Epigallocatechin-3-gallate (EGCG) Induces Cell Death of Oral Squamous Carcinoma Cells

박형목,이은경,정진,박봉수,박혜련,유미현 대한구강악안면병리학회 2009 대한구강악안면병리학회지 Vol.33 No.4

Green tea, derived from the plant Camellia sinensis, is one of the most common beverages consumed worldwide. Epigallocatechin-3-gallate (EGCG) is the most abundant and bioactive polyphenolic constituent in green tea. Understanding how intracellular signaling pathways respond to EGCG may provide a clue to the difference of cell responses and basis for usefulness of EGCG as a chemopreventive and/or chemotherapeutic agent. In the present study, we tried to check whether EGCG could be a useful agent in chemotherapeutic treatment of oral squamous carcinoma. Furthermore, we investigated which signaling pathway is involved in biologic activities of EGCG. EGCG induced the cell death of oral squamous carcinoma cells. Furthermore, it increased phosphorylation of Akt in serum-strarved oral squamous carcinoma cells. But, initial increase of Akt activation did not affect cell survival. Activities of Raf-1 and Erk showed inconsistent response to EGCG treatment, but Erk phosphorylation is consistent with Raf-1 activity in YD 10B cells. These changes of Raf-1 and Erk activity in EGCG treated cells were different depending on cell line type. Supposedly, the difference of cell component may affect the Raf-1 and Erk reactivity to EGCG treatment. Akt activation by EGCG is independent on activities of PDK1 and PTEN, and expression of bax and bcl-2 proteins were not changed by EGCG treatment. Therefore, EGCG treatment did not induce the apoptosis of YD 10B cell. On the other hand, vascular adhesion molecule (VCAM) was decreased by EGCG treatment, so it is possible that decrease of VCAM can play certain role in survival and/or cell death in EGCG treated cells.

(-)-Epigallocatechin-3-Gallate의 3T3-L1 세포에서 항산화 효소 활성 및 지방세포 분화 억제 효과

김영화(Younghwa Kim) 한국식품영양과학회 2017 한국식품영양과학회지 Vol.46 No.11

Epigallocatechin-3-gallate(EGCG)는 녹차에 함유된 flavonoid 계열 화합물로, 녹차의 대표적인 유효성분으로 알려져 있다. 본 연구는 3T3-L1 세포에서 EGCG의 항비만 작용 및 항산화 효소 활성을 평가하였다. EGCG는 지방세포 분화를 유의적으로 억제하였으며, 지방세포 분화와 관련된 단백질인 C/EBPα와 aP2의 발현을 감소시켰다. 동시에 EGCG는 GSH의 활성을 유도하였으며, 항산화 효소인 SOD, CAT, GPx 및 GR의 활성을 증가시켰다. EGCG는 지방세포 분화 동안 활성산소종의 형성을 감소시켰다. 체내 phase Ⅱ 효소인 HO-1과 GCLC는 EGCG에 의해 발현이 유도되었다. 따라서 EGCG는 3T3-L1 세포에서 항산화 효소의 활성화를 유도하며 동시에 지방세포로의 분화를 억제하는 것으로 나타났다. 또한, EGCG는 활성산소종의 생성을 억제하였으며, 이는 EGCG의 항산화 작용에 의한 지방세포 분화 억제 작용이 관련이 있는 것으로 판단된다. EGCG에 의한 항비만 효과와 항산화 작용을 연구한 본 연구는 녹차를 활용한 체지방 감소 효능을 지닌 식품 개발에 기초 자료로 활용될 수 있을 것으로 생각된다. Obesity contributes to the development of diseases, such as type Ⅱ diabetes, hypertension, coronary heart disease, and cancer. In addition, oxidative stress caused by reactive oxygen species (ROS) is recognized widely as a contributing factor in the development of chronic diseases. This study was examined the antioxidant and anti-adipogenic activities of epigallocatechin-3-gallate (EGCG) in 3T3-L1 preadipocytes. 3T3-L1 cells were differentiated with or without EGCG for 6 days. The production of glutathione (GSH) and the activities of the antioxidant enzymes, such as glutathione reductase (GR), glutathione peroxidase (GPx), catalase (CAT), and superoxide dismutase (SOD) were measured. EGCG inhibited significantly the lipid accumulation and the expression of adipogenic specific proteins including CCAAT/enhancer binding protein α and adipocyte fatty acid binding protein. The production of intracellular ROS was decreased significantly by EGCG in 3T3-L1 cells. EGCG increased the GSH production and the activities of GPx, GR, CAT, and SOD. Moreover, EGCG increased the protein expression of glutamate-cysteine ligase and heme oxygenase-1 in 3T3-L1 cells. These results suggest that EGCG increased the activity and expression of antioxidant enzymes and suppressed the lipid accumulation in 3T3-L1 cells. Therefore, the use of phytochemicals that can maintain the GSH redox balance in adipose tissue could be promising for reducing obesity.

