深板层内皮角膜移植术治疗大泡性角膜病变的实验研究
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摘要
目的
1.探讨一种稳定、持久的家兔角膜内皮损伤方法,从而建立适合评价深板层内皮角膜移植术(DLEK)治疗大泡性角膜病变(BK)的模型。
2.自行设计DLEK手术相关的器械,对家兔BK模型行经角膜-巩膜袋法的DLEK手术治疗,探讨手术技术方法,对临床效果进行评价。
3.观察小切口DLEK手术术后角膜地形图变化。方法
1.24只成年家兔,随机分为两组,每组12只,均行右眼手术。一组经角膜缘内1mm用30gauge空气注射套管针自穿刺口进入前房刮除全部后弹力层(DM)和内皮细胞,另一组单纯刮除内皮细胞。术后采用裂隙灯及活体生物显微镜观察角膜水肿混浊程度和角膜厚度,以角膜内皮镜行角膜内皮细胞计数,定期取标本行角膜内皮细胞染色及组织学检查。
2.32只右眼建立BK模型的家兔,随机分为两组,每组16只眼。一组经上方角膜缘切口制作板层角膜袋,采用自行设计的扁平环钻钻除一直径7.0mm角膜后板层组织,置换一直径7.25mm的带有健康DM膜和内皮的角膜后板层植片。另一组行穿透性角膜移植术(PK)。术后观察角膜透明性,角膜中央厚度、以及内皮细胞密度。定期取标本行角膜内皮染色及组织学检查。
3.20只眼部正常的成年家兔,随机分为两组。一组自上方角膜缘外1.0mm作一4-5mm巩膜隧道切口行小切口DLEK手术,
Objectives1. To establish a rabbit model with permanent endothelial damage, which is suitable to evaluate the therapeutic effects of corneal transplantation.2. To perform the deep lamellar endothelial keratoplasty (DLEK) through a scleraocorneal pocket incision on the rabbit model with self-made surgical instruments, and to assess the feasibility, therapeutic effect and complications of this transplantation method.3. To observe the corneal topography changes of the early stage after small-incision DLEK.Methods1. A total of 24 adult rabbits were assigned randomly into 2 groups. In one group (group Endo), a 30 gauge air injection cannula was inserted into the anterior chamber of the right eye viathe cornea, lmm central to the limbus, then the corneal endothelium were scratched while in the other group ( group DM+Endo), both the corneal endothelium and the Descmet's membrane was scraped off. The corneal cloudiness and edema were assessed by slit lamp, the corneal thickness measured by ultrasound biomicroscopy, and the endothelium observed and counted with specular microscopy. Corneal samples were examined by endothelium staining and HE staining at various time points. 2. Thirty-two rabbits with Bullous keratopathy (BK) were
randomly divided into two groups. In one group (group DLEK), a lamellar pocket was created through a superior limbus incision, a self-made specialized trephine was centered within the lamellar pocket, and a 7-mm trephination of the posterior stroma and endothelial tissue was performed, then a 7.25mm donor disk with healthy DM and endothelium was placed in the recipient bed. In the other group (group PK), standard penetrating keratoplasty (PK) was performed. Corneal clarity, central corneal thickness and endothelium density were evaluated.3. Twenty rabbits without eye diseases were randomized to two groups. In one group (group sDLEK), DLEK was performed through a 4.0-5.0mm scleral incision 1.0mm peripheral to the limbus; in the other group (group PK), PK was performed. Corneal power, astigmatism, corneal uniform index; corneal unspherical index and visual acuity were assessed by corneal topography.Results1. In group DM+Endo, the cornea remained cloudy and edematous for 6 months, and the magnitude of corneal thickness remained 3-4 times of that before the mechanical injury; In group Endo, the cornea became to clear by 1 month after the injury, and the corneal thickness and endothelium count was close to the normal level by 3 months after the injury.2. In group DLEK, the cornea began clearing at 2 weeks after the surgery in 14 of the 16 eyes, the corneal thickness became close to the normal level 3 months after surgery. The graft endothelial cell count at 6 months remained at normal range. The operation failed in 2 eyes, one because of the lamellar perforation, the other of anterior synechia. Fourteen of the 16 corneas also began clear at 2 weeks after surgery, two failed because of
