• Molecules and Cells
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• v.37 no.7
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• pp.562-567
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• 2014

Human Hertwig's epithelial root sheath/epithelial rests of Malassez (HERS/ERM) cells are epithelial remnants of teeth residing in the periodontium. Although the functional roles of HERS/ERM cells have yet to be elucidated, they are a unique epithelial cell population in adult teeth and are reported to have stem cell characteristics. Therefore, HERS/ERM cells might play a role as an epithelial component for the repair or regeneration of dental hard tissues; however, they are very rare population in periodontium and the primary isolation of them is considered to be difficult. To overcome these problems, we immortalized primary HERS/ERM cells isolated from human periodontium using SV40 large T antigen (SV40 LT) and performed a characterization of the immortalized cell line. Primary HERS/ERM cells could not be maintained for more than 6 passages; however, immortalized HERS/ERM cells were maintained for more than 20 passages. There were no differences in the morphological and immunophenotypic characteristics of HERS/ERM cells and immortalized HERS/ERM cells. The expression of epithelial stem cell and embryonic stem cell markers was maintained in immortalized HERS/ERM cells. Moreover, immortalized HERS/ERM cells could acquire mesenchymal phenotypes through the epithelial-mesenchymal transition via TGF-${\beta}1$. In conclusion, we established an immortalized human HERS/ERM cell line with SV40 LT and expect this cell line to contribute to the understanding of the functional roles of HERS/ERM cells and the tissue engineering of teeth.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.42 no.1
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• pp.20-30
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• 2016

Objectives: To validate a critical-size mandibular bone defect model in miniature pigs. Materials and Methods: Bilateral notch defects were produced in the mandible of dentally mature miniature pigs. The right mandibular defect remained untreated while the left defect received an autograft. Bone healing was evaluated by computed tomography (CT) at 4 and 16 weeks, and by micro-CT and non-decalcified histology at 16 weeks. Results: In both the untreated and autograft treated groups, mineralized tissue volume was reduced significantly at 4 weeks post-surgery, but was comparable to the pre-surgery levels after 16 weeks. After 16 weeks, CT analysis indicated that significantly greater bone was regenerated in the autograft treated defect than in the untreated defect (P=0.013). Regardless of the treatment, the cortical bone was superior to the defect remodeled over 16 weeks to compensate for the notch defect. Conclusion: The presence of considerable bone healing in both treated and untreated groups suggests that this model is inadequate as a critical-size defect. Despite healing and adaptation, the original bone geometry and quality of the pre-injured mandible was not obtained. On the other hand, this model is justified for evaluating accelerated healing and mitigating the bone remodeling response, which are both important considerations for dental implant restorations.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.14 no.1_2
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• pp.77-88
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• 1992

Recently, diabetic patients are increasing in the field of microvascular surgery. Diabetes melltius is known to be related to arterial damage, platelet malfunction and thrombus formation. After microvascular anastomosis, delayed repair and vascular occlusion occurred more frequently in diabetic state. This study was performed to investigate the patency rate and process of endothelial healing after microvascular anastomosis of femoral artery in diabetic rat by scanning electron microscope. The animals were divided into two groups, 20 diabetic-induced and 20 non-diabetic groups. Diabetes was induced with a injection of Streptozotocin(50mg/kg b.w., Sigma Chemical Co.) to tail vein. The results obtained were as follows: 1. Macroscopically, anastomotic site was intact except a few cases showed minimal inflammatory sign around the wound site. But the inflammatory change was frequently occurred in diabetic-induced group. 2. The patency rate was 95% (19/20) in non-diabetic group and 65% (13/20) in diabetic-induced group. 3. In the non-diabetic group, anstomotic region was mostly endothelized by the alignment along the long axis of vessel but stitchs were not covered with endothelial cells. The thichkening of vessel wall was not observed. 4. In the diabetic-induced group, anastomotic region was not endothelized but covered with blood cellular components and connective tissue instead of endothelial cells. The thickening of the vessel wall was prominent in some diabetic-induced rats. These results suggest that diabetes was related to delayed regeneration of endothelium of vessels after microsurgical anastomosis.