차 카테킨 EGCG (Epigallocatechin Gallate)의 구강세균에 대한 살균효과

유미옥,천재우,오계헌,Yu Mi-Ok,Chun Jae-Woo,Oh Kye-Heon 한국미생물학회 2004 미생물학회지 Vol.40 No.4

본 연구는 차 카테킨인 EGCG (epigallocatechin gallate)에 의한 구강세균의 살균효과를 조사하기 위하여 수행되었다. 초기 세포밀도 $10^{7}$ cell/ml의 대상 세균에 대한 항세균 활성은 EGCG 1ml당 5mg에서 조사하였다. 선택 또는 복합배지상에서 집락생성단위(colony-forming unit, CFU)에 근거하여 EGCG의 항세균 활성은 농도에 비례하는 것을 확인하였다. Streptococcus mutans와 Streptococcus sobrinus는 5mg/ml의 EGCG에서 8시간 이내에 완전히 살균되었다. Lactobacillus acidophilus와 Lactobacillus plantarum도 동일한 조건에서 각각 2시간과 4시간 이내에 살균되었다. 2.5 mg/ml의 EGCG로 처리한 사람의 타액에서 유래하는 총 구강세균의 생존율을 BHI 고체배지상에서 조사하였으며, 총 구강세균은 10시간 이내에 완전히 살균되었다. MS 고체배지상에서 S. mitis and S. salivarius의 집락은 감소되었으며 배양 12시간 이내에서는 집락은 관찰되지 않았다. 그 결과, EGCG는 입냄새와 치석의 원인이 되는 구강세균을 살균시키고 충치를 예방하는 자연적이며 효과적인 물질임이 입증되었다. The purpose of this work was to investigate the effect oftea catechin, epigallocatechin gallate (EGCG) on killing of oral bacteria. The antibacterial activity of 2.5 mg/ml and 5.0 mg/ml EGCG was investigated for target bacteria of which initial cell number was approximately adjusted to $10^{7}ml$. The antibacterial activity of EGCG was proportional to the concentration according to colony-forming unit(CFU) of target bacteria enumerating on selective and complex media. Streptococcus mutans and Streptococcus sobrinus at 5mg/ml EGCG were completely killed within 8 hrs. Lactobacillus plantarum and Lactobacillus acidophilus were also killed within 2 hrs and 4 hrs under the same conditions, respectively. Oral bacteria at 2.5 mg/ml EGCG were completely killed within 10 hr. Colony numvers of S. mitis and S. salivarius treated with 2.5 mg/ml EGCG were decreased on MS solid media and no colony was observed on the media within 12 hrs. In consequence, EGCG would be a natural and effective compound that kill oral bacteria being caused of bad breath, plaque and gingivitis, and for preventing and treating dental caries.

Epigallocatechin-3-gallate(EGCG)이 구강 편평세포암종 세포사를 유발하는 기전 연구

박형목,이은경,정진,박봉수,박혜련,유미현 대한구강악안면병리학회 2009 대한구강악안면병리학회지 Vol.33 No.4

Green tea, derived from the plant Camellia sinensis, is one of the most common beverages consumed worldwide. Epigallocatechin-3-gallate (EGCG) is the most abundant and bioactive polyphenolic constituent in green tea. Understanding how intracellular signaling pathways respond to EGCG may provide a clue to the difference of cell responses and basis for usefulness of EGCG as a chemopreventive and/or chemotherapeutic agent. In the present study, we tried to check whether EGCG could be a useful agent in chemotherapeutic treatment of oral squamous carcinoma. Furthermore, we investigated which signaling pathway is involved in biologic activities of EGCG. EGCG induced the cell death of oral squamous carcinoma cells. Furthermore, it increased phosphorylation of Akt in serum-strarved oral squamous carcinoma cells. But, initial increase of Akt activation did not affect cell survival. Activities of Raf-1 and Erk showed inconsistent response to EGCG treatment, but Erk phosphorylation is consistent with Raf-1 activity in YD 10B cells. These changes of Raf-1 and Erk activity in EGCG treated cells were different depending on cell line type. Supposedly, the difference of cell component may affect the Raf-1 and Erk reactivity to EGCG treatment. Akt activation by EGCG is independent on activities of PDK1 and PTEN, and expression of bax and bcl-2 proteins were not changed by EGCG treatment. Therefore, EGCG treatment did not induce the apoptosis of YD 10B cell. On the other hand, vascular adhesion molecule (VCAM) was decreased by EGCG treatment, so it is possible that decrease of VCAM can play certain role in survival and/or cell death in EGCG treated cells

