• Journal of Periodontal and Implant Science
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• v.31 no.2
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• pp.299-315
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• 2001

Application of membranes for guided tissue regeneration(GTR) have been confined to the subgingival barrier functions; however, many studies have provided evidence that some drugs, including tetracycline, initially can promote the growth of periodontal ligament or alveolar bone in peridontal therapy. Osseous regeneration in periodontal defects is increased by local administration of tetracycline due to its anti-collagenolytic effect, which enhances bone-forming ability via osteoblast cell chemotaxis and reduced bone resorption. The aim of this study was to evaluate effects of tetracycline loaded poly-L-lactide(PLLA) barrier membranes for guided bone regenerative potential. Tetracycline was incorporated into the PLLA membrane with the ratio 10% to PLLA by weight. Ability to guided bone regeneration of the membranes were tested by measuring new bone in the tibial defects($7{\times}10{\times}5\;mm^3$) of the beagle dog for 4,5, and 6 weeks. In control, drug-unloaded PLLA membranes were used in same size of defect. In histologic finding of the defect area, a few inflammatory cells were observed in both groups. These membrane were not perforated by connective tissue and maintained their mechanical integrity for the barrier function for 4-6 weeks. New bone formation was greater in defects covered by tetracycline-loaded membrane than in defects covered by drug- unloaded membranes. In bone regeneration guiding potential test, tetracycline-loaded membrane was more effective than drug- unloaded membranes(p<0.05). These results suggest that tetracycline-loaded PLLA membranes potentially enhance guided bone regenerative efficacy and might be a useful barrier for GTR in periodontal treatment.

• The korean journal of orthodontics
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• v.42 no.6
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• pp.307-317
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• 2012

Objective: The purpose of this study was to investigate the isolation and characterization of multipotent human periodontal ligament (PDL) stem cells and to assess their ability to differentiate into bone, cartilage, and adipose tissue. Methods: PDL stem cells were isolated from 7 extracted human premolar teeth. Human PDL cells were expanded in culture, stained using anti-CD29, -CD34, -CD44, and -STRO-1 antibodies, and sorted by fluorescent activated cell sorting (FACS). Gingival fibroblasts (GFs) served as a positive control. PDL stem cells and GFs were cultured using standard conditions conducive for osteogenic, chondrogenic, or adipogenic differentiation. Results: An average of $152.8{\pm}27.6$ colony-forming units was present at day 7 in cultures of PDL stem cells. At day 4, PDL stem cells exhibited a significant increase in proliferation (p < 0.05), reaching nearly double the proliferation rate of GFs. About $5.6{\pm}4.5%$ of cells in human PDL tissues were strongly STRO-1-positive. In osteogenic cultures, calcium nodules were observed by day 21 in PDL stem cells, which showed more intense calcium staining than GF cultures. In adipogenic cultures, both cell populations showed positive Oil Red O staining by day 21. Additionally, in chondrogenic cultures, PDL stem cells expressed collagen type II by day 21. Conclusions: The PDL contains multipotent stem cells that have the potential to differentiate into osteoblasts, chondrocytes, and adipocytes. This adult PDL stem cell population can be utilized as potential sources of PDL in tissue engineering applications.

• Restorative Dentistry and Endodontics
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• v.25 no.2
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• pp.170-179
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• 2000

The rh-BMP-4 is a subgroup of TGF-${\beta}$ superfamily. The application of rh-BMP in alveolar bony defect was reported to new alveolar bone and new cementum formation. For minimized complications following tooth replantation, a operator must replant a tooth fast at the pertinent position. This study was to evaluate the effect of rh-BMP-4 on periodontal regeneration and root resorption following tooth replantation in rats. The 50 Sprague-Dawley rats weighting about 130gm were used in this study. The animals were divided into three groups. Group 1 ; immediate replantation after extraction : Group 2 ; replantation stored teeth extraction of first molar, the removal of periodontal ligament with collagenase, and etching with citric acid : Group 3 ; replantation stored teeth with treated rh-BMP-4 in mesial root. Experimental animals were sacrificed 3, 7, 14 days after replantation by heart infusion. The maxillae were removed, fixed, demineralized, dehydrated, infiltrated and embedded with JB-4 mixture. For light microscopic observation, 5 micron sections were cut and stained with toluidine blue. The results of this study were as follows : 1. After experimental 3 days, all groups were observed dead space between periodontum and root. 2. After experimental 7 days, group 1 and group 3 were observed filling periodontal fibers between alveolar bone and root but group 2 were not. 3. After experimental 7 days, group 3 were observed appearance of attached cementoblast like cell on root surface. Group 1 were observed regular arrangement of fibroblasts and collagen fibers at ${\times}400$ observation. 4. After experimental 14 days, all group were observed filling periodontal fibers between alveolar bone and root. Group 1 were observed normal arrangement of periodontal fibers. Group 3 were observed less abnormal arrangement of periodontal fibers. Group 2 were not observed functional normal arrangement of periodontal fibers. 5. After experimental 14 days, group 2 and 3 were observed several root resorption and irregular root surface but group 1 were not. These results suggest that the rh-BMP-4 can stimulate cementogenesis and enhance to attach collagen fibers.