• Journal of Periodontal and Implant Science
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• v.49 no.4
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• pp.258-267
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• 2019

Purpose: Increased bone regeneration has been achieved through the use of stem cells in combination with graft material. However, the survival of transplanted stem cells remains a major concern. The purpose of this study was to evaluate the viability of transplanted mesenchymal stem cells (MSCs) at an early time point (24 hours) based on the type and form of the scaffold used, including type I collagen membrane and synthetic bone. Methods: The stem cells were obtained from the periosteum of the otherwise healthy dental patients. Four symmetrical circular defects measuring 6 mm in diameter were made in New Zealand white rabbits using a trephine drill. The defects were grafted with 1) synthetic bone (${\beta}$-tricalcium phosphate/hydroxyapatite [${\beta}-TCP/HA$]) and $1{\times}10^5MSCs$, 2) collagen membrane and $1{\times}10^5MSCs$, 3) ${\beta}-TCP/HA+collagen$ membrane and $1{\times}10^5MSCs$, or 4) ${\beta}-TCP/HA$, a chipped collagen membrane and $1{\times}10^5MSCs$. Cellular viability and the cell migration rate were analyzed. Results: Cells were easily separated from the collagen membrane, but not from synthetic bone. The number of stem cells attached to synthetic bone in groups 1, 3, and 4 seemed to be similar. Cellular viability in group 2 was significantly higher than in the other groups (P<0.05). The cell migration rate was highest in group 2, but this difference was not statistically significant (P>0.05). Conclusions: This study showed that stem cells can be applied when a membrane is used as a scaffold under no or minimal pressure. When space maintenance is needed, stem cells can be loaded onto synthetic bone with a chipped membrane to enhance the survival rate.

• BMB Reports
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• v.52 no.6
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• pp.409-414
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• 2019

Natural compounds isolated from medicinal herbs and plants have immense significance in maintaining bone health. Hydrolysable tannins have been shown to possess a variety of medicinal properties including antiviral, anticancer, and anti-osteoclastogenic activities. As a part of a study on the discovery of alternative agent against skeletal diseases, we isolated a hydrolysable tannin, 2-O-digalloyl-1,3,4,6-tetra-O-galloyl-${\beta}$-D-glucose (DTOGG), from Galla Rhois and examined the effect on osteoclast formation and function. We found that DTOGG significantly inhibited receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation by downregulating the expression of the key regulator in osteoclastogenesis as well as osteoclast-related genes. Analysis of RANKL/RANK signaling revealed that DTOGG impaired activation of $I{\kappa}B{\alpha}$ and p65 in the nuclear factor kappa-lightchain-enhancer of activated B cells (NF-${\kappa}B$) signaling pathway. Furthermore, DTOGG reduced bone resorbing activity of osteoclasts, compared to the vehicle-treated control. These results suggest that DTOGG could be a useful natural compound to manage osteoclast-mediated skeletal diseases.

• Journal of Periodontal and Implant Science
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• v.49 no.2
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• pp.90-104
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• 2019