1.探讨一种稳定、持久的家兔角膜内皮损伤方法,从而建立适合评价深板层内皮角膜移植术(DLEK)治疗大泡性角膜病变(BK)的模型。
2.自行设计DLEK手术相关的器械,对家兔BK模型行经角膜-巩膜袋法的DLEK手术治疗,探讨手术技术方法,对临床效果进行评价。
3.观察小切口DLEK手术术后角膜地形图变化。方法
1.24只成年家兔,随机分为两组,每组12只,均行右眼手术。一组经角膜缘内1mm用30gauge空气注射套管针自穿刺口进入前房刮除全部后弹力层(DM)和内皮细胞,另一组单纯刮除内皮细胞。术后采用裂隙灯及活体生物显微镜观察角膜水肿混浊程度和角膜厚度,以角膜内皮镜行角膜内皮细胞计数,定期取标本行角膜内皮细胞染色及组织学检查。
2.32只右眼建立BK模型的家兔,随机分为两组,每组16只眼。一组经上方角膜缘切口制作板层角膜袋,采用自行设计的扁平环钻钻除一直径7.0mm角膜后板层组织,置换一直径7.25mm的带有健康DM膜和内皮的角膜后板层植片。另一组行穿透性角膜移植术(PK)。术后观察角膜透明性,角膜中央厚度、以及内皮细胞密度。定期取标本行角膜内皮染色及组织学检查。
3.20只眼部正常的成年家兔,随机分为两组。一组自上方角膜缘外1.0mm作一4-5mm巩膜隧道切口行小切口DLEK手术,
Objectives1. To establish a rabbit model with permanent endothelial damage, which is suitable to evaluate the therapeutic effects of corneal transplantation.2. To perform the deep lamellar endothelial keratoplasty (DLEK) through a scleraocorneal pocket incision on the rabbit model with self-made surgical instruments, and to assess the feasibility, therapeutic effect and complications of this transplantation method.3. To observe the corneal topography changes of the early stage after small-incision DLEK.Methods1. A total of 24 adult rabbits were assigned randomly into 2 groups. In one group (group Endo), a 30 gauge air injection cannula was inserted into the anterior chamber of the right eye viathe cornea, lmm central to the limbus, then the corneal endothelium were scratched while in the other group ( group DM+Endo), both the corneal endothelium and the Descmet's membrane was scraped off. The corneal cloudiness and edema were assessed by slit lamp, the corneal thickness measured by ultrasound biomicroscopy, and the endothelium observed and counted with specular microscopy. Corneal samples were examined by endothelium staining and HE staining at various time points. 2. Thirty-two rabbits with Bullous keratopathy (BK) were
randomly divided into two groups. In one group (group DLEK), a lamellar pocket was created through a superior limbus incision, a self-made specialized trephine was centered within the lamellar pocket, and a 7-mm trephination of the posterior stroma and endothelial tissue was performed, then a 7.25mm donor disk with healthy DM and endothelium was placed in the recipient bed. In the other group (group PK), standard penetrating keratoplasty (PK) was performed. Corneal clarity, central corneal thickness and endothelium density were evaluated.3. Twenty rabbits without eye diseases were randomized to two groups. In one group (group sDLEK), DLEK was performed through a 4.0-5.0mm scleral incision 1.0mm peripheral to the limbus; in the other group (group PK), PK was performed. Corneal power, astigmatism, corneal uniform index; corneal unspherical index and visual acuity were assessed by corneal topography.Results1. In group DM+Endo, the cornea remained cloudy and edematous for 6 months, and the magnitude of corneal thickness remained 3-4 times of that before the mechanical injury; In group Endo, the cornea became to clear by 1 month after the injury, and the corneal thickness and endothelium count was close to the normal level by 3 months after the injury.2. In group DLEK, the cornea began clearing at 2 weeks after the surgery in 14 of the 16 eyes, the corneal thickness became close to the normal level 3 months after surgery. The graft endothelial cell count at 6 months remained at normal range. The operation failed in 2 eyes, one because of the lamellar perforation, the other of anterior synechia. Fourteen of the 16 corneas also began clear at 2 weeks after surgery, two failed because of
