体外模拟后发障的组织培养及药物防治研究
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摘要
白内障囊外摘除(ECCE)联合人工晶体(IOL)植入术是治疗各种白内障的主要手段，但该手术的主要术后并发症后发障可严重影响患者的视力。在手术后的1个月～5年，约有50％以上的病人出现后囊混浊(posterior capsule opacification，PCO)。而儿童白内障术后PCO的发生率几乎为100％。
目前的研究认为：PCO的形成是由于白内障囊外摘除术后血—房水屏障遭到破坏，导致外源性的细胞因子和炎症介质进入囊袋内，作用于前囊残留的晶状体上皮细胞上，使其发生成纤维细胞样改变，并向后囊不断增殖、迁徙，合成大量的胶原和细胞外基质，这些物质相互混杂，在后囊上形成膜状混浊物。该混浊物可引起后囊皱缩、变形，形成光散射，最终导致视力严重下降。
为了解PCO的形成机制，国外进行了多种体外模拟PCO的培养研究，为体外条件下观察PCO的形成过程和发生机制提供了优越的模型，而国内未见此方面的报道。此次实验我们欲进行牛晶状体后囊的组织培养，体外模拟PCO的形成，观察PCO形成过程中后囊晶状体上皮细胞的增殖分化规律以及血清对体外PCO模型的形成影响。普拉洛芬(Pranoprofen)属于非甾体类抗炎药物，它可明显改善白内障术后及眼前段非感染性炎症患者的主要症状和体征。但关于普拉洛芬对晶状体上皮细胞的抑制实验报道较少，因此，本次实验通过后囊的组织培养，进一步模拟普拉洛芬滴眼液滴眼后4小时在房水中的浓度，观察其对PCO的形成有无抑制作用。
郑州大学20()4届硕士学位论文
体外模拟后发障的组织培养及药物防治研究
一、实验方法
在无菌条件下，将分离的牛晶状体后囊细胞面朝上平铺于25ml培养瓶底
部，分别加入含O、10%、20%胎牛血清的DMEM培养液2.sml，将培养瓶竖直
放入37℃、5%coZ的培养箱中，培养液蒸发的水蒸汽可维持晶状体后囊表面湿
润，1小时后将培养瓶放平，使囊膜完全浸入培养液中，可见晶状体后囊已于
培养瓶的底部牢牢粘附，培养液不会使后囊卷曲、漂浮，再将培养瓶放入培养
箱中培养，每3天更换培养液1次，直到后囊晶状体上皮细胞达到融合。培养
的各阶段，在倒置显微镜下，用方格图计数板计数后囊晶状体上皮细胞的覆盖
率和融合时间，并用Giemsa染色和扫描电镜观察晶状体上皮细胞的形态。同时
在含0和2%胎牛血清的培养基中加入普拉洛芬，使其成为体内代谢4小时后
的房水内浓度(0.23m岁L)，进行体外培养，观察与对照组后囊上皮细胞融合
天数有无差异。
二、实验结果:
赤道部的晶状体上皮细胞向后囊中心部迅速增殖、迁徙，一般约需8一18
天可达到融合。细胞的增殖覆盖率与培养液中血清的含量呈正相关，融合时间
与血清含量呈负相关(P<0.05)。后囊晶状体上皮细胞发生成纤维细胞样改变，
并合成大量的细胞外基质后囊形成明显的混浊和皱缩。培养液中有无血清均可
出现后囊皱缩，且血清的含量越高，皱缩形成的数目越多，时间越短。对皱缩
区域的Giemsa染色和扫描电镜观察显示:各皱缩纹理排列几乎平行，皱缩边缘
有大量的晶状体上皮细胞聚集，且细胞的聚集方向与皱缩纹理的长轴平行。在
两种培养状态下，普拉洛芬在体内代谢4小时后的房水内浓度(0 23m留L)对
后囊上皮细胞的融合均有明显的抑制作用(P<0.01)。
三、讨论:
1.组织培养条件下的后囊能出现许多与体内PCO相似的改变，如晶状体
上皮细胞迅速地在后囊上增殖、纤维化，定向收缩使后囊形成皱缩，并分泌大
量的细胞外基质。它是体外模拟PCO形成和研究后发障发生机制的有效方法。
2.血清对体外PCO形成的影响说明:血一房水屏障破坏在PCO形成的
早期起到重要作用。后囊无血清组织培养结果显示:晶状体上皮细胞存在“自
分泌”机制，PCO的形成是由于“自分泌机制”和“旁分泌机制”共同作用的
郑州大学2004届硕士学位论文
体外模拟后发障的组织培养及药物防治研究
结果。
3.普拉洛芬是一种安全、有效的降低PCO发生率的药物，它可作为白内
障术后的常规用药。
In recent years, extracapsular cataract extraction (ECCE) and intraocular lens (IOL) implantation play the main role in cataract surgery. Posterior capsule opacification (PCO) , the major complication after ECCE, will affect the paients' visual acuity seriously. PCO occurs in up to 50% of paients between 1 month and 5 years after surgery., and occurs almost 100% in children.
