• Journal of Korean Dental Science
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• 제7권2호
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• pp.80-86
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• 2014

Purpose: Current common method of cytotoxicity evaluation for elastomeric impression materials use animal based cell lines, which the clinical relevance has been often questioned. Hence, the purpose of this study was to examine the difference in results with both human based and animal based fibroblast cell line. Materials and Methods: Three types of fibroblast cells were used in this study; conventional mouse fibroblasts of L929, human gingival fibroblasts (HGF-1), and immortalized human oral fibrobalsts (hTERT-hNOF). Test on extract and test by direct contact using different commercially available elastomeric impression materials were carried out according to the international standards. Result: There was significant difference in cell viability between types of fibroblasts cell used, where HGF-1 showed highest cell viability and L929 the lowest. Conclusion: Within the limitation of this study, careful consideration must be given when selecting the cells and interpreting the results for cytotoxicity evaluation of elastomeric impression materials, where use of human based cell lines such as hTERT-hNOF would be appropriate for both ease of cytotoxicity test and clinical relevance.

• 한국가축번식학회지
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• 제24권1호
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• pp.49-57
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• 2000

본 연구는 소 태아섬유아세포를 이용하여 핵이식 후 세포의 휴면처리, 세포의 passage 수 및 세포의 기원이 복제란의 발육에 미치는 영향을 검토하였다.3.57개 월령 한우 수컷 태아의 피부 및 간 조직에서 세포를 채취하여 체외에서 4∼6 회 계대배양 후 동결하였다가 핵이식에 사용하였다. 세포는 핵이식 전에 혈청기아처리를 하였으며, 대조구로는 활발히 분열 중의 무처리 세포를 사용하였다. Donor 세포는 미수정란의 탈핵세포질에 이식 후 전기융합 및 활성화를 실시하였고, 재구축배는 7∼9 일간 체외배양하여 발육농을 검토하였다. 배반포로 발육된 일부 복제란은 발정 7∼8 일째의 수란우에 이식하였다. 복제란의 배반포 발육율은 혈청기아 처리구 (25.3%)가 무처리구 (15.9%)에 비하여 유의적으로 높았으나 (P＜0.05), 세포의 passage 수 (4∼6회) 에 관계없이 23.1∼25.0%로 나타났고, 피부 및 간유래 복제란의 배반포 발육율도 23.8∼25.2% 로 두 조직세포 간에 차이가 없었다. 복제란의 이식 후 60일 및 120일에 수란우의 34.4% 및 15.6%가 각각 임신이 확인되었으며, 초자화동결된 태아 피부세포 복제란으로부터 1두의 수컷 송아지가 생산되었다. 본 연구의 결과는 복제란의 체외발육능이 세포의 휴면처리에 의하여 향상되었으나, 세포의 passage 수 (4∼6 회) 및 세포의 두 기원 (피부 및 간) 에 의해서는 영향을 받지 않으며, 태아 피부세포 유래 복제란으로부터 산자가 생산될 수 있음을 확증한다.

• Radiation Oncology Journal
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• 제24권3호
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• pp.179-184
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• 2006

목 적: 재조합 표피성장인자는(rhEGF) 다양한 표피와 상피 세포의 증식을 자극하는 것으로 알려져 있다. 본 연구에서는 방사선이 조사된 섬유아세포의 증식에 rhEGF의 효과를 알아보고자 하였다. 대상 및 방법: 인간에서 기원한 섬유아세포를 초대배양(primary culture)한 세포를 이용하였다. 대웅제약에서 유전자 재조합하여 대장균에서 발현하여 생산한 rhEGF를 제공 받아 사용하였다. 방사선 조사는 4 MV 선형가속기(CLINAC 600C, Varian, Palo Alto, CA, USA)를 이용하여 분당 2 Gy 내외의 선량률로 균일하게 조사하였다. 조사된 방사선량은 8 Gy이었다. 생존세포수는 trypan blue 염색법을 이용하였고, rhEGF에 의한 세포주기의 변화를 관찰하기 위하여 유세포 분석법을 시행하였다. 결 과: 4 Gy의 방사선을 조사한 후 7일째까지 생존 세포수를 trypan blue 염색법을 이용하여 측정한 결과 모든 rhEGF 농도(1.0 nM, 10 nM, 100 nM, 1,000 nM )에서 방사선 조사 단독군보다 생존세포 수가 많았다. 방사선을 조사하지 않은 섬유아세포에서 rhEGF를 10 nM처리한 후 FACS scan을 시행한 결과 세포주기 중에서 S기 비율이 증가하였다. 결 론: 방사선이 조사된 섬유아세포에서 rhEGF를 투여하면 rhEGF를 투여하지 않은 섬유아세포에 비해서 세포증식이 가속됨을 확인할 수 있었다.

