Three-Dimensional Matrix Stiffness Activates the Piezo1-AMPK-Autophagy Axis to Regulate the Cellular Osteogenic Differentiation

作者全名："Wu, Yanqiu; Xu, Xinxin; Liu, Fengyi; Jing, Zheng; Shen, Danfeng; He, Ping; Chen, Tao; Wu, Tianli; Jia, Hengji; Mo, Dingqiang; Li, Yuzhou; Zhang, He; Yang, Sheng"

作者地址："[Wu, Yanqiu; Xu, Xinxin; Liu, Fengyi; Jing, Zheng; Shen, Danfeng; He, Ping; Chen, Tao; Wu, Tianli; Jia, Hengji; Mo, Dingqiang; Li, Yuzhou; Zhang, He; Yang, Sheng] Chongqing Med Univ, Coll Stomatol, Chongqing 401147, Peoples R China; [Wu, Yanqiu; Xu, Xinxin; Liu, Fengyi; Jing, Zheng; Shen, Danfeng; He, Ping; Chen, Tao; Wu, Tianli; Jia, Hengji; Mo, Dingqiang; Li, Yuzhou; Zhang, He; Yang, Sheng] Chongqing Key Lab Oral Dis & Biomed Sci, Chongqing 400016, Peoples R China; [Wu, Yanqiu; Xu, Xinxin; Liu, Fengyi; Jing, Zheng; Shen, Danfeng; He, Ping; Chen, Tao; Wu, Tianli; Jia, Hengji; Mo, Dingqiang; Li, Yuzhou; Zhang, He; Yang, Sheng] Chongqing Municipal Key Lab Oral Biomed Engn Highe, Chongqing 400016, Peoples R China"

通信作者："Zhang, H; Yang, S (通讯作者)，Chongqing Med Univ, Coll Stomatol, Chongqing 401147, Peoples R China.; Zhang, H; Yang, S (通讯作者)，Chongqing Key Lab Oral Dis & Biomed Sci, Chongqing 400016, Peoples R China.; Zhang, H; Yang, S (通讯作者)，Chongqing Municipal Key Lab Oral Biomed Engn Highe, Chongqing 400016, Peoples R China."

来源：ACS BIOMATERIALS SCIENCE & ENGINEERING

ESI学科分类：MATERIALS SCIENCE

WOS号：WOS:001023142900001

JCR分区：Q2

影响因子：5.8

年份：2023

卷号：9

期号：8

开始页：4735

结束页：4746

文献类型：Article

关键词：Piezo1; extracellular matrix; AMPK; 3D cell culture; autophagy

摘要："Extracellular matrix (ECM) stiffness is a key stimulusaffectingcellular differentiation, and osteoblasts are also in a three-dimensional(3D) stiff environment during the formation of bone tissues. However,it remains unclear how cells perceive matrix mechanical stiffnessstimuli and translate them into intracellular signals to affect differentiation.Here, for the first time, we constructed a 3D culture environmentby GelMA hydrogels with different amino substitution degrees and foundthat Piezo1 expression was significantly stimulated by the stiff matrixwith high substitution; meanwhile, the expressions of osteogenic markersOSX, RUNX2, and ALP were also observably improved. Moreover, knockdownof Piezo1 in the stiff matrix revealed significant reduction of theabovementioned osteogenic markers. In addition, in this 3D biomimeticECM, we also observed that Piezo1 can be activated by the static mechanicalconditions of the stiff matrix, leading to the increase of the intracellularcalcium content and accompanied with a continuous change in cellularenergy levels as ATP was consumed during cellular differentiation.More surprisingly, we found that in the 3D stiff matrix, intracellularcalcium as a second messenger promoted the activation of the AMP-activatedprotein kinase (AMPK) and unc-51-like autophagy-activated kinase 1(ULK1) axis and modestly modulated the level of autophagy, bringingit more similar to differentiated osteoblasts, with increased ATPenergy metabolism consumption. Our study innovatively clarifies theregulatory role of the mechanosensitive ion channel Piezo1 in a staticmechanical environment on cellular differentiation and verifies theactivation of the AMPK-ULK1 axis in the cellular ATP energy metabolismand autophagy level. Collectively, our research develops the understandingof the interaction mechanisms of biomimetic extracellular matrix biomaterialsand cells from a novel perspective and provides a theoretical basisfor bone regeneration biomaterials design and application."

基金机构："National Natural Science Foundation of China [82171010, 82170936, 82001103]; Natural Science Foundation of Chongqing [cstc2021jcyj-jqX0028, CSTB2022NSCQ-MSX0925, CSTB2022NSCQ-MSX0098]; Chongqing Science and Health Joint Project [2020GDRC015]; Program for Youth Innovation in Future Medicine, Chongqing Medical University [W0079]; Scientific and Technological Research Program of Chongqing Municipal Education Commission [KJQN202200422, KJQN202200434]; China Postdoctoral Science Foundation [2022M720598]; Chongqing Scientific Research and Innovation Project for Postgraduates [CYB21188, CYB22216, CYS22358]"

基金资助正文："This work was supported by the National Natural Science Foundation of China (Grant Nos. 82171010, 82170936, and 82001103), the Natural Science Foundation of Chongqing (Grant Nos. cstc2021jcyj-jqX0028, CSTB2022NSCQ-MSX0925, and CSTB2022NSCQ-MSX0098), the Chongqing Science and Health Joint Project (Grant No. 2020GDRC015), the Program for Youth Innovation in Future Medicine, Chongqing Medical University (Grant No. W0079), the project supported by the Scientific and Technological Research Program of Chongqing Municipal Education Commission (Grant Nos. KJQN202200422 and KJQN202200434), the China Postdoctoral Science Foundation (No. 2022M720598), and the Chongqing Scientific Research and Innovation Project for Postgraduates (Grant Nos. CYB21188, CYB22216, and CYS22358)."