Researchse showed that the formation of PCO is because breakdown of the blood-aqueous barrier after ECCE, which lead to growth factor and inflammatory medium enter capsular bag and act on residual lens epithelial cells on the anterior capsule.The fibroblast-like cells migrate and proliferate rapidly on posterior capsule and secrect a lot of collagen and extracellular matrix. Opaciflcation occurs as a result of the formation of fibrotic membranes on the posterior capsule. Opaciflcation will result in the formation of wrinkle and distortion, which lead to light scattering and visual acuity decrease seriously
In order to find out the mechanism of PCO formation, a lot of model for
PCO in vitro had been established overseas that provided a good example to observe the process and mechanism of PCO formation ,but the report is seldom in domestic. In this research, tissue culture of lens epithelial cells on the posterior capsule to model posterior capsule opacification (PCO) will be erected, to observe the proliferation of cells and the affection of serum on model for PCO in vitro.the inhibition effects of pranoprofen on PCO were observed. Pranoprofen is a non-steroidal anti-inflammatory drug.In this research, we model pranoprofen whose concentration was like to that has metabolized for 4 hours in aqueous humor (0.23 mg/L) and observe the inhibition of pranoprofen on PCO formation.
1. Methed
To spread out the bovine lens posterior capsule with the cell layer upwards on the surface of a 25ml culture flask in sterile condition. DMEM with 0 ,10% and 20% fetal calf serum 2.5ml used as medium were added respectively. The flask was set on ites end in 37 5%CO incubator. The rising water vapour protected the tissue from drying, forming a wet chamber. After 1 hour the flask was laid down,so that the medium covered the epithelium and continue to culture.The medium was changed every 3 days until the confulence of lens epithelial cells on posterior capsule. The cell coverage and confluence time on posterior capsule were recorded with graticule by inverted microscope, and the cell morphology was observed by Giemsa staining and scanning electron-microscope. The lens epithelial cells on posterior capsule were cultured in medium with pranoprofen whose concentration was like to that has metabolized for 4 hours in aqueous humor (0.23 mg/L), and then observed the difference of confluence time with control group.
2. Result
The lens epithelial cells migrate and proliferate rapidly on posterior capsule from equatorial region to the center. Within 8 to 18 days the cells became confluent. The cell coverage was rising with the increase of serum concentration, and the cell confluence time was shortening with that (P < 0.05). The lens epithelial cells on posterior capsule make a fibroblast-like change and secrect a lot of collagen and extracellular matrix. The PCO and wrinkles were formed obviously. The wrinkles presence were not dependent upon serum concentration, but increased in munber and decreased in time with that. The morphology of wrinkles by Giemsa staining and scanning electron-microscope show: the wrinkles almostly paralleled with each other., there were closely packed cells along that, and the direction of packed cells were alinged mainly at right angles to the wrinkles. In two kind of culture condition, Pranoprofen whose concentration was like to that has metabolized for 4 hours in aqueous humor (0.23 mg/L) can suppress
the confluence of lens epithelial cells on posterior capsule (P < 0.01).
