兔眼微脉冲激光小梁成形术的实验研究
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摘要
目的：
1．通过动物实验了解微脉冲激光小梁成形术术后兔眼小梁网的组织病理和超微结构的变化，评价该激光的安全性和有效性。
2．通过比较小梁网的基质金属蛋白酶及其组织抑制剂的免疫组织化学的表达，探讨微脉冲激光小梁成形术对小梁网的作用，分析其可能的降眼压机制。
方法：
52只健康灰色家兔随机分为实验组32只，正常对照组20只。实验组动物于右眼激光后随机分为3天组、10天组、30天组、60天组，每组8只动物，右眼纳入治疗组，左眼纳入自身对照组，正常对照组动随机分成0天组、3天组、10天组、30天组、60天组，每组8眼。于术后1天、3天、7天、10天、30天、45天、60天行裂隙灯、眼底镜、眼压检查。术前及术后3天、10天、30天、60天行前房角镜，UBM、共聚焦显微镜、组织病理、超微结构和免疫组化观察。
结果：
1．治疗组眼压变化趋势与自身对照组和正常对照组之间有显著性差异，治疗组术后眼压低于自身对照组和正常对照组，结果有显著性差异，自身对照组和正常对照组之间眼压无显著性差异。
2．实验过程中治疗眼与对照眼裂隙灯、眼底检查、UBM检查、房角开放距离、角膜内皮细胞计数、内皮细胞形态无显著性差异。
3．治疗眼小梁网变化轻微，局部小梁板变薄。超微结构观察见胶原纤维断裂，包膜缺失，研究结束时有所恢复。
4．治疗眼MMP-2和TIMP-2增高，MMP-2增高幅度大于TIMP-2。
结论：
1．微脉冲激光小梁成形术采用的红外810nm激光，微脉冲模式，组织损伤轻微，术后前节安静，角膜上皮、内皮、房角开放距离无变化。与对照组相比，能够使眼压降低。
2．微脉冲激光小梁成形术超微结构观察部分小梁网胶原纤维断裂，包膜缺失，随着术后时间延长，小梁网重新恢复规整，胶原纤维包膜恢复，纤维束变细，小梁网眼变大，网眼内碎屑和红细胞减少。激光后MMP-2和TIMP-2表达均增高，但MMP-2增高幅度明显高于TIMP-2，可能与MDLT的降眼压机制有关。
3．微脉冲激光小梁成形术损伤小、无疼痛、固视配合好，激光参数的选择是治疗的关键。
Objective:
1. To observe histopathologic and ultrastructure of trabecular meshworkin rabbit after micropulse diode laser trabeculoplasty and to assessthe safety and variation of it.
2. The expression of MMP-2 and TIMP-2 in. trabecular meshwork wascompared before and after micropulse diode laser trabeculopiasty ,and theproper mechanisms of MDLT was analyzed.
Methods:
The rabbits were randomly divided into laser group and control group, lasergroup (32 rabbits) were divided into four groups according to the observeperiod ,the right eyes were performed MDLT. Control group(20 rabbits) weredivided into five groups according to the observe period. Rabbits wereobserved IOP, anterior chamber, corneal endothelisl cell density,histopathologic and ultrastructure of trabecular meshwork.
Results:
1. IOP at 3day,10day,30day,60day following laser treatment were remarkablelower than the control.
2. The changes of anterior chamber, anterior chamber angle open distance andcorneal endothelial cell density between each group could not bedetected.
3. The trabecular meshwork of Laser group was thinner and sometrabecular beam was cut , some lost its collagenous cover afterlaser, its could be recovered .
4. Expression of MMP-2 and TIMP-2 were increased in the laser group ,andexpression of MMP-2 was respectively higher than expression of TIMP-2.
Conclusions:
1. MDLT was found to be effective to lower IOP in rabbits and safetyto the eyes.
2. The holes of trabecular meshwork were enlarged, some trabecular beam was thinner after MDLT, the expression of MMP-2 wasrespectively higher than expression of TIMP-2. These all may be concernedwith the mechanism of MDLT.
