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

Bio-Design and Manufacturing 2022年第3期5卷 497-511页

作者： Pengtao Wang Xiaofang Wang Bo Wang Xian Li Zhengsong Xie Jie Chen Tasuku Honjo Xiaolin Tu Laboratory of Skeletal Development and Regeneration Institute of Life SciencesChongqing Medical UniversityChongqing 400016China Medical Data Science Academy Chongqing Medical UniversityChongqing 400016China Department of Immunology and Genomic Medicine Graduate School of MedicineKyoto UniversityYoshida Konoe-choSakyo-kuKyoto 606-8501Japan

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 hardmaterial 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 themineralization 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 RBPjκby Cre recombinase.These data indicate that osteocytic Dll4 activates RBPjκ-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.

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

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