• Journal of Periodontal and Implant Science
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• 제30권1호
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• pp.65-77
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• 2000

The purpose of this study was to evaluate the antimicrobial activity of Crassirhzimae rhizoma and its possible use as an oral antiseptics for prevention of periodontitis. Its antibacterial activity against periodontopathic microorganisms including Actinobacillus actiomycetem - comitans, Capnocytophaga ochracea, Streptococcus mutans, Porphyromonas gingivalis, Prevotella intermedia, Actinomyces viscosus, Fusobacterium nucleatumwas evaluated via modified stab culture method. The cytotoxicity against gingival fibroblasts and rat osteoblasts was investigated via [$^3H$]thymidine incorporation and cellular activity was investigated via MTT assay. Chlorhexidine was used as control group. Crassirhizomae rhizoma was prepared at concentrations of 0.2, 0.15, 0.1, 0.05%. Chlorhexidine was also prepared at the same concentration. Crassirhizomae rhizoma showed lower antimicrobial antivity against these microorganism than chlorhexidine, but this difference was not significant. And, Crassirhzomae rhizoma showed more cellular activity and less cytotoxicity than chlorhexidine on human gingival fibrablast and rat osteoblast. This study suggests that Crassirhzomae rhizoma might be a candidate for a safe oral antiseptic for the prevention and treatment of periodontal disease.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제22권2호
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• pp.123-132
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• 2000

$TGF-{\beta}_1$ is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to infection control. The objective of this study is to investigate production of $TGF-{\beta}$ in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of $TGF-{\beta}_1$ which may be responsible for infection control. The fibroblasts were originated from gingiva and facial dermis in 26 year-old male patient. In the presence of LPS($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$), SEB($0.01{\mu}g$, $0.l{\mu}g$, $1.0{\mu}g$) respectively, $cells(5{\times}10^3ml)$ were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, $cells(2.5{\times}10^5ml)$ were cultivated in EMEM with LPS(0.01, 0.1 and $1.0{\mu}g$), SEB(0.01, 0.1 and $1.0{\mu}g$) respectively and $LPS(0.1{\mu}g)$ and $SEB(0.1{\mu}g)$ in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and $TGF-{\beta}_1$ was assayed in duplicate. The results were as follows. 1. In gingival fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell Proliferation occurred very significantly since 3 days after incubation, compared with the control and the production of $TGF-{\beta}_1$ occurred very significantly at 1 day after incubation, compared with the control. 2. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of $TGF-{\beta}_1$ was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of $TGF-{\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of $TGF-{\beta}_1$ very significantly. The gingival and facial dermal fibroblasts have different phenotype each other The orchestrated understanding of fibroblast proliferation and $TGF-{\beta}_1$ production play an important part in host defense against the bacterial Infection and may prevent tissue necrosis such as necrotizing fasciitis and life-threatening syndrome such as multiple organ failure.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.279-289
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• 1995

The most important object of periodontal treatment is the perfect regeneration of destructed periodontal tissue. The healing of periodontal lesion is affected by several cells & factors, which result in formation of long juntional epithelium, root resorption, bony ankylosis or connective tissue attachment. And ideal healing is enhanced by epithilial exclusion or periodontal ligament cell activation. In this investigation, I studied the effect of Zizyphus Fructus extract which enhances biologic activity& collagen synthesis, on the chemotaxis & cell nature. The cells were obtained from interdental area & middle third area of the freshly extracted teeth for the orthodontic purpose. And they were fully incubated in${\alpha}-MEM$ solution containing $100{\mu]g/ml$ penicillin & $100{\mu]g/ml$ streptomycin followed by 6 generation incubation. The test cells were collected by trypsin-EDTA & centrifuge in the fully incubated cells, counted by Hernacyotmeter, incbated $5{\times}10^5/ml$ cells for 24 hours, re-incubated 24 hours in media containing natural extract and photographed. The cells were incubated for 4 hours in 48 well microchemotaxis chamber bisecting upper & lower chamber by 8ug/m pore polycarbonate membrane coating 5mg/ml gelatin solution. The migrated cells in microscope were counted, which meaned cell chemotaxis activity. The study had shown that the morphology of cell was spindle-shaped as the control group, and the subextract test groups were not significantly different. In gingival fibroblasts, the chemotaxis effect of PDGF was statistically significant compared to control group. The Zizyphus Fructus extract was more or less enhanced chemotaxis effect and in $1{\mu}g/ml$ concentration the chemotaxis effect was slightly elevated compared with $10{\mu}g/ml$ concentration. But, among the subextracts, it was not significantly defferent. In PDL cells, the chemotaxis effect of PDGF in statistically significant, and the zizyphus Fructus extract had shown the enhanced effect. The effect was slightly higher in $1{\mu}g/ml$ concentration than 10g/ml concentration,and no significance among the subextracts.