(-)-Epigallocatechin-3-gallate(EGCG)가 타액선 암세포의 제1형 교원질 부착 및 이동에 미치는 영향

윤효은,윤정훈,안상건 대한구강악안면병리학회 2011 대한구강악안면병리학회지 Vol.35 No.3

EGCG has inhibitory effect on a variety of cancers by inducing apoptosis and cell cycle arrest or inhibiting angiogenesis and metastasis. EGCG has been found to induce apoptosis in salivary gland carcinoma cells. But the potential anti-invasive effect of EGCG in salivary gland cancer has not been studied yet. The aim of this study is to evaluate the effect of EGCG on salivary gland adenocarcinoma SGT cell adhesion and migration to Type I collagen treatment. Western blot, adhesion and migration assay were performed to evaluate the impacts of EGCG on the expression of MMP-2/-9 and its upstream signaling molecules after treatment of type I collagen. SGT cell adhesion to type I collagen is significantly suppressed by EGCG. EGCG decreased expression of β1 integrin, phosphorylation of FAK, MMP-2/-9 compared with type I collagen treatment. In addition, EGCG inhibited the migration of SGT cells treated with type I collagen. These results suggest that EGCG could effectively inhibit the invasion and migration of human SGT cells by downregulating the expression of β1 integrin and MMP-2/-9 and phosphorylation of FAK, Akt, and Erk. Adhesion and migration to type I collagen of SGT cells can be influenced through EGCG.

EGCG의 Viral Hemorrhagic Septicemia Virus (VHSV) 감염에 의해 유도되는 산화스트레스 억제를 통한 세포 생존 효과

정은혜(Eun-Hye Jeong),조세영(Se-Young Cho),권요셉(Joseph Kwon),박근형(Keun-Hyung Park),문제학(Jae-Hak Moon),오명주(Myung-Ju Oh),김선재(Seon-Jae Kim),김두운(Duwoon Kim) 한국차학회 2014 한국차학회지 Vol.20 No.2

EGCG의 viral hemorrhagic septicemia virus (VHSV) 감염에 의해 유도되어지는 산화스트레스 억제를 통한 세포생존 효과를 알아보고자 EGCG 처리 시의 세포생존 활성, ROS 생성 저해, DNA fragmentation 저해 평가 등에 대한 실험을 수행하였다. VHSV를 포함하는 많은 바이러스들은 세포에 감염 시 ROS와 같은 산화적 스트레스를 유발한다. 100 μM 수준에서 EGCG는 다른 4종의 catechin류에 비해 무처리 대조구 대비 1.4배의 높은 세포생존 활성을 지니는 것으로 확인되었다. EGCG의 ROS 생성 감소 효과를 확인하고자 DCFH-DA를 세포 내에 잔존하는 활성산소종과 반응시켜 형광도를 측정한 결과 EGCG 비처리구에서는 VHSV에 의해 ROS가 1.7배 증가되었고, 100 μM의 EGCG를 12시간 처리 시 1.3배로 상대적으로 감소하는 것으로 나타났다. 바이러스에 의해 발생하는 ROS는 DNA fragmentation을 일으켜 세포 사멸을 유도하므로, 어류 세포인 FHM 세포를 이용하여 EGCG의 DNA fragmentation저해 효과를 확인해본 결과, 100 μM의 EGCG를 12시간 처리 시 비처리구에 비해 DNA의 fragmentation이 감소하는 것으로 나타났다. 이 결과들로부터, EGCG는 바이러스 감염에 의해 유발되어지는 산화적 스트레스를 억제함을 통해 세포 생존율을 증가시키는 것으로 확인되었다. Many viruses, including viral hemorrhagic septicemia virus (VHSV), initiate oxidative stresses such as generation of ROS, and the depletion of reduced thiols in virus infected cells. In this study, epigallocatechingallate (EGCG) showed the highest antiviral activities against the VHSV. The cell viability of EGCG was exposed to 1.4 times higher than any that of the other catechins at 100 μM compared to the control. The DNA fragmentation and ROS generation inhibition effect of EGCG in FHM cells, upon exposure to 0.2 moi VHSV for 42 h was examined. In regard to the cellular redox status and oxidative DNA damage to FHM cells infected with the VHSV, a treatment with 100 μM EGCG produced a 1.3-fold lower ROS generation and DNA fragmentation than the control, indicating that EGCG may increase the cell viability by suppressing the oxidative stress induced by a virus infection.

