{(R)}$ and the test group II were treated with PDGF and IGF and 4% methylcellulose gel with $Guidor^{(R)}$ coverage. Histological and histomorphometric analysis following 8 weeks of healing revealed the following results. 1. The new bone formation showed no statistically significant difference in all groups with $0.59{\pm}0.82mm$($14.03{\pm}19.60%$) for control, $0.70{\pm}0.39mm$($16.30{\pm}9.01%$) for group I, $0.87{\pm}0.76mm$($18.74{\pm}16.03%$) for group II. 2. The new cementum formation showed no statistically significant difference in all groups with $0.54{\pm}0.48mm$($l6.38{\pm}14.57%$) for control, $0.95{\pm}0.38mm$($23.43{\pm}9.30%$) for group I, $1.01{\pm}0.75mm$($22.10{\pm}16.ll%$) for gorup II. 3. The root resorption showed statistically significant differences betweenthe control group and all test groups(p<0.05) with $2.11{\pm}0.53mm$($52.93{\pm}12.32%$) for control, $0.63{\pm}0.27mm$($15.32{\pm}7.05%$) for group I, $0.89{\pm}0.33mm$ ($19.26{\pm}7.11%$) for group II. On the bases of these results, there were no statistically difference between treatment using resorbable membrane and resorbable membrane in conjunction with PDGF and IGF in the dehiscence defects, where it was difficult to maintain space. The use of membrane seemed to be more effective in the inhibition of root resorption.

• Journal of Periodontal and Implant Science
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• 제39권2호
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• pp.119-128
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• 2009

Purpose: The aim of this study was to evaluate the stemness of cells from canine dental tissues and bone marrow. Methods: Canine periodontal ligament stem cells (PDLSC), alveolar bone stem cells (ABSC) and bone marrow stem cells(BMSC) were isolated and cultured. Cell differentiations (osteogenic, adipogenic and chondrogenic) and surface antigens (CD146, STRO-1, CD44, CD90, CD45, CD34) were evaluated in vitro. The cells were transplanted into the subcutaneous space of nude mice to assess capacity for ectopic bone formation at 8 weeks after implantation. Results: PDLSC, ABSC and BMSC differentiated into osteoblasts, adipocytes and chondrocytes under defined condition. The cells expressed the mesenchymal stem cell markers differently. When transplanted into athymic nude mice, these three kinds of cells with hydroxyapatite /${\beta}$- tricalcium phosphate (HA/TCP) carrier showed ectopic bone formation. Conclusions: This study demonstrated that canine dental stem cells have stemness like bone marrow stem cells. Transplantation of these cells might be used as a therapeutic approach for dental stem cell-mediated periodontal tissue regeneration.

• Journal of Periodontal and Implant Science
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• 제38권sup2호
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• pp.317-324
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• 2008

Purpose: Aggregatibacter actinomycetemcomitans was associated with localized aggressive periodontitis, endocarditis, meningitis, and osteomyelitis. The cytolethal distending toxin (CDT) of A. actinomycetemcomitans was considered as a key factor of these diseases is composed of five open reading frames (ORFs). Among of them, An enzymatic subunit of the CDT, CdtB has been known to be internalized into the host cell in order to induce its genotoxic effect. However, CdtB can not be localized in host cytoplasm without the help of a heterodimeric complex consisting of CdtA and CdtC. So, some studies suggested that CdtC functions as a ligand to interact with GM3 ganglioside of host cell surface. The precise role of the CdtC protein in the mechanism of action of the holotoxin is unknown at the present time. The aim of this study was to generate recombinant CdtC proteins expression from A. actinomycetemcomitans, through gene cloning and protein used to investigate the function of Cdt C protein in the bacterial pathogenesis. Materials and Methods: The genomic DNA of A. actinomycetemcomitans Y4 (ATCC29522) was isolated using the genomic DNA extraction kit and used as template to yield cdtC genes by PCR. The amplifed cdtC genes were cloned into T-vector and cloned cdt C gene was then subcloned to pET28a expression vector. The pET28a-cdtC plasmid expressed in BL21 (DE3) Escherichia coli system. Diverse conditons were tested to opitimize the expression and purification of functional CdtC protein in E. coli. Results: In this study we reconstructed CdtC subunit of A. actinomycetemcomitans Y4 and comfirmed the recombinant CdtC expression by SDS-PAGE and Western Blotting. The expression level of the recombinant CdtC was about 2% of total bacterial proteins. Conclusion: The lab condition of procedure for the purification of functionally active recombinant CdtC protein is established. The active recombinant CdtC protein will serve to examine the role of CdtC proteins in the host recognition and enzyme activity of CDT and investigate the pathological process of A. actinomycetemcomitans in periodontal disease.

