• Journal of Periodontal and Implant Science
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• v.28 no.2
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• pp.235-247
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• 1998

Calcium sulfate has a long history of medical use as an implant material. The biocompatibiliry of the material has been clearly established. Bone ingrowth concomitant with resorption occurs rapidly with efficient conduction of bone from particle to particle. Calcium sulfate also has a potential for functioning as a good bamer membrane. The purpose of this study was to compare the biocompatibility of different types of calcium sulfate grafting materials including an expelimental calcium sulfate compound on periodontal ligament cells in vitro as a preliminary test towards the development of a more convenient and useful form of grafting material which could promote regeneration of periodontal tissue. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. cells were cultured in a.MEM culture medium containing 20% FBS, at $37^{\circ}C$ and 100% humidity, in a 5% CO2 incubator. Cells were cultured into 96 well culture plate $1{\times}104$ cells per well with $\alpha$-MEM and incubated for 24 hours. After discarding the medium, those cells were cultured in $\alpha$-MEM contained with 10% FBS alone (control group), in medcal-grade calcium sulfate(MGCS group), in plaster(plaster group), experimental calcium sulfate paste(CS paste group) for 1, 2, 3 day respectively. And then each group was characterized by examining of the cell counting, MTI assay, collagen synthesis. The results \vere as follows. 1. In the analysis of cell proliferation by cell counting, both medical-grdde calcium sulfate group and plaster group showed no stastically significant difference at day 1, 2, 3 accept for plaster group at day 1 compared to control group, but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.05). 2. In the analysis of cytotoxicity by MIT assay, both medical-grade calcium sJlfate group and plaster group showed no stastically significant difference compared to control group at day 1, 2, 3 but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.OS). 3. In the analysis of collagen synthesis by immunoblotting assay, high level was detected for medical-grade calcium sulfate group and plaster group at day 1, 2, 3 compared to CS paste group. On the basis of these results, medical-grade calcium sulfate and plaster was shown to possess biocompatibility whereas the CS paste had unfavourable outcome. This observation shows a need for modification of the materials contained in calcium sulfate paste.

• Journal of Periodontal and Implant Science
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• v.35 no.4
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• pp.949-959
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• 2005

이 연구는 임플란트를 식립하기를 원하는 전신건강상태가 양호하며 구강위생상태가 좋은 14명 환자(남자:8명, 여자:6명, 평균나이 : 44세)의 20개의 발치와 내에 흡수성 차폐막(BioMesh. Sam Yang Corporation, Korea)과 함께 탈회냉동 건조동종 골(dem-ineralized freezedried bone allografts, $250-500{\mu}m$. Pacific Coast Tissue Bank, U.S.A.)과 이종골(Bovine-Bone, Bio-Oss 0.25-1.0 mm, Geistlich, Biomaterials and Osteohealth, Switzerland)을 1:1(부피)로 혼합하여 이식한 후 그 치유양상을 관찰하고자 조직학적 및 면역조직화학적으로 평가하였다. 이직재가 탈락되는 것을 방지하기 위하여 발치한 후 1개윌이 경과된 후에 이식재와 차폐막을 위치시켰다. 표본제작을 위하여 이식술을 시행한 지 약 6개윌 후에 임플란트를 식립하기 직전 식립부위에서 trephine bur로 골을 채취하였는데, 20증례 중 7증례에서 임플란트를 식립하기 전에 차폐막이 노출되었다. 차폐막이 노출되지 않은 것을 대조군으로, 노출된 것을 실험군으로 설정하였다. 조직학적인 관찰을 위하여 통상적인 방법에 따라 탈회 표본을 제작하였고, alkaline phosphotase(ALP)틀 이용하여 면역조직화학적 염색을 시행한 후 골 형성 상태를 평가하여 다음과 같은 결과를 얻었다. 본 연구에서는 발치와내에서 골유도재생술 후 나타나는 치유 형태를 5가지 형태로 분류할 수 있었다. Type I, II와 III는 새로운 골 형성을 나타내지 않았고, 면역조직화학적 검사 시 ALP 음성 소견을 나타내었다. Type V는 새로운 골 형성과 ALP 양성 소견을 나타내었으나 염증, 괴사, 결합조직의 증식 등은 없었다. Type IV와 Type V의 차이는 결합조직의 증식여부로 구분되었다. 막이 노출되지않은 증례들 중 7 증례에서는 Type V의 치유 형태를, 2증례에서는 Type IV의 치유 형태를 나타내었다. 막이 노출되었던 증례에서는 Type I, II, III의 다양한 치유 형태를 나타내었다. 본 연구결과, 발치와 내에 골유도재생술을 시행한 후 차폐막의 노출 여부가 신생골 형성에 중요한 영향을 미칠 것으로 사료되며, 본 연구에서 분류한 치유 형태가 향후 골유도재생술 후의 결과 분석에 활용될 수 있을 것으로 사료된다.