• 대한치과보철학회지
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• 제30권2호
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• pp.247-257
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• 1992

The present study quantitates the in vitro cytotoxicity of a variety of denture base acrylic resins using cell culture techniques combined with image analysis to measure nuclear area and DNA contents. In this study, a comparison was made among direct curing, heat curing and microwave curing resins. The results obtained from this study were as follows : 1. Morphologically, cell process and nucleus became prominent but macroscopic difference according to the resins were nit observed. In addition, increased cellular density around the specimen were observed. 2. In DNA contents measurements, $S-G_2M$ phase cell was 15.47%, 14.58% in control and heat curing resin on 1st day and the others group $21.39\sim33.36%$ were measured. 3. Nuclear area and DNA contents were increased on 3rd day except DNA content of the microwave curing resin group. These results suggest that denture base acrylic resins stimulate gingival fibroblasts in vitro, especially stimulation of direct curing resin is larger and longer than the others.

• Journal of Korean Dental Science
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• 제13권1호
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• pp.35-41
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• 2020

Irritation fibroma is a reactive hyperplastic lesion caused by chronic stimuli with low intensity in the oral cavity. Irritation fibroma is common in middle-aged females but it may also occur at any age and sex. Clinical characteristics of irritation fibroma are similar to other reactive lesions or benign tumors, therefore, histological examination is essential to make an accurate diagnosis. This case report presents two cases of irritation fibroma occurred on the gingiva in young males. Two male patients in their 20s and 30s of age visited the clinic for the evaluation and treatment of painless gingival overgrowth in the anterior region. Clinically, the lesions were well-defined and firm, with similar color and texture to the adjacent normal gingiva. Excisional biopsy under local anesthesia was conducted with a scalpel, and the lesions were completely removed. Histopathologically, connective tissue consisting of dense collagen bundles, proliferation of fibroblasts and minor infiltrated inflammatory cells were observed. Based on the clinical and histopathological findings, the diagnosis of irritation fibroma was confirmed in both cases.

• Journal of Periodontal and Implant Science
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• 제32권3호
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• pp.685-695
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• 2002

황색 포도상구균은 급성 구강 감염에 있어서의 병원균이다. 그러나 그러한 황색 포도상구균의 병원성 기전은 완전히 이해되지 않았다. 이전 실험에서 황색 포도상구균의 단백질 A와 골격근의 액틴 필라멘트는 인체 상피 세포로의 황색 포도상구균의 침입에 관여한다. 구강 내 감염에 있어서의 황색 포도상구균의 병원성 기전을 조사하기 위해 인체의 치은 섬유모 세포에 대한 침입이 연구되고 있다. 급성 구강 감염을 가진 환자로부터 분리된 ATCC 25923 황색 포도상구균과 OPT 2 황색 포도상구균의 침입은 시간(0-120분)에 의존한다는 사실을 밝혀냈다. 60분을 초과하는 배양시간은 다시 배양된 균집락수 증가를 가져왔다. 배지에 접종한 세균의 숫자가 증가할 때 (100?10,000,000 cfu/ml/well), 직선적으로 증가한다. 단백질 A가 결핍된 Wood 46 황색 포도상구균의 침입은 단백질 A가 발현된 균주(ATCC 25923과 OPT 2)의 침입보다 훨씬 낮았다. 액틴 필라멘트의 합성을 방해하는 Cytochalasin D는 인체 치은 섬유모세포로 황색 포도상구균 (ATCC 25923과 OPT 2)이 침입하는 것을 방해한다. 이러한 결과는 구강내 감염을 일으키는 황색 포도상구균의 병원성 기전이 세포내 침입에 관여하고, 황색 포도상구균 단백질 A와 골격근의 액틴 필라멘트가 인체 치은 섬유모세포로의 황색 포도상구균의 침입 조절에 관여한다는 것을 보여준다.