3. Conclusion
(1) The posterior capsule in tissue culture shows many of the changes seen in vivo, including rapid cells growth and fibrosis, wrinkles, extracellul
目前的研究认为：PCO的形成是由于白内障囊外摘除术后血—房水屏障遭到破坏，导致外源性的细胞因子和炎症介质进入囊袋内，作用于前囊残留的晶状体上皮细胞上，使其发生成纤维细胞样改变，并向后囊不断增殖、迁徙，合成大量的胶原和细胞外基质，这些物质相互混杂，在后囊上形成膜状混浊物。该混浊物可引起后囊皱缩、变形，形成光散射，最终导致视力严重下降。
为了解PCO的形成机制，国外进行了多种体外模拟PCO的培养研究，为体外条件下观察PCO的形成过程和发生机制提供了优越的模型，而国内未见此方面的报道。此次实验我们欲进行牛晶状体后囊的组织培养，体外模拟PCO的形成，观察PCO形成过程中后囊晶状体上皮细胞的增殖分化规律以及血清对体外PCO模型的形成影响。普拉洛芬(Pranoprofen)属于非甾体类抗炎药物，它可明显改善白内障术后及眼前段非感染性炎症患者的主要症状和体征。但关于普拉洛芬对晶状体上皮细胞的抑制实验报道较少，因此，本次实验通过后囊的组织培养，进一步模拟普拉洛芬滴眼液滴眼后4小时在房水中的浓度，观察其对PCO的形成有无抑制作用。
郑州大学20()4届硕士学位论文
体外模拟后发障的组织培养及药物防治研究
一、实验方法
在无菌条件下，将分离的牛晶状体后囊细胞面朝上平铺于25ml培养瓶底
部，分别加入含O、10%、20%胎牛血清的DMEM培养液2.sml，将培养瓶竖直
放入37℃、5%coZ的培养箱中，培养液蒸发的水蒸汽可维持晶状体后囊表面湿
润，1小时后将培养瓶放平，使囊膜完全浸入培养液中，可见晶状体后囊已于
培养瓶的底部牢牢粘附，培养液不会使后囊卷曲、漂浮，再将培养瓶放入培养
箱中培养，每3天更换培养液1次，直到后囊晶状体上皮细胞达到融合。培养
的各阶段，在倒置显微镜下，用方格图计数板计数后囊晶状体上皮细胞的覆盖
率和融合时间，并用Giemsa染色和扫描电镜观察晶状体上皮细胞的形态。同时
在含0和2%胎牛血清的培养基中加入普拉洛芬，使其成为体内代谢4小时后
的房水内浓度(0.23m岁L)，进行体外培养，观察与对照组后囊上皮细胞融合
天数有无差异。
二、实验结果:
赤道部的晶状体上皮细胞向后囊中心部迅速增殖、迁徙，一般约需8一18
天可达到融合。细胞的增殖覆盖率与培养液中血清的含量呈正相关，融合时间
与血清含量呈负相关(P<0.05)。后囊晶状体上皮细胞发生成纤维细胞样改变，
并合成大量的细胞外基质后囊形成明显的混浊和皱缩。培养液中有无血清均可
出现后囊皱缩，且血清的含量越高，皱缩形成的数目越多，时间越短。对皱缩
区域的Giemsa染色和扫描电镜观察显示:各皱缩纹理排列几乎平行，皱缩边缘
有大量的晶状体上皮细胞聚集，且细胞的聚集方向与皱缩纹理的长轴平行。在
两种培养状态下，普拉洛芬在体内代谢4小时后的房水内浓度(0 23m留L)对
后囊上皮细胞的融合均有明显的抑制作用(P<0.01)。
三、讨论:
1.组织培养条件下的后囊能出现许多与体内PCO相似的改变，如晶状体
上皮细胞迅速地在后囊上增殖、纤维化，定向收缩使后囊形成皱缩，并分泌大
量的细胞外基质。它是体外模拟PCO形成和研究后发障发生机制的有效方法。
2.血清对体外PCO形成的影响说明:血一房水屏障破坏在PCO形成的
早期起到重要作用。后囊无血清组织培养结果显示:晶状体上皮细胞存在“自
分泌”机制，PCO的形成是由于“自分泌机制”和“旁分泌机制”共同作用的
郑州大学2004届硕士学位论文
体外模拟后发障的组织培养及药物防治研究
结果。
3.普拉洛芬是一种安全、有效的降低PCO发生率的药物，它可作为白内
障术后的常规用药。
In recent years, extracapsular cataract extraction (ECCE) and intraocular lens (IOL) implantation play the main role in cataract surgery. Posterior capsule opacification (PCO) , the major complication after ECCE, will affect the paients' visual acuity seriously. PCO occurs in up to 50% of paients between 1 month and 5 years after surgery., and occurs almost 100% in children.
