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Hassanpour K, Rezaei Kanavi M, Daftarian N, Samaeili A, Suri F, Pakravan M, Doozandeh A, Afsar Aski S, Fakhri M, Moghaddasi A, Ahmadieh H, Esfandiari H. The Inhibitory Effect of Connective Tissue Growth Factor Antibody on Postoperative Fibrosis in a Rabbit Model of Trabeculectomy. JOVR [Internet]. 2022Nov.24 [cited 2024Apr.19];17(4):486–496. Available from: https://knepublishing.com/index.php/JOVR/article/view/12300
https://doi.org/10.18502/jovr.v17i4.12300
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authors
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Kiana Hassanpour
Kiana Hassanpour
Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Mozhgan Rezaei Kanavi
Mozhgan Rezaei Kanavi
Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Narsis Daftarian
Narsis Daftarian
Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Azadeh Samaeili
Azadeh Samaeili
Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Fatemeh Suri
Fatemeh Suri
Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Mohammad Pakravan
Mohammad Pakravan
Ophthalmic Epidemiology Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Azadeh Doozandeh
Azadeh Doozandeh
Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Sasha Afsar Aski
Sasha Afsar Aski
Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Maryam Fakhri
Maryam Fakhri
Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Afrooz Moghaddasi
Afrooz Moghaddasi
Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Hamid Ahmadieh
Hamid Ahmadieh
Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Hamed Esfandiari
Hamed Esfandiari
Department of ophthalmology, Olmsted Medical Center, Rochester, MN, USA
Published Date
- Nov 24, 2022
The Inhibitory Effect of Connective Tissue Growth Factor Antibody on Postoperative Fibrosis in a Rabbit Model of Trabeculectomy
Abstract
Purpose: To compare the efficacy of subconjunctival injection of an anti-connective tissue growth factor antibody (anti-CTGF) versus mitomycin-C (MMC) and placebo in reducing scar formation in a rabbit model of trabeculectomy.
Methods: A total of 14 rabbits were included. Nine rabbits underwent trabeculectomy with subconjunctival injections of either anti-CTGF antibody, MMC, or balanced salt solution (BSS), each administered in three eyes, before peritomy. The anti-CTGF group received a repeated dose of the antibody five days after surgery. All nine rabbits were euthanized on day 14; the globes were stained with hematoxylin & eosin, Masson’s Trichrome, and immunohistochemistry for detecting alpha-smooth muscle (α-SMA) actin. RNA extraction was performed on five eyes of the remaining rabbits which included one eye without any surgery, one eye 5 hr after trabeculectomy without any injection, one eye five days after trabeculectomy without any injection, and two eyes five days after trabeculectomy with administration of MMC and BSS, respectively.
Results: The mean bleb area in the anti-CTGF, MMC, and control groups was 3.8 ± 1.45, 5.9 ± 1.4, and 3.5 ± 1.9 mm2, respectively. Collagenous tissue was found to occupy the bleb area by 13.7%, 13.5%, and 18.5%, respectively. This ratio was significantly higher in the BSS group (P = 0.04). The expression of CTGF mRNA after 5 hr and five days in eyes undergoing trabeculectomy were significantly more pronounced as compared to the unoperated eye. The mean H-SCORE of α-SMA-immune reactive cells calculated as the grade of staining multiplied by the percentage of immune stained cells was 14.6, 10.22, and 140.58 in the anti-CTGF, MMC, and control groups, respectively. While the control eyes had a significantly higher score (Ps < 0.001), the anti-CTGF and MMC groups were comparable (P = 0.87).
Conclusion: Based on the results of this animal study, the anti-CTGF antibody injection resulted in a significant reduction in collagenous tissue and myofibroblast cells after trabeculectomy.
Keywords:
Anti-connective Tissue Growth Factor, CTGF, Mitomycin-C, Trabeculectomy
References
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2. Bindlish R, Condon GP, Schlosser JD, D’Antonio J, Lauer KB, Lehrer R. Efficacy and safety of mitomycin-C in primary trabeculectomy: Five-year follow-up. Ophthalmology 2002;109:1336–1341.
3. Lockwood A, Brocchini S, Khaw PT. New developments in the pharmacological modulation of wound healing after glaucoma filtration surgery. Curr Opin Pharmacol 2013;13:65–71.
4. Rodriguez-Una I, Azuara-Blanco A, King AJ. Survey of glaucoma surgical preferences and post-operative care in the United Kingdom. Clin Exp Ophthalmol 2017;45:232– 240.
