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.

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>

2003년 국내 중증급성호흡기증후군 진료 현황 및 문제점 분석

이진수,김은실,정문현,백제중,정선화,안주희,최영화,이선희,고철우,김성범,김민자,박승철,기현균,송재훈,최상호,김양수,이상오,조용균,박영훈,정숙인,김연숙,이흥범,손창희,장성희,정희진,김우주 대한감염학회 2004 감염과 화학요법 Vol.36 No.3

목적 : 2002년 말 중국에서 SARS가 발생한 이후 국내에서도 2003년 10월까지 총 3명의 추정환자, 17명의 의심환자가 보고되었다. 향후 추가적인 SARS의 유행이 우려되는 상황에서, 그간의 SARS 환자 진료에 있어서의 실질적인 준비사항, 진료 현황 등에 대한 조사를 통해 문제점을 파악하여, 향후 더 나은 대비가 될 수 있도록 개선점을 제시하고자 하였다. 재료 및 방법 : SARS로 의심되는 환자를 진료 경험이 있는 병원의료진을 대상으로 2003년 10월에 설문조사를 실시하였다. 설문에는 SARS 환자 진료 시의 실질적인 조치, 진료 현황, 병실, 응급실 및 외래에서의 격리 시설과 준비사항, 보건당국의 관리와 지원에 관한 사항을 포함하였다. 결과 : 대상이 되는 22개 병원 중 17개(17/22, 77.2%) 병원이 설문에 응하였다. SARS 환자를 위한 격리실은 응급실, 외래, 일반병실 및 중환자실에서 각각 9개(9/17, 52.9%), 5개(5/17, 29.4%), 15개(15/16, 93.7%), 4개(4/16, 25%) 병원에서 음압처리가 되어있지 않은 일인실 혹은 다인실이 사용되었고, 1개(1/16, 6.3%) 병원에서만 일반병실에서 음압격리실이 운영되었다. 입원환자의 진찰 시 개인보호구의 착용은 거의 모든 의료기관에서 이루어졌다. 보건당국에서 SARS지정병원의 시설 등을 사전에 확인한 곳은 1곳(1/12, 8.3%)이였고, 14개 병원(14/15, 93.3%)에서는 보건당국에 의뢰한 검사결과를 통하 받지 못하였다. 결론 : 의료기관에서 SARS 환자용 격리실뿐만 아니라 기존의 격리실 설비 등이 미흡하였으며, 특히 중환자실 및 외래의 준비가 더욱 부족하였다. 보건당국의 의료기관에 대한 종합적인 지원이 부족하였고, 병원과의 원활한 연계가 잘 이루어지지 않았다. SARS 만이 아닌 격리를 필요로 하는 질환의 적절한 진료를 위해 향후 병원 시설의 정비와 정부차원에서의 보다 구체적이고 실질적인 대책마련이 필요하다. Background : There was an worldwide outbreak of the Severe Acute Respiratory Syndrome (SARS) originated from China in late 2002. During that period three cases of suspected SARS and 17 cases of probable SARS were reported in Korea. With the concerns about the reemergence of SARS-coV transmission, it is important to be prepared for any possibility. So, this study is aimed to analysis the past measures in managing SARS and propose the amendatory plans to improve the preparedness. Materials & Methods : Questionnaires were collected among clinicians with any experience in managing the probable or suspected SARS cases in Oct. 2003. 17 out of 22 hospitals responded to the questionnaire. The contents in the questionnaire were practical activities, personal equipments, response plans, isolation facilities in emergency centers, outpatient clinics, general wards and intensive care units, and relationship with the public health department. Results : The dedicated isolation rooms in emergency centers, outpatient clinics, general wards, and intensive care units were prepared in 9 (9/17, 52.9%), 5 (5/17, 29.4%), 15 (15/16, 93.7%), and 4 (4/16, 25.0%) hospitals, respectively. Except for one hospital that newly made negative pressure room for SARS, single or multi-bed rooms without airborne infection control were used in all the other hospitals. The personal precaution principles were kept quite well in general wards. Before the designation of SARS hospital by the public health department prior evalution to see if the hospital was suitable for managing SARS was conducted in only 1 (1/12, 8.3%) hospital. The results of laboratory diagnosis were reported back in 1 (1/15, 6.6%) hospital. Conclusions : The isolation facilities which can control airborne infection were almost deficient not only for SARS but also for other respiratory transmissible diseases. For the infection control of transmissible diseases including SARS, more investment is needed on medical facilities and comprehensive support from the public health department required.