Researchse showed that the formation of PCO is because breakdown of the blood-aqueous barrier after ECCE, which lead to growth factor and inflammatory medium enter capsular bag and act on residual lens epithelial cells on the anterior capsule.The fibroblast-like cells migrate and proliferate rapidly on posterior capsule and secrect a lot of collagen and extracellular matrix. Opaciflcation occurs as a result of the formation of fibrotic membranes on the posterior capsule. Opaciflcation will result in the formation of wrinkle and distortion, which lead to light scattering and visual acuity decrease seriously
In order to find out the mechanism of PCO formation, a lot of model for
PCO in vitro had been established overseas that provided a good example to observe the process and mechanism of PCO formation ,but the report is seldom in domestic. In this research, tissue culture of lens epithelial cells on the posterior capsule to model posterior capsule opacification (PCO) will be erected, to observe the proliferation of cells and the affection of serum on model for PCO in vitro.the inhibition effects of pranoprofen on PCO were observed. Pranoprofen is a non-steroidal anti-inflammatory drug.In this research, we model pranoprofen whose concentration was like to that has metabolized for 4 hours in aqueous humor (0.23 mg/L) and observe the inhibition of pranoprofen on PCO formation.
1. Methed
To spread out the bovine lens posterior capsule with the cell layer upwards on the surface of a 25ml culture flask in sterile condition. DMEM with 0 ,10% and 20% fetal calf serum 2.5ml used as medium were added respectively. The flask was set on ites end in 37 5%CO incubator. The rising water vapour protected the tissue from drying, forming a wet chamber. After 1 hour the flask was laid down,so that the medium covered the epithelium and continue to culture.The medium was changed every 3 days until the confulence of lens epithelial cells on posterior capsule. The cell coverage and confluence time on posterior capsule were recorded with graticule by inverted microscope, and the cell morphology was observed by Giemsa staining and scanning electron-microscope. The lens epithelial cells on posterior capsule were cultured in medium with pranoprofen whose concentration was like to that has metabolized for 4 hours in aqueous humor (0.23 mg/L), and then observed the difference of confluence time with control group.
2. Result
The lens epithelial cells migrate and proliferate rapidly on posterior capsule from equatorial region to the center. Within 8 to 18 days the cells became confluent. The cell coverage was rising with the increase of serum concentration, and the cell confluence time was shortening with that (P < 0.05). The lens epithelial cells on posterior capsule make a fibroblast-like change and secrect a lot of collagen and extracellular matrix. The PCO and wrinkles were formed obviously. The wrinkles presence were not dependent upon serum concentration, but increased in munber and decreased in time with that. The morphology of wrinkles by Giemsa staining and scanning electron-microscope show: the wrinkles almostly paralleled with each other., there were closely packed cells along that, and the direction of packed cells were alinged mainly at right angles to the wrinkles. In two kind of culture condition, Pranoprofen whose concentration was like to that has metabolized for 4 hours in aqueous humor (0.23 mg/L) can suppress
the confluence of lens epithelial cells on posterior capsule (P < 0.01).