녹차 카테킨, Epigallocathechin Gallate (EGCG)의 흰쥐췌장종양 선 세포 AR42J의 MAP Kinase 세포 신호전달 기전을 통한 Neurogenin 3 발현에 미치는 영향

김성옥(Kim Sung Ok),최원경(Choe Won Kyung) 韓國營養學會 2011 Journal of Nutrition and Health Vol.44 No.3

본 연구는 EGCG의 항 당뇨 활성기전으로 췌장종양 선세포 AR42J의 분화 및 내분비기능 개선에 미치는 영향과 그 세포 신호전달 기전을 확인하였다. 그 결과 첫째, AR42J 세포에 EGCG 처리 시 췌장종양 선세포의 세포증식이 농도 의존적으로 감소되었다. 둘째, 세포사멸 유도가 유의적으로 일어나지 않는 농도인 luM EGCG를 AR42J 세포에 처리한 결과 ngn 3, α-amylase, insulin은 EGCG처리 24시간에 mRNA, 단백질 발현증가를 나타내었고 48시간에 유의적 증가를 나타내었다. 셋째, EGCG 처리 시 ERK, JNK MAP Kinase 기전은 인산화 억제를 나타내었고 반면에 p38 기전의 인산화는 48시간에 유의적 증가를 하였다. 넷째, p38기전 저해제인 SB203580을 처리하여 EGCG가 MAP Kinase 기전중의 하나인 p38 기전 인산화 활성의 회복을 나타내어 ngn 3 발현을 위한 전사 신호전달 기전임을 다시 확인하였다. 따라서 녹차 생리활성 성분인 EGCG의 췌장종양 선 세포 AR42J 처리 결과 EGCG는 p38 MAP Kinase 기전 활성을 통해 췌장 선세포의 분화지표인 ngn 3 발현을 증가시키며 췌장내분비 기능 지표인 α-amylase, insulin 발현증가를 나타내어 세포의 내분비기능 개선에도 영향을 미치는 것으로 사료된다. Epigallocatechin gallate (EGCG), or epigallocatechin 3-gallate. is the ester of epigallocatcchin and gallic acid, and is a type of catec hin. EGCG may be therapeutic for many disorders including diabetic s and some types of cancer. Howev-er it is unknown whet her EGCG ca n induce transdifferentiation of pancreatic cells in pancreatitis. The aim of this study was to investigate the effects of EGCG on the expression of pancreatic regenerating related markers in pancreatic AR42J cells, a model of pancreatic progenitor cells. AR42J cells. differentiated with betacellulin and activin A, were cultured with/without EGCG in a time-dependent manner. Cell growth rate, levels of mRNA. and protein expression were examined with the MTT assay, quantitative PCR, and Western blots, respectively. The results showed that AR42J cell growth rates were inhibited by EGCG in a dose-dependent manner. mRNA and protein expression of amylase, insulin and neurogenin 3 (ngn 3) increased in AR42J cells treated with EGCG. Additionally, we demonstrated that the signal transduction pathway of mitogen-activated protein (MA P) kinase is active in EGCG-treated AR42J cells. ERK and JNK phosphorylation decreased in cells treated with EGCG but not p38 phosphorylation. Activation of the p38 MAP kinase pathway was confirmed by specific MAP kinase pathways inhibitors: U0126 for ERK, SP600126 for INK, and SB203580 for p38. Activated p38 phosphorylation was inhibited by the specific p38 inhibitor SB203580 but p38 phosphorylation was inhibit ed with increased EGCG treatment. The ERK and JNK MAP kinase pathways were not affected by EGCG treatment. Although further studies are needed, these result s suggest that EGCG affects the induction of pancreatic cell regeneration by increasing the ngn 3 protein and mRNA expression and activating the p38 MAP kinase pathway.

