Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells
With an aging population worldwide, research into bone metabolism and novel therapies for damaged and diseased bone is essential. Bone is a vascularized, dynamic tissue that undergoes a constant remodeling process mediated by osteocytes, osteoblasts and osteoclasts. In this study, a complex 3D in vi...
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Elsevier
2025-10-01
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590006425007240 |
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| author | Katharina Wirsig Nina Bürger Lisa Fleischhauer Nele Louisa Preuß Anne Bernhardt |
| author_facet | Katharina Wirsig Nina Bürger Lisa Fleischhauer Nele Louisa Preuß Anne Bernhardt |
| author_sort | Katharina Wirsig |
| collection | DOAJ |
| description | With an aging population worldwide, research into bone metabolism and novel therapies for damaged and diseased bone is essential. Bone is a vascularized, dynamic tissue that undergoes a constant remodeling process mediated by osteocytes, osteoblasts and osteoclasts. In this study, a complex 3D in vitro bone model combining these three main bone cell species with endothelial cells was developed. The different cell species were isolated from primary human tissue and spatially arranged using transwell inserts. Osteocytes differentiated from collagen-embedded osteoblasts, while osteoclasts simultaneously derived from peripheral blood mononuclear cells without receptor activator of NF-κB ligand (RANKL) supplementation. Different cultivation parameters were evaluated to define conditions that support differentiation and function of all cell types involved in quadruple culture. The cellular crosstalk in quadruple cultures stimulated osteoblast (ALP, BMP-2, IBSP, COL1A, VEGF) and osteocyte (SOST, DMP1) markers, while osteoclast (TRAP) and endothelial cell markers were reduced compared to respective mono- or co-cultures. Furthermore, mineralization was induced only in quadruple cultures, demonstrating the importance of signaling between the four cell types. This sophisticated human bone model provides a physiologically relevant culture system to study the complex crosstalk between bone cells, their precursors and endothelial cells during remodeling and vascularization. Moreover, it allows preclinical testing of bioactive factors, biomaterial extracts or drugs for translation into clinical practice without animal testing. |
| format | Article |
| id | doaj-art-e49c82c97a4c48f29b4def53b2f4eb7a |
| institution | DOAJ |
| issn | 2590-0064 |
| language | English |
| publishDate | 2025-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Materials Today Bio |
| spelling | doaj-art-e49c82c97a4c48f29b4def53b2f4eb7a2025-08-20T02:56:39ZengElsevierMaterials Today Bio2590-00642025-10-013410215410.1016/j.mtbio.2025.102154Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cellsKatharina Wirsig0Nina Bürger1Lisa Fleischhauer2Nele Louisa Preuß3Anne Bernhardt4Centre for Translational Bone, Joint- and Soft Tissue Research, Faculty of Medicine and University Hospital, TUD University of Technology, Fetscherstraße 74, 01307, Dresden, GermanyCentre for Translational Bone, Joint- and Soft Tissue Research, Faculty of Medicine and University Hospital, TUD University of Technology, Fetscherstraße 74, 01307, Dresden, GermanyCentre for Translational Bone, Joint- and Soft Tissue Research, Faculty of Medicine and University Hospital, TUD University of Technology, Fetscherstraße 74, 01307, Dresden, GermanyCentre for Translational Bone, Joint- and Soft Tissue Research, Faculty of Medicine and University Hospital, TUD University of Technology, Fetscherstraße 74, 01307, Dresden, GermanyCorresponding author.; Centre for Translational Bone, Joint- and Soft Tissue Research, Faculty of Medicine and University Hospital, TUD University of Technology, Fetscherstraße 74, 01307, Dresden, GermanyWith an aging population worldwide, research into bone metabolism and novel therapies for damaged and diseased bone is essential. Bone is a vascularized, dynamic tissue that undergoes a constant remodeling process mediated by osteocytes, osteoblasts and osteoclasts. In this study, a complex 3D in vitro bone model combining these three main bone cell species with endothelial cells was developed. The different cell species were isolated from primary human tissue and spatially arranged using transwell inserts. Osteocytes differentiated from collagen-embedded osteoblasts, while osteoclasts simultaneously derived from peripheral blood mononuclear cells without receptor activator of NF-κB ligand (RANKL) supplementation. Different cultivation parameters were evaluated to define conditions that support differentiation and function of all cell types involved in quadruple culture. The cellular crosstalk in quadruple cultures stimulated osteoblast (ALP, BMP-2, IBSP, COL1A, VEGF) and osteocyte (SOST, DMP1) markers, while osteoclast (TRAP) and endothelial cell markers were reduced compared to respective mono- or co-cultures. Furthermore, mineralization was induced only in quadruple cultures, demonstrating the importance of signaling between the four cell types. This sophisticated human bone model provides a physiologically relevant culture system to study the complex crosstalk between bone cells, their precursors and endothelial cells during remodeling and vascularization. Moreover, it allows preclinical testing of bioactive factors, biomaterial extracts or drugs for translation into clinical practice without animal testing.http://www.sciencedirect.com/science/article/pii/S2590006425007240In vitro bone modelOsteoblastsOsteocytesOsteoclastsEndothelial cellsCo-culture |
| spellingShingle | Katharina Wirsig Nina Bürger Lisa Fleischhauer Nele Louisa Preuß Anne Bernhardt Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells Materials Today Bio In vitro bone model Osteoblasts Osteocytes Osteoclasts Endothelial cells Co-culture |
| title | Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells |
| title_full | Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells |
| title_fullStr | Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells |
| title_full_unstemmed | Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells |
| title_short | Vascularized in vitro bone model as 3D quadruple culture with primary human osteoblasts, osteocytes, osteoclasts and endothelial cells |
| title_sort | vascularized in vitro bone model as 3d quadruple culture with primary human osteoblasts osteocytes osteoclasts and endothelial cells |
| topic | In vitro bone model Osteoblasts Osteocytes Osteoclasts Endothelial cells Co-culture |
| url | http://www.sciencedirect.com/science/article/pii/S2590006425007240 |
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