3. Conclusion
(1) The posterior capsule in tissue culture shows many of the changes seen in vivo, including rapid cells growth and fibrosis, wrinkles, extracellul
引文
1. Schaumberg DA, Dana MR, Christan WG, Glynn RJ et al. A systermatic overview of the incidence of posterior capsule opacification. Ophthalmology. 1998 Jul;105(7):1213～19
2. Kappelhof JP. Vrensen GF Jr, et al. The pathology of aftercataract; a minireview. Acta Ophthmd. 1992;Suppl 205:13-34
3. Manfred R. Tetz, Christoph Nimsgern. Posterior capsule opacification part 2:clinical findings. J Catract Refract Surg. 1999;25:1662～1674
4. Wilson ME, Pandey SK, Wernerl, et al. Pediatric cataract surgery; current techniques, complications and management. In :Agarwal S. Agarwal A, Sachder MS, et al, eds, phzcoemulsification, Laser Cataract Surgery and foldable IOLs, 2nd, et al, eds, phacoemulsification, Laser Cataract Surgery and Foldable IOLs, znd ed. Thorofare, NJ, Slack Inc. 2000;369～88
5. Jared Emery. Capsule opacification after cataract surgery. Curr Opin Ophthalmol. 1999; 10:73～80
6. David J. Spalton. Poserior capsule opacification after cataract surgery. Eye. 1999; 13:498～492
7. Eckart Bertelmann, Corina Kojetinsky, et al. Poserior capsule opacification and anteior capsule opacification. Curr Opin Ophthalmol. 2001;12:35～40
8. Julia M. Marcantoino, Gijs. J. M. Vrensen. Cell biology of Poserior capsule opacification. Eye. 1999;13:484～488
9. Liu CSC, Wormstone IM, Duncan G, et al. A study of human lens cell grown in vitro ;a model for posterior capsule opacification. Invest Ophthalmol Vis Sci. 1996;37:906
10. Nagamoto T, Bissen-Miyajima G, et al. A ring to support the capsule bag after cintinuous curvilinear capsulorhexis. J Catract Refract Surg. 1994;20:417
11. Saxby L, Rosen E, Boulton M, et al. Lens epithelial cell proliferation,
migration, and metaplasia following capsulorhexis. Br J Ophrhalmol. 1998;82:945
12．李秋明．添加晶状体皮质提高晶状体上皮细胞原代培养成功率．眼科研究 1997；15：5～7
13．李秋明，陆道炎，王丽天，等．Giemsa染色比色法检测晶状体上皮细胞增殖．眼科研究1996；14：86～89
14．李秋明，孟繁剑．晶状体前囊膜的立体培养研究．眼科研究．2003；21：276
15．鄂征主编．组织培养技术．第二版．北京：人民卫生出版社．1998：127～203
16. Bermbach G, Mayer U, Maumann GOH, et al. Human lens epithelial cells in tissue culture. Exp Eye Res. 1991; 52:113
17. Takahiro Ogawa, Mineko Yamamoto, Akiko Ogasawara, et al. Intraocular penetration of pranoprofen, a non-steroidal anti-inflammatroy drug. Folia Ophthalmo Jpn. 1986;37:829-834
18. Eleanor A, Blakely, Kathleen A, et al. Growth and differentiation of human lens epithelial cells in vitro on matrix. Invest Ophthalmol Vis Sci. 2000;41:3898-3907
19. Ria Beck, Barbara Nebe, et al. Inhibition of lens epithelial cells adhesion by the calcium antagonist mibefradil correlates with impaired integrin distribution and organization of the cytoskeleton. Graefe's Arch Clin Exp Ophthalmol. 2001 ;239:452-458
20. Wallentin N, Lundgren B, et al Development of posterior capsule opacification in the rabbit. Ophthalmic Res. 2002;34:14-22
21. Kumi Shirai, Yuka Okada, Shizuya Saika, et al. Expression of transcription factor AP-1 in Rat lens epithelial cells during wound repair. Exp Eye Res. 2001; 73:461-468
22. Nishi Okihiro. Poserior capsule opacification part 1: experimental investigations. J Cataract Refract Surg. 1999:25;106-117
23. McDonnell PJ, Rowen SL, Glaser BM, Sato M, et al. Posterior capsule
opacification;an in vitro model. Arch Ophthalmol. 1985; 103:1378-1381