• Macromolecular Research
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• 제10권3호
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• pp.158-167
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• 2002

In order to endow with new bioactive functionality from small intestine submucosa (SIS) powder as natural source to poly (L-lactide) (PLA) and poly (lactide-co-glycolide) (PLGA) synthetic biodegradable polymer, porous SIS/PLA and SIS/PLGA as natural/synthetic composite scaffolds were prepared by means of the solvent casting/salt leaching methods for the possibility of the application of tissue engineered bone and cartilage. A uniform distribution of good interconnected pores from the surface to core region was observed the pore size of 40~500 ${\mu}{\textrm}{m}$ independent with SIS amount using the solvent casting/salt leaching method. Porosities, specific pore areas as well as pore size distribution also were almost same. After the fabrication of SIS/PLA hybrid scaffolds, the wetting properties was greatly enhanced resulting in more uniform cell seeding and distribution. Five groups as PGA non-woven mesh without glutaraldehyde (GA) treatment, PLA scaffold without or with GA treatment, and SIS/PLA (Code No.3 ; 1 : 12 of salt content, (0.4 : 1 of SIS content, and 144 ${\mu}{\textrm}{m}$ of median pore size) without or with GA treatment were implanted into the back of nude mouse to observe the effect of SIS on the induction of cells proliferation by hematoxylin and eosin, and von Kossa staining for 8 weeks. It was observed that the effect of SIS/PLA scaffolds with GA treatment on bone induction are stronger than PLA scaffolds, that is to say, in the order of PLA/SIS scaffolds with GA treatment > PLA/SIS scaffolds without GA treatment > PGA nonwoven > PLA scaffolds only with GA treatment = PLA scaffolds only without GA treatment for the osteoinduction activity. The possible explanations are (1) many kinds of secreted, circulating, and extracellular matrix-bound growth factors from SIS to significantly affect critical processes of tissue development and differentiation, (2) the exposure of SIS to GA resulted in significantly calcification, and (3) peri-implant fibrosis due to covalent bonding between collagen molecule by crosslinking reaction. In conclusion, it seems that SIS plays an important role for bone induction in SIS/PLA scaffolds for the application of tissue engineering area.

• Journal of Periodontal and Implant Science
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• 제32권1호
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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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• 제32권1호
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• pp.143-160
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• 2002

타이태늄은 뛰어난 생체적합성과 적절한 물리적 성질을 바탕으로 치과 및 정형외과 영역의 매식재로 널리사용되어져 왔으며, 골과 매식재 사이의 골 융합 정도를 증가시킬 목적으로 물리, 화학적인 방법을 이용한 타이태늄의 표면처리에 관한 많은 연구들이 진행되어 왔다. 최근에는 부착단백질 또는 성장인자를 이용한 생체재료의 표면개질을 통하여 조직적합성 및 치유 능의 개선을 위한 시도들이 있어왔다. Fibronectin(FN)은 주요 세포외기질중의 하나로 생체 내 널리 분포하여 세포의 부착, 이동 및 증식에 관여하는 거대 당단백으로, RGD및 PHSRN 펩타이드 서열이 세포의 인테그린과 결합하여 세포의 활성을 조절하는 것으로 알려져 있다. 이 연구에서는 FN으로 처리된 타이태늄이 조골세포의 부착, 증식 및 분화에 미치는 영향과 이에 따른 석회화 정도에 미치는 영향을 관찰하여 부착분자를 이용한 타이태늄 표면개질의 효과를 규명하고자 하였다. 상업용 순수 타이태늄을 gold thiol법을 이용하여 표면처리 후, 혈장 FN(plasma FN, pFN)과 유전자재조합법을 이용하여 얻은 FN조각(FN type III 7-10, FNIII 7-10)을 피복한 시편을 실험군으로, 아무런 처리를 하지 않은것(smooth surface, SS)과 산 부식(Sandblasted and acid etched, SLA)처리된것을 대조군으로 이용하였다. 배양된 조골세포주(MC3T3-E1)를 사용하여 타이태늄 표면 처리에 따른 세포의 증식, 형태변화, 알칼리성 인산분해효소(ALPase) 생산 및 세포면역형광법을 이용한 분화정도를 시간 경과에 따라 관찰하였다. 조골세포증식의 경우 FNIII 7-10 처리군에서 pFN 처리군 및 대조군에 비해 시간경과에 따라 유의성있는 세포수의 증식이 관찰되었으며(p<0.05), ALPase 생성의 경우에도 FNIII 7-10 처리 군에서 아무 처리도 하지 않은 군에 비해 유의성 있게 높은 효소의 생성이 관찰되었다(p<0.05). 주사전자현미경을 이용한 세포의 형태관찰결과 아무 처리도 하지 않은 군에서는 마름모형태를 나타내었으며, 산 부식 처리된 군에서는 세포가 가시모양의 형태를 보인 반면 FN으로 처리된 두 군에서는 세포의 부착 및 펴짐이 매우 발달되어 있는 모습이 관찰되었다. 세포의 분화정도를 관찰하기 위하여 국소부착키나제(focal adhesion kinase, FAK), 및 actin stress fiber의 분포양상을 세포면역형광법을 이용하여 관찰한 결과 FN으로 표면처리된 두 군에서 아무런 처리도 하지않은 군 및 산 부식처리 한 군에 비해 프라크의 발현이 높게 나타났으며 잘 발달된 actin stress fiber의 소견을 나타내었다. 이 실험의 결과들은 gold thiol 법을 이용한 표면처리 후 FN부착을 통한 타이태늄의 표면개질이 조골세포의 부착, 증식 및 분화에 중요한 역할을 담당하여 석회화 정도를 촉진시키는 것을 보여주었으며, 이런 결과들은 더 짧은 FN조각을 이용한 다른 생체재료의 표면개질에 폭 넓게 응용할 수 있으리라 생각된다.

• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.41-49
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• 2002

The aim of this study was to compare periodontal conditions around mesially tipped molars by a tipping degree. Patients who had been consecutively treated at the Department of Periodontology, Chonbuk National University Hospital from October 1999 to August 2001 were assessed with radiographs taken at their molar regions. Of all molars investigated, 142 molars of 116 patients tipped mesially more than 10 degrees to the line perpendicular to an occlusal plane were selected for the study. The tipped molars were divided into 2 groups with a reference to a tipping degree, i.e., 66 slightly tipped(group 1 : <$30^{\circ}$) and 76 severely tipped molars (group 2 : ${\geq}30^{\circ}$). Probing depth(PD), plaque retention index(PRI) at mesial surfaces of tipped molars and tooth mobility(TM) were recorded at the clinical examination. Tipping degree(TD) and alveolar bony defect(ABD) at the mesial surface of the molars were measured in a radiograph. The results showed that no statistical differences were found between groups in all measured variables. In Pearson correlation analysis performed in the same group, a positive relationship was shown between PRI and PD in the group 1 and, in the group 2, between PRI and PD as well as PRI and ABD(p < 0.01). However, no statistically significant relationship was found between TD and all other variables in both groups. Within limitations of this study, it may be concluded that tipping degree did not seem to influence periodontal conditions, i.e., PD, ABD and TM of mesially tipped molars per se, but plaque presence/absence seemed to mainly affect the periodontal conditions of the tipped molars.

• Journal of Periodontal and Implant Science
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• 제30권4호
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• pp.707-727
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• 2000

매식된 인공치아의 성공을 위해서는 적절한 교합과 수동적 적합성을 갖는 보철물의 제작과 구강내 노출 직후부터의 세균성 치태조절이 요구된다. 본 연구는 전처리(passivation과 tridodecyl - methyl - ammonium chloride(TDMAC) 처리)가 다른 타이타늄 표 면에 chlorhexidine varnish와 테트라사이클린 을 도포시 약제의 방출역학을 알아보고 구강내 치태형성의 억제정도를 평가하기 위하여 시행 되었다. 이를 위해 방출용액으로 인산완충액 성분의 인조타액을 1일${\sim}$1개월간 매일 교환하여 약제농도를 측정하고 타이타늄 박막에 잔류한 약제 활성을 측정하였으며 항균제 도포한 타이타늄 원판을 부착한 장치를 구강내 위치시킨 1일${\sim}$3주 후 원판을 제거하여 주사전자현미경으로 세균 부착상을 관찰하였다. 테트라사이클린은 TDMAC 처리된 표면에서 $10{\sim}18$일까지 유효농도로 방출되었고 표면의 유효 항균 활성은 $3{\sim}4$주간 유지되었으며, chlorhexidine varnish 도포 시에는 TDMAC 전처리시 초기에 $3{\sim}7$일 간 증가한 유효 항균 활성을 방출하여 매식지대치 등에 이러한 항균제도포 시 매식치 주위환경에 항균활성 공급원으로 작용할 수 있음을 보였다. 주사현미경적 관찰시 모든 타이타늄 표면에서 구강내 위치 30분 후에는 세균이 부착되어 있지 않고 타액 단백질 성분에서 유래한 것으로 보이는 피막물질이 표면을 부분 또는 전면에 걸쳐 덮고 있었다. 구강내 노출 2시간 후 항균제 미도포 표본들에는 약간의 구균이 단층으로, $1{\sim}3$일 후에는 부분적으로 두꺼운 세균층을 형성하였고 7일 후에는 표면전체에 걸쳐 세균층이 덮여있었으며 주로 구균과 약간의 간균이 주종을 이루었다. 항균제 도포시 구강내 노출 1주일 이전까지는 미도포군에 비해 치태형성이 지연되는 경향을 보였지만 2주 이후에는 세균 수나 치태형성 양상이 유사하였다. 이 연구로부터 항균제 도포시 1주일 이전의 초기 치태형성을 감소시킬 수 있음을 알 수 있었으며 이러한 연구결과는 타이타늄 임프란트 지대치 표면에 항균제의 도포가 임상적으로 유용할 수 있음을 시사하였다.

• Journal of Periodontal and Implant Science
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• 제34권3호
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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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• 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.