3. MDLT was based upon sub-threshold level ,to choose the correct parameterswere very important.
1．通过动物实验了解微脉冲激光小梁成形术术后兔眼小梁网的组织病理和超微结构的变化，评价该激光的安全性和有效性。
2．通过比较小梁网的基质金属蛋白酶及其组织抑制剂的免疫组织化学的表达，探讨微脉冲激光小梁成形术对小梁网的作用，分析其可能的降眼压机制。
方法：
52只健康灰色家兔随机分为实验组32只，正常对照组20只。实验组动物于右眼激光后随机分为3天组、10天组、30天组、60天组，每组8只动物，右眼纳入治疗组，左眼纳入自身对照组，正常对照组动随机分成0天组、3天组、10天组、30天组、60天组，每组8眼。于术后1天、3天、7天、10天、30天、45天、60天行裂隙灯、眼底镜、眼压检查。术前及术后3天、10天、30天、60天行前房角镜，UBM、共聚焦显微镜、组织病理、超微结构和免疫组化观察。
结果：
1．治疗组眼压变化趋势与自身对照组和正常对照组之间有显著性差异，治疗组术后眼压低于自身对照组和正常对照组，结果有显著性差异，自身对照组和正常对照组之间眼压无显著性差异。
2．实验过程中治疗眼与对照眼裂隙灯、眼底检查、UBM检查、房角开放距离、角膜内皮细胞计数、内皮细胞形态无显著性差异。
3．治疗眼小梁网变化轻微，局部小梁板变薄。超微结构观察见胶原纤维断裂，包膜缺失，研究结束时有所恢复。
4．治疗眼MMP-2和TIMP-2增高，MMP-2增高幅度大于TIMP-2。
结论：
1．微脉冲激光小梁成形术采用的红外810nm激光，微脉冲模式，组织损伤轻微，术后前节安静，角膜上皮、内皮、房角开放距离无变化。与对照组相比，能够使眼压降低。
2．微脉冲激光小梁成形术超微结构观察部分小梁网胶原纤维断裂，包膜缺失，随着术后时间延长，小梁网重新恢复规整，胶原纤维包膜恢复，纤维束变细，小梁网眼变大，网眼内碎屑和红细胞减少。激光后MMP-2和TIMP-2表达均增高，但MMP-2增高幅度明显高于TIMP-2，可能与MDLT的降眼压机制有关。
3．微脉冲激光小梁成形术损伤小、无疼痛、固视配合好，激光参数的选择是治疗的关键。
Objective:
1. To observe histopathologic and ultrastructure of trabecular meshworkin rabbit after micropulse diode laser trabeculoplasty and to assessthe safety and variation of it.
2. The expression of MMP-2 and TIMP-2 in. trabecular meshwork wascompared before and after micropulse diode laser trabeculopiasty ,and theproper mechanisms of MDLT was analyzed.
Methods:
The rabbits were randomly divided into laser group and control group, lasergroup (32 rabbits) were divided into four groups according to the observeperiod ,the right eyes were performed MDLT. Control group(20 rabbits) weredivided into five groups according to the observe period. Rabbits wereobserved IOP, anterior chamber, corneal endothelisl cell density,histopathologic and ultrastructure of trabecular meshwork.
Results:
1. IOP at 3day,10day,30day,60day following laser treatment were remarkablelower than the control.
2. The changes of anterior chamber, anterior chamber angle open distance andcorneal endothelial cell density between each group could not bedetected.
3. The trabecular meshwork of Laser group was thinner and sometrabecular beam was cut , some lost its collagenous cover afterlaser, its could be recovered .
4. Expression of MMP-2 and TIMP-2 were increased in the laser group ,andexpression of MMP-2 was respectively higher than expression of TIMP-2.
Conclusions:
1. MDLT was found to be effective to lower IOP in rabbits and safetyto the eyes.
2. The holes of trabecular meshwork were enlarged, some trabecular beam was thinner after MDLT, the expression of MMP-2 wasrespectively higher than expression of TIMP-2. These all may be concernedwith the mechanism of MDLT.
3. MDLT was based upon sub-threshold level ,to choose the correct parameterswere very important.
引文
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[2] 病理学总论 武忠弼主编．第1版，北京：人民卫生出版社，1989：51
[3] Richard F. Targeting outflow facility in glaucoma management [J] Surv Ophthalmol 2003; 48(suppl1): 17～20
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[19] Parshley DE, Bradley JM et al. Early changes in matrix metalLoproteinases and inhibitors after in vitro laser treatment to the trabecular meshwork. [J] Cur Eye Res 1995; 14(6): 537～544
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[1] Finger PT, Packer S, Paglione RW, et al Thermoradiotherapy of choroidal melanoma[J]. Ophthalm ology, 1989, 96(9): 1384—1388.
