Elhabal Sammar Fathy, Faheem Ahmed Mohsen, Hababeh Sandra, Nelson Jakline, Elzohairy Nahla A, AbdelGhany Morsy Suzan Awad, Ewedah Tassneim M, Mousa Ibrahim S, Fouad Marwa A, Hamdan Ahmed Mohsen Elsaid
Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Modern University for Technology and Information (MTI), Mokattam, Cairo 11571, Egypt.
Department of Medical Biochemistry and Molecular Biology, Faculty of Medicine, Mansoura University, Mansoura 35516, Egypt.
Pharmaceutics. 2025 May 16;17(5):653. doi: 10.3390/pharmaceutics17050653.
Dry eye disease (DED), also known as "keratoconjunctivitis sicca", is a common chronic ocular surface disease accompanied by inflammation and diminished tear production. Bovine Lactoferrin (BLF), a multi-functional iron-binding glycoprotein found in tears, decreased significantly in patients with DED, used for the treatment of dry eye, conjunctivitis, and ocular inflammation. BLF has limited therapeutic efficacy due to poor ocular bioavailability. This study developed and optimized a BLF-loaded nanosuspension (BLF-NS) using the Box-Behnken Design (BBD). Optimized BLF-NS was then incorporated with polyvinyl pyrrolidone (PVP) and hydroxypropyl methyl cellulose (HPMC) dissolving microneedles (MNs). The formulations were characterized by Scanning and transmission microscopy, DSC, FTIR, ex vivo studies in corneal tissue from sheep and tested for its antibacterial and antifungal efficacy against Methicillin-Resistant (), , and , respectively. Moreover, they were tested for their Benzalkonium chloride (BCL) dry eye in a rabbit model. The optimized nanosuspension showed a vesicle size of (215 ± 0.45) nm, a Z.P (zeta potential) of (-28 ± 0.34) mV, and an Entrapment Efficiency (EE%) of (90 ± 0.66) %. The MNs were fabricated using a ratio of biodegradable polymers, PVP/HPMC. The resulting BLF-NS-MNs exhibited sharp pyramidal geometry with high mechanical strength, ensuring ocular insertion. In vitro release showed 95% lactoferrin release over 24 h, while ex vivo permeation achieved 93% trans-corneal delivery. In vivo, BLF-NS-MNs significantly reduced pro-inflammatory cytokines (TNF-α, IL-6, MMP-9, IL-1β, MCP-1) and upregulated antioxidant and anti-inflammatory genes (PPARA, SOD 1), restoring their levels to near-normal ( < 0.001). The nanosuspension combined with MNs has shown higher ocular tolerance against DED ensured by the Draize and Schirmer Tear Test.
干眼症（DED），也称为“角结膜干燥症”，是一种常见的慢性眼表疾病，伴有炎症且泪液分泌减少。牛乳铁蛋白（BLF）是一种在泪液中发现的多功能铁结合糖蛋白，在干眼症患者中显著减少，可用于治疗干眼症、结膜炎和眼部炎症。由于眼部生物利用度差，BLF的治疗效果有限。本研究采用Box-Behnken设计（BBD）开发并优化了一种载有BLF的纳米混悬液（BLF-NS）。然后将优化后的BLF-NS与聚乙烯吡咯烷酮（PVP）和羟丙基甲基纤维素（HPMC）溶解微针（MNs）相结合。通过扫描和透射显微镜、差示扫描量热法（DSC）、傅里叶变换红外光谱法（FTIR）对制剂进行表征，并在绵羊角膜组织中进行体外研究，分别测试其对耐甲氧西林金黄色葡萄球菌、白色念珠菌和烟曲霉的抗菌和抗真菌功效。此外，还在兔模型中测试了它们对苯扎氯铵（BCL）干眼症的效果。优化后的纳米混悬液显示囊泡大小为（215±0.45）nm，ζ电位（Z.P）为（-28±0.34）mV，包封率（EE%）为（90±0.66）%。微针采用可生物降解聚合物PVP/HPMC的比例制备。所得的BLF-NS-MNs呈现出具有高机械强度的尖锐金字塔形状，确保可插入眼部。体外释放显示24小时内乳铁蛋白释放率达95%，而体外渗透实现了93%的经角膜递送。在体内，BLF-NS-MNs显著降低促炎细胞因子（TNF-α、IL-6、MMP-9、IL-1β、MCP-1）并上调抗氧化和抗炎基因（PPARA、SOD 1），将其水平恢复至接近正常水平（P<0.001）。通过Draize和Schirmer泪液试验确保，纳米混悬液与微针相结合对干眼症表现出更高的眼部耐受性。
