Three-dimensional printed polycaprolactone-based scaffolds provide an advantageous environment for osteogenic differentiation of human adipose-derived stem cells.
The capacity of bone grafts to repair critical size defects can be greatly enhanced by the delivery of mesenchymal stem cells (MSCs). Adipose tissue is considered the most effective source of MSCs (ADSCs); however, the efficiency of bone regeneration using undifferentiated ADSCs is low. Therefore, this study proposes scaffolds based on polycaprolactone (PCL), which is widely considered a suitable MSC delivery system, were used as a three-dimensional (3D) culture environment promoting osteogenic differentiation of ADSCs. PCL scaffolds enriched with 5% tricalcium phosphate (TCP) were used. Human ADSCs were cultured in osteogenic medium both on the scaffolds and in 2D culture. Cell viability and osteogenic differentiation were tested at various time points for 42 days. The expression of RUNX2, collagen I, alkaline phosphatase, osteonectin and osteocalcin, measured by real-time polymerase chain reaction was significantly upregulated in 3D culture. Production of osteocalcin, a specific marker of terminally differentiated osteoblasts, was significantly higher in 3D cultures than in 2D cultures, as confirmed by western blot and immunostaining, and accompanied by earlier and enhanced mineralization. Subcutaneous implantation into immunodeficient mice was used for in vivo observations. Immunohistological and micro-computed tomography analysis revealed ADSC survival and activity toward extracellular production after 4 and 12 weeks, although heterotopic osteogenesis was not confirmed - probably resulting from insufficient availability of Ca/P ions. Additionally, TCP did not contribute to the upregulation of differentiation on the scaffolds in culture, and we postulate that the 3D architecture is a critical factor and provides a useful environment for prior-to-implantation osteogenic differentiation of ADSCs. Copyright © 2016 John Wiley & Sons, Ltd.
Department of Histology and Embryology, Centre for Biostructure Research, Medical University of Warsaw, Poland.|Centre for Preclinical Research and Technology, Poland.|Postgraduate School of Molecular Medicine, Medical University of Warsaw, Poland.
Sławomir
S
Rumiński
Faculty of Materials Science and Engineering, Warsaw University of Technology, Poland.
Barbara
B
Ostrowska
Faculty of Materials Science and Engineering, Warsaw University of Technology, Poland.
Jakub
J
Jaroszewicz
Department of Clinical Cytology, Centre of Postgraduate Medical Education, Warsaw, Poland.|Department of Anesthesiology and Intensive Care Medicine, Centre of Postgraduate Medical Education, Warsaw, Poland.
Tomasz
T
Skirecki
Department of Histology and Embryology, Centre for Biostructure Research, Medical University of Warsaw, Poland.
Krzysztof
K
Włodarski
Faculty of Materials Science and Engineering, Warsaw University of Technology, Poland.
Wojciech
W
Święszkowski
Department of Histology and Embryology, Centre for Biostructure Research, Medical University of Warsaw, Poland.|Centre for Preclinical Research and Technology, Poland.
Małgorzata
M
Lewandowska-Szumieł
D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D002113:Calcification, Physiologic / Q000187:drug effects; D002130:Calcium Phosphates / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects*; D010641:Phenotype; D011091:Polyesters / Q000494:pharmacology*; D066330:Printing, Three-Dimensional; D019919:Prosthesis Implantation; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D054457:Tissue Scaffolds / Q000737:chemistry*; D055114:X-Ray Microtomography
D002130:Calcium Phosphates; D011091:Polyesters; D015675:Osteocalcin; C016240:polycaprolactone; C018392:tricalcium phosphate
adipose-derived stem cells; bone tissue engineering; mesenchymal stem cells; osteoblasts; polycaprolactone