에탄올에 의하여 유도된 마우스 췌장 선포세포의 염증성 손상에서 췌장분비 효소의 활성 및 세포 재생관련 유전자들의 발현에 미치는 EGCG의 영향

김성옥(Sung Ok Kim),최영현(Yung Hyun Choi) 한국생명과학회 2013 생명과학회지 Vol.23 No.11

본 연구에서는 마우스 췌장의 선포세포(primary acinar cell)에서 에탄올에 의해 유도된 염증성 손상에 미치는 녹차의 생리활성 물질인 EGCG의 효과를 췌장분비 효소의 활성과 선포세포에서의 기능 회복 관련 마크로 알려진 유전자들(AMPK, RKIP 및 Reg1)의 발현 조절 측면에서 조사하였다. 대조군에 비해 에탄올이 처리된 세포에서는 α-amylase와 chymotrypsin의 활성이 증가되었지만, EGCG 처리에 의하여 그들 활성이 유의적으로 감소하였다. 세포 생존 및 보호와 관련 있는 AMPK 인산화 수준은 에탄올 단독 처리시 그 발현이 감소하였지만, EGCG 처리에 의하여 유의적으로 회복되었다. 세포 사멸과 세포독성 조절과 연관이 있는 RKIP 또한 그 발현 경향이 AMPK인산화 변화의 정도와 유사하였다. 또한 베타세포 재생에 관여하는 Reg1 발현은 에탄올이 처리된 선포세포에 EGCG를 처리하였을 경우 그 발현량이 유의적으로 증가하였다. 이상의 결과들을 종합하여, 항산화성 폴리페놀인 EGCG는 알코올성 췌장염으로 인한 세포의 손상이나 당뇨병 예방과 회복에 치료적 효과를 가질 수 있다고 제안한다. (-)-Epigallocatechin-3-gallate (EGCG), a green tea polyphenol, has been shown to have strong antibacterial, antiviral, antioxidant, anti-inflammatory, and chemopreventive effects. However it is unknown whether EGCG can recover alcohol-associated pancreatitis. The aim of this study was to investigate the effects of EGCG on pancreatic enzyme activities and the expressions of pancreatic regenerating related markers, such as adenosine monophosphate-activated protein kinase (AMPK), raf-1 kinase inhibitor protein (RKIP), and Regenerating gene 1 (Reg1), in mice pancreatic primary acinar cells. Our results revealed that activities of α-amylase and chymotrypsin were significantly increased in the cells treated with ethanol compared to the untreated control cells; however, the increased activities of both enzymes were markedly reduced by pretreatment with EGCG. Phosphorylation of AMPK and total expression of RKIP were decreased in the ethanol-treated primary acinar cells; however, these were both significantly increased in the EGCG-pretreated cells. In addition, when EGCG was treated, expression of Reg1 was markedly increased compared with that of the control or the ethanol-treated primary acinar cells, demonstrating that EGCG can modulate pancreatic regenerating related genes. Therefore, our findings suggest that EGCG may have therapeutic utility in the prevention or treatment of alcohol-associated pancreatitis.

녹차로부터 분리한 epigallocatechin-3-O-(3-O-methyl) gallate의 Helicobacter pylori 생육 억제 효과

반유진(Eu-Jin Ban),송은정(Eun-Jung Song),이동준(Dong-Jun Lee),문제학(Jae-Hak Moon),김선재(Seon-Jae Kim),박종환(Jong-Hwan Park),조정용(Jeong-Yong Cho) 한국차학회 2021 한국차학회지 Vol.27 No.1

