期刊文献+

紫外光-核黄素诱导羊膜胶原交联的实验研究 被引量:1

Crosslinking of human amniotic membrane by ultraviolet Ariboflavin
下载PDF
导出
摘要 背景研究证实,紫外光一核黄素光动力疗法可通过改变角膜的理化性质控制圆锥角膜的进展,羊膜移植已广泛用于眼表疾病,羊膜胶原成分与角膜相似,目前关于紫外光一核黄素诱导的胶原交联用于羊膜组织的研究尚少见。目的探讨紫外光一核黄素对冻存羊膜的作用。方法在知情同意条件下,获取并常规处理人羊膜组织,-80℃保存。制备成2mm×15mm大小,用简单随机法中的抽签法将其分为4个组,每组6片羊膜,前3组均给予紫外光一核黄素(质量分数0.1%)处理30rain(波长为370nm,紫外光功率分别为1、2、3mW/cm^2,光源距羊膜10mm),第4组不做任何处理,为对照组。利用微力材料试验机对处理后的各组羊膜分别进行生物力学测量,记录羊膜长度改变5%、10%、15%时各组样本所需的拉力值(mN)。用环钻钻取直径7mm的羊膜,用抽签法将其分为4个组,每组5片羊膜,各组处理情况同上,将处理后的各组羊膜浸于质量分数0.1%胶原蛋白酶I溶液中进行消化,对羊膜透明度进行评分,并记录羊膜完全溶解时间。关于羊膜的组织学观察,用抽签法将羊膜分为紫外光一核黄素组、核黄素组及对照组,每组3片羊膜,各组分别给予紫外光一核黄素(0.1%)处理30rain(波长为370nm,紫外光功率为3mW/cm^2,光源距羊膜10mm)、0.1%核黄素浸泡30min和生理盐水浸泡30min,透射电子显微镜下进行观察。结果羊膜长度改变5%、10%、15%时,对照组和1、2、3roW/era。紫外光组4个组所需拉力值差异均有统计学意义(F:3.411,P=0.037;F=9.927,P=0.001;F=11.118,P=0.000);交联后抗拉力性能明显增强,且紫外光功率越大,羊膜的抗拉性增加越明显,差异均有统计学意义(P〈0.05)。对照组羊膜完全酶解时间为(8.6±1.8)h,1、2、3mW/cm。紫外光组分别为(39.6±2.3)、(71.4±0.9)、(78.8±1.8)h,交联后抗酶解能力增强,且紫外光功率越大,羊膜的抗酶解能力增强越明显,差异有统计学意义(P〈0.01)。透射电子显微镜超微结构观察,交联后羊膜基质层胶原纤维密度增加、胶原间连接增多、胶原与上皮问的连接加强。结论紫外光-核黄素可使冻存羊膜发生胶原交联,从而使其组织结构发生变化,生物力学性质改变,抗酶解能力加强。 Background Studies confirmed that ultraviolet A (UVA)riboflavin photodynamic therapy can control keratoconus progresses by altering the physicochemical property of cornea. The collagen components of amniotic membrane transplantation is similar to that of cornea and amniotic membrane transplantation has been widely used to ocular surface reconstruction. However,the study on UVA riboflavin-induced-collagen crosslinking for amniotie tissue is less now. Objective This study was to investigate the role of UVAriboflavin on frozen-preserved human amniotie membrane. Methods Human amnions were obtained in informed consent and prepared into 2 mm×l5 mm pieces and were then divided into 4 groups using lottery method and 6 pieces for each group. The first 3 groups were treated with the photosensitizer riboflavin and UVA-irradiation ( wavelength :370 nm ; irradiation energy : 1,2 or 3 mW/cm^2 , distance :10 mm) for 30 minutes,and the untreated fourth group was as control group. Biomechanica] stressstrain test was performed using a microcomputercontrolled biomateria] tester and the stress(mN) was recorded when the strains were set to 5% , 10% and 15% . The 7 mm diameter of human amniotie membrane pieces were trephined and divided into 4 groups(5 pieces for each group) with the treated method as mentioned above, and then the buttons were exposed to 0. 1% collagenase I solution. The transparency was scored and the complete dissolving time was record. In histological evaluation, three groups (3 pieces for each group) of human amniotie membranes were treated using UVA-riboflavin( 3 mW/em2), 0. 1% riboflavin, normal saline for 30 minutes respectively and examined under the transmission electron microscopy. This study was performed under the permission of the Ethic Commission of Beijing Tongren Hospital. Results When the strain was 5% , 10% , 15% , the stress of control group and 1 , 2, 3 mW/cm2UVA group were statistically signifeantly different ( F = 3. 411 , P = 0. 037; F = 9. 927, P = 0. 001 ; F= 11. 118,P=0. 000). The tensile strength of human amniotic membrane cross-linked with UVA-riboflavin was statistically significantly increased in comparison to the control group( P〈0.05 ), and the tensile strength of human amniotie membrane became stronger as UVA power increased. The complete dissolve time was(8.6±1.8 ) hours for the control group, ( 39.6±2.3 ) hours for 1 mW/cm2 UVA group, ( 71.4± 0.9 ) hours for 2 mW/cm2 UVA group, (78.8± 1.8 ) hours for 3 mW/cm2 UVA group, showing the enhanced antienzyme ability of human amniotic membrane after cross-linking( P〈0.01 ). The collagen density in the UVAriboflavin treated group was increased,the connection among the collagen fibers as well as between the stroma and the epithelium became tighter than those of control group. Conclusions Collagen crosslinking with UVAriboflavin make the biomechanical strength and enzymatic resistance of human amniotic membrane enhance and ultrastructure change of human amniotic membrane.
出处 《中华实验眼科杂志》 CAS CSCD 北大核心 2012年第2期127-131,共5页 Chinese Journal Of Experimental Ophthalmology
基金 北京市自然科学基金资助项目(7112023)
关键词 羊膜 紫外光 核黄素 胶原交联 Amniotic membrane Ultraviolet A Riboflavin Collagen crosslinking
  • 相关文献

