Du Mengxing, Chen Chuanxiu, Zhou Yuxin, Song Yixuan, Wang Shan, Tang Shanshan, Yang Jia, Yang Xia, Liu Enli, Yu Yufeng
School of Basic Medical Sciences and Pharmacy, Shanxi Medical University, Taiyuan, 030001, China.
Shanxi Key Laboratory of Functional Proteins, Shanxi Jinbo Bio-Pharmaceutical Co., Ltd, Taiyuan, 030032, China.
Sci Rep. 2025 Jul 24;15(1):26907. doi: 10.1038/s41598-025-07879-6.
To improve the bioavailability and biological activity of recombinant type III humanized collagen (rhCol III), we optimized the preparation process of recombinant type III humanized collagen-loaded liposomes (rhCol III-LIPS) using the film dispersion and periplasmic extrusion methods, with the encapsulation rate as the response value. Then we analyzed the antioxidative activity of rhCol III-LIPS by determining the scavenging rate of free radicals. We investigated the protective effect of rhCol III-LIPS against HO-induced injuries in HaCaT cells by detecting cell vitality, contents of reactive oxygen species (ROS), malondialdehyde (MDA), and activities of antioxidases superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase dismutase (GSH-Px). The results showed, in condition of rhCol III-to-lipid ratio of 1:50, membrane-to-material ratio of 4:1, and ultrasound duration of 20 min, the encapsulation rate of the liposome reached 91.89%±0.59%. The free-radical scavenging rates for 1,1-diphenyl-2-picrohydrazine (DPPH), 2,2-l diazo-bisdiamine salt (ABTS), hydroxyl (OH), and superoxide (O) anions are 87.11% ± 2.54%, 93.72% ± 2.87%, 86.45% ± 1.62%, 76.67% ± 1.56% respectively. The protective effect on oxidation-damaged cells of rhCol III-LIPS was higher than rhCol III or blank liposomes (B-LIPS). In summary, rhCol III-LIPS has good physical stability and in-vitro antioxidative activity, effectively alleviating HO-induced oxidative injuries in HaCaT cells. Our study provides a more stable and efficient strategy for the application of rhCol.
为提高重组III型人源化胶原蛋白（rhCol III）的生物利用度和生物活性，我们采用薄膜分散法和周质挤压法，以包封率为响应值，优化了负载重组III型人源化胶原蛋白的脂质体（rhCol III-LIPS）的制备工艺。然后，通过测定自由基清除率分析rhCol III-LIPS的抗氧化活性。我们通过检测细胞活力、活性氧（ROS）、丙二醛（MDA）含量以及抗氧化酶超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH-Px）的活性，研究了rhCol III-LIPS对HaCaT细胞中HO诱导损伤的保护作用。结果表明，在rhCol III与脂质比例为1:50、膜与材料比例为4:1、超声处理时间为20分钟的条件下，脂质体的包封率达到91.89%±0.59%。对1,1-二苯基-2-苦基肼（DPPH）、2,2'-偶氮二（3-乙基苯并噻唑啉-6-磺酸）二铵盐（ABTS）、羟基（OH）和超氧阴离子（O）的自由基清除率分别为87.11%±2.54%、93.72%±2.87%、86.45%±1.62%、76.67%±1.56%。rhCol III-LIPS对氧化损伤细胞的保护作用高于rhCol III或空白脂质体（B-LIPS）。综上所述，rhCol III-LIPS具有良好的物理稳定性和体外抗氧化活性，能有效减轻HO诱导的HaCaT细胞氧化损伤。我们的研究为rhCol的应用提供了一种更稳定有效的策略。