• International Journal of Oral Biology
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• 제43권3호
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• pp.129-132
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• 2018

Enterococcus faecalis is a major causative agent of endodontic treatment failure. The purpose of this study was to investigate bactericidal effects of ethanol extract of Garcinia mangostana L. (mangosteen extract) on five strains of E. faecalis that were isolated from human oral cavities. The bactericidal effects of mangosteen extract were assessed by measurement of minimum bactericidal concentration (MBC) value. The cytotoxicity of mangosteen extract on immortalized human gingival fibroblasts, hTERT-hNOF, was determined based on cell counting method. The data revealed the MBC value of mangosteen extract against the E. faecalis strains was $4{\mu}g/ml$. Additionally, the cell viability of mangosteen extract on hTERT-hNOF was 83.7-89.1% at the 1 to $16{\mu}g/ml$. These findings indicated that mangosteen extract could be used as a root canal cleaner during management of endodontic treatment failure caused by E. faecalis.

• Natural Product Sciences
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• 제13권4호
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• pp.347-354
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• 2007

In this study, we investigated the potential of Resina Pini (RP) for anti-inflammatory and analgesic agents to treat inflammatory diseases such as gingivitis and periodontitis. Crude RP (RP1), recrystallized RP (RP2), and Ramus Mori Albae-treated RP (RP3), plus their respective water extracts (RP1-WE, RP2-WE and RP3-WE) were prepared for in vitro and in vivo tests. We couldn't find any signs of heavy metals pollution in all the RP samples. RP2-WE exhibited the highest viability of human gingival fibroblasts (HGF) and the strongest scavenging activity on superoxide anion. RP1, RP2 and RP3, RP2 showed potent scavenging activity on DPPH free radical. RP2-WE displayed a stronger inhibition on hyaluronidase (HAase) activity and RP3 also displayed potent HAase inhibition. RP2-WE, RP3-WE, RP3 and RP2 were reduced admirably the production of $PGE_2$ in HGF. In addition, RP2-WE and RP3-WE exhibited potent inhibitory activities on arachidonic acid-induced ear edema in mouse. Moreover, RP-2 prevented completely acetic acid-induced writhing by 100.0% and RP1, RP3, RP1-WE and RP2-WE also exhibited excellent protective activities against writhing. While aminopyrine, the positive control, showed 76.9% analgesic effect at the same dose. Taken together, these results suggest that recrystallized aqueous extract of Resina Pini could be a promising drug for the treatment of periodontal diseases.

• Restorative Dentistry and Endodontics
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• 제36권1호
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• pp.26-36
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• 2011