• Journal of Periodontal and Implant Science
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• 제26권4호
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• pp.841-858
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• 1996

Epidermal growth factor(EGF) is one of polypeptide growth factors. EGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of EGF on the human periodontal ligament cells and human gingival fibroblast cells that promote regeneration of periodntal tissue. The mitogenic effects of epidermal growth factor on human periodontal ligament cells and human gingival fibroblasts were evaluated by determining the incorporation of 5-Bromo-2'-deoxy-uridine into DNA of the cells in a dose dependent manner. The prepared cells were the primary cultured gingival fibroblast and periodontal ligament cells from humans, the fourth or sixth subpassages were used in the experiments. Cells were seeded in DMEM containing 10% FBS. 1, 10, 50, 100, $200{\eta}g/ml$ and epidermal growth factor were added to the quiescent cells for 24 hours, 48 hours and 72 hours. They were labeled with $10\{mu}l/200{\mu}l$ 5-Bromo-2'-deoxy-uridine for the last 6 hours of each culture. The results of the five determinants were presented as mean and S.D.. The results were as follows : The DNA synthetic activity of human gingival fibroblasts were increased dose dependently by epidermal growth factor at 24 hours, 48 hours and 72 hours. The mitogenic effects were similar at the 24 and 48 hours of epidermal growth factor, but the DNA synthetic activity of human gingival fibroblasts generally decreased at 72 hours. The DNA synthetic activity of human periodontal ligament cells were increased dose dependently by epidermal growth factor at 24 hours but the DNA synthetic activity decreased at $200{\eta}g/ml$ of each hour. Generally the maximum mitogenic effects were observed at the 48 hours application of epidermal growth factor. The DNA synthetic activity of human periodontal ligament cells generally decreased lower at 24, 72 hours than at 48 hours the application of epidermal growth factor. In the comparison of DNA synthetic activity between human gingival fibroblasts and human periodontal ligament cells, human periodontal ligament cells had slightly higher proliferation activity than human gingival fibroblasts for a longer time at the high dosage of the epidermal growth factor. In conclusion, epidermal growth factor have important roles in the stimulation of DNA synthesis in human periodontal ligament cells and human gingival fibroblasts, and thus may be useful for clinical applications in periodontal regenerative procedures.

• Toxicological Research
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• 제35권2호
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• pp.167-179
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• 2019