引文
1. Bourne WM, Kaufman HE. Specular microscopy of human corneal endothelium in vivo. Am J Ophthalmomol, 1976; 81: 319-323
2. Yee RW, Matsuda M, Schultz RO et al. Changes in the normal corneal endothelial cellular pattern as a function of age. Curr Eye Res, 1985; 4: 671-676
3. Gwin RM, Lerner I, Warren JK, et al. Decrease in canine corneal endothelial cell density and increase in corneal thickness as functions of age. Invest Ophthalmol Vis Sci, 1982; 22: 267-274
4. Laing RA, Sandstrom MM, Berrospi AR, et al. Changes in the corneal endothelium cellular as a function of age. Exp Eye Res, 1976; 22: 587-591
5. Maeno A, Naor J, Lee HM, Hunter WS, Rootman DS. Three decades of corneal transplantation: indications and patient characteristics. Cornea, 2000; 19: 7-11
6. Mamalis N, Anderson CW, Kreisler KR, Lundergan MK, Olson RJ. Changing trends in the indications for penetrating keratoplasty. Arch Ophthalmol, 1992; 110: 1409-1411
7. Lindquist TD, McGlothan JS, Rotkis WM, Chandler JW. Indications for penetrating keratoplasty: 1980-1988. Cornea, 1991; 10: 210-216
8. Sugar A, Sugar J. Techniques in penetrating keratoplasty: A quarter century of development. Cornea, 2000; 19: 603-610
9. Melles GRJ, Lander F, van Dooren BTH, Pels E, Beekjuis WH. Preliminary clinical results of posterior lamellar keratoplasty through a sclerocorneal pocket incision. Ophthalmology, 2000; 107: 1850-1857
10. Culbertson WW. Endothelial replacement: Flap approach. Ophthalmol Clin North Am, 2003; 16: 113-118
11. Terry MA. Endothelial replacement: the limbal pocket approach. Ophthalmol Clin North Am, 2003; 16: 103-112
12. Terry, MA. A new approach for endothelial transplantation: deep lamellar endothelial keratoplasty. Int Ophthalmol Clin, 2003; 43: 183-193
13. Binder RF, Binder HF. Regenerative process in the endothelium of the cornea. Arch Ophthalmol, 1957; 57: 11-16
14. Mprton PL, Ormsby HL. Healing of endothelium and Descemet's membrane of rabbit cornea. Am J Ophthalmol, 1958; 46: 62-69
15. Chi HH, Teng CC, Katzin HM. Healing process in the mechanical denudation of the corneal endothelium. Am J Ophthalmol, 1960; 46: 493-499
16. Mills NL, Donn A. Incorporation of tritium-labeled thymidine by rabbit corneal endothelium. Arch Ophthalmol, 1960; 64: 443-447
17. Bito LZ, Harding CV. Tritium retention by corneal endothelium after incorporation of H~3-thymidine. Arch Ophthalmol, 1961; 66: 553-559
18. Matsuda M, Sawa M, Edelhauser HF, et al. Cellular migration and morphology in corneal endothelial wound repair. Invest Ophthalmol Vis Sci, 1985; 26: 443-449
19. Olson LE, Marshall J, Rice NSC. Effects of ultrasound on the corneal endothelium. Ⅱ. The endotheliar repair process. Br J Ophthalmol, 1978; 62: 145-154
20. Matsuda M, Smelser GK. Endothelial healing in alkali burned corneas. Arch Ophthalmol, 1973; 89: 402-409
21. Chung JH, Fagerholm P. Endothelial healing in rabbit corneal alkali wounds. Acta Ophthalmol, 1987; 65: 648-656
22. Chi HH, Kelman CD,. Effect of freezing on ocular tissues: Ⅰ. Clinical and histologic study of corneal endothelium. Am J Ophthalmol, 1966; 61: 630-635
23. Faure JP, Kim YZ, Graf B. Formation of giant cells in the corneal endothelium during its regeneration after destruction by freezing. Exp Eye Res, 1971: 12: 6-12