[2] Pearson AR, Tanner V, Krightley SJ, Casswell AG What efect does laser photocoagulation have on driving visulfields in diabetics? Eye 1998; 12: 64-8
[3] Ulbig M, Arden G, Hamilton P. Color contrast sensitivity and pattern electroretinographic findings after diode and argon laser photocoagulation in diabetic retinopathy. Am J Ophthalmol, 1994; 117: 583-8
[4] Early treatment diabetic retinopathy study group. Photocoagulation for diabetic macular edema[J]. Int Ophthalmol Clinic, 1987, 27(4): 265—272
[5] Chong LP, Soriano D, Ramos AR. Sublethal laser damage to the retinal pigment epithelium by micro-pulse diode laser in primate eye (abstract). Invest Ophthalmol Vis Sci, 1996; 37: 5694
[6] Boutacoff TA, Buzawa D, Dorm G, Kelsoe W. Diode Iaser Micropulse minimal impact photocoagulation(DLMP)(unpublished data) . Iris Medical instruments. 1997
[7] Chong LP, Kohen L, et al. Selective RPE Damage by MicroPulse Diode Laser Photocoagulation. [J] Invest Ophthalmol Vis Sei,1992, 33(4): 722
[8] FribergTR .VlenketeshS . Alteration of pulse configuration afects the pain response during diode laser photocoagulation . [J] Lasers Surg Med 1995;16 : 3803
[9] Finger PT, Packer S, et al. Thermoradiotherapy of choroidal melanoma. [J] Ophthalmologr, 1989;96(9): 1384～1388.
[10] Early treatment diabetic retinopathy study group. Photocoagulation for diabetic macular edema. [J] Int Ophthalmol Clinic, 1987, 27(4):265～272.
[11] Kim SY1, Se nislo SR1 et al The Selective Effect of Micropulse Diode Laser Upon the Retina. [J] Invest Ophthalmol Vis Sci, 1996,37(3): 779
[12]Roider J, Michaud NA et al. Response of the retinal pigment epithelium to selective photocoagulation . [J] Arch Ophthalmol, 1992;110(12):1786—1792.
[13] Roider J, Michaud NA Histology of retinal lesions after continuous irradiation and selective micro-coagulation of the retinal pigment epithelium. [J]. Ophthalmologe, 1993;90(3): 274—8.
[14] Richardson PR, Boulton ME, et al. Immunocyto chemical study of retinal diode laser pbotocoagulation in the rat. [J] Br J Ophthalmol, 1996;80(12): 1092—1098.
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[18] Akduman L, 01k RJ. Subthreshold(invisible) modified grid diode laser photocoagulation in diffuse diabetic macular edema(DDME ). [J] Ophthalmic surg lasers , 1999;30(9): 705
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[20]Framme C,Brinkmann R, et al. Antofluorescence imagmg after selective RPE laser treatment in macular diseases and clinical outcome: a pilot study. [J]. Br J ophthalmol 2002, 86(10): 1099-1106
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[23] Friberg TR, Karatza EC. The treatment of macular disease using a micropulsed and continuous wave 810-nm diode, laser [J] Ophthalmology,1997,104(12): 2030—2038
[24] ulbig MW, Hamilton AM . comparative used of diode and argon laser for panretinal photocoagulation in diabetic retinopathy[J]. Ophthalmologe, 1993;90(5): 457—462.
[25] Akduman L, 01k RJ. Diode laser(810 nm) versus argon green (514 nm) modified grid photocoagulation for diffuse diabetic macular edema . [J] Ophthalmology;1997, 104(9): 1433～1441
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[1] Finger PT, Packer S, Paglione RW, et al Thermoradiotherapy of choroidal melanoma[J]. Ophthalm ology, 1989, 96(9): 1384—1388.
[2] Pearson AR, Tanner V, Krightley SJ, Casswell AG What efect does laser photocoagulation have on driving visulfields in diabetics? Eye 1998; 12: 64-8
[3] Ulbig M, Arden G, Hamilton P. Color contrast sensitivity and pattern electroretinographic findings after diode and argon laser photocoagulation in diabetic retinopathy. Am J Ophthalmol, 1994; 117: 583-8
[4] Early treatment diabetic retinopathy study group. Photocoagulation for diabetic macular edema[J]. Int Ophthalmol Clinic, 1987, 27(4): 265—272
[5] Chong LP, Soriano D, Ramos AR. Sublethal laser damage to the retinal pigment epithelium by micro-pulse diode laser in primate eye (abstract). Invest Ophthalmol Vis Sci, 1996; 37: 5694
[6] Boutacoff TA, Buzawa D, Dorm G, Kelsoe W. Diode Iaser Micropulse minimal impact photocoagulation(DLMP)(unpublished data) . Iris Medical instruments. 1997
[7] Chong LP, Kohen L, et al. Selective RPE Damage by MicroPulse Diode Laser Photocoagulation. [J] Invest Ophthalmol Vis Sei,1992, 33(4): 722
[8] FribergTR .VlenketeshS . Alteration of pulse configuration afects the pain response during diode laser photocoagulation . [J] Lasers Surg Med 1995;16 : 3803
[9] Finger PT, Packer S, et al. Thermoradiotherapy of choroidal melanoma. [J] Ophthalmologr, 1989;96(9): 1384～1388.