Ovarian cancer is the fifth main cause of pre-senescent death in women. Although chemotherapy is generally an efficient treatment, its side effects and the occurrence of chemotherapeutic resistance have prompted the need for alternative treatments. In this study, ${\alpha}$-mangostin and apigenin were evaluated as possible anticancer alternatives to the chemotherapeutic drug doxorubicin, used herein as a positive control. The ovarian adenocarcinoma cell line SKOV-3 (ATCC No. HTB77) was used as model ovarian cancer cells, whereas the skin fibroblast line CCD-986Sk (ATCC No. CRL-1947) and lung fibroblast line WI-38 (ATCC No. CCL-75) were used as model untransformed cells. Apigenin and doxorubicin inhibited the growth of SKOV-3 cells in a dose- and time-dependent manner. After 72 hr exposure, doxorubicin was mostly toxic to SKOV-3 cells, whereas apigenin was toxic to SKOV-3 cells but not CCD-986Sk and WI-38 cells. ${\alpha}$-Mangostin was more toxic to SKOV-3 cells than to CCD-986Sk cells. A lower cell density, cell shrinkage, and more unattached (floating round) cells were observed in all treated SKOV-3 cells, but the greatest effects were observed with ${\alpha}$-mangostin. With regard to programmed cell death, apigenin caused early apoptosis within 24 hr, whereas ${\alpha}$-mangostin and doxorubicin caused late apoptosis and necrosis after 72 hr of exposure. Caspase-3 activity was significantly increased in ${\alpha}$-mangostin-treated SKOV-3 cells after 12 hr of exposure, whereas only caspase-9 activity was significantly increased in apigenin-treated SKOV-3 cells at 24 hr. Both ${\alpha}$-mangostin and apigenin arrested the cell cycle at the $G_2/M$ phase, but after 24 and 48 hr, respectively. Significant upregulation of BCL2 (apoptosis-associated gene) and COX2 (inflammation-associated gene) transcripts was observed in apigenin- and ${\alpha}$-mangostin-treated SKOV-3 cells, respectively. ${\alpha}$-Mangostin and apigenin are therefore alternative options for SKOV-3 cell inhibition, with apigenin causing rapid early apoptosis related to the intrinsic apoptotic pathway, and ${\alpha}$-mangostin likely being involved with inflammation.

• 대한세포병리학회지
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• 제4권1호
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• pp.70-73
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• 1993

Giant cell tumor(GCT) occurs very unusually in the rib(less than 1% of GCT). We present the cytologic features of GCT of the rib. It showed multiple cellular clusters composed of characteristic, benign looking osteoclast-like multinucleated giant cells and fibroblast-like mononuclear cells. The multinucleated giant cells contained numerous nuclei (average, 30 to 40 per cell, which were closely packed. The nuclei in giant cells were remarkably uniform and round to oval. The mononuclear, neoplastic stromal cells were elongated and spindle-shaped. There was no cytologically malignant portion in the tumor.

• 한국환경성돌연변이발암원학회지
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• 제11권2호
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• pp.99-106
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• 1991

수용체를 통한 저밀도 지방단백질(LDL)의 세포내 이동과 함께 일어나는 지용성 돌연변이원인 벤조피렌의 세포내 이동에 관하여 조사하였다. [$^3$H]로 표지된 벤조피 렌의 Hep 2 세포내 이동을 조사한 결과, 배지에 LDL을 첨가한 경우. LDL이 첨가되지 않은 경우보다 3배나 많은 양의 벤조피렌이 세포내로 이동하여 세포내 물질과 결합하였음을 관찰하였다. 그러나 10$\mu$M의 황산구리로 LDL의 아포단백질을 변형 시킨 후 변형된 LDL을 배지에 첨가하고 벤조피렌의 세포내 이동을 조사한 건과, 정상 LDL이 첨가된 경우보다 벤조피렌의 세포내 이동이 현저히 감소하였다. 이 결과는 LDL을 이동체로 이용한 벤조피렌의 세포내 이동이 세포의 수용체와 LDL의 아포 단백질의 인식과정을 거친다는 사실을 뒷받침한다고 여겨진다.

• 대한의생명과학회지
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• 제18권2호
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• pp.165-168
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• 2012

Hox proteins containing DNA-binding homedomain act as transcription factors important for anteroposterior body patterning during vertebrate embryogenesis. However, the precise mechanisms by which signal pathways are transduced to regulate the Hox gene expression are not clear. In the course of an attempt to isolate an upstream regulatory factor(s) controlling Hox genes, protein kinase B alpha (Akt1) has been identified as a putative regulator of Hox genes through in silico analysis (GEO profile). In the Gene Expression Omnibus (GEO) dataset GDS1784 at the NCBI (National Center for Biotechnology Information) site, Hox genes were differentially expressed depending on the presence or absence of Akt1. Since it was not well known how Akt1 regulates the specific Hox genes, whose transcription was reported to be regulated by epigenetic modifications such as histone acetylation, methylation etc., the expression of Gcn5, a histone acetyltransferase (HAT), was analyzed in wild type (WT) as well as in $Akt1^{-/-}$ mouse embryonic fibroblast (MEF) cells. RT-PCR analysis revealed that the amount of Gcn5 mRNA was similar in both WT and $Akt1^{-/-}$ MEFs. However, the protein level of Gcn5 was significantly increased in $Akt1^{-/-}$ MEF cells. The half life of Gcn5 was 1 hour in wild type whereas 8 hours in $Akt1^{-/-}$ MEF. These data all together, indicate that Gcn5 is post-transcriptionally down-regulated and the protein stability is negatively regulated by Akt1 in MEF cells.