• Journal of Periodontal and Implant Science
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• v.37 no.4
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• pp.791-803
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• 2007

The purpose of this study was to evaluate exophytically vertical bone formation in residual ridge of the beagle dog by the concept of guided bone regeneration with a titanium reinforced e-PTFE membrane combined with irradiated cancellous human bone. Twelve male beagle dogs(mean age 1.5 years and mean weight 12kg) were used for this study. The alveolar ridges after extraction of all mandibular premolars were surgically and horizontally removed. At 8 weeks after extractions, full-thickness flap was reflected and cortical bone was removed with round bur and copious irrigation. Rectangular parallelepiped(10mm in length, 5mm in width, and 4mm in height) bended with titanium-reinforced e-PTFE(TR e-PTFE) membrane was placed on the decorticated alveolar ridge, fixed with metal pins and covered with full-thickness flap and assigned as a control group. Test groups ere treated with TR e-PTFE membrane filled with irradiated cancellous human bone. Of twelve beagle dogs, four control dogs and four test dogs without membrane exposure to oral cavity were sacrificed at 8 and 16 weeks respectively. The surgical sites were dissected out, fixed in 4% buffered formaldehyde, dyed using a Villanueva staining technique, and processed for embedding in plastic resin. The cutting and grinding methods were routinely processed for histologic and histomophometric analyis of exophytic bone formation as well as statistical analysis. The results of this study were as follows: 1. Exophytic bone formation in the both of experimental groups was increased respectively after surgery from 23.40% at 8 weeks to 46.26% at 16 weeks in the control groups, from 40.23% at 8 weeks to 47.11% at 16 weeks in the test groups(p<0.05). 2. At 8 weeks after surgery, exophytic bone formation was made 40.23% in the test groups and 33.40% in the control groups. Exophytic bone formation was significantly made in the test group more than in the control group. At 16 weeks after surgery, exophytic bone formation was made 44.11% in the test groups and 46.26% in the control groups. Exophytic bone formation was made in the test groups more than in the control groups, but there was no statistically significant differences. 3. The membrane was fixed with metal pins to closely contact it to the bone surface. So, collapse and deviation of the membrane could be prevented and in growth of connective tissue also could be blocked from the periphery of the membrane. On the basis of these findings, wee suggest that intraoral experimental model for exophytic bone formation may be effective to evaluate the effect of bone graft material. And it indicates that combined use of membrane and ICB graft material is more effective than use of membrane only for exophytic bone formation.

• Journal of Periodontal and Implant Science
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• v.37 no.4
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• pp.655-669
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• 2007

Reactive oxygen species (ROS) have been implicated in the pathogenesis of various diseases. And vitamin C has shown a protective effect for the tissues. The aim of this study was to evaluate the effects of $H_2O_2$ and ascorbic acid on matrix metalloproteinase-1 (MMP-1), tissue inhibitor of metalloproteinase (TIMP: TIMP-1, TIMP-2), Type 1 collagen, fibronectin, and PDLs22 level in human periodontal ligament fibroblasts (hPDLF) via reverse transcription-polymerase chain reaction (RT-PCR). hPDLF was obtained from a healthy periodontium and cultured in Dulbecco's modified Eagles's medium plus 10% fetal bone serum. The concentration of ascorbic acid in hPDLF was $50{\mu}g/ml$, and that of $H_2O_2$ in hPDLF was 0.03% and 0.00003%. Ascorbic acid only, $H_2O_2$ only and mixture of ascorbic acid and $H_2O_2$ were applied with hPDLF for 1-, 3-, and 30-min. respectively. The gene expression of MMP-1-, TIMP-1-, TIMP-2-, Type 1 collagen-, fibronectin-, and PDLs22-mRNA in hPDLF was analysed via RT-PCR. The results were as follows; 1. hPDLF in response to 30-min. incubation with 0.03% $H_2O_2$ did not show any gene expression. 2. In all the experimental groups, the gene expression of fibronectin mRNA showed the decreased tendency compared to control. 3. In all the experimental groups, the gene expression of TIMP-1 mRNA showed the tendency similar to control. 4. hPDLF in response to 30-min. incubation with 0.03% $H_2O_2$ and ascorbic acid increased mRNA induction for MMP-1. 5. In all the experimental groups, hPDLF increased mRNA induction for PDLs22, collagen type 1, and TIMP-2 compared to control. Within the limited experiments, $H_2O_2$ and ascorbic acid increased mRNA induction for PDLs22, collagen type 1, TIMP-2 in hPDLF. More research will be needed in order to confirm the relative importance of the different roles of ROS and antioxidants in hPDLF from a periodontal regeneration or repair standpoint.

