Reconstruction of Rabbit Corneal Epithelium on Lyophilized Amniotic Membrane Using the Tilting Dynamic Culture Method

Ahn, Jae-Il,Lee, Doo-Hoon,Ryu, Yang-Hwan,Jang, In-Keun,Yoon, Mun-Young,Shin, Youn Ho,Seo, Young-Kwon,Yoon, Hee-Hoon,Kim, Jae-Chan,Song, Kye-Yong,Yang, Eun-Kyung,Kim, Ki-Ho,Park, Jung-Keug Blackwell Publishing Inc 2007 Artificial Organs Vol.31 No.9

<P>Abstract: </P><P>Rabbit corneal epithelium was reconstructed using tilting dynamic culture with a self-manufactured, amniotic membrane (AM) supporter and a lyophilized amniotic membrane (LAM). Rabbit corneal epithelial (RCE) cells were cultured and cryopreserved after isolation from the limbus. The second- and third-passage RCE cells were plated onto the epithelial side of the LAM of Ahn's AM supporter. Two days later, the air–liquid interface culture was maintained with third-passage RCE cells for 6 days and second-passage corneal epithelial cells for 9 days. The average viability of thawed RCE cells, assessed using trypan blue dye exclusion, was 77.42%. The reconstructed corneal epithelium was characterized by histological (hematoxylin and eosin) and immunohistochemical staining (proliferating cell nuclear antigen) for light microscopy, and by reverse transcriptase-polymerase chain reaction, glucose assay, and transmission electron microscopy. The basal layer of the reconstructed corneal epithelium was well formed, and the epithelium was tightly constructed due to the increase in cell proliferation and differentiation caused by the tilting dynamic culture, as opposed to static culture. Tilting dynamic culture was useful for the reconstruction of the epithelium using easily damaged epithelial cells and resulted in more stratum cell layers. Moreover, cytokeratin (CK3) mRNA expression in tilting dynamic cultured third-passage RCE cells seeded onto AM was greater than in static cultured third-passage RCE cells. The morphology of the reconstructed corneal epithelium on LAM by tilting dynamic culture for 9 days resembled that of the skin epidermis. This was thought to be because the tilting dynamic culture not only accelerated the proliferation and differentiation of cells by physical or mechanical stimulation, but also ensured that the supply of medium was delivered to the basal cells more efficiently. Thus, the reconstruction of the corneal epithelium using LAM and tilting dynamic culture was considered to be a good in vitro model for autologous or allogeneic transplantation of corneal epithelium and skin epidermis in patients with damaged epithelia. </P>

40세 이하 관상동맥 질환자에서 장기적 임상 관찰

이승욱,정명호,박종철,박형욱,조상희,김원,김계훈,이상현,강경태,조장현,김남호,김건형,안영근,조정관,박종춘,강정채 全南大醫大 2001 전남의대학술지 Vol.37 No.2

食用油脂類에 對한 免疫生物學的 硏究

朴炳哲,文宰奎,朴榮吉,金杏順,安年衡,金度勳,金正勳,安榮根,李相根,鮮于演 圓光大學校 1990 論文集 Vol.24 No.2

食用油脂는 健康維持에 必須的인 營養素이다. 그러나 食用油脂는 脂肪을 構成하는 脂肪酸의 種類와 攝取하는 量에 따라 健康을 保衛하는 必須的인 食品의 成分이 되기도 하고 有害한 影響을 끼칠 수도 있어서 脂質代謝와 關連된 疾病 뿐만아니라 老化를 促進하고, 乳房癌, 大腸癌等의 癌發生과도 關連된 것으로 밝혀져 있다. 本 硏究는 우리生活環境에서 食用으로 接할 수 있는 10種의 油脂에 對한 免疫生物學的인 活性에서 究明한 結果 對照群인 참기름食餌群에 比해 올리브유食用群은 全般的으로 體液性 및 細胞性免疫을 有意性있게 增加시켰으나, 大食細胞의 活性과 末梢循環白血球數는 有意性없는 減少를 보였고, 動物性 油脂인 牛脂食餌群과 植物性 油脂인 옥수수기름食餌群, 들기름食餌群, 米糠油食餌群, 大豆油食餌群, 菜種油食餌群 및 고추씨기름食餌群에서는 全般的으로 體液性 및 細胞性免疫, 大食細胞의 活性 및 末梢循環白血球數를 有意性있게 減少시켰다. This study was performed to investigate the effects of edible oils and fats on the immunobiological responses in ICR male mice. Ten groups of experimental diets, such as sesame oil diet, beef tallow diet, lard oil diet, olive oil diet, corn oil diet, perilla oil diet, rice polishing oil diet, soybeen oil diet, rape seed oil diet, and red pepper seed oil diet were fed adlibitum to the ICR male mice for 27 days. The results of this study were summarized as followings. 1. Both humoral and cellular immune responses, phagocyte activity, and number of leukocytes in sesame oil group were increased. However, the increasing rate of body weight and the weight retios of spleen and thymus to body were generally decreased. 2. Hemagglutination titer(HA) and rosette forming cell(RFC) of the beef tallow group and the lard group were significantly lower than the sesame oil group(P<0.01), but the lard group significantly increased Arthus reaction and delayed type hypersensitivity(DTH)(P<0.01). 3. Arthus reaction, DTH, and RFC of the olive oil group were significantly higher than the sesame oil group(P<0.01). 4. DTH of the corn oil group was significantly higher than the sesame oil group(P<0.01) whereas phagochyte activity was significantly lower(P<0.01). But no statistically significant differences were found between the two groups in the humoral immune response. 5. HA, PFC, DTH, RFC, phagocyte activity, and number of leukocytes of the perilla oil group and the rice polishing oil group were significantly lower than the sesame oil group(P<0.01). 6. Both humoral and cellular immune responses, phagocyte activity, and number of leukocytes of the soybeen oil group, the rape seed oil group, and the red pepper oil group were significantly lower than the sesame oil group(P<0.01).

