Department of Pharmaceutical Sciences, Faculty of Pharmacy, Universitas Airlangga, Campus C UNAIR Mulyorejo, Surabaya 60115, Indonesia; Stem Cell Research and Development Center, Universitas Airlangga, 2nd Floor Institute of Tropical Disease Building, Campus C UNAIR Mulyorejo Surabaya 60115, Indonesia; Skin and Cosmetic Technology Centre of Excellence, Faculty of Pharmacy, Universitas Airlangga, Campus C UNAIR Mulyorejo, Surabaya 60115, Indonesia.
Master Program of Pharmaceutical Sciences, Faculty of Pharmacy, Universitas Airlangga, Campus C UNAIR Mulyorejo, Surabaya 60115, Indonesia.
Eur J Pharm Biopharm. 2024 Nov;204:114482. doi: 10.1016/j.ejpb.2024.114482. Epub 2024 Sep 13.
Microneedles offer a promising solution to enhancing dermal delivery of amniotic mesenchymal stem cell metabolite product (AMSC-MP), which contains hydrophilic protein components with high molecular weight, for the purposes of skin rejuvenation and improving human health. This study aimed to evaluate the physicochemical characteristics and in vivo efficacy of AMSC-MP-loaded microneedle patches for effectively regenerating skin tissues in UV-aging induced mice. Dissolving microneedle patches, composed of polyvinyl alcohol with an MW of 9-10 kDa and polyvinylpyrrolidone with an MW of 56 kDa, were fabricated using the double-casting method at three AMSC-MP concentrations: i.e., 30 % (MN30), 25 % (MN25), and 20 % (MN20). The microneedles patches were then evaluated for morphological, mechanical resistance, and insertion properties. An ex vivo release study was also conducted using the Franz cell method, and in vivo efficacy and irritation were then determined through collagen density scores, fibroblast cell counts, and skin irritation studies of UV-aging induced mice. The AMSC-MP microneedles displayed a pyramidal shape with 500 µm sharp tips. Mechanical testing revealed that MN30 achieved its deepest insertion into Parafilm® M (447.44 ± 37.21 µm), while MN25 achieved its deepest insertion into full-thickness porcine skin (717.92 ± 25.40 µm). The study revealed a controlled EGF release for up to 24 h, with MN20 exhibiting the highest deposition (55.94 ± 12.34 %). These findings demonstrate the successful penetration of microneedles through the stratum corneum and viable epidermis. Collagen density scores and fibroblast cell counts were significantly higher in all microneedle formulations than the control, with MN30 having the highest values. Inflammatory cell counts indicated minimal presence suggesting non-irritation in the in vivo study. Dissolving microneedle patches exhibited favorable characteristics and efficiently delivered AMSC-MP with minimal potential for irritation, providing potential technology for delivering biological anti-aging agents for the purposes of fostering skin regeneration.
微针为提高羊膜间充质干细胞代谢产物（AMSC-MP）的透皮传递提供了一种很有前途的解决方案，AMSC-MP 含有高相对分子质量的亲水性蛋白成分，用于皮肤年轻化和改善人类健康。本研究旨在评估负载 AMSC-MP 的微针贴片的理化特性和体内功效，以有效再生 UV 老化诱导的小鼠皮肤组织。使用 MW 分别为 9-10 kDa 的聚乙烯醇和 MW 为 56 kDa 的聚乙烯吡咯烷酮，通过双铸法制备微针贴片，包括 3 种 AMSC-MP 浓度：30%（MN30）、25%（MN25）和 20%（MN20）。然后评估微针贴片的形态、机械阻力和插入特性。还使用 Franz 细胞法进行了体外释放研究，并通过胶原密度评分、成纤维细胞计数以及 UV 老化诱导的小鼠皮肤刺激研究来确定体内功效和刺激作用。AMSC-MP 微针呈 500 µm 尖顶的金字塔形。机械测试表明，MN30 能够最深地插入 Parafilm® M（447.44±37.21 µm），而 MN25 能够最深地插入全厚猪皮（717.92±25.40 µm）。研究表明，EGF 可控制释放长达 24 h，MN20 的沉积量最高（55.94±12.34%）。这些发现表明微针成功穿透了角质层和有活力的表皮。所有微针制剂的胶原密度评分和成纤维细胞计数均显著高于对照组，MN30 组的值最高。在体内研究中，炎性细胞计数表明存在的炎症细胞很少，提示微针无刺激性。溶解微针贴片表现出良好的特性，可有效传递 AMSC-MP，且潜在刺激性最小，为传递生物抗衰老剂以促进皮肤再生提供了潜在技术。