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.3
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• pp.395-402
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• 2005

In considering the healing process of injured periodontal tissue, healing rate would be influenced by the cellular activity of periodontal fibroblasts(PDLFs). In addition, the reattachment among PDLFs should be induced for healing process. The purpose of this study was to evaluate the effects of epidermal growth factor(EGF) on the proliferation and attachment of PDLFs and to verify the efficacy of EGF as a storage media or a pre-replantation conditioner of traumatically avulsed tooth. Human recombinant epidermal growth factor(hrEGF) and human periodontal fibroblasts from first premolar were prepared. At first, MTT assay was done to evaluate the toxic effect on human periodontal fibroblast and the maximum cellular growth of EGF. Cellular proliferation rate was then compared between control group and 10ng/ml EGF added group. Also, western blot was done to evaluate the expression of fibronectin in both groups. The results were as follows: 1. From MTT assay, EGF showed no toxic effect on PDL fibroblasts. The highest proliferation was shown at 10ng/ml EGF. 2. In 10ng/ml EGF added group, the degree of proliferation of PDLFs was significantly higher than that in control group. 3. Fibronectin expression of EGF added group was also significantly higher than that of control group. From this study we could conclude that EGF enhanced the regeneration rate of periodontal fibroblast, which could be used as a pretreatment agent or a storage media for traumatically avulsed teeth.

• Korean Journal of Medicinal Crop Science
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• v.1 no.2
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• pp.129-136
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• 1993

For the clonal proliferation of Gentiana scabra Bunge. which is one of the medicinaland ornamental plant, establishment multiplication of shoot through tissue culture technique and transplantation into soil were carried out. The shoot proliferation increased on the MS medium containing 0.5mg/l NAA and 0.5mg/l BAP. Optimum pH for shoot growth was pH 5.9, consequently MS medium supplemented with 2g/l activated charcoal was most effective for plant growth. There are two types of somaclonal variants, tall type was 63% and dwarf type was 37%.

• Journal of Dental Rehabilitation and Applied Science
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• v.36 no.3
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• pp.145-157
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• 2020

Purpose: This study was to determine the possible effects of trichloroacetic acid (TCA) and epidermal growth factor (EGF) through cell survival genes of the PI3K-AKT signaling pathway when applying an hydrophobically modified glycol chitosan (HGC)-based nanocontrolled release system to human gingival fibroblasts in oral soft tissue regeneration. Materials and Methods: An HGC-based nano-controlled release system was produced, followed by the loading of TCA and EGF. The group was divided into control (CON), TCA-loaded nano-controlled release system (EXP1), and the TCA- and EGF- individually loaded nano-controlled release system (EXP2). A total for 29 genes related to the PI3K-AKT signaling pathway were analyzed after 48h of culture in human gingival fibroblasts. Real-time PCR, 1- way ANOVA and multiple regression analysis were performed. Results: Cell survival genes were significantly upregulated in EXP1 and EXP2. From multiple regression analysis, ITGB1 was determined to be the most influential factor for AKT1 expression. Conclusion: The application of TCA and EGF through the HGC-based nano-controlled release system can up-regulate the cell survival pathway.

• Journal of Chest Surgery
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• v.29 no.11
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• pp.1182-1190
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• 1996

The best treatment of congenital or acquired tracheal stenosis is resection and end to end anastomosis. Various prosthetic material and tissue graft replacement can be considered when the stenotic segment is too long, but their uses are still limited due to many serious complications. The present study examined the effect of immunosuppression and cryopreserved allograft trachea after intraperitoneal omental implantation for evaluation of the possibility of tracheal transplantation. Thirty tracheal segments were harvested from fifteen donor Wistar rats. Among them eighteen segments were implanted immediately(group I, II, III) and twelve segments were used for cryopreservation(group IV, V). Heterotopical intraperitoneal implantation was performed in five groups of rats(n=6); Group I was Wistar syngeneic controls and received no immunosuppression. Group II and III were those of Sprague-Dawley recipients, the former receiving no immunosuppression and the latter receiving immunosuppression(Cyclosporin A 15mg/kg/day, Methylprednisolone 2mg/kg/day). Group IV and V were groups of Sprague-Dawley recipients, the former receiving immunosuppression and the latter receiving no Immunosuppression. After 28 days, rats were sacrificed and the tracheal segments were histologically evaluated. Epithelial thickness was significantly decreased in group II, IV. Epithelial regeneration score was also significantly decreased in II. All rats maintained well their round tracheal contour. In conclusion; I) trachea could be preserved for a long time with cryo method, 2) epithelium could regenerate fully with omentopexy in cryopreserved trachea, 3) immunosuppresion was not necessary with cryopreserved trachea.