연구목적: 본 연구는 in vitro 상에서 치수, 치은, 치주인대 세포를 neuropeptide (substance P, Calcitonin gene related peptides (CGRP))및 inflammatory cytokine (TNF-$\alpha$)으로 자극 시 matrix metalloproteinase (MMPs)의 생성 및 발현을 관찰한 것으로 치아와 치아 주변 조직에 염증이 존재할 경우 neuropeptide 및 inflammatory cytokine과 치아 경조직 혹은 치조골의 remodeling에 중요한 역할을 하는 matrix metalloproteinase (MMPs)와의 관계를 규명하고자 하였다. 연구 재료 및 방법: 시편으로는 우식이 없는 건전한 제3대구치(n = 10)를 사용하였으며, 발거 후 즉시 Phosphate buffered saline (PBS)에 보관하고 치아에서 치은과 치주인대 조직을 채취하였다. 치아를 종축으로 절단하고 치수 조직을 채취하여 조각으로 분리한 후 시편을 PBS에서 세 번 세척하였다. Plate에 치수, 치은, 치주인대 시편 조각을 위치시켜 Dulbecco's Modified Eagle Medium (DMEM)을 첨가하여 세포를 배양하였다. 치수, 치은, 치주인대 세포를 culture dish에서 confluence에 도달할 때 까지 배양하여 Fetal Bovine Serum (FBS)가 포함되지 않은 배지로 교환하여, $37^{\circ}C$에서 24시간 동안 배양한 후 Phosphate buffered saline (PBS)로 1회 세척하고 Substance P ($10^{-8}\;M$, $10^{-5}\;M$)가 포함된 배양액과 Mock (배양액만 포함됨)으로 4시간, 24시간동안 자극하였다. CGRP ($10^{-6}\;M$)을 함유한 배양액 및 TNF-$\alpha$(2 ng/mL)를 포함한 배양액으로 각각 24시간동안 세포를 자극하였다. 각각 다른 농도의 TNF-$\alpha$(2 ng/mL, 10 ng/mL, 100 ng/mL)를 포함한 배양액으로 24시간 동안 세포를 자극 한 후 RNase protection assay 및 Enzyme linked immunosorbent assay를 시행하였다. 결과: SP와 CGRP는 치수, 치은, 치주인대 세포의 MMPs발현에 관여 하지 않았다. TNF-$\alpha$로 24시간 자극 시 치수, 치은, 치주인대 세포에서 MMP-1,-12, -13의 발현을 증가시켰다. 반면, TNF-$\alpha$는 치수, 치은, 치주인대 세포들에서 TIMP-3의 발현을 감소시켰다. 서로 다른 농도의 TNF-$\alpha$(2 ng/mL, 10 ng/mL, 100 ng/mL)로 24시간 자극 시 MMP-1과 MMP-13의 발현이 증가하였다. 결론: 치아와 치아 주변 조직에 염증이 존재 시 TNF-$\alpha$가 증가함에 따라 치수, 치은, 치주인대로부터의 치아의 경조직 혹은 치조골의 remodeling에 관여하는 matrix metalloproteinase (MMPs)가 중요한 역할을 하는 것으로 사료된다.

• Journal of Periodontal and Implant Science
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• 제37권1호
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• pp.35-44
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• 2007

Periodontal ligament (PDL) is the connective tissue located between the tooth root and alveolar bone. In a previous study, PDLs22 was isolated as a PDL-specific gene by using subtractive hybrid-ization between cultured PDL fibroblasts and gingival fibroblasts. It was also suggested that PDLs22 plays important roles in the development, differentiation and maintenance of periodontal tissues. However, little is known about functional study of PDLs22 using recombinant protein in PDL fibroblast differentiation and periodontium formation. In this study, in order to produce the PDLs22 recombinat protein, PDLs22 expression vector were constructed and expressed its protein in various host cell and temperature conditions. The results were as follows: 1. PDLs22 protein was not strongly expressed In the induction system using pRSET-PDLs22 construct. 2. When the BL21(DE3) pLysS was used as a expression host, PDLS22 protein was strongly ex-pressed in the induction system using pHCEIIBNd-PDLs22 construct. 3. The PDLs22 protein was recognized at a molecular weight of 28 kDa in western blots. 4. Almost of the expressed PDLs22 protein was not soluble and observed like as inclusion body. 5. The protein solubility was not improved after modification of induction time and temperature during PDLs22 protein production. In this study, the system for the PDLs22 protein production was connstructed. However, the re-results suggest that further studies will be needed to produce the considerable amount of PDLs22 re-combinat protein, which can use for the periodontal regeneration.