Eczacioglu Numan, Postina Annika, Ebert Melanie, Laffleur Flavia, Kali Gergely, Seybold Anna, Bernkop-Schnürch Andreas
Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, Innsbruck 6020, Austria; Department of Bioengineering, Karamanoğlu Mehmetbey University, 70110 Karaman, Türkiye.
Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80/82, Innsbruck 6020, Austria.
Acta Biomater. 2025 Aug;202:545-558. doi: 10.1016/j.actbio.2025.07.027. Epub 2025 Jul 12.
This study aimed to compare the performance of dry reverse micelles (dRMs) and wet reverse micelles (wRMs) in self-emulsifying drug delivery systems (SEDDS). Reverse micelles (RMs) were formed using cationic, amphoteric, and non-ionic surfactants with various model dyes incorporated into these RMs. Reverse critical micellar concentration (rCMC), entrapment efficiency (EE), and partition coefficients (logD) were determined. SEDDS were loaded with RMs, and their size distributions, polydispersity index (PDI), and zeta potential were evaluated. Cytotoxicity levels, cellular uptake, and membrane permeability of Caco-2 cells were tested. Furthermore, horseradish peroxidase (HRP) loaded SEDDS-RMs were administered to rats, and plasma HRP concentration and bioavailability were determined. Sorbitan monooleate-based dRMs exhibited an rCMC of 0.95 % and 0.6 % for wRMs, with R values of 4.7 and 43.2, respectively. The lower the hydrophilic lipophilic balance (HLB) of the oily phase, the lower the water uptake capacity of reverse micelles. Incorporating methylene blue into dRMs resulted in a logD of 1.56 and an EE of 97 %, compared to 0.59 and 74 % in wRMs. Cytotoxicity studies revealed over 90 % cell survival at concentrations up to 0.5 % for SEDDS-dRMs, while SEDDS-wRMs caused complete cell death at concentrations exceeding 0.4 %. Transwell diffusion studies showed that SEDDS-dRMs and SEDDS-wRMs enhanced membrane permeability by 7.5- and 13.5-fold, respectively. SEDDS containing HRP-loaded dry and wet reverse micelles provided an oral bioavailability of 11.2 % and 7.9 % in rats, respectively. Conclusion: Due to their ability to achieve higher logD, enhance encapsulation efficiency, and improve the oral bioavailability of a model protein, dRMs are favored over wRMs in SEDDS. STATEMENT OF SIGNIFICANCE: Although several strategies have been proposed to improve the bioavailability of peptide-based drugs, challenges related to stability and oral bioavailability still remain. Reverse micelles, which are nanoscale structures, have great potential with advantages such as high encapsulation efficiency, protection against enzymatic degradation, and increased permeability across biological membranes. This study presents a comparative evaluation of dry and wet reverse micelles in self-emulsifying drug delivery systems. The findings highlight that reverse micelles are a promising nanocarrier system for enhancing drug solubility, stability, and cellular permeability, ultimately contributing to more effective oral drug delivery strategies. This advancement could significantly pave the way for optimizing reverse micelle-based formulations for biomedical applications, especially in protein and peptide therapeutics.
本研究旨在比较干反胶束（dRMs）和湿反胶束（wRMs）在自乳化药物递送系统（SEDDS）中的性能。使用阳离子、两性离子和非离子表面活性剂形成反胶束（RMs），并将各种模型染料掺入这些RMs中。测定了反临界胶束浓度（rCMC）、包封率（EE）和分配系数（logD）。将RMs负载到SEDDS中，并评估其粒径分布、多分散指数（PDI）和zeta电位。测试了Caco-2细胞的细胞毒性水平、细胞摄取和膜通透性。此外，将负载辣根过氧化物酶（HRP）的SEDDS-RMs给予大鼠，并测定血浆HRP浓度和生物利用度。基于脱水山梨醇单油酸酯的dRMs的rCMC为0.95%，wRMs为0.6%，R值分别为4.7和43.2。油相的亲水亲油平衡（HLB）越低，反胶束的吸水能力越低。将亚甲蓝掺入dRMs中时，logD为1.56，EE为97%，而在wRMs中分别为0.59和74%。细胞毒性研究表明，对于SEDDS-dRMs，在浓度高达0.5%时细胞存活率超过90%，而SEDDS-wRMs在浓度超过0.4%时导致细胞完全死亡。Transwell扩散研究表明，SEDDS-dRMs和SEDDS-wRMs分别使膜通透性提高了7.5倍和13.5倍。含有负载HRP的干、湿反胶束的SEDDS在大鼠中的口服生物利用度分别为11.2%和7.9%。结论：由于dRMs能够实现更高的logD、提高包封效率并改善模型蛋白的口服生物利用度，因此在SEDDS中dRMs比wRMs更受青睐。重要性声明：尽管已经提出了几种策略来提高肽类药物的生物利用度，但与稳定性和口服生物利用度相关的挑战仍然存在。反胶束作为纳米级结构，具有很大的潜力，具有高包封效率、防止酶降解以及增加跨生物膜通透性等优点。本研究对自乳化药物递送系统中的干、湿反胶束进行了比较评估。研究结果突出表明，反胶束是一种有前途的纳米载体系统，可用于提高药物溶解度、稳定性和细胞通透性，最终有助于实现更有效的口服给药策略。这一进展可为优化基于反胶束的生物医学应用制剂，特别是蛋白质和肽类治疗药物，显著铺平道路。