24. Renard G, Hirsh M, Faure JP, et al. Ultrastructural study on the regeneration of the corneal endothelial cells. Jpn J Ophthalmol, 1976; 20: 243-251
25. Khodadoust AA, Green K. Physiological function of regenerating endothelium. Invest Ophthalmol Vis Sci, 1976; 15: 96-103
26. Minkowski JS. Bartels SP. delori BF, et al. Corneal endothelial function and structure following cryo-injury in the rabbit. Invest Ophthalmol Vis Sci, 1984; 25: 1416-1425
27. Van Horn DL, Sendele DD, Seideman S et, al. Regenerative capacity of the corneal endothelium in rabbi and cat. Invest Ophthalmol Vis Sci, 1977; 17: 597-613
28. Yee RW, Geroski DH, Mamoru M, et al. Correlation of corneal endothelial pump site density barrier function and morphology in wound repair. Invest Ophthalmol Vis Sci, 1985; 26: 1191-1201
29. Capella JA. Regeneration of endothelium in diseased and injured corneas. Am J Ophthalmol, 1972; 74: 810-817
30. Hirsh M, Renard G, Faure JP, et al. Formation of intracellular spaces and junctions in regenerating rabbit corneal endothelium. Exp Eye Res, 1976; 23: 385-391
31. Maurice D. Perlman M. Permanent destruction of the corneal endothelium in rabbits. Invest Ophthalmol Vis Sci, 1977; 16: 646-649
32. Tuft SJ, William KA, Coster DJ. Endothelial repair in the rat cornea. Invest Ophthalmol Vis Sci, 1986; 27: 1199-1207
33. Ilmonen M, Lehtosalo JI, Vitanen J, et al. Initial healing of the posterior corneal surface following perforating trauma in guinea pig: A scanning electron microscope study. Acta Ophthalmol, 1984; 62: 787-796
34. Van Horn DL, Sendele DD, Seideman S. Regeneration capacity of the corneal endothelium in rabbit and cat. Invest Ophthalmol Vis Sci, 1977; 16: 597-613
35. Ogita Y, Higuchi S, Kari K, at al. Wound healing of the endothelium of the living cat cornea: A specular microscopic study. Jpn J Ophthalmol, 1981; 25: 326-332
2. Yee RW, Matsuda M, Schultz RO et al. Changes in the normal corneal endothelial cellular pattern as a function of age. Curr Eye Res, 1985; 4: 671-676
3. Gwin RM, Lerner I, Warren JK, et al. Decrease in canine corneal endothelial cell density and increase in corneal thickness as functions of age. Invest Ophthalmol Vis Sci, 1982; 22: 267-274
4. Laing RA, Sandstrom MM, Berrospi AR, et al. Changes in the corneal endothelium cellular as a function of age. Exp Eye Res, 1976; 22: 587-591
5. Maeno A, Naor J, Lee HM, Hunter WS, Rootman DS. Three decades of corneal transplantation: indications and patient characteristics. Cornea, 2000; 19: 7-11
6. Mamalis N, Anderson CW, Kreisler KR, Lundergan MK, Olson RJ. Changing trends in the indications for penetrating keratoplasty. Arch Ophthalmol, 1992; 110: 1409-1411
7. Lindquist TD, McGlothan JS, Rotkis WM, Chandler JW. Indications for penetrating keratoplasty: 1980-1988. Cornea, 1991; 10: 210-216
8. Sugar A, Sugar J. Techniques in penetrating keratoplasty: A quarter century of development. Cornea, 2000; 19: 603-610
9. Melles GRJ, Lander F, van Dooren BTH, Pels E, Beekjuis WH. Preliminary clinical results of posterior lamellar keratoplasty through a sclerocorneal pocket incision. Ophthalmology, 2000; 107: 1850-1857
10. Culbertson WW. Endothelial replacement: Flap approach. Ophthalmol Clin North Am, 2003; 16: 113-118
11. Terry MA. Endothelial replacement: the limbal pocket approach. Ophthalmol Clin North Am, 2003; 16: 103-112