Purpose: The aim of this study was to evaluate clinical and radiographic changes and the survival rate after periodontal surgery using deproteinized bovine bone mineral (DBBM) with 10% collagen or DBBM with a collagen membrane in endo-periodontal lesions. Methods: A total of 52 cases (41 patients) with at least 5 years of follow-up were included in this study. After scaling and root planing with or without endodontic treatment, periodontal regenerative procedures with DBBM with 10% collagen alone or DBBM with a collagen membrane were performed, yielding the DBBM + 10% collagen and DBBM + collagen membrane groups, respectively. Changes in clinical parameters including the plaque index, bleeding on probing, probing pocket depth, gingival recession, relative clinical attachment level, mobility, and radiographic bone gains were evaluated immediately before periodontal surgical procedures and at a 12-month follow-up. Results: At the 12-month follow-up after regenerative procedures, improvements in clinical parameters and radiographic bone gains were observed in both treatment groups. The DBBM + 10% collagen group showed greater probing pocket depth reduction ($4.52{\pm}1.06mm$) than the DBBM + collagen membrane group ($4.04{\pm}0.82mm$). However, there were no significant differences between the groups. Additionally, the radiographic bone gain in the DBBM + 10% collagen group ($5.15{\pm}1.54mm$) was comparable to that of the DBBM + collagen membrane group ($5.35{\pm}1.84mm$). The 5-year survival rate of the teeth with endo-periodontal lesions after periodontal regenerative procedures was 92.31%. Conclusions: This study showed that regenerative procedures using DBBM with 10% collagen alone improved the clinical attachment level and radiographic bone level in endo-periodontal lesions. Successful maintenance of the results after regenerative procedures in endo-periodontal lesions can be obtained by repeated oral hygiene education within strict supportive periodontal treatment.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.4
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• pp.277-286
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• 2006

In the present study, I investigated the effects of N-methyl-D-aspartate (NMDA), arachidonic acid (AA), and Nitric Oxide Synthase Inhibitor (NOS-I), alone or in combination, on the viability of cultured primary normal human oral keratinocytes (NHOK). Specifically, we examined whether AA and NOS-I could protect primary NHOK from glutamate cytotoxicity. The purpose of this study was therefore the preliminary study for the examination of the interaction between these agents and NHOK in order to elucidate the mechanisms by which epithelial growth and regeneration are regulated. NHOK were obtained from gingival tissue of 20 individuals aged 20 to 29, and third passage (P3) cells were used for this study. Cell viability was measured by the MTT assay. NMDA and NNA, a calcium dependent NOS inhibitor, induced an initial increase in cell number, which subsequently decreased by the $7^{th}$ day. Low concentration of AA ($0.5\;{\mu}M$ & $1\;{\mu}M$) induced an increase in cell number while high concentrations of AA ($5\;{\mu}M$ & $10\;{\mu}M$) induced a decrease in cell number. The decrease in cell number induced by NMDA at the $7^{th}$ day was abolished by the addition of low concentrations of AA ($0.5\;{\mu}M$ & $1\;{\mu}M$) or NOS inhibitors. Low concentrations of AA ($0.5\;{\mu}M$ & $1\;{\mu}M$) or NOS inhibitors may protect the NHOK from NMDA induced cytotoxicity. These reactions might be related to the NMDA receptor in the cell and the change of the intracellular calcium ion concentration.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.26 no.1
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• pp.24-39
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• 2000

Various methods and graft materials have been used to fill in the defect adjacent to the implants and considered as clinically acceptable. But it is not clear whether the regenerated bone increases the implant-bone contact and supports the implant. The purpose of this study is to evaluate regenerated bone surrounding implants using bone morphogenetic protein(BMP) and demineralized freeze-dried bone(DFDB), and the interfaces between implants and regenerated bone. bBMP was extracted and partially purified from the bovine bone matrix using heparine chromatography. Demineralized freeze-dried bone was made from the dog. Inactive insoluble collagenous bone matrix(IBM) of dog was used as carrier of bBMP. Interfaces of titanium coated epoxy resin implants were processed for demineralized section for transmission electron microscopy(TEM) and those of screw type implants were for nondemineralized section for light and fluoromicroscopic examination. Implants were inserted in the inferior border of mandible of adult dogs and artificial bony defects($3{\times}3{\times}4mm$) were made at the mesial and distal side of implants. Defects were filled with BMP(BMP group) and DFDB(DFDB group). For the fluoromicroscopic examination, the fluorescent dyes(oxytetracycline, calcein green, alizarin red) were injected 2, 4, 6, 8, 12 weeks after implantation. The experimental animals were sacrificed at the 6th and the 12th week and their mandible were extirpated and processed for examination with light microscopy, fluoromicroscopy and TEM. The obtained results were as follows : 1. By the light microscopic findings, the defects were filled with woven bone at the 6th week and compact bone at the 12th week, and the osseointegrations were seen in both groups. There was no histological difference between them. 2. On the basis of the histomorphometric analysis, BMP group(6th week: 40.25%, 12th week: 56.04%) had higher bony contact ratio than DFDB group(38.37%, 42.63%). There was significant difference between two groups at the 12th week(p<0.05). 3. The amount of bone formation in BMP group was more prominent than in DFDB group. Significant difference was noted among two groups at the 6th and the 8th week(p<0.05). 4. By the transmission electron microscopic findings, $0.4-2{\mu}m$ soft tissue layer was found in adjacent to the interfaces and over the collagen fibrils of bone at the 6th week. However, about 100nm amorphous layer was noted at the interface or collagen fibrils directly extended to the titanium surface at the 12th week. There was no significant difference between two groups. 5. These results suggest that BMP and DFDB can be used as good graft materials in the regeneration of bone adjacent to implant, and BMP is more valuable as a bone inducer than DFDB.