부분 치수절단술을 이용한 복잡 치관파절의 치료 : 증례보고

안병덕,김영재,장기택,이상훈,김종철,한세현,김정욱 大韓小兒齒科學會 2008 大韓小兒齒科學會誌 Vol.35 No.2

혼합 치열기 및 초기 영구치열기의 소아,청소년에서 외상에 의한 치관 파절은 빈번하게 발생한다. 치관파절은 치수의 노출 유무에 따라 단순 치관파절,복잡 치관파절로 분류된다. 치수가 노출된 복잡 치관파절의 경우 치수에 대한 고려 및 처치가 필수적인데,초기 영구치열기에는 전치부 치근의 발육이 완전히 이루어지지 않았기 때문에 완성된 치근을 가지는 치아보다 치료가 복잡하다. 치료의 목적은 적절한 치수 치료를 통해 치근의 완성을 도모하는 것으로 이를 위해 직접 치수복조술,부분 치수절단술,치경부 치수절단술의 방법을 이용한다. 이 중 부분 치수절단술은 성공률도 높으며 치경부 치수를 보존하기 때문에 여러 장점을 지니는 치료방법이다. 복잡 치관파절을 주소로 내원한 환자에서 부분 치수절단술을 시행하여 양호한 결과를 나타내어 이를 보고하는 바이다. Traumatic crown fracture in mixed dentition or early permanent dentition is relatively common. Crown fracture is classified into simple or complicated crown fracture by the presence of pulp exposure. The condition of pulp must be considered in treatment of crown fractures with pulp exposure. Treatment of immature crown-fractured incisor with pulp exposure is more complex because of its incomplete root formation. Pulp capping, partial pulpotomy, cervical pulpotomy can be used for continuous root development in immature crown-fractured incisor with pulp exposure. The success rate of partial pulpotomy is very high and there are several advantages of partial pulpotomy because the cell-rich coronal pulp tissue is preserved. This paper reports 2 cases of crown-fractured permanent incisors with pulp exposure that had been treated by partial pulpotomy successfully.

마우스에 있어서 Isoniazid와 Rifampicin의 免疫毒性에 對한 酒精 및 高麗人蔘의 影響

안영근,김정훈,유항묵,강영세 圓光大學校 1988 論文集 Vol.22 No.2

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 報告된 바 있다. 한편 酒精의 長期間 暴露는 肝毒性과 T-lymphocyte 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 倂用投與時 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 倂用投與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 ginseng extracts reduced or removed the decline of humoral and cellular immune responses and macrophage activity by immunotoxicity of Rifampicin and INAH.

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.