5. Group C-TS. A phase III study of subconjunctival human anti–transforming growth factor β2 monoclonal antibody (CAT-152) to prevent scarring after first-time trabeculectomy. Ophthalmology 2007;114:1822–30.e2.
6. Abreu JG, Ketpura NI, Reversade B, De Robertis E. Connective-tissue growth factor (CTGF) modulates cell signalling by BMP and TGF-β. Nat Cell Biol 2002;4:599.
7. Adler SG, Schwartz S, Williams ME, Arauz-Pacheco C, Bolton WK, Lee T, et al. Phase 1 study of anti-CTGF monoclonal antibody in patients with diabetes and microalbuminuria. Clin J Am Soc Nephrol 2010;5:1420– 1428.
8. Esson DW, Neelakantan A, Iyer SA, Blalock TD, Balasubramanian L, Grotendorst GR, et al. Expression of connective tissue growth factor after glaucoma filtration surgery in a rabbit model. Invest Ophthalmol Vis Sci 2004;45:485–491.
9. Fan W-H, Pech M, Karnovsky MJ. Connective tissue growth factor (CTGF) stimulates vascular smooth muscle cell growth and migration in vitro. Eur J Cell Biol 2000;79:915–923.
10. Lu H, Kojima K, Battula VL, Spong S, Canizales M, Lock RB, et al. Targeting connective tissue growth factor (CTGF) in acute lymphoblastic leukemia preclinical models: Anti- CTGF monoclonal antibody attenuates leukemia growth. Ann Hematol 2014;93:485–492.
11. Wang Q, Usinger W, Nichols B, Gray J, Xu L, Seeley TW, et al. Cooperative interaction of CTGF and TGF-β in animal models of fibrotic disease. Fibrogenesis Tissue Repair 2011;4:4.
12. Yuan HP, Li XH, Yang BB, Shao ZB, Yan LP. [Expression of connective tissue growth factor after trabeculectomy in rabbits]. Zhonghua Yan Ke Za Zhi 2009;45:168–174.
13. Wang JM, Hui N, Fan YZ, Xiong L, Sun NX. Filtering bleb area and intraocular pressure following subconjunctival injection of CTGF antibody after glaucoma filtration surgery in rabbits. Int J Ophthalmol 2011;4:480–483.
14. Browne JG, Ho SL, Kane R, Oliver N, Clark AF, O’Brien CJ, et al. Connective tissue growth factor is increased in pseudoexfoliation glaucoma. Invest Ophthalmol Vis Sci 2011;52:3660–3666.
15. Fleenor DL, Shepard A, Jacobson N, Pang I-H, Clark AF. Agents which regulate, inhibit, or modulate the activity and/or expression of connective tissue growth factor (CTGF) as a unique means to both lower intraocular pressure and treat glaucomatous retinopathies/optic neuropathies. Google Patents; 2008.
16. Shepard AR, Pang I-H. RNAi inhibition of CTGF for treatment of ocular disorders. Google Patents; 2009.
17. Taylor AW. Primary open-angle glaucoma: A transforming growth factor-β pathway–mediated disease. Am J Pathol 2012;180:2201–2204.
18. Motevasseli T, Daftarian N, Kanavi MR, Ahmadieh H, Bagheri A, Hosseini SB, et al. Ocular safety of intravitreal connective tissue growth factor neutralizing antibody. Curr Eye Res 2017;42:1194–1201.
19. Fedchenko N, Reifenrath J. Different approaches for interpretation and reporting of immunohistochemistry analysis results in the bone tissue–a review. Diagnostic Pathol 2014;9:221.
20. Esfandiari H, Pakravan M, Loewen NA, Yaseri M. Predictive value of early postoperative IOP and bleb morphology in Mitomycin-C augmented trabeculectomy. F1000Research 2017;6.
21. Mead AL, Wong TT, Cordeiro MF, Anderson IK, Khaw PT. Evaluation of anti-TGF-β2 antibody as a new postoperative anti-scarring agent in glaucoma surgery. Invest Ophthalmol Vis Sci 2003;44:3394–3401.
22. Seibold LK, Sherwood MB, Kahook MY. Wound modulation after filtration surgery. Surv Ophthalmol 2012;57:530–550.
23. Wipff P-J, Rifkin DB, Meister J-J, Hinz B. Myofibroblast contraction activates latent TGF-β1 from the extracellular matrix. J Cell Biol 2007;179:1311–1323.
