结合组织学评分的牛角膜浑浊和渗透性方法研究
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- 英文论文题名：Research on bovine corneal opacity and permeability test integrating histopathological evaluation
- 论文作者：陈彧 ; 喻欢 ; 程树军 ; 谈伟君 ; 秦瑶
- 英文论文作者：Chen Yu ; Yu Huan ; Cheng Shu-jun ; Tan Wei-jun ; Qin Yao ; Guangdong Pharmaceutical University ; Guangdong Inspection and Quarantine Technology Center ; Sun Yat-sen University
- 年：2016
- 作者机构：广东药学院;广东出入境检验检疫局技术中心;中山大学;
- 论文关键词：眼刺激 ; 替代方法 ; 牛角膜浑浊及渗透实验 ; 组织学
- 英文论文关键词：eye irritation ; alternative methods ; BCOP ; histopathology
- 会议召开时间：2016-06-15
- 会议录名称：第十一届中国化妆品学术研讨会论文集
- 英文会议录名称：The Proceedings of the 11th Seminar on Cosmetic China
- 语种：中文
- 分类号：R99
- 学会代码：ZGXE
- 会议名称：第十一届中国化妆品学术研讨会
- 会议地点：中国上海
- 主办单位：中国香料香精化妆品工业协会
- 学会名称：中国香料香精化妆品工业协会
- 页数：8
- 文件大小：1208k
- 原文格式：O
- 会议级别：全国
摘要
为了优化替代眼刺激动物试验的牛角膜浑浊及渗透实验(BCOP)方法,提高其适用范围和预测性,本研究选取新鲜牛眼角膜,直接暴露液体化学受试物作用10 min,后孵育2 h和荧光素钠处理90min,浊度仪检测角膜浑浊度和酶标仪检测荧光素钠渗透性,同时将角膜常规固定和制备组织学切片,观察组织学病理变化并评分,基于上述3个指标建立预测模型。结果显示,将角膜浑浊度和渗透性2个定量指标,与角膜上皮组织、基质组织和内皮细胞的组织学变化评分进行组合,可对轻微刺激到中等刺激的范围进行准确细分。研究表明,结合组织学观察的优化BCOP方法可提高其预测性,适用于眼刺激机制研究和产品评估。
To refine the alternative method for eye irritation of Bovine Corneal Opacity and Permeability(BCOP),and to improve its applicable range and predictability.The fresh bovine corneas were obtained and directly exposed to liquid chemical substances for 10 min and then post-incubate for 2h,following fluorescein sodium is treated to corneas.The cornea opacity was recorded using the opacitometer,permeability was determined using the spectrophotometer and the histopathological changes of the cornea were observed and graded after routinely fixed and paraffin embedding.Finally,the predictive models based on the above three indexes were established.The results showed that besides the corneal opacity and permeability,integrated histopathological changes of epithelium,stroma and endothelium,which would be accurately distinguished the range of mild irritation to moderate irritation.In conclusion that refined BCOP method combined with histological observation could improve its predictability and benefit for eye irritation mechanism study and products evaluation.
To refine the alternative method for eye irritation of Bovine Corneal Opacity and Permeability(BCOP),and to improve its applicable range and predictability.The fresh bovine corneas were obtained and directly exposed to liquid chemical substances for 10 min and then post-incubate for 2h,following fluorescein sodium is treated to corneas.The cornea opacity was recorded using the opacitometer,permeability was determined using the spectrophotometer and the histopathological changes of the cornea were observed and graded after routinely fixed and paraffin embedding.Finally,the predictive models based on the above three indexes were established.The results showed that besides the corneal opacity and permeability,integrated histopathological changes of epithelium,stroma and endothelium,which would be accurately distinguished the range of mild irritation to moderate irritation.In conclusion that refined BCOP method combined with histological observation could improve its predictability and benefit for eye irritation mechanism study and products evaluation.
引文
[1]程树军,焦红.实验动物替代方法原理与应用[M].北京:科学出版社,2010:285-304.
[2]OECD.OECD guideline for testing of chemicals No.437:Bovine Corneal Opacity and Permeability Test Method for Identifying i)Chemicals Inducing Serious Eye Damage and ii)Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage[S].OECD Paris,2013,1-27.
[3]EPA BRD-Final Report.Background Review Document of an In Vitro Approach for EPA Toxicity Labeling of Anti-Microbial Cleaning Products:EPA Toxicity Labeling Background Review Document[R].USA:EPA,2009.
