Ansari Ramsha, Farooq Uzma, Ali Ahsan, Abbas Mohd Haider, Rehman Hamidullah Chaudhary Abdur, Jain Pooja, Mirza Mohd Aamir, Iqbal Zeenat
Department of Pharmaceutics, School of Pharmaceutical Education and Research (SPER), Jamia Hamdard, New Delhi 110062, India.
Department of Pharmaceutics, School of Pharmaceutical Education and Research (SPER), Jamia Hamdard, New Delhi 110062, India.
Int J Pharm. 2025 May 15;676:125559. doi: 10.1016/j.ijpharm.2025.125559. Epub 2025 Apr 4.
Ageing produces structural alteration of skin tissue when exposed to UV radiation, pollution and smoking, resulting in functional decline and mortality of the cells. The proposed study aimed at developing a commercially scalable fennel oil (FO) based metformin-loaded nanoemulsion (MTF-NE) incorporated in the hydrogel (MTF-HYD-NEG) providing anti-ageing effect.
The scalable development of phytocompound incorporated nanoemulsion was done using a multi-step process involving excipient screening, optimization of nanoemulsion and thermodynamic stability assessment. The MTF-NE was developed using aqueous titration method containing Tween 80 as the surfactant and Transcutol P as the co-surfactant. The optimized nanoemulsion was incorporated into a hydrogel to form MTF-HYD-NEG. The formulation was characterized based on globule size, polydispersity index (PDI), zeta potential, spreadability, viscosity, and permeability. In vitro drug release and ex vivo permeation studies were conducted to assess sustained drug release. Confocal laser scanning microscopy (CLSM) was used to evaluate skin penetration. The in vivo studies were conducted using UV-B radiation induced mice models and old mice models to assess the anti-ageing efficacy through histopathological analysis, immunohistochemistry (MMP-1 expression), and biochemical assays (GSH, SOD, CAT, and MDA levels).
The characterization of the optimized MTF-NE exhibited a monodispersed pattern with an average globule size of 261.1 nm, PDI of 0.1305 and zeta potential of -4.4 mV. The MTF-NE was then converted into a hydrogel. The permeability of the MTF-HYD-NEG was assessed by in vitro release and ex vivo permeation studies that demonstrated a sustained release effect of Metformin (MTF) with the value of 71.565 ± 4.995 % over 24 hours and 4.503 ± 2.669 mg/cm over 8 hours of study. Additionally, the release profile was further validated by the confocal microscopy through the mice's skin tissue. The mean intensity was observed to be 53.472 which was retained at 4.472 at 50 µm depth of permeation demonstrating the better penetration of the nanocarrier. The data displayed by histopathology and MMP-1 expression by immunohistochemistry showed recuperation in the dermal tissue of the mice model seen in the post-treatment group of MTF-HYD-NEG in comparison to the control group and conventional treatment groups and depletion in MMP-1 production, improved dermal collagen and diminished wrinkles.
The results of the MTF-HYD-NEG formulation displayed enhanced permeability with a prolonged effect, depicting a reduction in MMP-1 expression, which could serve as a promising anti-ageing and antioxidant preparation attributed to the presence of a fennel oil base.
衰老会使皮肤组织在暴露于紫外线辐射、污染和吸烟环境时发生结构改变，导致细胞功能衰退和死亡。本研究旨在开发一种具有商业可扩展性的、基于茴芹油（FO）的载二甲双胍纳米乳剂（MTF-NE），并将其掺入水凝胶中制成MTF-HYD-NEG，以提供抗老化效果。
采用多步骤工艺进行植物化合物纳米乳剂的可扩展性开发，包括辅料筛选、纳米乳剂优化和热力学稳定性评估。MTF-NE采用水相滴定法制备，以吐温80为表面活性剂，二乙二醇单乙醚为助表面活性剂。将优化后的纳米乳剂掺入水凝胶中形成MTF-HYD-NEG。通过球粒大小、多分散指数（PDI）、zeta电位、铺展性、粘度和渗透性对该制剂进行表征。进行体外药物释放和离体渗透研究以评估药物的持续释放。采用共聚焦激光扫描显微镜（CLSM）评估皮肤渗透性。使用紫外线B辐射诱导的小鼠模型和老年小鼠模型进行体内研究，通过组织病理学分析、免疫组织化学（MMP-1表达）和生化测定（谷胱甘肽、超氧化物歧化酶、过氧化氢酶和丙二醛水平）评估抗老化效果。
优化后的MTF-NE表征显示为单分散模式，平均球粒大小为261.1nm，PDI为0.1305，zeta电位为-4.4mV。然后将MTF-NE转化为水凝胶。通过体外释放和离体渗透研究评估MTF-HYD-NEG的渗透性，结果表明二甲双胍（MTF）具有持续释放效果，在24小时内释放值为71.565±4.995%，在8小时研究中释放值为4.503±2.669mg/cm。此外，共聚焦显微镜通过小鼠皮肤组织进一步验证了释放曲线。观察到平均强度为53.472，在渗透深度为50μm时保留在4.472，表明纳米载体具有更好的渗透性。组织病理学数据和免疫组织化学显示的MMP-1表达表明，与对照组和传统治疗组相比，MTF-HYD-NEG治疗后组的小鼠模型真皮组织有所恢复，MMP-1产生减少，真皮胶原蛋白改善，皱纹减少。
MTF-HYD-NEG制剂的结果显示出增强的渗透性和延长的效果，表明MMP-1表达降低，由于存在茴芹油基质，这可能是一种有前景的抗老化和抗氧化制剂。