24. Lipson KE, Wong C, Teng Y, Spong S. CTGF is a central mediator of tissue remodeling and fibrosis and its inhibition can reverse the process of fibrosis. Fibrogenesis Tissue Repair 2012;5:S24.
25. Iyer P, Maddala R, Pattabiraman PP, Rao PV. Connective tissue growth factor–mediated upregulation of neuromedin U expression in trabecular meshwork cells and its role in homeostasis of aqueous humor outflow. Invest Ophthalmol Vis Sci 2012;53:4952–4962.
26. Kuespert S, Junglas B, Braunger BM, Tamm ER, Fuchshofer R. The regulation of connective tissue growth factor expression influences the viability of human trabecular meshwork cells. J Cell Mol Med 2015;19:1010–1020.
27. Junglas B, Kuespert S, Seleem AA, Struller T, Ullmann S, Bösl M, et al. Connective tissue growth factor causes glaucoma by modifying the actin cytoskeleton of the trabecular meshwork. Am J Pathol 2012;180:2386–2403.
28. Schild C, Trueb B. Mechanical stress is required for highlevel expression of connective tissue growth factor. Exp Cell Res 2002;274:83–91.
29. Daftarian N, Bayeghi O, Rezaei Kanavi M, Ahmadieh H. Effects of intravitreal connective tissue growth factor neutralizing antibody on the epiretinal membrane formation; an experimental study. Invest Ophthalmol Vis Sci 2019;60:5812.
30. Igarashi A, Okochi H, Bradham D, Grotendorst GR. Regulation of connective tissue growth factor gene expression in human skin fibroblasts and during wound repair. Mol Cell Biol 1993;4:637–645.
31. Daniels JT, Occleston NL, Crowston JG, Khaw PT. Effects of antimetabolite induced cellular growth arrest on fibroblastfibroblast interactions. Exp Eye Res 1999;69:117–127.
32. Occleston NL, Daniels JT, Tarnuzzer RW, Sethi KK, Alexander RA, Bhattacharya SS, et al. Single exposures to antiproliferatives: Long-term effects on ocular fibroblast wound-healing behavior. Invest Ophthalmol Vis Sci 1997;38:1998–2007.
33. Sherwood MB. A sequential, multiple-treatment, targeted approach to reduce wound healing and failure of glaucoma filtration surgery in a rabbit model (an American Ophthalmological Society thesis). Trans Am Ophthalmol Soc 2006;104:478–492.
34. Yu-Wai-Man C, Khaw PT. Developing novel anti-fibrotic therapeutics to modulate post-surgical wound healing in glaucoma: big potential for small molecules. Expert Rev Ophthalmol 2015;10:65–76.
35. Daftarian N, Rohani S, Kanavi MR, Suri F, Mirrahimi M, Hafezi-Moghadam A, et al. Effects of intravitreal connective tissue growth factor neutralizing antibody on choroidal neovascular membrane-associated subretinal fibrosis. Exp Eye Res 2019;184:286–295.
36. Yamanaka O, Saika S, Ikeda K, Miyazaki K-i, Kitano A, Ohnishi Y. Connective tissue growth factor modulates extracellular matrix production in human subconjunctival fibroblasts and their proliferation and migration in vitro. Jpn J Ophthalmol 2008;52:8–15.
37. Li B, Wang JH-C. Fibroblasts and myofibroblasts in wound healing: Force generation and measurement. J Tissue Viability 2011;20:108–120.
38. Darby IA, Laverdet B, Bonté F, Desmoulière A. Fibroblasts and myofibroblasts in wound healing. Clin Cosmet Investig Dermatol 2014;7:301.
39. Kuiper EJ, Roestenberg P, Ehlken C, Lambert V, van Treslong-de Groot HB, Lyons KM, et al. Angiogenesis is not impaired in connective tissue growth factor (CTGF) knockout mice. J Histochem Cytochem 2007;55:1139–1147.
40. Khaw PT, Sherwood MB, MacKay SL, Rossi MJ, Schultz G. Five-minute treatments with fluorouracil, floxuridine, and mitomycin have long-term effects on human Tenon’s capsule fibroblasts. Arch Ophthalmol 1992;110:1150–1154.
2. Bindlish R, Condon GP, Schlosser JD, D’Antonio J, Lauer KB, Lehrer R. Efficacy and safety of mitomycin-C in primary trabeculectomy: Five-year follow-up. Ophthalmology 2002;109:1336–1341.