초등학교 수학교육에서 계산기 활용에 대한 문제의 유형과 교수ㆍ학습 모형 개발

안병곤,김용태,임해경,류근봉 광주교육대학교 초등교육연구소 2000 初等敎育硏究 Vol.15 No.1

This paper aims to enhance students' interest in the use of calculators in mathematics education and promote their use of calculators in real-life situations. Towards these ends, problem types and instructional models developed for the efficient utilization of calculators. The instructional models focus on teaching mathematics relying on the path through which expert teachers have gone through to gain relevant knowledge. By developing problem types and instructional models suitable for calculator use, we can contribute to a better attainment of instructional goals in mathematics education. The instructional models and problem types will aid teachers in making decisions about instructional development plan and basic features of instructional activities. The use of a new medium will also lead to increased interest and confidence in learning, thus contributing to the enhancement of students' ego.

마우스에 있어서 Olive Oil의 食餌가 免疫反應에 미치는 影響

안영근,박병철,김정훈,이상근,박영길 원광대학교 식품약품안전성연구소 1991 食品藥品安全性硏究 Vol.4 No.-

This study was performed to investigate the effect of olive oil diet on the immune response in ICR male mice. Experimental diets of 4 groups were fed ad libitum to the ICR male mice for 27 days. The results of this study were summarized as followings: 1. 10% Olive oil diet group as compared with the control diet group significantly decreased liver weight rate but significantly increased hemagglutination titer (HA), Arthus reaction, delayed type hypersensitivity reaction (DTH), rosette forming cell (RFC), and phagocyte activity. 2. 20% Olive oil diet group as compared with the control diet group significantly increased body weight gain, liver weight rate, and HA but significantly decreased Arthus reaction, DTH, RFC, phagocyte activity, and peripheral circulating white blood cell (WBC). 3. 30% Olive oil diet group as compared with the control diet group significantly increased liver weight rate but significantly decreased body weight gain, Arthus reaction, plaque forming cell (PFC), DTH, RFC, phagocyte activity, and WBC. The results showed that the increase of olive oil doses significantly decreased humoral and cellular immune responses, phagocyte activity, and WBC.

A Comparison of Lyophilized Amniotic Membrane with Cryopreserved Amniotic Membrane for the Reconstruction of Rabbit Corneal Epithelium

Ahn Jae-Il,Jang In-Keun,Lee Doo-Hoon,Seo Young-Kwon,Yoon Hee-Hoon,Shin Youn-Ho,Kim Jae-Chan,Song Kye-Yong,Lee Hee-Gu,Yang Eun-Kyung,Kim Ki-Ho,Park Jung-Keung The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3

Many researchers have employed cryopreserved amniotic membrane (CAM) in the treatment of a severely damaged cornea, using corneal epithelial cells cultured on an amniotic membrane (AM). In this study, two Teflon rings were made for culturing the cells on the LAM and CAM, and were then used to support the AM, which is referred to in this paper as an Ahn's AM supporter. The primary corneal epithelial cells were obtained from the limbus, using an ex-plantation method. The corneal epithelium could be reconstructed by culturing the third­passage corneal epithelial cells on the AM. A lyophilized amniotic membrane (LAM) has a higher rate of graft take, a longer shelf life, is easier to store, and safer, due to gamma irradiation, than a (AM. The corneal epithelium reconstructed on the LAM and (AM, supported by the two­Teflon rings, was similar to normal corneal epithelium. However, the advantages of the LAM over that of the (AM make the former more useful. The reconstruction model of the corneal epithelium, using AM, is considered as a good in vitro model for transplantation of cornel epithelium into patients with a severely damaged cornea.

