Abstract
The purpose of the current study was to evaluate the usefulness of adipose-derived stem cells (ASCs) for bone injury therapy. Lipoaspirates were collected from the abdomen regions of 17 healthy female donors (mean age 49 ± 6 years) using Coleman technique or Body-jet liposuction. In the present study, the primary objective was the in vitro characteristics of human ASCs. The secondary objective was the optimization of the cell seeding process on 3D-printed scaffolds using polycaprolactone (PCL) or polycaprolactone covered with tricalcium phosphate (PCL + 5% TCP). Biological evaluation of human ASC showed high efficiency of isolation obtaining a satisfying amount of homogeneous cell populations. Results suggest that ASCs can be cultured in vitro for a long time without impairing their proliferative capacity. Growth kinetics shows that the highest number of cells can be achieved in passage 5 and after the 16th passage; there is a significant decrease of cell numbers and their proliferative potential. The percentage of colony forming units from the adipose stem cells is 8% ± 0.63% (p < 0.05). It was observed that the accumulation of calcium phosphate in the cells in vitro, marked with Alizarin Red S, was increased along with the next passage. Analysis of key parameters critically related to the cell seeding process shows that volume of cell suspension and propagation time greatly improve the efficiency of seeding both in PCL and PCL + 5% TCP scaffolds. The cell seeding efficiency did differ significantly between scaffold materials and cell seeding methods (p < 0.001). Increased seeding efficiency was observed when using the saturation of cell suspension into scaffolds with additional incubation. Alkaline phosphatase level production in PCL + 5% TCP scaffold was better than in PCL-only scaffold. The study results can be used for the optimization of the seeding process and quantification methods determining the successful implementation of the preclinical model study in the future tissue engineering strategies.
Highlights
Regenerating or replacing bone defects is an important research field in tissue engineering
Cultures of adipose-derived stem cells (ASCs) at passage 3 were analyzed for the expression of cell surface markers
Representative histograms show results of ASC staining for indicated surface markers from one representative donor (Figure 3)
Summary
Regenerating or replacing bone defects is an important research field in tissue engineering. Current methods for surgical treatment of fractures and bone defects primarily use metal implants, and autologous and allogeneic bone grafts still represent the gold standard for bone repair. Mesenchymal stem cells derived from adipose tissue are promising cell source for bone lesion repair [1]. This is important for the optimization of methods aimed at isolation, characterization, expansion, and evaluation of differentiation potential [2]. These parameters ensure the quality of stem cells and the safety of their use. There are conflicting reports on the effect of donor age on adipose human mesenchymal stem
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