Hassanpour Kiana, Kanavi Mozhgan Rezaei, Daftarian Narsis, Samaeili Azadeh, Suri Fatemeh, Pakravan Mohammad, Doozandeh Azadeh, Aski Sasha Afsar, Fakhri Maryam, Moghaddasi Afrooz, Ahmadieh Hamid, Esfandiari Hamed
Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Ophthalmology, Imam Hossein Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
J Ophthalmic Vis Res. 2022 Nov 29;17(4):486-496. doi: 10.18502/jovr.v17i4.12300. eCollection 2022 Oct-Dec.
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.
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.
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 mm, 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 ( = 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 (s 0.001), the anti-CTGF and MMC groups were comparable ( = 0.87).
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.
比较结膜下注射抗结缔组织生长因子抗体（抗CTGF）、丝裂霉素C（MMC）和安慰剂在兔小梁切除术模型中减少瘢痕形成的疗效。
共纳入14只兔子。9只兔子在切开球结膜前，对每只兔子的三只眼睛分别进行小梁切除术，并结膜下注射抗CTGF抗体、MMC或平衡盐溶液（BSS）。抗CTGF组在术后5天重复注射一次抗体。所有9只兔子在第14天安乐死；取出眼球，用苏木精-伊红染色、Masson三色染色和免疫组织化学检测α平滑肌（α-SMA）肌动蛋白。对其余兔子的五只眼睛进行RNA提取，其中包括一只未做手术的眼睛、一只小梁切除术后5小时未注射任何药物的眼睛、一只小梁切除术后5天未注射任何药物的眼睛，以及两只小梁切除术后5天分别注射MMC和BSS的眼睛。
抗CTGF组、MMC组和对照组的平均滤过泡面积分别为3.8±1.45、5.9±1.4和3.5±1.9平方毫米。胶原组织在滤过泡面积中所占比例分别为13.7%、13.5%和18.5%。BSS组的这一比例显著更高（P = 0.04）。与未手术的眼睛相比，小梁切除术后5小时和5天眼睛中CTGF mRNA的表达明显更显著。抗CTGF组、MMC组和对照组中，以染色等级乘以免疫染色细胞百分比计算的α-SMA免疫反应性细胞的平均H评分分别为14.6、10.22和140.58。虽然对照组的评分显著更高（P < 0.001），但抗CTGF组和MMC组相当（P = 0.87）。
基于这项动物研究的结果，小梁切除术后注射抗CTGF抗体可显著减少胶原组织和成肌纤维细胞。
