Department of Physical and Quantum Chemistry, Faculty of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland.
Department of Molecular and Cellular Biology, Faculty of Pharmacy, Wroclaw Medical University, Wroclaw, Poland.
Int J Nanomedicine. 2024 May 23;19:4701-4717. doi: 10.2147/IJN.S450181. eCollection 2024.
Numerous failures in melanoma treatment as a highly aggressive form of skin cancer with an unfavorable prognosis and excessive resistance to conventional therapies are prompting an urgent search for more effective therapeutic tools. Consequently, to increase the treatment efficiency and to reduce the side effects of traditional administration ways, herein, it has become crucial to combine photodynamic therapy as a promising therapeutic approach with the selectivity and biocompatibility of a novel colloidal transdermal nanoplatform for effective delivery of hybrid cargo with synergistic effects on melanoma cells.
The self-assembled bilosomes, co-stabilized with L-α-phosphatidylcholine, sodium cholate, Pluronic P123, and cholesterol, were designated, and the stability of colloidal vesicles was studied using dynamic and electrophoretic light scattering, also provided in cell culture medium (Dulbecco's Modified Eagle's Medium). The hybrid compounds - a classical photosensitizer (Methylene Blue) along with a complementary natural polyphenolic agent (curcumin), were successfully co-loaded, as confirmed by UV-Vis, ATR-FTIR, and fluorescent spectroscopies. The biocompatibility and usefulness of the polymer functionalized bilosome with loaded double cargo were demonstrated in vitro cyto- and phototoxicity experiments using normal keratinocytes and melanoma cancer cells.
The in vitro bioimaging and immunofluorescence study upon human skin epithelial (A375) and malignant (Me45) melanoma cell lines established the protective effect of the PEGylated bilosome surface. This effect was confirmed in cytotoxicity experiments, also determined on human cutaneous (HaCaT) keratinocytes. The flow cytometry experiments indicated the enhanced uptake of the encapsulated hybrid cargo compared to the non-loaded MB and CUR molecules, as well as a selectivity of the obtained nanocarriers upon tumor cell lines. The phyto-photodynamic action provided 24h-post irradiation revealed a more significant influence of the nanoplatform on Me45 cells in contrast to the A375 cell line, causing the cell viability rate below 20% of the control.
As a result, we established an innovative and effective strategy for potential metastatic melanoma treatment through the synergism of phyto-photodynamic therapy and novel bilosomal-origin nanophotosensitizers.
黑色素瘤是一种侵袭性很强的皮肤癌，预后不良，对常规疗法有较强的耐药性，导致其治疗失败率居高不下。因此，为了提高治疗效率，降低传统给药方式的副作用，迫切需要寻找更有效的治疗手段。在此，将光动力疗法作为一种很有前途的治疗方法与新型胶体经皮纳米平台的选择性和生物相容性相结合，以有效递送电荷，并对黑色素瘤细胞产生协同作用，这一点变得至关重要。
本研究设计了自组装的双分子层囊泡，并用 L-α-磷脂酰胆碱、胆酸钠、泊洛沙姆 P123 和胆固醇共同稳定。在细胞培养液（Dulbecco's Modified Eagle's Medium）中研究胶体囊泡的稳定性，使用动态光散射和电泳光散射进行研究。通过紫外-可见分光光度法、衰减全反射傅里叶变换红外光谱和荧光光谱证实了成功共载了两种混合化合物 - 经典光敏剂（亚甲蓝）和互补的天然多酚试剂（姜黄素）。体外细胞毒性和光毒性实验证明了聚合物功能化载双载药双分子层囊泡的生物相容性和实用性，该实验使用了正常角质形成细胞和黑色素瘤癌细胞。
体外生物成像和免疫荧光研究在人皮肤上皮（A375）和恶性（Me45）黑色素瘤细胞系上建立了聚乙二醇化双分子层囊泡表面的保护作用。在细胞毒性实验中也证实了这一作用，同时在人皮肤角质形成细胞（HaCaT）上也进行了测定。通过流式细胞术实验表明，与未负载的 MB 和 CUR 分子相比，负载混合载药的纳米载体的摄取能力增强，并且对肿瘤细胞系具有选择性。光照后 24 小时的植物光动力作用显示，与 A375 细胞系相比，纳米载体对 Me45 细胞的影响更为显著，导致细胞存活率低于对照的 20%。
总之，通过植物光动力疗法和新型双分子层囊泡来源的纳米光敏剂的协同作用，我们建立了一种治疗潜在转移性黑色素瘤的创新而有效的策略。