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.4
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• pp.657-661
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• 2005

Various chemotherapeutic agents have been recommended for pulpotomy of primary teeth, and there are formocresol, ferric sulfate, and calcium hydroxide. Of those, formocresol has fixation effect of pulp tissue and high clinical success rate, so it is most commonly used agent. But formocresol has strong cytotoxic effects, thus many articles reported displacement and loss of permanent successor, amelogenesis imperfecta, mutation by general absorption, possibility of cancer induction. Recently, it has been reported that leakage by imperfect temporary sealing when FC-soaked cotton was inserted into the root canal caused necrosis of surrounding tissues. and that necrosis of alveolar bone related to the use of excessive formocresol. In this case, 2nd primary molar of upper left jaw was treated using formocresol in local clinic, but extracted because of lasting pain. Furthermore, symptoms didn't disappear so patient was refered to us. The patient was 8-year-old male, had foul odor from oral cavity and circular alveolar bone necrosis around the permanent successor' crown. Thus sequestrectomy was operated and observed through 19 months after operation, we found normal root development of permanent successor but no complete regeneration of alveolar bone defect and attached gingiva. Lesion of periodontal tissues by formocresol is irreversible, so we have to confirm the indication in using formocresol and pay attention to complete temporary sealing.

• Restorative Dentistry and Endodontics
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• v.35 no.1
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• pp.5-12
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• 2010

The purpose of this study was to evaluate the pulp tissue reaction to direct pulp capping of mechanically exposed beagle dogs' pulp with several capping materials. A total of 36 teeth of 2 healthy beagle dongs were used. The mechanically exposed pulps were capped with one of the followings: (1) Mineral Trioxide Aggregate (MTA: $ProRoot^{(R)}$ MTA. Dentsply, Tulsa, USA), (2) Clearfil SE Bond (Dentin adhesive system: Kuraray, Osaka, Japan), (3) Ultra-Blend (Photo-polymerized Calcium hydroxide: Ultradent, South Jordan, USA), (4) Dycal (Quick setting Calcium hydroxide: LD Caulk Co., Milford, USA) at 7, 30, and 90 days before sacrificing. The cavities were restored with Z350 flowable composite resin (3M ESPE, St. Paul. MN, USA). After the beagle dogs were sacrificed, the extracted teeth were fixed, decalcified, prepared for histological examination and stained with HE stain. The pulpal tissue responses to direct pulp capping materials were assessed. In MTA calcium hydroxide, and photo-polymerized calcium hydroxide groups, initial mild inflammatory cell infiltration, newly formed odontoblast-like cell layer and hard tissue bridge formation were observed. Compared with dentin adhesive system, these materials were biocompatible and good for pulp tissue regeneration. In dentin adhesive system group, severe inflammatory cell infiltration, pulp tissue degeneration and pulp tissue necrosis were observed. It seemed evident that application of dentin adhesive system in direct pulp capping of beagle dog teeth cannot lead to acceptable repair of the pulp tissue with dentine bridge formation.