Three-dimensional printed polycaprolactone-based scaffolds provide an advantageous environment for osteogenic differentiation of human adipose-derived stem cells

Abstract

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.

Comparison of adipose stem cells sources from various locations of rat body for their application for seeding on polymer scaffolds

“…We observed that after 14 d of culturing ASCs on PCL þ 5%TCP, osteogenic differentiation increased compared to undifferentiated ASCs cultured in standard culture medium. The results thus obtained are consistent with those of previous studies reporting an increase in ALP activity after 14 d of ASC differentiation on scaffolds [37][38][39][40][50][51][52][53]. We also observed that modification of scaffolds can influence the degree of cell differentiation.…”

Donor Site Location Is Critical for Proliferation, Stem Cell Capacity, and Osteogenic Differentiation of Adipose Mesenchymal Stem/Stromal Cells: Implications for Bone Tissue Engineering

“…The supplementation of the culture medium with platelet lysate or platelet-rich plasma also improved the osteogenic differentiation of Ad-MSCs [ 14 , 22 , 23 ]. Also, the cultivation on three-dimensional scaffolds further improved the osteogenic properties of Ad-MSCs [ 24 , 25 ], amongst others, through the control of cell agglomeration [ 26 ]. Furthermore, co-cultivation with endothelial cells, vascular cells, or osteoblastic cells improved the osteogenic differentiation of Ad-MSCs [ 27 , 28 , 29 ].…”

Processing of (Co)Poly(2-oxazoline)s by Electrospinning and Extrusion from Melt and the Postprocessing Properties of the (Co)Polymers

“…Matrices with strictly defined parameters can also be obtained using a variety of innovative 3D printing technologies [21][22][23]. One well-known method is the extrusion of polymers from melt.…”