参考文献19

  • 1Kim JC,Tseng SC.Transplantation of preserved human amniotic membrance for surface reconstruction in severely damaged rabbit corneas[J].Cornea,1995,14:473-484.
  • 2Lee JH,Wee WE,Chung ES,et al.Development of a newly designed double-fixed Seoul-type keratoprosthesis[J].Arch Ophthalmol,2000,1 18:1673-1678.
  • 3潘红卫,陈建苏.紫外光核黄素交联治疗圆锥角膜的研究进展[J].眼科研究,2008,26(5):397-400. 被引量:14
  • 4Wollensak G,Aurich H,Pham DT,et al.Hydration behaviour of porcine cornea crosslinked with riboflavin and ultraviolet A[J].J Cataract Refract Surg,2007,33:516-521.
  • 5Spoerl E,Wollensak G,Dittert DD,et al.Thermomechanical behavior of collagen-cross-linked porcine cornea[J].Ophthalmologica,2004,218:136-140.
  • 6Kohlhaas M,Spoerl E,Schilde T,et al.Biomechanical evidence of the distribution of cross-links in corneas treated with riboflavin and ultraviolet A light[J].J Cataract Refract Surg,2006,32:279-283.
  • 7Spoerl E,Wollensak G,Seiler T.Increased resistance of crosslinked cornea against enzymatic digestion[J].Curr Eye Res,2004,29:35-40.
  • 8Wollensak G,Wilsch M,Spoerl E,et al.Collagen fiber diameter in the rabbit cornea after collagen cross-linking by riboflavin/UVA[J].Cornea,2004,23:503-507.
  • 9Wollensak G,Spoerl E,Wilsch M,et al.Keratocyte apoptosis after corneal collagen cross-linking using riboflavin/UVA treatment[J].Cornea,2004,23:43-49.
  • 10Wollensak G,Spoerl E,Wilsch M,et al.Endothelial cell damage after riboflavin-ultraviolet A treatment in the rabbit[J].J Cataract Refract Surg,2003,29:1786-1790.

