紫外线诱导下的真皮成纤维细胞亚型改变及硅酸锌生物陶瓷对光损伤皮肤的修复作用

Ultraviolet-induced changes in dermal fibroblast subtypes and repair of photodamaged skin by zinc silicate bioceramics

目的紫外线B(Ultraviolet B,UVB)对皮肤的损害包括破坏皮肤屏障功能、引发皮肤疾病以及促进恶性肿瘤的发生。真皮层的成纤维细胞(Fibroblasts,Fb)可分为真皮乳头层成纤维细胞(Papillary fibroblasts,Fp)与真皮网状层成纤维细胞(Reticular fibroblasts,Fr),在皮肤再生过程中扮演重要角色。本研究阐明光损伤会导致不同亚型Fb的改变,并针对这一靶点,寻找一种可以修复光损伤的治疗策略。方法构建皮肤光损伤小鼠模型,采集生物样本进行HE和Masson染色评估组织结构,DHE染色检测组织中活性氧(Reactive oxygen species,ROS)水平,γH2AX染色标记DNA损伤的细胞,荧光染色Lrig1及Dlk1定性定量不同Fb亚型。另外,UVB诱导下培养Fb,流式细胞术检测Fb的表面标志物,qPCR检测胶原纤维合成分泌相关mRNA表达。在体外试验及体内光损伤模型中,分别应用ZnCS配制的离子溶液及装载ZnCS的微针贴片干预,进行Lrig1和DHE、γH2AX荧光染色。结果急性光损伤皮肤真皮层炎症细胞浸润,胶原纤维排列紊乱,ROS表达水平在表皮及真皮浅层显著增加,伴有真皮浅层Fp的DNA损伤更甚。与对照小鼠相比,光损伤皮肤真皮层Lrig1+Fb显著减少,Dlk1+Fb没有明显差异。Fb经UVB辐照后,流式检测示Fb中Lrig1+Fb比例明显减少,同时COL1A1表达下调,MMP3和MMP9表达上调;最后,硅酸盐生物陶瓷(ZnCS)促进Fp的增殖,ZnCS干预后,光损伤皮肤的ROS生成和损伤情况明显减轻。结论本研究首次发现Fp减少与光损伤进展有关;同时,对Fp增殖有促进作用的ZnCS对光损伤皮肤具有显著修复作用。

Objective Ultraviolet B(UVB)radiation damages the skin by disrupting the skin barrier function,causing various skin diseases,and facilitating the formation of malignant tumors.Fibroblasts in the dermis are categorized into papillary fibroblasts(Fp)and reticular fibroblasts(Fr),crucial for skin regeneration.The present study elucidates that photodamage leads to alterations in different subtypes of Fb,and addresses this phenomenon to find a therapeutic strategy that can repair photodamage.Methods A mouse model of skin photodamage was established and biological specimens were collected for subsequent analyses,including HE and Masson staining to evaluate tissue structure,DHE staining to detect ROS levels in tissues,γH2AX staining to identify cells with DNA damage,immunofluorescence staining for Lrig1 and Dlk1 to identify distinct Fb subtypes.Furthermore,UVB was used to induce and culture fibroblasts,followed by flow cytometry to detect surface markers,and qPCR to assess mRNA expression related to collagen synthesis and secretion.Lrig1 and DHE,ZnCS-configured ionic solutions and microneedle patch interventions loaded with ZnCS.Results In photodamaged skin,the dermis exhibited infiltrative inflammatory cells and disrupted collagen fiber structure with reduced content,accompanied by a significant increase in ROS expression throughout the epidermis and superficial dermis,accompanied by more DNA damage in superficial dermal fibroblasts.Compared to the control group,there was a marked reduction in Lrig1+Fb cells within the photodamaged skin dermis,while no significant difference was detected in Dlk1+Fb cells.Following UVB irradiation of fibroblasts,flow cytometry analysis revealed a substantial decrease in the proportion of Lrig1+Fb cells among the fibroblasts.Additionally,downregulation of COL1A1 expression was noted,alongside upregulation of MMP3 and MMP9 expressions.Finally,silicate bioceramics(ZnCS)promoted Fp proliferation,and ROS generation and damage in photodamaged skin were significantly reduced after intervention with ZnCS.Conclusion Fp reducing is associated with the progression of photodamage to photoaging.Meanwhile,ZnCS,which has a promoting effect on Fp proliferation,exerts a significant repairing effect on photodamaged skin.