Alginate 배양내에서 Transglutaminase가 연골막세포에 미치는 영향

박정선,공차배,김충현,안현정,정미현,성상철,김희중,조현철,이상훈,이명철 대한정형외과연구학회 2003 대한정형외과연구학회지 Vol.6 No.2

蔘茸扶正湯의 放射線 照射로 損傷된 組織 恢復 및 造血促進 效果

崔昇勳,安圭錫,金正洙 대한동의병리학회 1997 동의생리병리학회지 Vol.11 No.2

현재 서양의학에서 활용되고 있는 항암 요법으로는 外科處置, 化學療法, 放射線療法 및 免疫療法 等이 있으며, 그 중에서는 抗癌劑에 의한 화학요법과 방사선요법이 가장 많이 응용되고 있다. 그런데 이들은 癌細胞뿐만 아니라 正常細胞까지도 殺傷함으로써 腫瘍細胞를 억제하는 동시에 骨髓造血器, 消化系統 및 免疫機能을 포함하는 인체의 정상적인 기능을 손상시키는 문제점으로 그 사용에 제한을 받고 있다. 따라서, 본 연구는 蔘茸扶正湯의 항암요법으로 손상된 조직의 회복 및 조혈촉진 효과를 규명하기 위해 생쥐를 대상으로 하여 실험하였다. 그 결과, 생쥐 비장세포에 대한 增殖效果, 造血促進因子의 分泌能, 방사선에 대한 임파구의 防禦效果, 방사선에 대한 조혈세포 防禦效果, 방사선(9Gy)을 조사받은 생쥐의 生存率 등에서 유의성있는 결과를 얻어내어 아래와 같이 보고하는 바이다. Radiotherapy is an irreplaceable method of cancer treatment. But it has several side effects, especially damages to the hemopoietic and Immune system. Therefore radioprotectors are required to treat cancer successfully. A lot of Herbs and Herbal prescriptions are reported to have radioprotective effects. Above all, those to support the healthy energy and strengthen the body resistance are found more effective. This study was performed to evaluate the radioprotective effects of prescription Shenrong Fuzheng Tang(S. F. T), which consists of 16 kinds of herbs. We investigated proliferation of murine splenocytes, secretion of colony-stimulating-factors(CSFs), immunocompetence after irradiation in-vitro, and Endogenous spleen colony assay, survival assay in-vivo. When splenocytes were cultured with Shenrong Fuzheng Tang (S.F.T) (500㎍/㎖), proliferation was enhanced 5.7 times compared to control cultured with medium alone(P〈0.05) and, showed highest proliferation at 4th day after incubation. In order to evaluate stimulation of hemopoiesis of Shenrong Fuzheng Tang(S.F.T), the supernatant of splenocytes cultured with optimal concentration of Shenrong Fuzheng Tang(S.F.T) was used to measure CSFs secretion. The result showed enhanced secretion of colony-stimulating-factors(CSFs) compared to control(P〈0.05). To evaluate the protective effect of lymphocytes from irradiation, proliferation of splenocytes stimulated by LPS and ConA after incubation with Shenrong Fuzheng Tang(S.F.T) for 24h Prior to Irradiation(1∼3 Gy) was measured. The results showed higher proliferation of Shenrong Fuzheng Tang(S.F.T.) treated cells than that of non-treated cells. And percentage increases of irradiated splenocytes per non-irradiated splenocytes were also higher in Shenrong Fuzheng Tang(S. F. T)-treated cells than control. Endogenous spleen colony assay, to evaluate the protection of hemopoietic cells form irradiation, showed increased number of colonies(p=0.03) in Shenrong Fuzheng Tang(S.F.T) treated murine spleen(10.3±1.9) compared to non-treated murine spleen(3.4±0.8). Survival time of mice irradiated with lethal dose of γ-ray(9Gy) was prolonged in Shenrong Fuzheng Tang(S.F.T) treated group prior to irradiation as compared to non-treated group. According to these results we can suggest that prescription Shenrong Fuzheng Tang(S.F.T) has radioprotective effects and be used to protect the hemopoietic and immune system from damages of anti-cancer radiotherapy.

고위험 유방암환자에서 CPB(Cyclophosphamide, Cisplatin, BCNU) 고용량 화학요법과 자가조혈모세포이식술 : A Phase II Multi-Center Trial

