Iftode Loredana, Cadinoiu Anca Niculina, Raţă Delia Mihaela, Atanase Leonard Ionuț, Vochiţa Gabriela, Rădulescu Luminița, Popa Marcel, Gherghel Daniela
Faculty of Medicine, "Grigore T. Popa" University of Medicine and Pharmacy, 700115 Iasi, Romania.
"Cristofor Simionescu" Faculty of Chemical Engineering and Environmental Protection, "Gheorghe Asachi" Technical University, 700050 Iasi, Romania.
Int J Mol Sci. 2025 Jan 22;26(3):922. doi: 10.3390/ijms26030922.
Liposomes are intensively used as nanocarriers for biology, biochemistry, medicine, and in the cosmetics industry and their non-toxic and biocompatible nature makes these vesicles attractive systems for biomedical applications. Moreover, the conjugation of specific ligands to liposomes increases their cellular uptake and therapeutic efficiency. Considering these aspects, the aim of the present study was to obtain new formulations of cationic liposomes coated with dual-peptide functionalized carboxymethyl chitosan (CMCS) for the treatment of inner ear diseases. In order to achieve efficient active targeting and ensuring a high efficacy of the treatment, CMCS was functionalized with Tet1 peptide, to target specific ear cells, and TAT peptide, to ensure cellular penetration. Furthermore, dexamethasone phosphate was loaded as a model drug for the treatment of ear inflammation. The infrared spectroscopy confirmed the functionalization of CMCS with the two specific peptides. The mean diameter of the uncovered liposomes varied between 167 and 198 nm whereas the CMCS-coated liposomes ranged from 179 to 202 nm. TEM analysis showed the spherical shape and unilamellar structure of liposomes. The release efficiency of dexamethasone phosphate after 24 h from the uncoated liposomes was between 37 and 40% and it appeared that the coated liposomes modulated this release. The obtained results demonstrated that the liposomes are hemocompatible since, for a tested concentration of 100 µg/mL, the liposome suspension had a lysis of erythrocytes lower than 2.5% after 180 min of incubation. In addition, the peptide-functionalized CMCS-coated liposomes induced a non-significant effect on the viability of normal V79-4 cells after 48 h, at the highest doses. Values of 71.31% were recorded (CLCP-1), 77.28% (CLCP-2) and 74.36% (CLCP-3), correlated with cytotoxic effects of 28.69%, 22.72%, and 25.64%.
脂质体被广泛用作生物学、生物化学、医学以及化妆品行业的纳米载体，其无毒且具有生物相容性，使得这些囊泡成为生物医学应用中具有吸引力的系统。此外，将特定配体与脂质体偶联可增加其细胞摄取和治疗效率。考虑到这些方面，本研究的目的是获得用双肽功能化羧甲基壳聚糖（CMCS）包被的阳离子脂质体新制剂，用于治疗内耳疾病。为了实现有效的主动靶向并确保治疗的高效性，CMCS用Tet1肽进行功能化以靶向特定耳细胞，并用TAT肽进行功能化以确保细胞穿透。此外，负载磷酸地塞米松作为治疗耳部炎症的模型药物。红外光谱证实了CMCS与两种特定肽的功能化。未包被的脂质体平均直径在167至198nm之间，而CMCS包被的脂质体直径范围为179至202nm。透射电子显微镜分析显示脂质体呈球形且为单层结构。未包被的脂质体在24小时后磷酸地塞米松的释放效率在37%至40%之间，且似乎包被的脂质体调节了这种释放。获得的结果表明脂质体具有血液相容性，因为在测试浓度为100μg/mL时，脂质体悬浮液在孵育180分钟后红细胞裂解率低于2.5%。此外，在最高剂量下，肽功能化的CMCS包被的脂质体在48小时后对正常V79 - 4细胞的活力没有显著影响。记录的值分别为71.31%（CLCP - 1）、77.28%（CLCP - 2）和74.36%（CLCP - 3），细胞毒性效应分别为28.69%、22.72%和25.64%。