• Proceedings of the Korean Vacuum Society Conference
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• 2010.02a
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• pp.44-45
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• 2010

We are currently conducting studies on culturing and biocompatibility assessment of various cells such as neural stem cells and induced pluripotent stem cells(IPS cells) on carbon nanotube (CNT), on nerve regeneration electrodes, and on silicon wafers with a focus on developing nerve integrated CNT based bio devices for interfacing with living organisms, in order to develop brain-machine interfaces (BMI). In addition, we are carried out the chemical modification of carbon nanotube (mainly SWCNTs)-based bio-nanosensors by the plasma ion irradiation (plasma activation) method, and provide a characteristic evaluation of a bio-nanosensor using bovine serum albumin (BSA)/anti-BSA binding and oligonucleotide hybridization. On the other hand, the researches in the case of "novel plasma" have been widely conducted in the fields of chemistry, solid physics, and nanomaterial science. From the above-mentioned background, we are conducting basic experiments on direct irradiation of body tissues and cells using a micro-spot atmospheric pressure plasma source. The device is a coaxial structure having a tungsten wire installed inside a glass capillary, and a grounded ring electrode wrapped on the outside. The conditions of plasma generation are as follows: applied voltage: 5-9 kV, frequency: 1-3 kHz, helium (He) gas flow: 1-1.5 L/min, and plasma irradiation time: 1-300 sec. The experiment was conducted by preparing a culture medium containing mouse fibroblasts (NIH3T3) on a culture dish. A culture dish irradiated with plasma was introduced into a $CO_2$-incubator. The small animals used in the experiment involving plasma irradiation into living tissue were rat, rabbit, and pick and are deeply anesthetized with the gas anesthesia. According to the dependency of cell numbers against the plasma irradiation time, when only He gas was flowed, the growth of cells was inhibited as the floatation of cells caused by gas agitation inside the culture was promoted. On the other hand, there was no floatation of cells and healthy growth was observed when plasma was irradiated. Furthermore, in an experiment testing the effects of plasma irradiation on rats that were artificially given burn wounds, no evidence of electric shock injuries was found in the irradiated areas. In fact, the observed evidence of healing and improvements of the burn wounds suggested the presence of healing effects due to the growth factors in the tissues. Therefore, it appears that the interaction due to ion/radicalcollisions causes a substantial effect on the proliferation of growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), and transforming growth factor (TGF) that are present in the cells.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.82-87
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• 2007

Five dogs with renal failure were referred to the Veterinary Medical Teaching Hospital at Kangwon National University. These dogs had the common history of consumption of Pedigree dry dog food produced in Thailand plant for over 1 month. The dogs showed anorexia, emaciation, vomiting, and polydipsia/polyuria. And in one severely affected dog, bloody diarrhea and hypothermia were seen. The remarkable clinicopathological signs were high value of BUN and creatinine. In some dogs, GGT, phosphorus and lipase were increased. However, no significant changes of complete blood count were found. In urinalysis, hematuria, low specific gravity urine, proteinuria, and calcium oxalate-like crystals were observed. Two severely affected dogs were died. The remained dogs were recovered gradually after change of dog food and supportive therapy. Pathological findings were seen typically in kidneys. Renal atrophy, congestion of the glomerular capillary, and diffuse degeneration, necrosis, dystrophic calcification and regeneration in the tubular epithelium were seen. Yellowish brown fluorolucent laminated materials or particles were quite often found in the lumina of the necrotizing renal tubules of cortex and medulla. Proliferation of fibrous tissue in the interstitium was also seen. By the mycotoxin analysis of the Pedigree dry dog food, ochratoxin A (OTA) and citrinin were detected as much as the concentration of 372.8 ppb and 8.3 ppb, respectively. The final diagnosis of renal failure caused by OTA and citrinin toxicosis was made on the basis of history takings, clinical signs, clinicopathological and pathological findings, and analysis of mycotoxins.