Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow

Abstract Adipose-derived stem cells (ASCs) provide an ample, easily accessible source of multipotent cells, an alternative to bone marrow-derived stromal cells (BMSCs), capable of differentiating into osteoblasts. However, the osteogenic potential of ASCs is reportedly lower than that of BMSCs and p...

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Main Authors: Karolina Truchan, Barbara Zagrajczuk, Katarzyna Cholewa-Kowalska, Anna Maria Osyczka
Format: Article
Language:English
Published: BMC 2025-03-01
Series:Journal of Biological Engineering
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Online Access:https://doi.org/10.1186/s13036-025-00491-2
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author Karolina Truchan
Barbara Zagrajczuk
Katarzyna Cholewa-Kowalska
Anna Maria Osyczka
author_facet Karolina Truchan
Barbara Zagrajczuk
Katarzyna Cholewa-Kowalska
Anna Maria Osyczka
author_sort Karolina Truchan
collection DOAJ
description Abstract Adipose-derived stem cells (ASCs) provide an ample, easily accessible source of multipotent cells, an alternative to bone marrow-derived stromal cells (BMSCs), capable of differentiating into osteoblasts. However, the osteogenic potential of ASCs is reportedly lower than that of BMSCs and protocols to effectively differentiate ASCs into osteoblasts are in high demand. Here, we present novel strategies for effective osteogenic differentiation of human ASCs by combining their culture on bioactive growth surfaces with their treatment with specific supplements in osteogenic medium and application of fluid shear stress. Human ASCs were cultured on PLGA-based composites containing 50 wt% sol-gel bioactive glasses (SBGs) from the SiO2-CaO±P2O5 system, either unmodified or modified with 5 wt% ZnO or SrO. The osteogenic medium was supplemented with recombinant human bone morphogenetic protein 2 (BMP-2), MEK1/2 kinase inhibitor (PD98059) and indirect Smurf1 inhibitor (Phenamil). Fluid shear stress was applied with a standard horizontal rocker. ASC culture on SBG-PLGA composites along with the osteogenic medium supplements enhanced the expression of both early and late osteogenic markers. Modification of SBG with either SrO or ZnO further enhanced osteogenic gene expression compared to ASCs cultured on composites containing unmodified SBGs. Notably, the application of fluid shear stress synergistically strengthened the osteogenic effects of bioactive composites and medium supplements. We also show that the presented culture strategies can drive ASCs toward osteoblastic cells in a 3-day culture period and provide mineralizing osteoblasts through a short, 7-day ASC preculture on bioactive composites. Our results also indicate that the applied osteogenic treatment leads to the phosphorylation of β-catenin and CREB or the COX-2 expression. We believe the presented strategies are feasible for rapid ASC differentiation to early osteoblasts or mineralizing osteoblastic cells for various potential cell-based bone regeneration therapies. Graphical Abstract
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spelling doaj-art-2beabd42157e436dba9d3faa27c2a81d2025-08-20T02:56:12ZengBMCJournal of Biological Engineering1754-16112025-03-0119111910.1186/s13036-025-00491-2Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flowKarolina Truchan0Barbara Zagrajczuk1Katarzyna Cholewa-Kowalska2Anna Maria Osyczka3Department of Cell Biology and Imaging, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian UniversityDepartment of Glass Technology and Amorphous Coatings, Faculty of Materials Science and Ceramics, AGH University of Science and TechnologyDepartment of Glass Technology and Amorphous Coatings, Faculty of Materials Science and Ceramics, AGH University of Science and TechnologyDepartment of Cell Biology and Imaging, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian UniversityAbstract Adipose-derived stem cells (ASCs) provide an ample, easily accessible source of multipotent cells, an alternative to bone marrow-derived stromal cells (BMSCs), capable of differentiating into osteoblasts. However, the osteogenic potential of ASCs is reportedly lower than that of BMSCs and protocols to effectively differentiate ASCs into osteoblasts are in high demand. Here, we present novel strategies for effective osteogenic differentiation of human ASCs by combining their culture on bioactive growth surfaces with their treatment with specific supplements in osteogenic medium and application of fluid shear stress. Human ASCs were cultured on PLGA-based composites containing 50 wt% sol-gel bioactive glasses (SBGs) from the SiO2-CaO±P2O5 system, either unmodified or modified with 5 wt% ZnO or SrO. The osteogenic medium was supplemented with recombinant human bone morphogenetic protein 2 (BMP-2), MEK1/2 kinase inhibitor (PD98059) and indirect Smurf1 inhibitor (Phenamil). Fluid shear stress was applied with a standard horizontal rocker. ASC culture on SBG-PLGA composites along with the osteogenic medium supplements enhanced the expression of both early and late osteogenic markers. Modification of SBG with either SrO or ZnO further enhanced osteogenic gene expression compared to ASCs cultured on composites containing unmodified SBGs. Notably, the application of fluid shear stress synergistically strengthened the osteogenic effects of bioactive composites and medium supplements. We also show that the presented culture strategies can drive ASCs toward osteoblastic cells in a 3-day culture period and provide mineralizing osteoblasts through a short, 7-day ASC preculture on bioactive composites. Our results also indicate that the applied osteogenic treatment leads to the phosphorylation of β-catenin and CREB or the COX-2 expression. We believe the presented strategies are feasible for rapid ASC differentiation to early osteoblasts or mineralizing osteoblastic cells for various potential cell-based bone regeneration therapies. Graphical Abstracthttps://doi.org/10.1186/s13036-025-00491-2Adipose-derived stem cellsOsteoinductionBMP-2PhenamilERK inhibitorZinc
spellingShingle Karolina Truchan
Barbara Zagrajczuk
Katarzyna Cholewa-Kowalska
Anna Maria Osyczka
Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow
Journal of Biological Engineering
Adipose-derived stem cells
Osteoinduction
BMP-2
Phenamil
ERK inhibitor
Zinc
title Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow
title_full Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow
title_fullStr Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow
title_full_unstemmed Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow
title_short Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow
title_sort rapid osteoinduction of human adipose derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow
topic Adipose-derived stem cells
Osteoinduction
BMP-2
Phenamil
ERK inhibitor
Zinc
url https://doi.org/10.1186/s13036-025-00491-2
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AT barbarazagrajczuk rapidosteoinductionofhumanadiposederivedstemcellsgrownonbioactivesurfacesandstimulatedbychemicallymodifiedmediaflow
AT katarzynacholewakowalska rapidosteoinductionofhumanadiposederivedstemcellsgrownonbioactivesurfacesandstimulatedbychemicallymodifiedmediaflow
AT annamariaosyczka rapidosteoinductionofhumanadiposederivedstemcellsgrownonbioactivesurfacesandstimulatedbychemicallymodifiedmediaflow