Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao, People's Republic of China.
Int J Nanomedicine. 2024 Nov 1;19:11163-11179. doi: 10.2147/IJN.S480800. eCollection 2024.
Fungal keratitis is a serious blinding eye disease. Traditional drugs used to treat fungal keratitis commonly have the disadvantages of low bioavailability, poor dispersion, and limited permeability.
To develop a new method for the treatment of fungal keratitis with improved bioavailability, dispersion, and permeability.
Zeolitic Imidazolate Framework-8 (ZIF-8) was formed by zinc ions and 2-methylimidazole linked by coordination bonds and characterized by Scanning electron microscopy (SEM), X-ray diffraction (XRD), and Zeta potential. The safety of ZIF-8 on HCECs and RAW 264.7 cells was detected by Cell Counting Kit-8 (CCK-8). Safety evaluation of ZIF-8 on mice corneal epithelium was conducted using the Draize corneal toxicity test. The effects of ZIF-8 on fungal growth, biofilm formation, and hyphae structure were detected by Minimal inhibit concentration (MIC), crystal violet staining, Propidium Iodide (PI) testing, and calcofluor white staining. The anti-inflammatory effects of ZIF-8 on RAW 246.7 cells were evaluated by Quantitative Real-Time PCR Experiments (qPCR) and Enzyme-linked immunosorbent assay (ELISA). Clinical score, Colony-Forming Units (CFU), Hematoxylin-eosin (HE) staining, and immunofluorescence were conducted to verify the therapeutic effect of ZIF-8 on C57BL/6 female mice with fungal keratitis.
In vitro, ZIF-8 showed outstanding antifungal effects, including inhibiting the growth of over 90% at 64 μg/mL, restraining the formation of biofilm, and destroying cell membranes. In vivo, treatment with ZIF-8 reduced corneal fungal load and mitigated neutrophil infiltration in fungal keratitis, which effectively reduced the severity of keratitis in mice and alleviated the infiltration of inflammatory factors in the mouse cornea. In addition, ZIF-8 reduces the inflammatory response by downregulating the expression of pro-inflammatory cytokines TNF-α, IL-6, and IL-1β after infection in vivo and in vitro.
ZIF-8 has a significant anti-inflammatory and antifungal effect, which provides a new solution for the treatment of fungal keratitis.
真菌性角膜炎是一种严重的致盲眼病。传统的抗真菌药物普遍存在生物利用度低、分散性差和通透性有限等缺点。
开发一种提高生物利用度、分散性和通透性的治疗真菌性角膜炎的新方法。
采用锌离子和 2-甲基咪唑通过配位键连接形成沸石咪唑酯骨架-8（ZIF-8），并通过扫描电子显微镜（SEM）、X 射线衍射（XRD）和 Zeta 电位对其进行表征。采用细胞计数试剂盒（CCK-8）检测 ZIF-8 对 HCECs 和 RAW 264.7 细胞的安全性。通过 Draize 角膜毒性试验评估 ZIF-8 对小鼠角膜上皮的安全性。采用最小抑菌浓度（MIC）、结晶紫染色、碘化丙啶（PI）检测和钙荧光白染色检测 ZIF-8 对真菌生长、生物膜形成和菌丝结构的影响。通过实时定量 PCR 实验（qPCR）和酶联免疫吸附试验（ELISA）评估 ZIF-8 对 RAW 246.7 细胞的抗炎作用。采用临床评分、集落形成单位（CFU）、苏木精-伊红（HE）染色和免疫荧光验证 ZIF-8 对真菌性角膜炎 C57BL/6 雌性小鼠的治疗效果。
体外实验结果表明，ZIF-8 表现出优异的抗真菌作用，在 64μg/mL 时可抑制超过 90%的真菌生长，抑制生物膜形成，并破坏细胞膜。体内实验结果表明，ZIF-8 治疗可降低真菌性角膜炎小鼠角膜真菌负荷和中性粒细胞浸润，有效减轻小鼠角膜炎的严重程度，减轻小鼠角膜中炎症因子的浸润。此外，ZIF-8 通过下调 TNF-α、IL-6 和 IL-1β等促炎细胞因子在体内和体外感染后的表达，降低炎症反应。
ZIF-8 具有显著的抗炎和抗真菌作用，为治疗真菌性角膜炎提供了新的解决方案。
