具有光热抗菌活性的聚多巴胺修饰藻蓝蛋白纳米颗粒促进小鼠伤口皮肤愈合

doi:10.12122/j.issn.1673-4254.2025.09.15

摘要/Abstract

摘要：

目的评估聚多巴胺修饰藻蓝蛋白纳米颗粒（PDA@PC NPs）在光热、抗菌和促进创面修复的效果。方法采用“一锅法”以藻蓝蛋白（C-PC）和盐酸多巴胺为原料制备PDA@PC NPs纳米颗粒并进行表征。体外实验：考察808 nm激光照射后PDA@PC NPs纳米颗粒的升温效果；采用CCK-8法检测PDA@PC NPs的生物相容性；采用平板菌落计数法评估PDA@PC NPs的光热抗菌性能。体内实验：选取SPF级6~8周的BALB/c雄性小鼠，使用无菌打孔器在小鼠脊柱两侧对称建立2个全层皮肤创面（1.0 cm×1.0 cm），并在每个创面均匀接种200 μL金黄色葡萄球菌悬液，分为对照组、PDA@PC NPs组和PDA@PC NPs光照治疗组，于第0、7、14天对创面进行拍照后计算创面愈合率，比较组织形态学变化，进而评估创面愈合情况。结果体外实验结果显示，PDA@PC NPs在质量浓度高达500 μg/mL下显示低细胞毒性；PDA@PC NPs显示出显著的光热性能，并且经过808 nm激光照射后呈现出极强的光热抗菌效果（P<0.001）。体内实验显示，治疗7、14 d后，PDA@PC NPs组、PDA@PC NPs光照组创面恢复较快，其中PDA@PC NPs光照组更为明显（P<0.001）。HE和Masson染色显示，PDA@PC NPs组和PDA@PC NPs光照组的创面观察到大量的肉芽组织形成和胶原沉积，其中以PDA@PC NPs光照组最明显。结论 PDA@PC NPs具备良好的光热抗菌作用，并可有效促进伤口愈合。

关键词: 聚多巴胺, 藻蓝蛋白, 光热治疗, 伤口愈合

Abstract:

Objective To evaluate the photothermal and antibacterial activities of polydopamine-modified phycocyanin nanoparticles (PDA@PC NPs) and their capacity for promoting wound healing. Methods PDA@PC NPs were synthesized from phycocyanin (C-PC) and dopamine hydrochloride using a one-pot method. The photothermal activity of the nanoparticles was assessed in vitro by 808 nm laser irradiation, their biocompatibility was evaluated using CCK-8 assay, and their photothermal antibacterial activity by plate colony counting. In adult male BALB/c mice, two symmetrical full-thickness skin wounds (1.0 cm ×1.0 cm) were created on both sides of the spine, and 200 μL of Staphylococcus aureus suspension was inoculated into the wounds. The mice were divided into control group, PDA@PC NPs group, and PDA@PC NPs with laser irradiation group, and wound healing rates and histomorphological changes in the wound tissues were evaluated on days 0, 7 and 14 after modeling. Results The synthesized PDA@PC NPs exhibited no obvious cytotoxicity up to a concentration of 500 μg/mL and showed strong photothermal and antibacterial activities in response to 808 nm laser irradiation. In the mouse models, the size of the infected skin wounds showed substantial reduction at 7 and 14 days in PDA@PC NPs group and PDA@PC NPs with laser irradiation group, and the mean wound healing rate was faster in the latter group. HE staining and Masson's trichrome staining revealed extensive granulation tissue formation and collagen deposition on the wound surfaces in both of the treatment groups, and these changes were more obvious in the PDA@PC NPs with laser irradiation group. Conclusion PDA@PC NPs possess excellent photothermal and antibacterial activities and can effectively promote wound healing in mice.

Key words: polydopamine, phycocyanin, photothermal therapy, wound healing

张晨, 许智, 李想, 何彭翼翔, 曲凯林, 宁奇, 金奕玏, 杨宿睿, 吴旭. 具有光热抗菌活性的聚多巴胺修饰藻蓝蛋白纳米颗粒促进小鼠伤口皮肤愈合[J]. 南方医科大学学报, 2025, 45(9): 1959-1966.

Chen ZHANG, Zhi XU, Xiang LI, Pengyixiang HE, Kailin QU, Qi NING, Yile JIN, Surui YANG, Xu WU. Polydopamine-modified phycocyanin nanoparticles with photothermal antimicrobial activity promote skin wound healing in mice[J]. Journal of Southern Medical University, 2025, 45(9): 1959-1966.

图/表 6

图1 PDA@PC NPs水中的DLS(A), PDA@PC NPs电镜图(B), PDA@PC NPs的红外光谱图(C)

Fig.1 DLS of PDA@PC NPs in water (A), electron microscopy image of PDA@PC NPs (B), and its infrared spectra (C).

图2 紫外吸收光谱和光热表征

Fig.2 Ultraviolet absorption spectrum and photothermal characterization of the nanoparticles. A: UV-vis spectra of PDA@PC NPs with different concentrations. B: Mass absorption coefficient of PDA@PC NPs. C: Temperature curves of PDA@PC NPs with varying concentrations under 808 nm laser irradiation at 2.0 W/cm2. D: Temperature curves of PDA@PC NPs (500 μg/mL) under 808 nm laser irradiation at different power densities. E: Thermostability of PDA@PC NPs(500 μg/mL) in water exposed to 808 nm laser (2.5 W/cm2) for 4 on/off cycles. F: Photothermal conversion profiles of PDA@PC NPs. G: In vitro photothermal images of centrifuge tubes filled with various concentrations of PDA@PC NPs.

图3 体外细胞实验, 用不同浓度的PDA@PC NPs处理的L929细胞在24 h(A)和48 h(B)的细胞活力

Fig.3 Cell viability of L929 cells treated with PDA@PC NPs at different concentrations for 24 h (A)and 48 h (B).

图4 不同处理的抗菌能力评价

Fig.4 Evaluation of antimicrobial ability of the nanoparticles. A: Plate colony growth. B: Staphylococcus aureus colony formation and bacterial survival rate. C: E. coli colony formation and bacterial survival rate. ***P<0.001.

图5 小鼠创面愈合情况

Fig.5 Representative images of wound healing in mice (A), the healing rates (B), and simulated images of the healing processes (C). ***P<0.001, ****P<0.0001.

图6 小鼠创面组织病理学变化

Fig.6 Histopathological changes in the wound tissues of the mice. A: HE staining results. B: Masson staining results.

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