Background: Corneal neovascularization (NV) is a process of abnormal vessel growth into the transparent cornea from the limbus and can disturb the light passing through the cornea, resulting in vision loss or even blindness. The use of nanomedicine as an effective therapeutic formulation in ophthalmology has led to higher drug bioavailability and a slow drug release rate. In this research, we designed and explored the feasibility of a new nanomedicine, gp91 ds-tat (gp91) peptide-encapsulated gelatin nanoparticles (GNP-gp91), for inhibiting corneal angiogenesis.
Methods: GNP-gp91 were prepared by a two-step desolvation method. The characterization and cytocompatibility of GNP-gp91 were analyzed. The inhibition effect of GNP-gp91 on HUVEC cell migration and tube formation was observed by an inverted microscope. The drug retention test in mouse cornea was observed by in vivo imaging system, fluorescence microscope, and DAPI/TAMRA staining. Finally, the therapeutic efficacy and evaluation of neovascularization-related factors were conducted through the in vivo corneal NV mice model via topical delivery.
Results: The prepared GNP-gp91 had a nano-scale diameter (550.6 nm) with positive charge (21.7 mV) slow-release behavior (25%, 240hr). In vitro test revealed that GNP-gp91 enhanced the inhibition of cell migration and tube formation capacity via higher internalization of HUVEC. Topical administration (eyedrops) of the GNP-gp91 significantly prolongs the retention time (46%, 20 min) in the mouse cornea. In chemically burned corneal neovascularization models, corneal vessel area with a significant reduction in GNP-gp91 group (7.89%) was revealed when compared with PBS (33.99%) and gp91 (19.67%) treated groups via every two days dosing. Moreover, GNP-gp91 significantly reduced the concentration of Nox2, VEGF and MMP9 in NV’s cornea.
Conclusion: The nanomedicine, GNP-gp91, was successfully synthesized for ophthalmological application. These data suggest that GNP-gp91 contained eyedrops that not only have a longer retention time on the cornea but also can treat mice corneal NV effectively delivered in a low dosing frequency, GNP-gp91 eyedrops provides an alternative strategy for clinical ocular disease treatment in the culture.
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中文翻译：

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功能性肽载明胶纳米颗粒作为角膜新生血管治疗的滴眼液

背景：角膜新生血管（NV）是异常血管从角膜缘长入透明角膜的过程，可以干扰通过角膜的光线，导致视力下降甚至失明。纳米药物作为眼科有效治疗制剂的使用导致了更高的药物生物利用度和缓慢的药物释放速率。在这项研究中，我们设计并探索了一种新的纳米药物 gp91 ds-tat (gp91) 肽包封明胶纳米颗粒 (GNP-gp91) 用于抑制角膜血管生成的可行性。
方法：GNP-gp91 通过两步去溶剂化法制备。分析了 GNP-gp91 的表征和细胞相容性。倒置显微镜观察GNP-gp91对HUVEC细胞迁移和管腔形成的抑制作用。通过体内成像系统、荧光显微镜和DAPI/TAMRA染色观察药物在小鼠角膜中的滞留试验。最后，通过局部递送的体内角膜NV小鼠模型进行了治疗效果和新生血管形成相关因子的评价。
结果：制备的 GNP-gp91 具有纳米级直径 (550.6 nm) 和正电荷 (21.7 mV) 缓释行为 (25%, 240hr)。体外试验表明，GNP-gp91 通过更高的 HUVEC 内化增强对细胞迁移和管形成能力的抑制。GNP-gp91 的局部给药（滴眼液）显着延长了在小鼠角膜中的保留时间（46%，20 分钟）。在化学烧伤角膜新生血管模型中，与每两天给药一次的 PBS (33.99%) 和 gp91 (19.67%) 治疗组相比，GNP-gp91 组 (7.89%) 的角膜血管面积显着减少。此外，GNP-gp91 显着降低了 NV 角膜中 Nox2、VEGF 和 MMP9 的浓度。
结论：成功合成了纳米药物 GNP-gp91 用于眼科应用。这些数据表明，GNP-gp91 滴眼液不仅在角膜上具有更长的保留时间，而且可以以低给药频率有效地治疗小鼠角膜 NV，GNP-gp91 滴眼液为临床眼部疾病治疗提供了一种替代策略。文化。
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