[10] Early treatment diabetic retinopathy study group. Photocoagulation for diabetic macular edema. [J] Int Ophthalmol Clinic, 1987, 27(4):265～272.
[11] Kim SY1, Se nislo SR1 et al The Selective Effect of Micropulse Diode Laser Upon the Retina. [J] Invest Ophthalmol Vis Sci, 1996,37(3): 779
[12]Roider J, Michaud NA et al. Response of the retinal pigment epithelium to selective photocoagulation . [J] Arch Ophthalmol, 1992;110(12):1786—1792.
[13] Roider J, Michaud NA Histology of retinal lesions after continuous irradiation and selective micro-coagulation of the retinal pigment epithelium. [J]. Ophthalmologe, 1993;90(3): 274—8.
[14] Richardson PR, Boulton ME, et al. Immunocyto chemical study of retinal diode laser pbotocoagulation in the rat. [J] Br J Ophthalmol, 1996;80(12): 1092—1098.
[15] Pollack JS, Kim JE, et al, Tissue effects of subclinical diode laser treatment of the retina. [J] Arch Ophthalmol, 1998; 116(12): 1633—1639
[16] Funatsu H, Wilson CA, et al Comparative study of the effects of argon and diode laser photocoagulation on retinal oxygenation. [J]. Graefes Arch Clin Exp Ophthalmol, 1997;235(3): 168—175
[17]KimSY, SanisloSR, etal. The selective effect of micmpulse diode laser upon the retina. [J] Invest ophthalmol Vis Sci, 1996;37(3):s779.
[18] Akduman L, 01k RJ. Subthreshold(invisible) modified grid diode laser photocoagulation in diffuse diabetic macular edema(DDME ). [J] Ophthalmic surg lasers , 1999;30(9): 705
[19] Roider J, Brinkmann R, et al. Subthreshold(retinal pigment epithelium )photocoagulatian in macular disease: a pilot study.[J]. Br J ophthalmol 2000;84(1): 40—47
[20]Framme C,Brinkmann R, et al. Antofluorescence imagmg after selective RPE laser treatment in macular diseases and clinical outcome: a pilot study. [J]. Br J ophthalmol 2002, 86(10): 1099-1106
[21] Early Treatment Diabetic Retinopathy Study Research Group. Early photocoagulation for diabetic retinopathy . ETDRS report 9. [J] Ophthalmology 1991: 98 : 757-85
[22] Moorman CM Hamilton AM Clinical Applications of the MicroPulse Dioder laser. [J] Eye, 1999; 13:145— 150
[23] Friberg TR, Karatza EC. The treatment of macular disease using a micropulsed and continuous wave 810-nm diode, laser [J] Ophthalmology,1997,104(12): 2030—2038
[24] ulbig MW, Hamilton AM . comparative used of diode and argon laser for panretinal photocoagulation in diabetic retinopathy[J]. Ophthalmologe, 1993;90(5): 457—462.
[25] Akduman L, 01k RJ. Diode laser(810 nm) versus argon green (514 nm) modified grid photocoagulation for diffuse diabetic macular edema . [J] Ophthalmology;1997, 104(9): 1433～1441
[26] Sinclair SH, Alaniz R, et al Laser treatment Of diabetic macular edema :comparison of ETDRS-level treatment with threshold-level treatment by using high-contrast discriminat central visual field testing. [J]. Semin Ophthalmol, 1999;14(4):214-222.
[27] Rodanant N, Friberg TR, et al. Predictors of drusen reduction after subthreshold infrared (810nm) diode laser macular grid photocoagulation for nonexudative age-related macular degeneration.[J]Am J Ophthalmol 2002; 134(4):577—585.
[28]01k RJ, Friberg TR etal. Therapeutic benefits of infrared (810nm) diode laser macular grid photocoagulation in prophylactic treatment of nonexudative age-related macular degeneration:two years result of a randomized pilot study. [J] Ophthalmology, 2000;107(6): 1019—1021.
[29] Roider J, Wirbelauer C, et al. Variability of RPE reaction in two cases after selective RPE laser effects in prophylactic treatmeng of drusen. [J] Graefe' s Arch Clin Exp Ophthalmol. 1999;237(1): 45～50
[30] Youn J, Cox TA, et al. A Clinical comparison of Transscleral Cyclophotocoagulation with Neodymium: YAG and Semiconductor Diode Lasers. Am J Ophthalmol. 1998; 126: 640～7