12. Terry, MA. A new approach for endothelial transplantation: deep lamellar endothelial keratoplasty. Int Ophthalmol Clin, 2003; 43: 183-193
13. Binder RF, Binder HF. Regenerative process in the endothelium of the cornea. Arch Ophthalmol, 1957; 57: 11-16
14. Mprton PL, Ormsby HL. Healing of endothelium and Descemet's membrane of rabbit cornea. Am J Ophthalmol, 1958; 46: 62-69
15. Chi HH, Teng CC, Katzin HM. Healing process in the mechanical denudation of the corneal endothelium. Am J Ophthalmol, 1960; 46: 493-499
16. Mills NL, Donn A. Incorporation of tritium-labeled thymidine by rabbit corneal endothelium. Arch Ophthalmol, 1960; 64: 443-447
17. Bito LZ, Harding CV. Tritium retention by corneal endothelium after incorporation of H~3-thymidine. Arch Ophthalmol, 1961; 66: 553-559
18. Matsuda M, Sawa M, Edelhauser HF, et al. Cellular migration and morphology in corneal endothelial wound repair. Invest Ophthalmol Vis Sci, 1985; 26: 443-449
19. Olson LE, Marshall J, Rice NSC. Effects of ultrasound on the corneal endothelium. Ⅱ. The endotheliar repair process. Br J Ophthalmol, 1978; 62: 145-154
20. Matsuda M, Smelser GK. Endothelial healing in alkali burned corneas. Arch Ophthalmol, 1973; 89: 402-409
21. Chung JH, Fagerholm P. Endothelial healing in rabbit corneal alkali wounds. Acta Ophthalmol, 1987; 65: 648-656
22. Chi HH, Kelman CD,. Effect of freezing on ocular tissues: Ⅰ. Clinical and histologic study of corneal endothelium. Am J Ophthalmol, 1966; 61: 630-635
23. Faure JP, Kim YZ, Graf B. Formation of giant cells in the corneal endothelium during its regeneration after destruction by freezing. Exp Eye Res, 1971: 12: 6-12
24. Renard G, Hirsh M, Faure JP, et al. Ultrastructural study on the regeneration of the corneal endothelial cells. Jpn J Ophthalmol, 1976; 20: 243-251
25. Khodadoust AA, Green K. Physiological function of regenerating endothelium. Invest Ophthalmol Vis Sci, 1976; 15: 96-103
26. Minkowski JS. Bartels SP. delori BF, et al. Corneal endothelial function and structure following cryo-injury in the rabbit. Invest Ophthalmol Vis Sci, 1984; 25: 1416-1425
27. Van Horn DL, Sendele DD, Seideman S et, al. Regenerative capacity of the corneal endothelium in rabbi and cat. Invest Ophthalmol Vis Sci, 1977; 17: 597-613
28. Yee RW, Geroski DH, Mamoru M, et al. Correlation of corneal endothelial pump site density barrier function and morphology in wound repair. Invest Ophthalmol Vis Sci, 1985; 26: 1191-1201
29. Capella JA. Regeneration of endothelium in diseased and injured corneas. Am J Ophthalmol, 1972; 74: 810-817
30. Hirsh M, Renard G, Faure JP, et al. Formation of intracellular spaces and junctions in regenerating rabbit corneal endothelium. Exp Eye Res, 1976; 23: 385-391
31. Maurice D. Perlman M. Permanent destruction of the corneal endothelium in rabbits. Invest Ophthalmol Vis Sci, 1977; 16: 646-649
32. Tuft SJ, William KA, Coster DJ. Endothelial repair in the rat cornea. Invest Ophthalmol Vis Sci, 1986; 27: 1199-1207
33. Ilmonen M, Lehtosalo JI, Vitanen J, et al. Initial healing of the posterior corneal surface following perforating trauma in guinea pig: A scanning electron microscope study. Acta Ophthalmol, 1984; 62: 787-796
34. Van Horn DL, Sendele DD, Seideman S. Regeneration capacity of the corneal endothelium in rabbit and cat. Invest Ophthalmol Vis Sci, 1977; 16: 597-613
35. Ogita Y, Higuchi S, Kari K, at al. Wound healing of the endothelium of the living cat cornea: A specular microscopic study. Jpn J Ophthalmol, 1981; 25: 326-332