组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2024, Vol. 20 ›› Issue (5): 528-.

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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

  

  • 出版日期:2024-10-08 发布日期:2024-11-25

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

  • Online:2024-10-08 Published:2024-11-25

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摘要：

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。

关键词:

Abstract:

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，
γH2AX fluorescence staining was respectively performed in in vitro assays and in vivo photodamage models by applying 
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.