Cell Ratio-Dependent Osteoblast–Endothelial Cell Crosstalk Promoting Osteogenesis–Angiogenesis Coupling via Regulation of Microfluidic Perfusion and Paracrine Signaling

Osteogenesis–angiogenesis coupling, a dynamic and coordinated interaction between skeletal and vascular cells, is essential for fracture healing. However, the effects of these cell ratios and their interactions under microfluidic perfusion and paracrine signaling on osteogenesis–angiogenesis couplin...

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Main Authors: Yuexin Wang, Shu Chen, Wenwen Fan, Sixian Zhang, Xi Chen
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Micromachines
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Online Access:https://www.mdpi.com/2072-666X/16/5/539
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author Yuexin Wang
Shu Chen
Wenwen Fan
Sixian Zhang
Xi Chen
author_facet Yuexin Wang
Shu Chen
Wenwen Fan
Sixian Zhang
Xi Chen
author_sort Yuexin Wang
collection DOAJ
description Osteogenesis–angiogenesis coupling, a dynamic and coordinated interaction between skeletal and vascular cells, is essential for fracture healing. However, the effects of these cell ratios and their interactions under microfluidic perfusion and paracrine signaling on osteogenesis–angiogenesis coupling have rarely been reported. In this study, dynamic and static models of osteogenesis–angiogenesis coupling were developed and the osteogenic and angiogenic effects of the two models were compared. Static co-cultures of MC3T3-E1 and bEnd.3 cells in Transwell inserts showed a cell ratio-dependent reciprocal relation: a ratio of 1:1 (MC3T3-E1:bEnd.3) favored osteogenesis, whereas a ratio of 2:1 (MC3T3-E1:bEnd.3) promoted angiogenesis. On that basis, we developed an osteogenesis–angiogenesis coupling chip based on microfluidic technology. The microfluidic perfusion within the chip further enhanced the mineralizing effect of osteoblasts and the angiogenic effect of endothelial cells, respectively, and increased the secretion of vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2) compared to the static Transwell insert model. The results suggest that the microfluidic chip enhanced the potential of osteogenesis–angiogenesis coupling mediated by paracrine signaling. Overall, the chip is not only a powerful model for understanding bone–vascular interaction but also a scalable platform for high-throughput drug screening and personalized therapy development for fractures.
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spelling doaj-art-e9105db8a9d04fe7bb3a2785b9bd09fd2025-08-20T03:14:29ZengMDPI AGMicromachines2072-666X2025-04-0116553910.3390/mi16050539Cell Ratio-Dependent Osteoblast–Endothelial Cell Crosstalk Promoting Osteogenesis–Angiogenesis Coupling via Regulation of Microfluidic Perfusion and Paracrine SignalingYuexin Wang0Shu Chen1Wenwen Fan2Sixian Zhang3Xi Chen4Key Laboratory for Ultrafine Materials of Ministry of Education, Frontiers Science Center for Materiobiology and Dynamic Chemistry, Engineering Research Center for Biomedical Materials of Ministry of Education, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, ChinaKey Laboratory for Ultrafine Materials of Ministry of Education, Frontiers Science Center for Materiobiology and Dynamic Chemistry, Engineering Research Center for Biomedical Materials of Ministry of Education, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, ChinaKey Laboratory for Ultrafine Materials of Ministry of Education, Frontiers Science Center for Materiobiology and Dynamic Chemistry, Engineering Research Center for Biomedical Materials of Ministry of Education, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, ChinaKey Laboratory for Ultrafine Materials of Ministry of Education, Frontiers Science Center for Materiobiology and Dynamic Chemistry, Engineering Research Center for Biomedical Materials of Ministry of Education, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, ChinaKey Laboratory for Ultrafine Materials of Ministry of Education, Frontiers Science Center for Materiobiology and Dynamic Chemistry, Engineering Research Center for Biomedical Materials of Ministry of Education, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, ChinaOsteogenesis–angiogenesis coupling, a dynamic and coordinated interaction between skeletal and vascular cells, is essential for fracture healing. However, the effects of these cell ratios and their interactions under microfluidic perfusion and paracrine signaling on osteogenesis–angiogenesis coupling have rarely been reported. In this study, dynamic and static models of osteogenesis–angiogenesis coupling were developed and the osteogenic and angiogenic effects of the two models were compared. Static co-cultures of MC3T3-E1 and bEnd.3 cells in Transwell inserts showed a cell ratio-dependent reciprocal relation: a ratio of 1:1 (MC3T3-E1:bEnd.3) favored osteogenesis, whereas a ratio of 2:1 (MC3T3-E1:bEnd.3) promoted angiogenesis. On that basis, we developed an osteogenesis–angiogenesis coupling chip based on microfluidic technology. The microfluidic perfusion within the chip further enhanced the mineralizing effect of osteoblasts and the angiogenic effect of endothelial cells, respectively, and increased the secretion of vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2) compared to the static Transwell insert model. The results suggest that the microfluidic chip enhanced the potential of osteogenesis–angiogenesis coupling mediated by paracrine signaling. Overall, the chip is not only a powerful model for understanding bone–vascular interaction but also a scalable platform for high-throughput drug screening and personalized therapy development for fractures.https://www.mdpi.com/2072-666X/16/5/539osteoblastendothelial cellosteogenesis–angiogenesis couplingmicrofluidic cell cultureorgan-on-a-chipin vitro model
spellingShingle Yuexin Wang
Shu Chen
Wenwen Fan
Sixian Zhang
Xi Chen
Cell Ratio-Dependent Osteoblast–Endothelial Cell Crosstalk Promoting Osteogenesis–Angiogenesis Coupling via Regulation of Microfluidic Perfusion and Paracrine Signaling
Micromachines
osteoblast
endothelial cell
osteogenesis–angiogenesis coupling
microfluidic cell culture
organ-on-a-chip
in vitro model
title Cell Ratio-Dependent Osteoblast–Endothelial Cell Crosstalk Promoting Osteogenesis–Angiogenesis Coupling via Regulation of Microfluidic Perfusion and Paracrine Signaling
title_full Cell Ratio-Dependent Osteoblast–Endothelial Cell Crosstalk Promoting Osteogenesis–Angiogenesis Coupling via Regulation of Microfluidic Perfusion and Paracrine Signaling
title_fullStr Cell Ratio-Dependent Osteoblast–Endothelial Cell Crosstalk Promoting Osteogenesis–Angiogenesis Coupling via Regulation of Microfluidic Perfusion and Paracrine Signaling
title_full_unstemmed Cell Ratio-Dependent Osteoblast–Endothelial Cell Crosstalk Promoting Osteogenesis–Angiogenesis Coupling via Regulation of Microfluidic Perfusion and Paracrine Signaling
title_short Cell Ratio-Dependent Osteoblast–Endothelial Cell Crosstalk Promoting Osteogenesis–Angiogenesis Coupling via Regulation of Microfluidic Perfusion and Paracrine Signaling
title_sort cell ratio dependent osteoblast endothelial cell crosstalk promoting osteogenesis angiogenesis coupling via regulation of microfluidic perfusion and paracrine signaling
topic osteoblast
endothelial cell
osteogenesis–angiogenesis coupling
microfluidic cell culture
organ-on-a-chip
in vitro model
url https://www.mdpi.com/2072-666X/16/5/539
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