비접촉식 경면현미경을 이용한 20대의 각막내피세포 분석

경다슬,김효진 대한시과학회 2023 대한시과학회지 Vol.25 No.3

목적 : 건강한 20대를 대상으로 각막내피세포의 형태학적 분석을 실시하고, 양안, 성별, 굴절교정수술 여부, 콘택트렌즈 착용여부, 코로나바이러스-19의 감염에 따른 차이를 알아보고자 하였다. 방법 : 충청남도 일개 대학교에서 20대 남녀 87명(174안)을 대상으로 하였다. 각막내피세포의 형태학적 분석은 비접촉식 경면현미경을 이용하여 검사하였고, 각막내피세포의 밀도, 내피세포의 다형성, 육각형비율, 그리고중심각막두께를 분석하였다. SPSS 18.0 프로그램을 이용하여 양안, 성별, 그리고 그 외에 조사한 변수에 따른 각막내피세포의 밀도, 다형성 및 육각형 비율을 측정하였다. 결과 : 우안과 좌안에서 각막내피세포의 밀도는 각각 2,977.7±238.7 cells/mm2와 3,008.9±236.7 cells/mm2(p=0.056)이었고, 남성과 여성은 각각 2,970.9±237.9 cells/mm2와 2,983.6±241.8 cells/mm2로 통계적으로 유의한 차이가 없었다(p=0.807). 각막내피세포의 다형성과 육각형 비율도 양안, 성별, 굴절교정수술 여부. 코로나바이러스-19 감염 유무에 따라 유의한 차이가 없었다(p>0.050). 그러나 각막내피세포의 다형성과 육각형 비율은 콘택트렌즈 착용 군과 대조군에서 각각 30.7±4.5%와 61.9±5.4% 그리고 28.2±4.0%와 64.7±4.7% 를 보였다(각각 p=0.018와 p=0.029). 결과 : 한국 20대 성인을 대상으로 분석한 각막내피세포 밀도의 평균값은 2,993.3±237.5 cells/mm2이었고, 양안과 성별에 따른 차이는 없었다. Purpose : Morphological analysis of corneal endothelial cell was conducted in young adults Korean in their 20s, and differences according both eyes, sex, refractive surgery, contact lenses, COVID-19 infection were investigated. Methods : A total of 87 (174 eyes) men and women in their 20s were enrolled in a university in Chungcheongnam-do. Morphological analysis of corneal endothelial cell was examined using a noncontact specular microscopey, and corneal endothelial cell density, endothelial cell polymorphism, hexagonal ratio, and central corneal thickness were analyzed. The SPSS 18.0 program was used to compare differences in corneal endothelial cell density, pleomorphism, and hexagonal ratio according to both eyes, sex, and other variables investigated. Results : The density of corneal endothelial cell was 2,977.7±238.7 cells/mm2 in the right eye and 3,008.9±236.7 cells/mm2 in the left eye (p=0.056). Men had 2,970.9±237.9 cells/mm2 and women had 2,983.6±241.8 cells/mm2, and there was no statistically significant difference (p=0.807). Polymorphism and hexagonal ratio of corneal endothelial cell were also not significantly different according to both eyes, sex, refractive surgery, COVID-19 infection (p>0.050). However, the polymorphism and hexagonal ratio of corneal endothelial cell according to whether or not they wore contact lenses were 30.7±4.5% and 61.9±5.4% in the contact lens wearing group and 28.2±4.0%, and 64.7±4.7% in the control group (p=0.018 and p=0.029, respectively). Conclusion : The average value of corneal endothelial cell density analyzed in Korean in their 20s was 2,993.3±237.5 cells/mm2, and there was no difference between both eyes and sex.