24. Marie-Jose B. R. Tassignon, Veva De Groot, et al. Bag-in-the-lens implantation of intraocular lenses. J Cataract Refract Surg. 2002;28:1182-1188
25. Meacock WR, Spalton DJ, Stanford MR, et al. Role of cytokines in the pathogenesis of posterior capsule opacification. Br J Ophthalmol. 2000 Mar;84(3):332
26. Gordon-Thomoson, C. de, IonghRU., Hales, A. M. Chamberlian, C. G. Mcavoy, J. W. et al. Differential cataractogenic potency of TGF-β 1, TGF-β2, and TGF-β3 and their expression in the post natal rat eye. Invest Ophthalmol Vis Sci. 1998;39:1399-1409
27. Chamberlian, C. G. Mcavoy, J. W. et al. Induction of lens fiber differentiation by acidic and basic fibroblast growth factor (FGF). Growth Factor. 1, 125-134
28. Pande, M. Spalton, D. J. Marshall, J. et al. Continuous curvilinear capsulorhexis and intraocular lens biocompatibility. J Cataract Refract Surg. 1996; 22:89-97
29．李秋明，陆道炎，王丽天，等．晶状体上皮细胞在体外的分化规律及血清和眼组织对其增殖的影响．中华眼科杂志 1997；33：139～42
30. Wormstone IM, Liu CSC, RakicJ-M, et al. Human lens epithelial cell proliferation in a protein-free medium. Invest Ophthalmolo Vis Sci. 1997;38:396-404
31. Wormstone I. M., Katia D. R. T., McMahon G., Tamiya Shigeo, et al. FGF: an autocrine regulator of human lens cell growth independent of added stimuli. Invest Ophthalmolo Vis Sci. 2001;42:1305
32. Wormstone I. M., Shigeo Tamiya, Julia M. Marcantonio, and John R. Reddan Hepatocyte, et al. Growth factor function and c-Met expression in human lens epithelial cells. Invest Ophthalmolo Vis Sci. 2000;41:4216
33. Wormstone I. M., Shigeo Tamiya, Anderson I., Duncan G, et al. TGF-β 2-
induced matrix modification and cell transdifferentiation in the human lens capsular bag. Invest Ophthalmolo Vis Sci. 2002;43:2301
34. Wormstone IM. Posterior capsule opacification: a cell biological perspective. Invest Ophthalmolo Vis Sci. 2002;74:337-347.
35. Kurosaka D, Kato K, Nagamoto T, Negishi K, et al. Growth Factors influence contractility and alpha-smooth muscle actin expression in bovine lens epithelial cells. Invest Ophthalmol Vis Sic. 1995 Jul;36(8):1701
36. Daijiro Kurosaka, Katsuhiko Kato, et al. Presence of a-smooth muscle actin expression in lens epithelial cells of aphakic rabbit eyes. Br J Ophthalmol. 1996; 80:906-909
37. Daijiro Kurosaka, Katsuhiko Kato, et al. Exreacellular matrixes influence a-smooth muscle actin expression in cultureed porcine lens epithelial cells. Current Eye Research. 1999; 19:260-263
38. Maruyanma Y, Teraswa M, Anami K, et al. Pharmacological studiea of 2 (5h (1) benzopyrano (2, 3b) pyeidin 7 yl) propionic acid (y 8004). Antiinflammatory, analgesic and antipyretic activities (Japanese). Folia Pharmacol Jpn. 1975, 71:857～62
39. Ogawa T, Ogasawara A., Yamamono Y, et al. Anti-inflammatory activity of pranoprofen ophthalmic solution on rat experimental conjunctivitis. Folia Pharmacol Jpn. 1982, 33:1244～50
40. Tobe T, Hagihara M, Tsuboi K, et al Effects of prostaglansins synthetase inhibitor for inflammation after intraocular lens implantation comparison between diclofenac sodium and pranoprofen eye drops. Jpn J Clin Ophthalmol. 1992;46:571～9
41. Ohashi K, Komatsu A, Omine N et al. Value of pranoprofen ophthalmic solution after early withdrawal of topical corticosteroid in uveitis. Jpn J Clin Ophthalmol. 1989, 43:1961～7
42. Ogawa T, Ogasawara A. et al. Effects of pranoprofen on rabbit uveitis. Journal of the Eye. 1988, 5:1334～41
43．赵家良，黎晓新．普拉洛芬滴眼液治疗白内障术后炎症和非感染性眼前段炎症的临床评价．眼科研究．2000：18：560～564
44．孟繁剑，李秋明．苦参碱、双氯芬酸钠、普拉洛芬对于晶状体上皮细胞增殖的影响．郑州大学2003届硕士学位论文(未发表)．