최정혜,안명주,조은경,신동복,이재훈,이문희,김철수 대한조혈모세포이식학회 2001 대한조혈모세포이식학회지 Vol.6 No.2

배경: 림프절 전이가 많은 고위험군 유방암환자에서 CPB(cyclophosphamide, cisplatin, BCNU)를 이용한 고용량화학요법 및 자가조혈모세포이식술이 무병생존율에 미치는 영향과 안전성을 규명하기 위하여 연구를 시행하였다. 방법: 1998년 2000년 까지 한양대학병원, 가천의대병원과 인하대학병원에서 4개 이상 림프절을 침범한 유방암으로 근치적 절제술 시행 후 고용량 화학요법과 자가조혈모세포이식을 시행 받은 13명의 환자를 대상으로 하였고 대조군은 4개 이상 림프절을 침범한 유방암으로 보조화학요법을 받은 환자들 중 39명을 무작위로 선택하여 비교분석하였다. 고용량 화학요법은 제-6일에 BCNU 450 mg/m², 제-5일부터 3일까지 cyclophosphamide 2.0 g/m²와 cisplatin 55 mg/m²를 각각 투여하였다. 결과: 채집된 단핵구수의 중앙값은 8.5 x 10^(8)/kg (범위 2.44-17.90)이었고 CD34양성세포수의 중앙값은 11.0 x 10^(6)/kg (범위 0.8-78.35)이었다. 모든 환자가 이식 후 중앙치 10.0일(범위 8-15)에 말초혈액 과립구가 500 /mm³에 도달하였고 중앙치 9.0일(범위 0-18)에 혈소판이 20,000 /mm³에 도달하였다. 고용량 화학요법을 시행 받은 환자군의 추적기간의 중앙값은 24.6 개월 (범위 15.5-40.5)이었으며 총 13명의 환자 중 1명이 뇌전이로 인하여 사망하였으며 3명(23.1%)에서 재발이 관찰되었다. 대조군은 재발율이 25.6%이었고 무병생존율은 고용량 화학요법을 시행 받은 군(70.5%)과 대조군(78.8%) 사이에서 통계학적으로 의미 있는 차이를 나타내지 않았다 (P=0.744). 고용량 화학요법을 시행 받은 환자 중 10명 76.9%에서 발열이 관찰되었고 WHO 기준 3도 및 4도의 부작용은 혈액학적 부작용이 6예, 소화기적 부작용이 6예에서 관찰되었다. 결론: 고위험군 유방암 환자에서 CPB를 이용한 고용량화학요법 및 자가조혈모세포이식술은 안전하게 적용할 수 있는 치료의 한 방법이나 고용량 화학요법의 효과에 대해서는 앞으로 발표될 대규모 3상 연구의 결과를 주시해야 할 것으로 생각된다. Background: Despite the improvement of prognosis by multimodality therapy for the treatment of breast cancer, primary breast cancer patients with multiple involved axillary lymph node still have a high risk of recurrence. Methods: Between January 1998 and May 2000, we treated 13 women with histologically proven stage II or III breast cancer with 4 or more axillary lymph nodes at the time of primary mastectomy with high-dose chemotherapy and autologous stem cell transplantation. Results: The median age was 45 years (range: 32-57) and the median number of involved lymph node was 13 (range: 6-31). Ten patients were premenopausal state and 40 percent were positive hormone receptor. Seven patients received radiation therapy. Patients were treated with high-dose CPB (cyclophosphamide 2 g/m² D-5--3, cisplatin 55 mg/m² D-5--3, BCNU 450 mg/m² D-6) with autologous peripheral blood stem cell transplantation after 2-4 cycles of adjuvant chemotherapy (CAF, CEF or AC). We compared those patients with age and other clinical factors-matched 39 patients who received conventional adjuvant chemotherapy. The median follow-up for surviving patients who received high-dose chemotherapy was 25.6 months. The relapse rate and 2-year relapse-free survival was 23.1%, 25.6% and 70.5%, 78.8% for high-dose and conventional dose chemotherapy, respectively, but there was no significant difference between two groups (P>0.05). Conclusion: Further follow-up and additional studies are required to resolve the role of high-dose chemotherapy with autologous stem cell transplantation in high-risk breast cancer.

악골 외상성 골 낭의 임상 및 병리조직학적 분석

윤정훈,김도경,안상건,김수관 大韓顎顔面成形再建外科學會 2003 Maxillofacial Plastic Reconstructive Surgery Vol.25 No.1

We reviewed a series of 22 traumatic bone cysts of the jaws experienced during the 21-year period between 1982 and 2002 at the Chosum University Dental Hospital. The study group consisted of 13 females (59.1%) and 9 males (40.9%). The patient's mean age was 25.2 years, with peak incidence occuring in the first and second decade. Most traumatic bone cyst was located in the posterior mandible. Radiographically, well-defined round radiolucent (9 cases) or scalloped appearance (8 cases) were the predominant type, whereas 2 cases were ill-defined. A provisional clinical diagnosis was varied, only 9 of which were correct. Histopathologically, all the biopsy specimen composed of loose fibrous connective tissue accompanied by inflammatory reaction (13 cases), giant cells (9 cases). In addition, calcifications (6 cases) and new bone formation (5 cases) in the cyst wall were also observed. Taken together, correlative analysis of histopathologic and clinico-radiographic features is essential for definitive diagnosis and management.