3D printing of osteocytic Dll4 integrated with PCL for cell fate determination towards osteoblasts in vitro

作者全名："Wang, Pengtao; Wang, Xiaofang; Wang, Bo; Li, Xian; Xie, Zhengsong; Chen, Jie; Honjo, Tasuku; Tu, Xiaolin"

作者地址："[Wang, Pengtao; Wang, Xiaofang; Wang, Bo; Xie, Zhengsong; Chen, Jie; Tu, Xiaolin] Chongqing Med Univ, Inst Life Sci, Lab Skeletal Dev & Regenerat, Chongqing 400016, Peoples R China; [Li, Xian] Chongqing Med Univ, Med Data Sci Acad, Chongqing 400016, Peoples R China; [Honjo, Tasuku] Kyoto Univ, Grad Sch Med, Dept Immunol & Genom Med, Sakyo Ku, Yoshida Konoe Cho, Kyoto 6068501, Japan"

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：BIO-DESIGN AND MANUFACTURING

ESI学科分类：　

WOS号：WOS:000784032600001

JCR分区：Q1

影响因子：7.9

年份：2022

卷号：　

期号：　

开始页：　

结束页：　

文献类型：Article; Early Access

关键词：Integrated 3D printing; PCL scaffold; Cell-laden hydrogel; Osteocytic Dll4; Bone marrow stromal cell; Osteoblast differentiation; Cell viability in hard material; RBPj kappa; Notch signaling

摘要："Since 3D printed hard materials could match the shape of bone, cell survival and fate determination towards osteoblasts in such materials have become a popular research target. In this study, a scaffold of hard material for 3D fabrication was designed to regulate developmental signal (Notch) transduction guiding osteoblast differentiation. We established a polycaprolactone (PCL) and cell-integrated 3D printing system (PCI3D) to reciprocally print the beams of PCL and cell-laden hydrogel for a module. This PCI3D module holds good cell viability of over 87%, whereas cells show about sixfold proliferation in a 7-day culture. The osteocytic MLO-Y4 was engineered to overexpress Notch ligand Dll4, making up 25% after mixing with 75% stromal cells in the PCI3D module. Osteocytic Dll4, unlike other delta-like family members such as Dll1 or Dll3, promotes osteoblast differentiation and the mineralization of primary mouse and a cell line of bone marrow stromal cells when cultured in a PCI3D module for up to 28 days. Mechanistically, osteocytic Dll4 could not promote osteogenic differentiation of the primary bone marrow stromal cells (BMSCs) after conditional deletion of the Notch transcription factor RBPj kappa by Cre recombinase. These data indicate that osteocytic Dll4 activates RBPj kappa-dependent canonical Notch signaling in BMSCs for their oriented differentiation towards osteoblasts. Additionally, osteocytic Dll4 holds a great potential for angiogenesis in human umbilical vein endothelial cells within modules. Our study reveals that osteocytic Dll4 could be the osteogenic niche determining cell fate towards osteoblasts. This will open a new avenue to overcome the current limitation of poor cell viability and low bioactivity of traditional orthopedic implants. [GRAPHICS] ."

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

基金资助正文："The authors thank Professor Dr. Linda Bonewald for her kindly providing the MLO-Y4 cell line and appreciate Dr. Jun Li for his statistical analysis of data. This work is supported by the National Natural Science Foundation of China (Nos. U1601220, 82072450, and 81672118), Chongqing Science and Technology Commission-Basic Science and Frontier Technology Key Project (No. cstc2015jcyjBX0119), and Chongqing Medical University Intelligent Medicine Research Project (No.ZHYX202115)."