45. Nishi O, et al. Disruption of the blood-aqueous barrior by residual lens spithelial cells after intraocular lens implantation. Ophthalmic Surg. 1992;23:325
2. Kappelhof JP. Vrensen GF Jr, et al. The pathology of aftercataract; a minireview. Acta Ophthmd. 1992;Suppl 205:13-34
3. Manfred R. Tetz, Christoph Nimsgern. Posterior capsule opacification part 2:clinical findings. J Catract Refract Surg. 1999;25:1662～1674
4. Wilson ME, Pandey SK, Wernerl, et al. Pediatric cataract surgery; current techniques, complications and management. In :Agarwal S. Agarwal A, Sachder MS, et al, eds, phzcoemulsification, Laser Cataract Surgery and foldable IOLs, 2nd, et al, eds, phacoemulsification, Laser Cataract Surgery and Foldable IOLs, znd ed. Thorofare, NJ, Slack Inc. 2000;369～88
5. Jared Emery. Capsule opacification after cataract surgery. Curr Opin Ophthalmol. 1999; 10:73～80
6. David J. Spalton. Poserior capsule opacification after cataract surgery. Eye. 1999; 13:498～492
7. Eckart Bertelmann, Corina Kojetinsky, et al. Poserior capsule opacification and anteior capsule opacification. Curr Opin Ophthalmol. 2001;12:35～40
8. Julia M. Marcantoino, Gijs. J. M. Vrensen. Cell biology of Poserior capsule opacification. Eye. 1999;13:484～488
9. Liu CSC, Wormstone IM, Duncan G, et al. A study of human lens cell grown in vitro ;a model for posterior capsule opacification. Invest Ophthalmol Vis Sci. 1996;37:906
10. Nagamoto T, Bissen-Miyajima G, et al. A ring to support the capsule bag after cintinuous curvilinear capsulorhexis. J Catract Refract Surg. 1994;20:417
11. Saxby L, Rosen E, Boulton M, et al. Lens epithelial cell proliferation,
migration, and metaplasia following capsulorhexis. Br J Ophrhalmol. 1998;82:945
12．李秋明．添加晶状体皮质提高晶状体上皮细胞原代培养成功率．眼科研究 1997；15：5～7
13．李秋明，陆道炎，王丽天，等．Giemsa染色比色法检测晶状体上皮细胞增殖．眼科研究1996；14：86～89
14．李秋明，孟繁剑．晶状体前囊膜的立体培养研究．眼科研究．2003；21：276
15．鄂征主编．组织培养技术．第二版．北京：人民卫生出版社．1998：127～203
16. Bermbach G, Mayer U, Maumann GOH, et al. Human lens epithelial cells in tissue culture. Exp Eye Res. 1991; 52:113
17. Takahiro Ogawa, Mineko Yamamoto, Akiko Ogasawara, et al. Intraocular penetration of pranoprofen, a non-steroidal anti-inflammatroy drug. Folia Ophthalmo Jpn. 1986;37:829-834
18. Eleanor A, Blakely, Kathleen A, et al. Growth and differentiation of human lens epithelial cells in vitro on matrix. Invest Ophthalmol Vis Sci. 2000;41:3898-3907
19. Ria Beck, Barbara Nebe, et al. Inhibition of lens epithelial cells adhesion by the calcium antagonist mibefradil correlates with impaired integrin distribution and organization of the cytoskeleton. Graefe's Arch Clin Exp Ophthalmol. 2001 ;239:452-458
20. Wallentin N, Lundgren B, et al Development of posterior capsule opacification in the rabbit. Ophthalmic Res. 2002;34:14-22
21. Kumi Shirai, Yuka Okada, Shizuya Saika, et al. Expression of transcription factor AP-1 in Rat lens epithelial cells during wound repair. Exp Eye Res. 2001; 73:461-468
22. Nishi Okihiro. Poserior capsule opacification part 1: experimental investigations. J Cataract Refract Surg. 1999:25;106-117
23. McDonnell PJ, Rowen SL, Glaser BM, Sato M, et al. Posterior capsule
opacification;an in vitro model. Arch Ophthalmol. 1985; 103:1378-1381
24. Marie-Jose B. R. Tassignon, Veva De Groot, et al. Bag-in-the-lens implantation of intraocular lenses. J Cataract Refract Surg. 2002;28:1182-1188
25. Meacock WR, Spalton DJ, Stanford MR, et al. Role of cytokines in the pathogenesis of posterior capsule opacification. Br J Ophthalmol. 2000 Mar;84(3):332
