壳聚糖-β-环糊精/透明质酸杂化水凝胶抗皮肤光老化作用的实验研究

Chitosan- β -cyclodextrin/hyaluronic acid hybrid hydrogel against skin photoaging: An experimental study

  • 摘要: 背景 光老化是皮肤长期暴露于紫外线所致的慢性损伤过程。最终导致皮肤松弛、皱纹加深、屏障功能下降,因此开发对抗皮肤光老化的新型材料具有重要临床意义。目的 针对高分子量透明质酸难以透皮吸收的难题,本研究旨在构建一种新型壳聚糖-β-环糊精/透明质酸杂化水凝胶(chitosan-β-cyclodextrin/hyaluronic acid hybrid hydrogel,CS-β-CD-HAs),以实现HA的无创高效递送,并系统评估其在动物体内对抗皮肤光老化的功效及潜在机制。方法 制备CS-β-CD-HAs,并利用X-射线衍射(X-ray Diffraction,XRD)、傅里叶变换红外光谱(fourier transform infrared spectroscopy,FTIR)FTIR、扫描电子显微镜(scanning electron microscopy,SEM)、流变学等技术对其进行全面理化表征;9 只SD大鼠随机分为空白对照组、CS-β-CD-HAs 组及超纯水组,每组3 只,分别测定皮肤水分含量以评估其保湿能力;20 只SD 大鼠随机分为空白对照组(Control)、光老化模型组(Model)、水凝胶外用组(CS-β-CD-HAs)与HA注射组(INJ-HAs),每组5 只,通过UVA+UVB递增照射大鼠背部皮肤建立光老化模型,从大体观察及光老化外观评分、组织病理学(H&E、天狼猩红染色)、基因表达(RT-qPCR检测Col-1、Col-3、Elastin、Mmp-3)及生化指标如羟脯氨酸(hydroxyproline,Hyp)含量、超氧化物歧化酶(superoxide dismutase,SOD)活性、丙二醛(malondialdehyde,MDA)含量等多维度评估治疗效果。结果 CS-β-CD-HAs呈现稳定的三维多孔网络结构,具有良好的溶胀性、自愈合特性及优异的透皮性能。细胞实验表明,CS-β-CD-HAs可显著促进人皮肤成纤维细胞(human skin fibroblast,HSF)增殖与迁移,具有良好的细胞相容性。动物实验表明,CS-β-CD-HAs无创外用可提高皮肤水合能力(P<0.001),与光老化模型组相比,CS-β-CD-HAs组可改善光老化皮肤外观评分(P<0.001),减轻了真皮胶原流失(P<0.001),上调Col-1、Col-3 和Elastin mRNA 表达(P<0.05),抑制Mmp-3 mRNA 表达(P<0.01),提升Hyp 含量(P<0.05),增强SOD活性(P<0.05)并降低MDA含量(P<0.05),且与注射同分子量透明质酸相比,两种给药方式抗光老化效果无明显统计学差异。结论 本研究证实CS-β-CD-HAs 是一种高效的非侵入性HA递送材料,能够通过维持细胞外基质稳态与减轻氧化应激损伤来有效改善皮肤光老化。

     

    Abstract: Background Developing innovative non-invasive formulations against skin photoaging holds considerable importance. Objective To address the challenge of poor transdermal absorption of high-molecular-weight hyaluronic acid (HA), this study aimed to develop a novel chitosan-β-cyclodextrin/hyaluronic acid hybrid hydrogel (CS-β-CD-HAs) for the non-invasive and efficient delivery of HA, and to evaluate its efficacy against skin photoaging in vivo.Methods CS-β-CD-HAs were prepared and characterized using XRD, FTIR, SEM, and rheology. 9 SD rats were randomly divided into a blank control group, a CS-β-CDHAs group, and an ultrapure water group to assess skin hydration capacity. 20 SD rats were randomly divided into a control group (Control), a photoaging model group (Model), a hydrogel topical application group (CS-β-CD-HAs), and an HA injection group (INJ-HAs). A photoaging model was established on the dorsal skin of rats by incremental exposure to UVA and UVB irradiation. The therapeutic effects were evaluated through macroscopic observation and photoaging score, histopathology (H&E and Sirius red staining), gene expression (RT-qPCR analysis of Col-1, Col-3, Elastin, and Mmp-3 mRNA), and biochemical assays (Hyp content, SOD activity, MDA content). Results The successfully prepared CS- β -CD-HAs exhibited a stable three-dimensional porous network structure with good swelling capacity, self-healing properties, and excellent transdermal performance. Cell experiments showed that CS-β-CD-HAs significantly promoted the proliferation and migration of human skin fibroblasts (HSFs), indicating good cytocompatibility. Animal experiments demonstrated that topical application of CS-β-CD-HAs significantly enhanced skin hydration capacity (P < 0.001). Compared to the Model group, CS-β-CD-HAs treatment improved the macroscopic photoaging score (P <0.001), alleviated dermal collagen loss (P < 0.001), upregulated mRNA expression of Col-1, Col-3, and Elastin (P < 0.05), suppressed Mmp-3 mRNA expression (P < 0.01), increased Hyp content (P < 0.05), enhanced SOD activity (P < 0.05), and reduced MDA levels (P < 0.05). No statistically significant difference in anti-photoaging efficacy was observed between topical application of CS-β-CD-HAs and injection of the same molecular weight HA.Conclusion CS-β-CD-HAs prove to be an efficient non-invasive HA delivery material, effectively ameliorating skin photoaging by maintaining extracellular matrix homeostasis and mitigating oxidative stress damage.

     

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