CHIR99021-Treated Osteocytes with Wnt Activation in 3D-Printed Module Form an Osteogenic Microenvironment for Enhanced Osteogenesis and Vasculogenesis

作者全名："Luo, Yisheng; Liu, Yangxi; Wang, Bo; Tu, Xiaolin"

作者地址："[Luo, Yisheng; Liu, Yangxi; Wang, Bo; Tu, Xiaolin] Chongqing Med Univ, Inst Life Sci, Lab Skeletal Dev & Regenerat, Chongqing 400016, Peoples R China"

通信作者："Tu, XL (通讯作者)，Chongqing Med Univ, Inst Life Sci, Lab Skeletal Dev & Regenerat, Chongqing 400016, Peoples R China."

来源：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES

ESI学科分类：CHEMISTRY

WOS号：WOS:000955387500001

JCR分区：Q1

影响因子：4.9

年份：2023

卷号：24

期号：6

开始页：　

结束页：　

文献类型：Article

关键词：osteocyte; osteogenic microenvironment; COOME; bone marrow stromal cells; CHIR99021

摘要："Finding a bone implant that has high bioactivity that can safely drive stem cell differentiation and simulate a real in vivo microenvironment is a challenge for bone tissue engineering. Osteocytes significantly regulate bone cell fate, and Wnt-activated osteocytes can reversely regulate bone formation by regulating bone anabolism, which may improve the biological activity of bone implants. To achieve a safe application, we used the Wnt agonist CHIR99021 (C91) to treat MLO-Y4 for 24 h, in a co-culture with ST2 for 3 days after withdrawal. We found that the expression of Runx2 and Osx increased, promoted osteogenic differentiation, and inhibited adipogenic differentiation in the ST2 cells, and these effects were eliminated by the triptonide. Therefore, we hypothesized that C91-treated osteocytes form an osteogenic microenvironment (COOME). Subsequently, we constructed a bio-instructive 3D printing system to verify the function of COOME in 3D modules that mimic the in vivo environment. Within PCI3D, COOME increased the survival and proliferation rates to as high as 92% after 7 days and promoted ST2 cell differentiation and mineralization. Simultaneously, we found that the COOME-conditioned medium also had the same effects. Therefore, COOME promotes ST2 cell osteogenic differentiation both directly and indirectly. It also promotes HUVEC migration and tube formation, which can be explained by the high expression of Vegf. Altogether, these results indicate that COOME, combined with our independently developed 3D printing system, can overcome the poor cell survival and bioactivity of orthopedic implants and provide a new method for clinical bone defect repair."

基金机构："National Natural Science Foundation of China [U1601220, 81672118, 82072450]; Chongqing Science and Technology Commission-Basic Science and Frontier Technology Key Project [cstc2015jcyjBX0119]"

基金资助正文："This research was funded by the National Natural Science Foundation of China (U1601220, 81672118, 82072450), Chongqing Science and Technology Commission-Basic Science and Frontier Technology Key Project (cstc2015jcyjBX0119)."