• 한국재료학회지
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• 제20권7호
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• pp.392-400
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• 2010

The electrospinning process was established as a promising method to fabricate nano and micro-textured scaffolds for tissue engineering applications. A BCP-loaded PCL micro-textured scaffold thus can be a viable option. The biocompatibility as well as the mechanical properties of such scaffold materials should be optimized for this purpose. In this study, a composite scaffold of poly ($\varepsilon$-caprolactone) (PCL)-biphase calcium phosphate (BCP) was successfully fabricated by electrospinning. EDS and XRD data show successful loading of BCP nano particles in the PCL fibers. Morphological characterization of fibers shows that with a higher loaded BCP content the fiber surface was rougher and the diameter was approximately 1 to 7 ${\mu}m$. Tensile modulus and ultimate tensile stress reached their highest values in the PCL- 10 wt% BCP composite. When content of nano ceramic particles was low, they were dispersed in the fibers as reinforcements for the polymer matrix. However, at a high content of ceramic particles, the particles tend to agglomerate and lead to decreasing tensile modulus and ultimate stress of the PCL-BCP composite mats. Therefore, the use of nano BCP content for distribution in fiber polymer using BCP for reinforcement is limited. Tensile strain decreased with increasing content of BCP loading. From in vitro study using MG-63 osteoblast cells and L-929 fibroblast like cells, it was confirmed that electrospun PCL-BCP composite mats were biocompatible and that spreading behavior was good. As BCP content increased, the area of cell spreading on the surface of the mats also increased. Cells showed the best adherence on the surface of composite mats at 50 wt% BCP for both L-929 fibroblast-like cells and MG-63 osteoblast cell. PCL- BCP composites are a promising material for application in bone scaffolds.

• 대한한의학회지
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• 제39권1호
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• pp.63-74
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• 2018

Objectives: Periodontitis is an inflammatory disease with the destruction of periodontal ligament, alveolar bone loss and inflammation of gingva, leading to teeth loss. Eclipta prostrata L. (Ecliptae Herba) has been used to treat the inflammatory disease as a Korean traditional medicine. The aim of this study is to investigate the effects of E. prostrata L. on periodontitis. Methods: E. prostrata L. was extracted with water and lyophilized. The aqueous extract of E. prostrata L. (EP) was topically applied to the periodontal lesion for 2 weeks. To induce the periodontitis, a 3-0 nylon ligature was placed around the cervix of the lower first molar in rat. Rats were divided into 3 groups (n = 7); NL group (non-ligatured and non-treated), L group (ligatured and vehicle-treated) and EP group (ligatured and EP-treated). After sacrifice, the mandibles was dissected and stained with methylene blue solution to analyze the alveolar bone loss. The expression of MMP-9 was determined in gingival tissues. To confirm the effect of EP on recovery of gingiva, mRNA expressions of type I pro-collagen and MMP-9 levels were investigated in LPS-treated HS68 fibroblast cells. In addition, inflammatory mediators were evaluated in LPS-treated RAW264.7 cells. Results: Alveolar bone loss was significantly inhibited by EP treatment. The mRNA expression of MMP-9 was attenuated in rats treated with EP. In addition, treatment with EP increased the expression of type I pro-collagen, while the expression of MMP-9 was decreased in LPS-stimulated HS68 fibroblast cells. Furthermore, EP down-regulated the LPS-induced IL-6, $TNF-{\alpha}$, COX-2 and iNOS production in RAW264.7 cells. Conclusions: These results suggest that EP have ameliorative effects on periodontitis through inhibiting alveolar bone loss and modulating the inflammatory mediators. Therefore, E. prostrata L. may be an alternative on patients with periodontitis.