수력을 이용한 수정체유화술이 각막내피세포에 미치는 영향

강승범,길소연,정성근.Seung Bum Kang. M.D.. So Yeon Gil. M.D.. Sung Kun Chung. M.D. 대한안과학회 2007 대한안과학회지 Vol.48 No.3

Purpose: To access the effect of phacoemulsification using the AquaLase? (Alcon Laboratories, TX, U.S.A.) on corneal endothelial cell. Methods: This study comprised 47 eyes of 43 patients having phacoemulsification with the AquaLase?. Various corneal endothelial cell parameters, including corneal endothelial cell density, corneal thickness and morphologic change, were evaluated up to 2 months postoperatively. Results: Preoperative mean corneal endothelial cell density was 2818±282 cell/mm2. Postoperative mean corneal endothelial cell density was 2682±390 cell/mm2 at 1 day, 2701±263 cell/mm2 at 1 month and 2721±396 cell/mm2 at 2 month. Corneal endothelial cell loss was 4.74%, 4.00% and 3.55% at postoperative 1 day, 1 month and 2 months respectively. There was no statistically significant difference between preoperative and postoperative corneal endothelial cell density (p>0.05). The corneal endothelial cell loss was not increased even though the phacoemulsification time with the AquaLase? was increased (p>0.05). Preoperative mean corneal thickness was 559.4±37.4 ?m. Postoperative mean corneal thickness was 586.9±35.1 ?m at 1 day, 573.3±30.1 ?m at 1 month and 554.9±26.7 ?m at 2 month. There was a significant increase in corneal thickness at postoperative 1 day (p=0.001). However, there was no statistically significant difference of corneal thickness between preoperative and postoperative 1 month and 2 month (p>0.05). Conclusions: The phacoemulsification with the AquaLase? dose not cause the significant corneal endothelial cell density and corneal thickness change postoperatively.

접이식 홍채고정안내렌즈삽입술 후 각막내피세포 밀도의 5년간 변화

남명식(Myungsik Nam),강성용(Sung Yong Kang),김선웅(Sun Woong Kim) 대한안과학회 2021 대한안과학회지 Vol.62 No.4

목적: 접이식 홍채고정안내렌즈(Artiflex; Ophtec BV, Groningen, The Netherlands) 삽입 수술 후 5년간 각막내피세포 밀도의 변화를 보고하고, 전방부피와 전방깊이의 영향을 분석하였다. 대상과 방법: 본 연구는 2009년 3월부터 2014년 3월까지 근시교정을 위한 접이식 유수정체 홍채고정렌즈(Artiflex) 수술을 받은 88명, 146안이 포함되었다. 정상인 20세, 25세, 30세 각각 100명의 각막내피세포 밀도를 측정 후 연령에 따른 각막내피세포 변화를 추정한 모방대조군으로 설정하여 수술 후 변화와 비교하였다. 렌즈와 각막내피까지의 거리 및 전방부피, 전방깊이를 4군으로 나눈 후 각막내피세포 밀도의 변화를 비교하였다. 결과: 각막내피세포 밀도의 평균값은 수술 전, 수술 후 1년, 3년, 5년이 각각 3,038.8 ± 223.2, 2,985.3 ± 235.5, 2,905.1 ± 231.3, 2,814.9 ± 250.5 cells/mm2로 나타났다(각각 1.73%, 4.36%, 7.31% 감소, p<0.001). 이는 수술을 받지 않은 20세와 25세군의 2.2%의 차이, 25세와 30세군의 1.2% 차이와 비교하여 통계적으로 유의한 감소였다(p<0.001). 귀쪽의 렌즈와 각막내피 거리와 전방부피가 작은 군에서 각막내피세포 밀도의 감소가 다른 군에 비해 유의하게 컸다. 결론: 접이식 홍채고정렌즈 삽입 수술 후 5년간 각막내피세포 밀도의 유의한 감소가 있으며, 렌즈와 각막내피까지의 거리와 전방부피가 적을수록 각막내피감소가 크게 나타났다. Purpose: To investigate changes in corneal endothelial cell density over 5 years after implantation of Artiflex lenses (Ophtec BV, Groningen, The Netherlands) and to explore the influence of anterior chamber depth and volume on changes in endothelial cell density. Methods: This study included 146 eyes of 88 patients implanted with foldable phakic iris-fixation intraocular (Artiflex) lenses for myopia correction from March 2009 to March 2014. To simulate chronological changes in endothelial cell density with age, we collected normal endothelial cell density data from 100 healthy subjects aged 20, 25, or 30 years, and compared the changes among these groups at 5 years after surgery. Changes in corneal endothelial cell density were compared among four groups to which subjects were assigned based on the distance from the intraocular lens to the corneal endothelium, and the volume and depth of the preoperative anterior chamber. Results: The mean endothelial cell density, pre-operatively and at 1, 3, and 5 years post-operatively, was 3,038.8 ± 223.2, 2,985.3 ± 235.5, 2,905.1 ± 231.3, and 2,814.9 ± 250.5 cells/mm2, respectively (1.73%, 4.36%, and 7.31% reduction, respectively, vs. pre-operatively, p < 0.001). These differences were greater than those observed among different age groups (2.2% difference between 20- and 25-year-olds, and 1.2% difference between 25- and 30-year-olds, p < 0.001). A shorter distance from the lens to the endothelium and smaller anterior chamber volume were associated with greater decreases in endothelial cell density following surgery. Conclusions: Corneal endothelial cell density significantly decreased during a 5-year follow-up after Artiflex lens implantation. The distance from the intraocular lens to the corneal endothelium, and the anterior chamber volume, were negatively correlated with the rate at which endothelial cell density decreased over time.