26. Gordon-Thomoson, C. de, IonghRU., Hales, A. M. Chamberlian, C. G. Mcavoy, J. W. et al. Differential cataractogenic potency of TGF-β 1, TGF-β2, and TGF-β3 and their expression in the post natal rat eye. Invest Ophthalmol Vis Sci. 1998;39:1399-1409
27. Chamberlian, C. G. Mcavoy, J. W. et al. Induction of lens fiber differentiation by acidic and basic fibroblast growth factor (FGF). Growth Factor. 1, 125-134
28. Pande, M. Spalton, D. J. Marshall, J. et al. Continuous curvilinear capsulorhexis and intraocular lens biocompatibility. J Cataract Refract Surg. 1996; 22:89-97
29．李秋明，陆道炎，王丽天，等．晶状体上皮细胞在体外的分化规律及血清和眼组织对其增殖的影响．中华眼科杂志 1997；33：139～42
30. Wormstone IM, Liu CSC, RakicJ-M, et al. Human lens epithelial cell proliferation in a protein-free medium. Invest Ophthalmolo Vis Sci. 1997;38:396-404
31. Wormstone I. M., Katia D. R. T., McMahon G., Tamiya Shigeo, et al. FGF: an autocrine regulator of human lens cell growth independent of added stimuli. Invest Ophthalmolo Vis Sci. 2001;42:1305
32. Wormstone I. M., Shigeo Tamiya, Julia M. Marcantonio, and John R. Reddan Hepatocyte, et al. Growth factor function and c-Met expression in human lens epithelial cells. Invest Ophthalmolo Vis Sci. 2000;41:4216
33. Wormstone I. M., Shigeo Tamiya, Anderson I., Duncan G, et al. TGF-β 2-
induced matrix modification and cell transdifferentiation in the human lens capsular bag. Invest Ophthalmolo Vis Sci. 2002;43:2301
34. Wormstone IM. Posterior capsule opacification: a cell biological perspective. Invest Ophthalmolo Vis Sci. 2002;74:337-347.
35. Kurosaka D, Kato K, Nagamoto T, Negishi K, et al. Growth Factors influence contractility and alpha-smooth muscle actin expression in bovine lens epithelial cells. Invest Ophthalmol Vis Sic. 1995 Jul;36(8):1701
36. Daijiro Kurosaka, Katsuhiko Kato, et al. Presence of a-smooth muscle actin expression in lens epithelial cells of aphakic rabbit eyes. Br J Ophthalmol. 1996; 80:906-909
37. Daijiro Kurosaka, Katsuhiko Kato, et al. Exreacellular matrixes influence a-smooth muscle actin expression in cultureed porcine lens epithelial cells. Current Eye Research. 1999; 19:260-263
38. Maruyanma Y, Teraswa M, Anami K, et al. Pharmacological studiea of 2 (5h (1) benzopyrano (2, 3b) pyeidin 7 yl) propionic acid (y 8004). Antiinflammatory, analgesic and antipyretic activities (Japanese). Folia Pharmacol Jpn. 1975, 71:857～62
39. Ogawa T, Ogasawara A., Yamamono Y, et al. Anti-inflammatory activity of pranoprofen ophthalmic solution on rat experimental conjunctivitis. Folia Pharmacol Jpn. 1982, 33:1244～50
40. Tobe T, Hagihara M, Tsuboi K, et al Effects of prostaglansins synthetase inhibitor for inflammation after intraocular lens implantation comparison between diclofenac sodium and pranoprofen eye drops. Jpn J Clin Ophthalmol. 1992;46:571～9
41. Ohashi K, Komatsu A, Omine N et al. Value of pranoprofen ophthalmic solution after early withdrawal of topical corticosteroid in uveitis. Jpn J Clin Ophthalmol. 1989, 43:1961～7
42. Ogawa T, Ogasawara A. et al. Effects of pranoprofen on rabbit uveitis. Journal of the Eye. 1988, 5:1334～41
43．赵家良，黎晓新．普拉洛芬滴眼液治疗白内障术后炎症和非感染性眼前段炎症的临床评价．眼科研究．2000：18：560～564
44．孟繁剑，李秋明．苦参碱、双氯芬酸钠、普拉洛芬对于晶状体上皮细胞增殖的影响．郑州大学2003届硕士学位论文(未发表)．
45. Nishi O, et al. Disruption of the blood-aqueous barrior by residual lens spithelial cells after intraocular lens implantation. Ophthalmic Surg. 1992;23:325