Xu Yidan, Zhang Yi, Wang Hao, Tang Huimiao, Zeng Wanxin, Wen Xiang
Department of Dermatology, West China Hospital, Sichuan University, Chengdu, China.
Laboratory of Dermatology, Clinical Institute of Inflammation and Immunology, Frontiers Science Center for Disease-related Molecular Network, West China Hospital, Sichuan University, Chengdu, China.
Lasers Surg Med. 2025 Aug;57(6):528-543. doi: 10.1002/lsm.70033. Epub 2025 May 30.
Microneedle radiofrequency (MRF) is a promising skin rejuvenation treatment. However, the mechanisms underlying its effects on extracellular matrix (ECM) remodeling remain unclear. This study aimed to investigate the immediate histological effects of MRF under varying settings, its short-term impact on collagen and elastin synthesis, and the roles of fibroblasts and adipose-derived stem cells (ADSCs).
Porcine abdominal skin was treated with an MRF device containing 49 insulated microneedles using varying energy parameters (8-12 W; 100-300 ms). Immediate histological responses to treatment were evaluated through hematoxylin and eosin (H&E) staining. Short-term changes in collagen and elastin synthesis at Days 7 and 28 posttreatment were assessed via picrosirius red and Victoria blue staining. Additionally, expression and distribution of ECM remodeling-related proteins (MMPs, TGF-β, EGF, Ki67) and ADSCs were analyzed by multiplex immunohistochemistry (mIHC) and western blot analysis.
H&E staining revealed thermal coagulation zones in the dermis immediately after MRF treatment, with zone size increasing with higher power and longer pulse durations (p < 0.05). By Day 28, Collagen I and III densities and organization significantly improved, with the Collagen I/III ratio rising to 7.05 ± 1.21 in the treatment area (p < 0.01) and 3.90 ± 0.37 in the surrounding dermis (p < 0.001). Elastic fibers also showed increased density. mIHC staining demonstrated significant upregulation of MMP-1, MMP-3, and MMP-13 expression in treated and surrounding dermal regions by Day 7 (p < 0.01); however, by Day 28, MMP-1, MMP-9, and MMP-13 expression significantly decreased (p < 0.05), whereas MMP-3 remained elevated. Furthermore, expression levels of TGF-β, EGF, and Ki67 significantly increased by Day 28 (p < 0.05). mIHC analysis of the fibroblast marker FSP-1 coexpression, along with Western blot analysis of Collagen I, Collagen III, MMP-1, MMP-3, TGF-β, and EGF, revealed similar trends. Notably, significant expression of ADSC markers was detected at Day 7 posttreatment (p < 0.01).
MRF predominantly promotes the synthesis of Collagen I and Collagen III, increasing the Collagen I/III ratio, and regulates the expression of MMP-1, MMP-3, MMP-9, TGF-β, and EGF. These factors collectively drive fibroblast activation, migration, and ECM remodeling. These changes are indicative of the potential for MRF to support skin regeneration and rejuvenation. Preliminary findings suggest that ADSCs may contribute to these regenerative processes.
微针射频（MRF）是一种很有前景的皮肤年轻化治疗方法。然而，其对细胞外基质（ECM）重塑作用的潜在机制仍不清楚。本研究旨在探讨不同参数设置下MRF的即时组织学效应、其对胶原蛋白和弹性蛋白合成的短期影响，以及成纤维细胞和脂肪来源干细胞（ADSCs）的作用。
使用含有49根绝缘微针的MRF设备，以不同的能量参数（8 - 12W；100 - 300ms）处理猪腹部皮肤。通过苏木精和伊红（H&E）染色评估治疗后的即时组织学反应。在治疗后第7天和第28天，通过苦味酸天狼星红和维多利亚蓝染色评估胶原蛋白和弹性蛋白合成的短期变化。此外，通过多重免疫组织化学（mIHC）和蛋白质印迹分析，分析ECM重塑相关蛋白（基质金属蛋白酶、转化生长因子-β、表皮生长因子、Ki67）和ADSCs的表达及分布。
H&E染色显示，MRF治疗后立即在真皮层出现热凝固区，区域大小随功率增加和脉冲持续时间延长而增大（p < 0.05）。到第28天，I型和III型胶原蛋白的密度及排列显著改善，治疗区域的I/III型胶原蛋白比例升至7.05 ± 1.21（p < 0.01），周围真皮层为3.90 ± 0.37（p < 0.001）。弹性纤维密度也增加。mIHC染色显示，到第7天，治疗及周围真皮区域的基质金属蛋白酶-1、基质金属蛋白酶-3和基质金属蛋白酶-13表达显著上调（p < 0.01）；然而，到第28天，基质金属蛋白酶-1、基质金属蛋白酶-9和基质金属蛋白酶-13表达显著下降（p < 0.05），而基质金属蛋白酶-3仍保持升高。此外，到第28天，转化生长因子-β、表皮生长因子和Ki67的表达水平显著增加（p < 0.05）。对成纤维细胞标志物FSP-1共表达的mIHC分析，以及对I型胶原蛋白、III型胶原蛋白、基质金属蛋白酶-1、基质金属蛋白酶-3、转化生长因子-β和表皮生长因子的蛋白质印迹分析，显示出相似趋势。值得注意的是，在治疗后第7天检测到ADSC标志物的显著表达（p < 0.01）。
MRF主要促进I型和III型胶原蛋白的合成，提高I/III型胶原蛋白比例，并调节基质金属蛋白酶-1、基质金属蛋白酶-3、基质金属蛋白酶-9、转化生长因子-β和表皮生长因子的表达。这些因素共同驱动成纤维细胞的激活、迁移和ECM重塑。这些变化表明MRF具有支持皮肤再生和年轻化的潜力。初步研究结果表明，ADSCs可能有助于这些再生过程。