섬유주 각막 이행부위의 형태학적 특성: 편평조직표본 및 미세구조

구오섭,고명규,O Sub Koo,Myung Kyoo Ko 대한안과학회 2007 대한안과학회지 Vol.48 No.5

Purpose: To evaluate the morphological characteristics of the transitional zone between the corneal endothelium and the trabecular meshwork by flat preparation and electron microscopy. Methods: The materials comprised 12 eyes examined by the flat preparation and 7 eyes by the electron microscopy. The specimens were derived from the transitional tissue between the corneal endothelium and the trabecular meshwork. The specimens in the flat preparation were stained with hematoxylin-eosin and examined by light microscopy. The specimens for scanning electronic microscopy (SEM) and in transmission electronic microscopy (TEM) were examined through routine processes. Results: In the specimens examined by the flat preparation, unlike peripheral corneal endothelial cells, the endothelial cell nuclei in the transitional zone were overlapped and morphologically oval. On SEM, unlike typical hexagonality and tight interdigitation of corneal endothelial cells, the endothelial cells in the transitional zone were partially successive, spaced intercellularly, and morphologically irregular. On TEM, the endothelial cells in the transitional zone were partially successive. Conclusions: The loss of cell-cell contact of endothelial cells in the transitional zone may lead to the potential proliferation capacity of endothelial cells in the transitional zone under specific conditions. Therefore, further studies on the proliferation capacity of endothelial cells in the transitional zone are needed together with more research on cell biology.

진동을 이용한 수정체유화술과 수정체혼탁정도와의 관계 및 각막에 미치는 영향

길소연,강승범,이승희,정성근,So Yeon Gil,Seung Bum Kang,Seung Hee Lee,Sung Kun Chung 대한안과학회 2006 대한안과학회지 Vol.47 No.12

Purpose: To access the effect of phacoemulsification, using Neosonix? (Alcon Laboratories, TX, USA), on the cornea and lens opacity. Methods: This study involved 42 eyes of 37 patients who underwent phacoemulsification using the Neosonix from December 2005 through March 2006. We measured the density and the thickness of lenses using a Scheimpflug camera and evaluated the correlation between the density and the phaco time. To evaluate the effects of Neosonix? on the corneal endothelium, we measured various corneal endothelial cell parameters, including corneal endothelial cell density, corneal thickness, and morphologic changes up to two months postoperatively. Results: The preoperative mean values of lens density for the anterior cortex, nucleus, and posterior cortex were 105.86±34.99 CCT, 82.17±35.14 CCT, and 77.88±52.98 CCT, respectively. The mean phacoemulsification time was 48.67±29.79 seconds and increased significantly, according to the increase of the densities of the lens nucleus and cortex (p<0.01). Postoperative mean corneal endothelial cell density was 2741±474 cell/mm2 at one day, 2757±447 cell/mm2 at one month, and 2765±422 cell/mm2 at two months. Corneal endothelial cell loss was 4.39%, 3.83%, and 3.56% at one day, one month, and two months postoperatively, respectively. There was no statistically significant difference between the preoperative and postoperative values of corneal endothelial cell density (p>0.05). Conclusions: The phacoemulsification time using oscillation device significantly increased as the density of the lens cortex and nucleus increased, and there was no statistically significant difference between the preoperative and postoperative values of corneal endothelial cell density.

Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells

Haeeun Shin(Haeeun Shin),Joon Ki Min(Joon Ki Min),Na Rae Kim(Na Rae Kim),Kyoung Yul Seo(Kyoung Yul Seo),Hee Seung Chin(Hee Seung Chin),Soyoung Lee(Soyoung Lee),Ji Won Jung(Ji Won Jung) 대한안과학회 2023 Korean Journal of Ophthalmology Vol.37 No.1

Purpose: Human corneal endothelial progenitor cells (HCEPs), which has been selectively isolated and differentiated into human corneal endothelial cells (HCECs), are crucial for repairing corneal endothelial damage. In this study, we evaluated the roles of a Rho-assisted kinase (ROCK) inhibitor, Y-27632, on the isolation and expansion of HCEPs, and assessed the in vitro effects of different concentrations of Y-27632 on the differentiated HCEPs. Methods: HCEPs were isolated and expanded in a medium with and without 10μM Y-27632, and then differentiated into HCECs in a medium with fetal bovine serum. The characteristics of HCEPs and differentiated HCEPs were confirmed by immunofluorescence staining. The proliferation, viability, morphology, and wound-healing ability of differentiated HCEPs were assessed in the presence of different concentrations of Y-27632. Results: Y-27632 enabled the isolation and expansion of HCEPs from the corneal endothelium. The differentiated HCEPs showed an optimal increase in proliferation and survival in the presence of 10μM Y-27632. As the concentration of Y-27632 increased, differentiated HCEPs became elongated, and actin filaments were redistributed to the periphery of cells. Y-27632 also caused a concentration-dependent enhancement in the wound-healing ability of differentiated HCEPs. Conclusions: Y-27632 enabled the isolation and expansion of HCEPs. It also enhanced the proliferation, viability, and migration of differentiated HCEPs.

녹내장환자에서 중심각막내피세포 검사의 중요성

오태훈,장동진,김재우,문정일,김현승,Tae Hoon Oh,MD,Dong Jin Chang,MD,Jae Woo Kim,MD,Jung Il Moon,MD,PhD,Hyun Seung Kim,MD,PhD 대한안과학회 2012 대한안과학회지 Vol.53 No.1

Purpose: To evaluate the possible causes of decreased central corneal endothelial cell count in glaucoma patients and to acknowledge the importance of central corneal endothelial cell count evaluation. Methods: This retrospective case-control study included 60 glaucomatous eyes that were being treated with topical anti- glaucoma drugs, 30 eyes that underwent laser iridotomy or trabeculectomy, and 60 control eyes. Intraocular pressure, duration of topical anti-glaucoma medications, mean number of topical anti-glaucoma drugs, and preoperative central corneal endothelial cells were analyzed. Results: The numbers of central corneal endothelial cells in the topical anti-glaucoma medication group, the surgical group and the control group were 2681.30 ± 355.33, 2435.57 ± 646.81 and 2822.08 ± 330.17 cells/mm2, respectively. The numbers in the topical anti-glaucoma medication group and the surgical group were significantly lower than that of the control group. The number of central corneal endothelial cells was significantly low in patients with longer duration of disease and in those taking a greater number of topical anti-glaucoma drugs (p = 0.003, p = 0.010) Conclusions: In glaucoma patients, corneal endothelial cell count is decreased by glaucoma surgery, laser therapy, long-term or multiple topical anti-glaucoma medications. When performing cataract surgery in glaucoma patients, preoperative corneal endothelial evaluation is essential. J Korean Ophthalmol Soc 2012;53(1):37-42