3. Lockwood A, Brocchini S, Khaw PT. New developments in the pharmacological modulation of wound healing after glaucoma filtration surgery. Curr Opin Pharmacol 2013;13:65–71.
4. Rodriguez-Una I, Azuara-Blanco A, King AJ. Survey of glaucoma surgical preferences and post-operative care in the United Kingdom. Clin Exp Ophthalmol 2017;45:232– 240.
5. Group C-TS. A phase III study of subconjunctival human anti–transforming growth factor β2 monoclonal antibody (CAT-152) to prevent scarring after first-time trabeculectomy. Ophthalmology 2007;114:1822–30.e2.
6. Abreu JG, Ketpura NI, Reversade B, De Robertis E. Connective-tissue growth factor (CTGF) modulates cell signalling by BMP and TGF-β. Nat Cell Biol 2002;4:599.
7. Adler SG, Schwartz S, Williams ME, Arauz-Pacheco C, Bolton WK, Lee T, et al. Phase 1 study of anti-CTGF monoclonal antibody in patients with diabetes and microalbuminuria. Clin J Am Soc Nephrol 2010;5:1420– 1428.
8. Esson DW, Neelakantan A, Iyer SA, Blalock TD, Balasubramanian L, Grotendorst GR, et al. Expression of connective tissue growth factor after glaucoma filtration surgery in a rabbit model. Invest Ophthalmol Vis Sci 2004;45:485–491.
9. Fan W-H, Pech M, Karnovsky MJ. Connective tissue growth factor (CTGF) stimulates vascular smooth muscle cell growth and migration in vitro. Eur J Cell Biol 2000;79:915–923.
10. Lu H, Kojima K, Battula VL, Spong S, Canizales M, Lock RB, et al. Targeting connective tissue growth factor (CTGF) in acute lymphoblastic leukemia preclinical models: Anti- CTGF monoclonal antibody attenuates leukemia growth. Ann Hematol 2014;93:485–492.
11. Wang Q, Usinger W, Nichols B, Gray J, Xu L, Seeley TW, et al. Cooperative interaction of CTGF and TGF-β in animal models of fibrotic disease. Fibrogenesis Tissue Repair 2011;4:4.
12. Yuan HP, Li XH, Yang BB, Shao ZB, Yan LP. [Expression of connective tissue growth factor after trabeculectomy in rabbits]. Zhonghua Yan Ke Za Zhi 2009;45:168–174.
13. Wang JM, Hui N, Fan YZ, Xiong L, Sun NX. Filtering bleb area and intraocular pressure following subconjunctival injection of CTGF antibody after glaucoma filtration surgery in rabbits. Int J Ophthalmol 2011;4:480–483.
14. Browne JG, Ho SL, Kane R, Oliver N, Clark AF, O’Brien CJ, et al. Connective tissue growth factor is increased in pseudoexfoliation glaucoma. Invest Ophthalmol Vis Sci 2011;52:3660–3666.
15. Fleenor DL, Shepard A, Jacobson N, Pang I-H, Clark AF. Agents which regulate, inhibit, or modulate the activity and/or expression of connective tissue growth factor (CTGF) as a unique means to both lower intraocular pressure and treat glaucomatous retinopathies/optic neuropathies. Google Patents; 2008.
16. Shepard AR, Pang I-H. RNAi inhibition of CTGF for treatment of ocular disorders. Google Patents; 2009.
17. Taylor AW. Primary open-angle glaucoma: A transforming growth factor-β pathway–mediated disease. Am J Pathol 2012;180:2201–2204.
18. Motevasseli T, Daftarian N, Kanavi MR, Ahmadieh H, Bagheri A, Hosseini SB, et al. Ocular safety of intravitreal connective tissue growth factor neutralizing antibody. Curr Eye Res 2017;42:1194–1201.
19. Fedchenko N, Reifenrath J. Different approaches for interpretation and reporting of immunohistochemistry analysis results in the bone tissue–a review. Diagnostic Pathol 2014;9:221.
20. Esfandiari H, Pakravan M, Loewen NA, Yaseri M. Predictive value of early postoperative IOP and bleb morphology in Mitomycin-C augmented trabeculectomy. F1000Research 2017;6.
21. Mead AL, Wong TT, Cordeiro MF, Anderson IK, Khaw PT. Evaluation of anti-TGF-β2 antibody as a new postoperative anti-scarring agent in glaucoma surgery. Invest Ophthalmol Vis Sci 2003;44:3394–3401.