조혈모세포이식 환자에서의 기계 환기의 위험 인자 : Assessment of Risk Factors

안중경,이홍기,황정혜,박세훈,이효락,송서영,이순일,박준오,김기현,김원석,정철원,임영혁,강원기,박근칠 대한조혈모세포이식학회 2002 대한조혈모세포이식학회지 Vol.7 No.1

연구배경: 조혈모세포이식을 시행한 환자에서 집중 치료 시 예후가 매우 나쁜 것으로 알려져 있고, 기계 환기는 강력한 사망 예측 인자로서 보고되고 있다. 그러나 현재까지 이식 환자에서 기계 환기의 위험 인자를 밝힌 연구는 매우 드물다. 따라서 기계 환기를 시행한 조혈모세포이식 환자의 임상적 특징을 살펴보고 기계 환기에 대한 위험 인자를 알아보고자 하였다. 방법: 삼성서울병원에서 조혈모세포이식을 시행한 환자 중 기계 환기를 시행한 23명과 기계 환기를 시행하지 않은 142명을 대상으로 후향적 연구를 시행하였다. 기계 환기에 대한 위험 인자의 여부에 관해서는 chi-square 또는 Fisher's exact 검정을 시행하였으며 기계 환기에 대한 각 위험 인자의 영향에 관해서는 다중로지스틱 회귀분석을 시행하였다. 결과: 기계 환기를 시행한 23명의 환자 중 30일째 생존자는 1명이었고 6개월째 생존율은 0%였다. 생존자와 사망자 모두에서 다기관 기능부전이 관찰되었으며, APACHE II 점수와 SAPS II 점수에 의한 예측 사망률은 각각 56%, 59%였다. 조혈모세포이식 후 기계 환기의 위험 인자로는 선행 질환이 혈액질환, 부분불일치 동종이식, 간정맥폐색성질환, 이식 전 질병 상태가 재발하였거나 약물에 반응하지 않는 경우였다. 다중로지스틱 회귀분석 결과 기계 환기의 위험 인자로 통계적으로 유의한 차이를 나타낸 것은 단지 부분불일치 동종이식을 시행한 경우였다. 결론: 현재까지 조혈모세포이식 환자에서 집중 치료의 역할에 대해서는 확실히 정립되어 있지는 않으며, 이식 환자에서 기계 환기는 강력한 사망 예측 인자이다. 따라서, 조혈모세포이식을 시행한 환자에서 기계 환기의 위험 인자와 불량한 예후 인자를 고려하여 기계 환기 여부에 대한 신중한 결정을 내려야 한다. Background: Respiratory failure requiring mechanical ventilation is a frequent, critical complication of hematopoietic stem cell transplantation (HSCT). Patients who require mechanical ventilatory support after HSCT generally have a very poor prognosis. Mechanical ventilation in HSCT recipients is a strong predictive factor of mortality. The objectives of this study are to describe clinical characteristics of HSCT recipients undergoing mechanical ventilation and to identify the risk factors for mechanical ventilatory support after HSCT. Methods: We performed a retrospective chart review of all patients >15 yrs old who received HSCT at Samsung Medical Center and subsequently required mechanical ventilatory support between 1996 and 2001. Results: Thirty-day mortality rate in HSCT recipients undergoing mechanical ventilation was 95.6%. The mean predictive mortality rates of APAHCE II score and SAPS II score were 56% and 59%, respectively. Reasons for mechanical ventila-tion were sepsis (47.8%) followed by fungal infection (13%) and diffuse alveolar hemorrhage (8.7%). Univariate analysis identified relapsed or refractory diseases at HSCT, hematologic disease, hepatic venoocclusive disease and allogeneic or HLA-mismatched transplant as significant risk factors for mechanical ventilation. On multivariate logistic regression analysis, only allogeneic mismatched transplant remained significant. Conclusion: Overall outcome of HSCT recipients undergoing mechanical ventilation is very poor. Therefore, the risk factors and the poor prognostic factors for mechanical ventilation should be taken into account in making further treatment decision for HSCT recipients requiring mechanical ventilation.