• Journal of the korean academy of Pediatric Dentistry
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• v.50 no.3
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• pp.347-359
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• 2023

Objective: This study aimed to evaluate the suitability of polydeoxyribonucleotides (PDRN) as a storage medium for avulsed teeth. Materials and Methods: The viability of human periodontal ligament (PDL) cells stored in Hank's balanced salt solution and PDRN solutions (concentrations, 10, 25, 50, and 100 ㎍/mL) and tap water was measured using the Cell Counting Kit-8 and Live/Dead assays. In addition, Nitric oxide detection and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to evaluate the anti-inflammatory effect of PDRN. Results: The viability of PDL cells stored in a 100 ㎍/mL PDRN solution was significantly higher than that of cells stored in the other solutions (p < 0.01). Furthermore, cells stored in 100 ㎍/mL PDRN solution demonstrated a significantly reduced NO production (p < 0.0001), and cells stored in 50 and 100 ㎍/mL PDRN solutions expressed significantly lower levels of tumor necrosis factor α, interleukin (IL) -4, IL-6, and IL-10 (p < 0.01) compared to cells stored in HBSS. Conclusion: The PDRN solution exhibited cell-preserving and anti-inflammatory effects on the PDL cells. The findings of this study can serve as a basis for further experiments directed at the development of an effective storage medium for avulsed teeth.

• Restorative Dentistry and Endodontics
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• v.32 no.5
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• pp.419-425
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• 2007

The aim of this study was to evaluate the radiopacity and cytotoxicity of three resin-based (AH 26, EZ fill and AD Seal), a zinc oxide-eugenol-based (ZOB Seal), and a calcium hydroxide-based (Sealapex) root canal sealers. Specimens, 10 mm in diameter and 1 mm in thickness, were radiographed simultaneously with an aluminum step wedge using occlusal films, according to ISO 6876/2001 standards. Radiographs were digitized, and the radiopacity of sealers was compared to the different thicknesses of the aluminum step wedge, using the Scion image software. Using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the cytotoxicity of each material was determined in immortalized human periodontal ligament (IPDL) cells. The results demonstrated that EZ fill was the most radiopaque sealer, while Sealapex was the least radiopaque (p < 0.05). AH 26, AD Seal and ZOB Seal presented intermediate radiopacity values. All the materials evaluated, except for Sealapex, presented the minimum radiopacity required by ISO standards. The cell viabilities of resin-based root canal sealers were statistically higher than that of other type of root canal sealers through the all experimental time. Further, EZ fill showed statistically lower cell viability in 24 and 48 hours compared to AD Seal and in 72 hours compared to all other resin-based root canal sealers. However, there was no correlation between the radiopacity and cytotoxicity of three resin-based root canals sealers (p > 0.05). These results indicate that resin-based root canal sealer is more biocompatible and has advantage in terms of radiopacity.

• Restorative Dentistry and Endodontics
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• v.35 no.4
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• pp.285-294
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• 2010

The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa), group 8 (low-temperature preservation at $0^{\circ}C$ under no additional pressure), group 9 (low-temperature preservation at $-5^{\circ}C$ under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$ water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 7 ($0^{\circ}C$/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance). By the results of this study, low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa suggest the possibility for long term preservation of teeth.

• Restorative Dentistry and Endodontics
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• v.34 no.4
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• pp.356-363
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• 2009

The purpose of this study was to evaluate the viability of periodontal ligament cells in rat teeth using slow cryo-preservation method under pressure by means of MTT assay and WST-1 assay. Eighteen teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both sides of the first and second maxillary molars were extracted as atraumatically as possible under Tiletamine anesthesia. The experimental groups were group 1 (Immediate control), group 2 (Cold preservation at $4^{\circ}C$for 1 week), group 3 (Slow freezing), group 4 (Slow freezing under pressure of 3 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$water bath, then MTT assay and WST-1 assay were processed. One way ANOVA and Tukey method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 4 showed significantly higher viability of periodontal ligament cells than group 2 and 3 (p < 0.05), but showed lower viability than immediate control group. By the results of this study, slow cryo-preservation method under pressure suggests the possibility for long term cryo-preservation of the teeth.

• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.593-607
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• 1999

치주인대세포는 치주인대의 유지와 개조에 있어서 중요한 역할을 담당하는 섬유아세포성 세포로서, 세포에 가해진 여러가지 조건에 따라 다양한 표현형의 변화를 나타내는 것으로 알려져 있다. 기계적 응력은 치주인대세포의 세포증 식과 밀접히 연관되어 있는 것으로 알려져 있으며, 이는 세포주기 조절인자들의 발현을 증가 시킴으로써 이루어질 것으로 생각되나 그 자세 한 작용기전은 알려져 있지 않다. 그러므로 이 연구의 목적은 기계적 응력이 사람 치주인대세 포의 세포증식과 세포주기 조절인자의 발현에 미치는 영향을 연구하기 위하여 사람 치주인대 세포에 기계적 응력을 가한 후 세포증식을 관찰하고 , 세포주기조절인자들인 p 53 , $p21^{WAF1/CIP1}$ cyclin-dependent kinases(cdks), cyclins 및 proliferating cell nuclear antigen(PCNA)의 단백질 발현 변화를 연구하였다. 본 연구에 사용한 사람 치주인 대세포는 교정치료를 목적으로 발거한 건전한 사람 소구치의 치주인대로부터 explantation culture하여 얻은 후 계대배양을 시행하여 제6 계대의 세포를 사용하였다. 배양한 사람 치주인 대세포를 55-mm Petriperm dish당 $1{\times}10^4$ 개를 분주하고, dish당 1kg의 기계적 응력을 가하면서 12일동안 세포배양을 시행하였다. 사람 치주인대세포의 세포증식은 기계적 응력을 가한 후 8-12일 사이에 현저히 증가하였으며, PCNA 단백질의 발현은 기계적 응력을 가한 후 6-10일 사이에 현저히 증가하였다. 또한 기계적 응력은 사람 치주 대세포의 cdk4, cdk6, cdk2 및 cyclin D1 단백질의 발현을 다소 증가 시켰으나, p53 및 $p21^{WAF1/CIP1}$ 단백질의 발현은 큰 변화가 없었다. 이상의 결과 서 기계적 응력은 사람 치주인대세포 의 p53 및 $p21^{WAF1/CIP1}$ 단백질 발현의 변화 없이 cdks 단백질 발현을 증가시킴으로써 세포증식을 증가시키는 것으로 생각된다.

• Journal of Periodontal and Implant Science
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• v.41 no.4
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• pp.192-200
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• 2011

Purpose: The aim of this study is to compare the gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow for characterization of dental stem cells. Methods: We employed GeneChip analysis to the expression levels of approximately 32,321 kinds of transcripts in 5 samples of bone-marrow-derived mesenchymal stem cells (BMSCs) (n=1), periodontal ligament stem cells (PDLSCs) (n=2), and dental pulp stem cells (DPSCs) (n=2). Each cell was sorted by a FACS Vantage Sorter using immunocytochemical staining of the early mesenchymal stem cell surface marker STRO-1 before the microarray analysis. Results: We identified 379 up-regulated and 133 down-regulated transcripts in BMSCs, 68 up-regulated and 64 down-regulated transcripts in PDLSCs, and 218 up-regulated and 231 down-regulated transcripts in DPSCs. In addition, anatomical structure development and anatomical structure morphogenesis gene ontology (GO) terms were over-represented in all three different mesenchymal stem cells and GO terms related to blood vessels, and neurons were over-represented only in DPSCs. Conclusions: This study demonstrated the genome-wide gene expression patterns of STRO-$1^+$ mesenchymal stem cells derived from dental tissues and bone marrow. The differences among the expression profiles of BMSCs, PDLSCs, and DPSCs were shown, and 999 candidate genes were found to be definitely up- or down-regulated. In addition, GOstat analyses of regulated gene products provided over-represented GO classes. These data provide a first step for discovering molecules key to the characteristics of dental stem cells.

• Journal of Periodontal and Implant Science
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• v.30 no.4
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• pp.869-885
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• 2000

Fibroblasts are major cellular components of gingiva and periodontal ligament. They regulate the healing process after surgery or injury. Recently, many natural medicines, whose advantages are less side effects and possibility of long-term use, have been studied for their capacity, their anti-bacterial and anti-inflammatory effects and regenerative potential of periodontal tissues. Sophorae radix have been traditionally used as an anti-bacterial and antiinflammatory drug in oriental medicine. The purpose of present study was to investigate the effects of Sophorae radix extract on cell cycle progression and its molecular mechanism in human gingival fibroblasts. Sophorae radix extracts($100{\mu}g/ml$) notably increased cell proliferation and cell activity in the human gingival fibroblasts as compared to non-supplemented controls. There was an increase in the S phase and a decrease in the G1 phase in $100{\mu}g/ml$ of Sophorae radix extracts group as compared to non-supplemented controls. The level of cyclin E and cdk 2 protein in test group was higher than that of control groups. But that of cyclin D, cdk 4, and cdk 6 was not distinguished from controls. The level of p53 protein in test group was lower than that of controls, whereas that of p21 was not different. The level of pRB protein in test group was higher than that of controls, whereas that of p16 was lower. These results indicate that the increase of cell proliferation by Sophorae radix extracts may be due to the increased expression of cyclin E and cdk 2, and the decreased expression of p53 and p16 in human gingival fibroblasts.