각막내피세포의 형태학적 비교 분석

김진숙 ; 이인학 김천대학교 2010 김천대학교 논문집 Vol.30 No.-

The purpose of the study was to investigate the correlations between corneal endothelial cell shape according to sex, age and right and left eyes measured by non-contact specular microscope. There is no significant difference in endothelial cell density according to sex and right and left eyes. Endothelial cell was classified to octagon (A8) from triangle (A3) according to cell shape. The percentage of A6 shape cells (hexagonality) was higher than another shape cells in endothelial cell. The percentage of A5 and A6 cells are statistically significant in the right and left ener eye(p=0.046, p=0.020). The distribution of endothelial cell shape was shown no statistically significant correlation with sex in the right and left eyes, except A4 and A7 in the right eye. The distribution of A4 and A8 shape cells was shown asignificant correlation with age in right eye. Therefore, the result which morphometric comparative analysis of the corneal endothelial cell according to sex, age and right and left eyes has shown a variety of cell shape having a significant correlation according to each facts

Screening and Characterization of Drugs That Protect Corneal Endothelial Cells Against Unfolded Protein Response and Oxidative Stress

Kim, Eun Chul,Toyono, Tetsuya,Berlinicke, Cynthia A.,Zack, Donald J.,Jurkunas, Ula,Usui, Tomohiko,Jun, Albert S. Association for Research in Vision and Ophthalmolo 2017 Investigative Ophthalmology & Visual Science Vol.58 No.2

<P><B>Purpose</B></P><P>To screen for and characterize compounds that protect corneal endothelial cells against unfolded protein response (UPR) and oxidative stress.</P><P><B>Methods</B></P><P>Bovine corneal endothelial cells (BCECs) were treated for 48 hours with 640 compounds from a Food and Drug Administration (FDA)-approved drug library and then challenged with thapsigargin or H<SUB>2</SUB>O<SUB>2</SUB> to induce UPR or oxidative stress, respectively. Cell viability was measured using the CellTiter-Glo survival assay. Selected “hits” were subjected to further dose-response testing, and their ability to modulate expression of UPR and oxidative stress markers was assessed by RT-PCR, Western blot, and measurement of protein carbonyl and 8-hydroxydeoxyguanosine (8-OHdG) adducts in immortalized human corneal endothelial cells (iHCECs).</P><P><B>Results</B></P><P>Forty-one drugs at 20 μM and 55 drugs at 100 μM increased survival of H<SUB>2</SUB>O<SUB>2</SUB>-challenged cells, and 8 drugs at 20 μM and 2 drugs at 100 μM increased survival of thapsigargin-challenged cells, compared with untreated control cells. Nicergoline, ergothioneine, nimesulide, oxotremorine, and mefenamic acid increased survival of both H<SUB>2</SUB>O<SUB>2</SUB>- and thapsigargin-challenged cells. Oxotremorine altered DNA damage inducible 3 (<I>CHOP</I>) gene expression, glucose-regulated protein 78 kDa (GRP78) and activating transcription factor 4 (ATF4) protein expression, and protein carbonyl and 8-OHdG levels. Mefenamic acid altered GRP78 protein expression and protein carbonyl and 8-OHdG levels.</P><P><B>Conclusions</B></P><P>Oxotremorine and mefenamic acid are potential survival factors for corneal endothelial cells under UPR and oxidative stress. The described assay can be further expanded to screen additional drugs for potential therapeutic effect in corneal endothelial diseases such as Fuchs' endothelial corneal dystrophy.</P>

토끼각막내피세포의 배양에 대한 성장인자의 효과

차홍원 울산대학교 의과대학 1995 울산의대학술지 Vol.4 No.1

In order to investigate the effects of epidermal growth factor(EGF), fibroblast growth factor(FGF) and transforming growth factor-β(TGF-β), rabbit corneal endothelial cells were cultured. After adding growth factors individually or in combination, DNA synthesis of corneal endothelial cells was measured. EGF and FGF increased DNA synthesis of corneal endothelial cells at all concentration tested(0.03ng/ml-300ng/ml) compared with the control. The maximum increase of DNA synthesis was shown at the concentration of 0.3-10ng/ml of EGF or 10-30ng/ml of FGF. TGF-β reduced DNA synthesis of corneal endothelial cells at all concentration tested (0.03ng/ml-10ng/ml). DNA synthesis of corneal endothelial cells was more increased when EGF and FGF were added in combination. TGF-β inhibited DNA synthesis induced by EGF or FGF when added at the same time. These results indicate that combination of EGF and FGF has maximum effect in DNA synthesis increase of rabbit corneal endothelial cells.