마우스에 있어서 Cimetidine , Ranitidine 및 Famotidine이 免疫反應에 미치는 影響

안영근,김정훈,이상근 한국환경독성학회 1990 환경독성보건학회지 Vol.5 No.2

Experiments were performed on mice to investigate the influences of cimetidine, ranitidine and famotidine on the immune response. Immune response were evaluated by antibody, Arthus reaction (Arthus), delayed type hypersensitivity (DTH), rosette forming cell (RFC), phagocyte activity and whit( blood cell (WBC) in mice, sensitized and challenged with sheep red blood cells (SRB"). The weight of liver, spleen and thymus were measured. Following results obtained in this experiment. 1) The administration of cimetidine as compared to normal group significantly decreased Arthus, Hemagglutinin titer (HA), RFC, DTH, WBC and phagocyte activity, but increased the activity of serum albumin. 2) The administration of ranitidine as compared to normal group decreased RFC and HA. 3) The administration of Famotidine as compared to normal group decreased DTH and RFC, and significantly decreased HA, Arthus and serum protein. 4) The administration of ranitidine and famotidine decreased more humoral immune response than cellular immune response, but the administration of cimetidine significantly decreased humoral and cellular immune response, WBC and phagocyte activity.

마우스에 있어서 Cimetidine, Ranitidine 및 Famotidine이 免疫反應에 미치는 影響

안영근,김정훈,이상근 원광대학교 식품약품안전성연구소 1991 食品藥品安全性硏究 Vol.4 No.-

Expreiments were preformed on mice to investgate the influences of cimetidine, ranitidine and famotidine on the immune response. Immune response were evaluated by antibody. Arthus reaction(Arthus), delayed type hypersensitivity(DTH), rosette forming cell(RFC), phagocyte activity and white blood cell (WBC) in mice, sentitized and challenged with sheep red blood cells (SRB). The weight of liver, spleen and thymus were measured. Following results obtained in this experiment. 1) The administration of cimetidine as compared to normal group significantly decreased Arthus, Hemagglutinin titer(HA), RFC, DTH, WBC and phagocyte activity, but increased the activity of serum albumin. 2) The administration of ranitidine as compared to normal group decreased RFC and HA. 3) The administration of Famotidine as compared to normal group decreased DTH and RFC, and significantly decreased HA, Arthus and serum protein. 4) The administration of ranitidine and famotidine decreased more humoral immune response than cellular immune response, but the administration of cimetidine significantly decreased humoral and cellular immune response, WBC and phagocyte activity.

마우스에 있어서 Isoniazid와 Rifampicin의 免疫毒性에 대한 酒淸 및 高麗人蔘의 影響

안영근,김정훈,유항묵,강영세 원광대학교 식품약품안전성연구소 1987 食品藥品安全性硏究 Vol.1 No.-

結核의 一般的인 藥物療法으로 利用되고 있는 Isonicotinic acid hydrazide(INAH)와 Rifampicin의 倂用投與는 結核의 治療效果를 改善시킬 수 있는 反面에 INAH와 Rifampicin의 長期 倂用投與가 肝毒性과 免疫抑制作用을 出現시킨다고 新告된 바 있다. 한편 酒精의 長期間 暴露는 肝毒性과 T-Iymphocyte機能抑制 및 體液性 免疫에 손상을 가져 온다고 알려진 바 있고 高麗人蔘은 細胞性 및 體液性 免疫 修飾作用이 있으며 特히 石油 ether 分劃은 그 作用이 强하여 細胞性 및 體液性 免疫 macrophage의 活性 등을 현저하게 亢進시킨다고 本人 等이 밝힌 바 있다. 本 硏究의 結果 INAH와 Rifampicin倂用與時 이들 藥物에 의한 免疫 毒性에 對하여 酒精 및 高麗人蔘을 倂用하였을 때 INAH와 Rifampicin은 細胞性 및 體液性 免疫을 低下시켰으며 酒精의 倂用投餘는 免疫 毒性을 增加하였으나 高麗人蔘의 倂用投與는 細胞性 및 體液性 免疫과 大食細胞의 活性을 增加시킴을 밝혔다. 1. Rifampicin and INAH exposure decreased relative spleen weight, humoral and cellular immune responses and macrophage activity. 2. Alcohol combined administration significantly increased the cellular immunotoxicity of Rifampicin and INAH. Especially, 4% alcohol combined administration significantly suppressed the cellular immune response and macrophage activity. 3. Generally, Korean giseng extracts reduced of removed the decline of humoral and cellular immune responses and macrophage activity by immunotoxicity of Rifampicin and INAH.