Low-intensity pulsed ultrasound promotes cell viability of hUSCs in volumetric bioprinting scaffolds via PI3K/Akt and ERK1/2 pathways.

Jiahui Chen, Yuanchao Li, Xiaoqi Dai, Mei Huang, Meiling Chen, Yifei Zhan, Yaochuan Guo, Yuxuan Du, Liuqiang Li, Meiqin Liu, Maofang Huang, Jun Bian, Dehui Lai
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Abstract

The study aimed to investigate the impact of low-intensity pulsed ultrasound (LIPUS) on human urinary-derived stem cells (hUSCs) viability within three-dimensional (3D) cell-laden gelatin methacryloyl (GelMA) scaffolds. hUSCs were integrated into GelMA bio-inks at concentrations ranging from 2.5% to 10% w/v and then bioprinted using a volumetic-based method. Subsequent exposure of these scaffolds to LIPUS under varying parameters or sham irradiation aimed at optimizing the LIPUS treatment. Assessment of hUSCs viability employed Cell Counting Kit-8 (CCK8), cell cycle analysis, and live&dead cell double staining assays. Additionally, Western blot analysis was conducted to determine protein expression levels. With 3D bio-printed cell-laden GelMA scaffolds successfully constructed, LIPUS promoted the proliferation of hUSCs. Optimal LIPUS conditions, as determined through CCK8 and live&dead cell double staining assays, was achieved at a frequency of 1.5 MHz, a spatial-average temporal-average intensity (ISATA) of 150 mW cm-2, with an exposure duration of 10 min per session administered consecutively for two sessions. LIPUS facilitated the transition from G0/G1 phase to S and G2/M phases and enhanced the phosphorylation of ERK1/2 and PI3K-Akt. Inhibition of ERK1/2 (U0126) and PI3K (LY294002) significantly attenuated LIPUS-induced phosphorylation of ERK1/2 and PI3K-Akt respectively, both of which decreased the hUSC viability within 3D bio-printed GelMA scaffolds. Applying a LIPUS treatment at an ISATA of 150 mW cm-2promotes the growth of hUSCs within 3D bio-printed GelMA scaffolds through modulating ERK1/2 and PI3K-Akt signaling pathways.

低强度脉冲超声通过PI3K/Akt和ERK1/2途径促进体积生物打印支架中hUSCs的细胞活力。
目的: 本研究旨在调查低强度脉冲超声(LIPUS)对三维细胞负载明胶甲基丙烯酰(GelMA)支架中人尿源性干细胞(hUSCs)存活率的影响。随后将这些支架暴露于不同参数下的 LIPUS 或假辐照,目的是优化 LIPUS 处理。采用细胞计数试剂盒-8(CCK8)、细胞周期分析和活死细胞双重染色法评估 hUSCs 的存活率。结果: 随着三维生物打印细胞负载 GelMA 支架的成功构建,LIPUS 促进了 hUSCs 的增殖。通过CCK8和活死细胞双重染色试验确定的最佳LIPUS条件是:频率为1.5 MHz,空间-时间-平均强度(ISATA)为150 mW/cm2,每次照射时间为10分钟,连续照射两次。LIPUS 可促进细胞从 G0/G1 期向 S 期和 G2/M 期过渡,并增强 ERK1/2 和 PI3K-Akt 的磷酸化。抑制ERK1/2(U0126)和PI3K(LY294002)可分别显著减轻LIPUS诱导的ERK1/2和PI3K-Akt磷酸化,这两种抑制都会降低三维生物打印凝胶MA支架中的hUSC活力。 结论: 在150 mW/cm2的ISATA条件下应用LIPUS处理,可通过调节ERK1/2和PI3K-Akt信号通路,促进三维生物打印GelMA支架中的hUSC生长。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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