22. Seibold LK, Sherwood MB, Kahook MY. Wound modulation after filtration surgery. Surv Ophthalmol 2012;57:530–550.
23. Wipff P-J, Rifkin DB, Meister J-J, Hinz B. Myofibroblast contraction activates latent TGF-β1 from the extracellular matrix. J Cell Biol 2007;179:1311–1323.
24. Lipson KE, Wong C, Teng Y, Spong S. CTGF is a central mediator of tissue remodeling and fibrosis and its inhibition can reverse the process of fibrosis. Fibrogenesis Tissue Repair 2012;5:S24.
25. Iyer P, Maddala R, Pattabiraman PP, Rao PV. Connective tissue growth factor–mediated upregulation of neuromedin U expression in trabecular meshwork cells and its role in homeostasis of aqueous humor outflow. Invest Ophthalmol Vis Sci 2012;53:4952–4962.
26. Kuespert S, Junglas B, Braunger BM, Tamm ER, Fuchshofer R. The regulation of connective tissue growth factor expression influences the viability of human trabecular meshwork cells. J Cell Mol Med 2015;19:1010–1020.
27. Junglas B, Kuespert S, Seleem AA, Struller T, Ullmann S, Bösl M, et al. Connective tissue growth factor causes glaucoma by modifying the actin cytoskeleton of the trabecular meshwork. Am J Pathol 2012;180:2386–2403.
28. Schild C, Trueb B. Mechanical stress is required for highlevel expression of connective tissue growth factor. Exp Cell Res 2002;274:83–91.
29. Daftarian N, Bayeghi O, Rezaei Kanavi M, Ahmadieh H. Effects of intravitreal connective tissue growth factor neutralizing antibody on the epiretinal membrane formation; an experimental study. Invest Ophthalmol Vis Sci 2019;60:5812.
30. Igarashi A, Okochi H, Bradham D, Grotendorst GR. Regulation of connective tissue growth factor gene expression in human skin fibroblasts and during wound repair. Mol Cell Biol 1993;4:637–645.
31. Daniels JT, Occleston NL, Crowston JG, Khaw PT. Effects of antimetabolite induced cellular growth arrest on fibroblastfibroblast interactions. Exp Eye Res 1999;69:117–127.
32. Occleston NL, Daniels JT, Tarnuzzer RW, Sethi KK, Alexander RA, Bhattacharya SS, et al. Single exposures to antiproliferatives: Long-term effects on ocular fibroblast wound-healing behavior. Invest Ophthalmol Vis Sci 1997;38:1998–2007.
33. Sherwood MB. A sequential, multiple-treatment, targeted approach to reduce wound healing and failure of glaucoma filtration surgery in a rabbit model (an American Ophthalmological Society thesis). Trans Am Ophthalmol Soc 2006;104:478–492.
34. Yu-Wai-Man C, Khaw PT. Developing novel anti-fibrotic therapeutics to modulate post-surgical wound healing in glaucoma: big potential for small molecules. Expert Rev Ophthalmol 2015;10:65–76.
35. Daftarian N, Rohani S, Kanavi MR, Suri F, Mirrahimi M, Hafezi-Moghadam A, et al. Effects of intravitreal connective tissue growth factor neutralizing antibody on choroidal neovascular membrane-associated subretinal fibrosis. Exp Eye Res 2019;184:286–295.
36. Yamanaka O, Saika S, Ikeda K, Miyazaki K-i, Kitano A, Ohnishi Y. Connective tissue growth factor modulates extracellular matrix production in human subconjunctival fibroblasts and their proliferation and migration in vitro. Jpn J Ophthalmol 2008;52:8–15.
37. Li B, Wang JH-C. Fibroblasts and myofibroblasts in wound healing: Force generation and measurement. J Tissue Viability 2011;20:108–120.
38. Darby IA, Laverdet B, Bonté F, Desmoulière A. Fibroblasts and myofibroblasts in wound healing. Clin Cosmet Investig Dermatol 2014;7:301.
39. Kuiper EJ, Roestenberg P, Ehlken C, Lambert V, van Treslong-de Groot HB, Lyons KM, et al. Angiogenesis is not impaired in connective tissue growth factor (CTGF) knockout mice. J Histochem Cytochem 2007;55:1139–1147.
40. Khaw PT, Sherwood MB, MacKay SL, Rossi MJ, Schultz G. Five-minute treatments with fluorouracil, floxuridine, and mitomycin have long-term effects on human Tenon’s capsule fibroblasts. Arch Ophthalmol 1992;110:1150–1154.