二级参考文献24

  • 1Colin J, Cochener B, Savary G, et al. Correcting keratoconus with intracorneal rings [J]. J Cataract Refract Surg,2000,26 ( 8 ) : 1117 - 1122.
  • 2Wollensak G,Spoerl E, Seiler T. Riboflavin/ultraviolet-a-induced collagen crosslinking for the treatment of keratoconus [ J ]. Am J Ophthalmol, 2003,135 (5) : 620 - 627.
  • 3Spoerl E, Mrochen M, Sliney D, et al. Safety of UVA-riboflavin crosslinking of the cornea [ J ]. Cornea,2007,26 (4) : 385 - 389.
  • 4Wollensak G,Iomdina E, Dittert DD, et al. Wound healing in the rabbit cornea after corneal collagen cross-linking with riboflavin and UVA [ J ]. Cornea,2007,26 ( 5 ) : 600 - 605.
  • 5Kohlhaas M, Spoerl E, Schilde T, et al. Biomechanical evidence of the distribution of cross-links in corneastreated with riboflavin and ultraviolet A light [ J ]. J Cataract Refract Surg, 2006,32 ( 2 ) : 279 - 283.
  • 6Wollensak G, Spoerl E, Seiler T. Stress-strain measurements of human and porcine corneas after riboflavin-ultraviolet-A-induced cross-linking [ J]. J Cataract Refract Surg,2003,29(9) : 1780 - 1785.
  • 7Wollensak G, Wilsch M, Spoed E, et aL Collagen fiber diameter in the rabbit cornea after collagen crosslinking by riboflavin/UVA [J]. Cornea, 2004,23 ( 5 ) : 503 - 507.
  • 8Wollensak G,Aurich H, Pham DT, et al. Hydration behavior of porcine cornea crosslinked with riboflavin and ultraviolet A[ J]. J Cataract Refract Surg,2007,33 (3) : 516 - 521.
  • 9Spoerl E, Wollensak G, Seller T. Increased resistance of crosslinked cornea against enzymatic digestion[ J]. Curr Eye Res ,2004,29( 1 ) : 35 -40.
  • 10Wollensak G, Spoerl E, Wilsch M, et al. Keratocyte apoptosis after corneal collagen cross-linking using riboflavin/UVA treatment[J]. Cornea,2004, 23 (1):43-49.

共引文献13

同被引文献35

  • 1Kok YO,Tan GF,Loon SC.Review:keratoconus in Asia[J].Cornea,2012,31:581-593.
  • 2Spoerl E,Seiler T.Techniques for stiffening the cornea[J].J Refract Surg,1999,15:711-713.
  • 3Knox Cartwright NE,Tyrer JR,Marshall J.In vitro quantification of the stiffening effect of corneal cross-linking in the human cornea using radial shearing speckle pattern interferometry[J].J Refract Surg,2012,28:503-508.
  • 4Poli M,Cornut PL,Balmitgere T,et al.Prospective study of corneal collagen cross-linking efficacy and tolerance in the treatment of keratoconus and corneal ectasia:3-year results[J].Cornea,2013,32:583-590.
  • 5Goldich Y,Marcovich AL,Barkana Y,et al.Clinical and corneal biomechanical changes after collagen cross-linking with riboflavin and UV irradiation in patients with progressive keratoconus:results after 2 years of follow-up[J].Cornea,2012,31:609-614.
  • 6Henriquez MA,Izquierdo L Jr,Bernilla C,et al.Riboflavin/ Ultraviolet A corneal collagen cross-linking for the treatment of keratoconus:visual outcomes and Scheimpflug analysis[J].Cornea,2011,30:281-286.
  • 7Kolli S,Aslanides IM.Safety and efficacy of collagen crosslinking for the treatment of keratoconus[J].Expert Opin Drug Saf,2010,9:949-957.
  • 8Gkika M,Labiris G,Kozobolis V.Corneal collagen cross-linking using riboflavin and ultraviolet-A irradiation:a review of clinical and experimental studies[J].Int Ophthalmol,2011,31:309-319.
  • 9Spoerl E,Hoyer A,Pillunat LE,et al.Corneal cross-linking and safety issues[J].Open Ophthalmol J,2011,5:14-16.
  • 10Schumacher S,Oeftiger L,Mrochen M.Equivalence of biomechanical changes induced by rapid and standard corneal cross-linking,using riboflavin and ultraviolet radiation[J].Invest Ophthalmol Vis Sci,2011,52:9048-9052.

引证文献1

二级引证文献7

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部