[4]Furukawa M,Sakakibara T,Itoh K,et al.Histopathological evaluation of the ocular-irritation potential of shampoos,make-up removers and cleansing foams in the bovine corneal opacity and permeability assay[J].Toxicol Pathol,2015,28(4):243-8.
[5]秦瑶,程树军,黄健聪,等.整合CAMVA和BCOP方法检测化妆品的眼刺激性[J].中国比较医学杂志,2014,24(6):78-82.
[6]Niessen CM.Tight junctions/adherens junctions:basic structure and function[J].Invest Dermatol,2007,127(11):2525-2532.
[7]Scott L,Eskes C,Hoffinann S,et al.A proposed eye irritation testing strategy to reduce and replace in vivo studies using Bottom-Up and Top-Down approaches[J].Toxicol in Vitro,2010,24(1):1-9.
[8]Hayashi K,Mori T,Abo T,et al.A tiered approach combining the short time exposure(STE)test and the bovine corneal opacity and permeability(BCOP)assay for predicting eye irritation potential of chemicals[J].Toxicol Sci,2012,37(2):269-280.
[9]Wilson,S.L,Ahearne,M,Hopkinson,A.An overview of current techniques for ocular toxicitytesting[J].Toxicology,2015,327,32-46.
[10]Schrage A,Kolle S.N,Moreno MC,et al.The bovine corneal opacity and permeability test in routine ocular irritation testing and its improvement within the limits of OECD test guideline 437[J].Altern Lab Anim,2011,39(1):37-53.
[11]Kolle SN,Kandarova H,Wareing B,et al.In-house validation of the EpiOcular(TM)eye irritation test and its combination with the bovine corneal opacity and permeability test for the assessment of ocular irritation[J].Altern Lab Anim,2011,39(4):365-387.
[12]黄健聪,秦瑶,程树军,等.牛角膜浑浊渗透试验方法预测眼刺激性的研究[J].中国卫生检验杂志,2014,24(14):1980-1983.
[2]OECD.OECD guideline for testing of chemicals No.437:Bovine Corneal Opacity and Permeability Test Method for Identifying i)Chemicals Inducing Serious Eye Damage and ii)Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage[S].OECD Paris,2013,1-27.
[3]EPA BRD-Final Report.Background Review Document of an In Vitro Approach for EPA Toxicity Labeling of Anti-Microbial Cleaning Products:EPA Toxicity Labeling Background Review Document[R].USA:EPA,2009.
[4]Furukawa M,Sakakibara T,Itoh K,et al.Histopathological evaluation of the ocular-irritation potential of shampoos,make-up removers and cleansing foams in the bovine corneal opacity and permeability assay[J].Toxicol Pathol,2015,28(4):243-8.
[5]秦瑶,程树军,黄健聪,等.整合CAMVA和BCOP方法检测化妆品的眼刺激性[J].中国比较医学杂志,2014,24(6):78-82.
[6]Niessen CM.Tight junctions/adherens junctions:basic structure and function[J].Invest Dermatol,2007,127(11):2525-2532.
[7]Scott L,Eskes C,Hoffinann S,et al.A proposed eye irritation testing strategy to reduce and replace in vivo studies using Bottom-Up and Top-Down approaches[J].Toxicol in Vitro,2010,24(1):1-9.
[8]Hayashi K,Mori T,Abo T,et al.A tiered approach combining the short time exposure(STE)test and the bovine corneal opacity and permeability(BCOP)assay for predicting eye irritation potential of chemicals[J].Toxicol Sci,2012,37(2):269-280.
[9]Wilson,S.L,Ahearne,M,Hopkinson,A.An overview of current techniques for ocular toxicitytesting[J].Toxicology,2015,327,32-46.
[10]Schrage A,Kolle S.N,Moreno MC,et al.The bovine corneal opacity and permeability test in routine ocular irritation testing and its improvement within the limits of OECD test guideline 437[J].Altern Lab Anim,2011,39(1):37-53.
[11]Kolle SN,Kandarova H,Wareing B,et al.In-house validation of the EpiOcular(TM)eye irritation test and its combination with the bovine corneal opacity and permeability test for the assessment of ocular irritation[J].Altern Lab Anim,2011,39(4):365-387.
[12]黄健聪,秦瑶,程树军,等.牛角膜浑浊渗透试验方法预测眼刺激性的研究[J].中国卫生检验杂志,2014,24(14):1980-1983.