Epigallocatechin-3-O-gallate (EGCG)는 차의 대표적인 유용성분 중 하나로, 항산화, 항암, 그리고 항염증등 다양한 건강기능성과 더불어 Helicobacter pylori의 생육을 억제하는 효과도 우수하다고 알려져 있다. 그러나, EGCG에 methyl기가 결합된 형태인 epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3 Me)에 대한 H. pylori의생육 억제 효과는 보고된 바가 없다. 본 연구에서는 H. pylori P1WT 및 SS1 균주를 대상으로 차로부터 분리한EGCG3 Me의 항균 활성을 평가하였다. EGCG3 Me은 octadecylsilane-medium performance liquid chromatography를 이용하여 가루녹차(Camellia sinensis var. sinensis cv. Benifuuki)의 70% ethanol 추출물로부터분리되었으며, 그 구조는 ESI-MS, 1D-, 그리고 2D-NMR 분석을 통하여 확인되었다. 가루녹차로부터 분리된EGCG3 Me는 EGCG와 거의 유사하게 농도 의존적으로 H. pylori P1WT와 SS1 균주에 대한 생육 억제 효과를나타내었다. 특히, 두 균주 모두 50 그리고 100 μM의 EGCG 및 EGCG3 Me 처리에 의해 성장이 크게 억제되었으며, 100 μM 농도의 경우 P1WT와 SS1 균주의 성장을 매우 높은 수준으로 억제하였음을 관찰할 수 있었다. EGCG3 Me의 H. pylori 생육 억제 효과는 EGCG와 거의 유사하였다. 이상의 결과로부터 EGCG3 Me는 EGCG 와 더불어 H. pylori 생육 억제 효과를 갖는 차의 유용 소재로 활용이 기대된다. Epigallocatechin-3-O-gallate (EGCG), a major bioactive compound in green tea, has been shown to strongly inhibit the growth of Helicobacter pylori (H. pylori) apart from its other multiple health benefits. However, no study on the anti-Helicobacter pylori activity of epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3 Me), a unique O-methylated form of EGCG, is currently available. The present study aimed to compare the growth inhibitory activities of EGCG and EGCG3 Me against H. pylori strains P1WT and SS1. EGCG3 Me was isolated from a 70% ethanol extract of green tea (Camellia sinensis var. sinensis cv. Benifuuki) by using octadecylsilane-medium performance liquid chromatography. Its structure was confirmed by ESI-MS and 1D- and 2D-NMR experiments. Both EGCG and EGCG3 Me inhibited the growth of H. pylori strains P1WT and SS1 in a dose-dependent manner. The antibacterial activity of EGCG3 Me was similar to that of EGCG. EGCG is well-known to be a potential anti-H. pylori agent. These results indicate that EGCG3 Me may be a valuable therapeutic agent against H. pylori infection.

Acute Cytotoxicity Testing of Polyhexamethylene-biguanide (PHMG) and Epigallocatechin-gallate (EGCG) Mixture on the Cultured Human Corneal Epithelial Cell

Nam-youl Kim,Koon-Ja Lee 대한시과학회 2018 대한시과학회지 Vol.20 No.4

목적: 항균제 polyhexamethylene Biguanide(PHMB), epigallocatechin gallate(EGCG) 및 PHMB/EGCG 혼합물의 각막상피세포(primary human corneal epithelial cells, HCEpiCs)에 대한 급성독성을 평가하고자 하였다. 방법: 각막상피세포를 0.00001~0.005% PHMB, 0.001~5% EGCG 및 0.00005% PHMB/0.05% EGCG 혼합물이 각각 포함된 배양액에서 30분, 60분, 120분 및 240분 동안 배양하였다. 배양한 각막상피세포를 고정한 다음 Draq 5로 염색하고 공초점현미경과 ImageXpress UltraTM를 이용하여 세포형태를 관찰하여 세포 생존율과 세포자살(apoptosis)을 비교하였다. 결과: 배양된 각막상피세포는 0.00005% 이하의 PHMB 농도 및 0.05% 이하의 EGCG 농도에서는 세포 독성이 나타나지 않았다. 0.00005% PHMB/0.05% EGCG 혼합물이 포함된 배양액에서 급성 세포독성은 관찰되지 않았으나 240분 배양시킨 경우에는 손상된 각막상피세포 수가 증가하고 생존 세포의 수는 감소하였다. 결론: 항균 시너지 효과를 갖는다고 보고된 0.00005% PHMB/0.05% EGCG 혼합물의 경우 각막상피세포에 대한 급성독성은 없었으나 만성효과에 대해서는 추가적인 연구가 필요할 것으로 생각된다. Purpose : To evaluate the acute cytotoxic effect of polyhexamethylene biguanide (PHMB), epigallocatechin gallate (EGCG) and PHMB/EGCG mixture on the cultured human corneal epithelial cell (HCEpiC). Methods : HCEpiCs were cultured in the media of HCEpiC containing 0.00001~0.005% PHMB, 0.001~5% EGCG and 0.00005% PHMB/0.05% EGCG mixture respectively for 30, 60, 120 and 240 min. Cultured HCEpiCs were fixed and stained with Draq5 and cell viability and apoptosis were evaluated using confocal microscope and ImageXpress UltraTM. Results : Cultured HCEpiC did not show cytotoxic effect at below 0.00005% PHMB and below 0.05% EGCG concentration. In the media containing 0.00005% PHMB/0.05% EGCG, acute cytototoxic effect was not found, whereas damaged HCEpiCS were increased and survival cells were decreased in the media incubated for 240 min. Conclusion : The mixture of 0.00005% PHMB/0.05% EGCG showed non acute cytotoxic effect on the cultured HCEpiCs, however it is needed to investigate its chronic cytotoxic effect.