MiR-221-inhibited adipose tissue-derived mesenchymal stem cells bioengineered in a nano-hydroxy apatite scaffold

Abstract

The repair of skeletal defects is the main goal of bone tissue engineering. Recent literature highlighted various regulatory roles of microRNAs in stem cell fate determination. In addition, the role of porous hydroxyapatite/polycaprolacton (nHA/PCL) as a bioactive scaffold which enhances adipose tissue-derived mesenchymal stem cells (AT-MSCs) growth and osteogenic differentiation has been proved. The aim of the present study was to investigate the synergistic potential of both down-regulating miR-221 and nHA/PCL scaffold seeding in osteogenic potential of AT-MSCs. After isolation and characterization of AT-MSCs, the transfection of anti-miR-221 was performed into the cells using lipofectamine 2000 and the transfected cells were seeded into a synthesized nHA/PCL scaffold. The DAPI staining confirmed the presence of AT-MSCs on nHA/PCL scaffold. Quantitative expression of osteoblast marker genes, Runx2, and osteocalcin of the transfected cells in the scaffold were evaluated. Interestingly, significant upregulation of transcribed Runx2 and osteocalcin genes (P < 0.01) were observed in miR-221-inhibited nHA/PCL seeded cells. Also, alkaline phosphatase activity (ALP) was significantly higher (P < 0.01) in miR-221-inhibited AT-MSCs seeded on nHA/PCL than those seeded on nHA/PCL or transfected with anti-miR-221, individually. The results of this combination suggest a valuable method for enhancing osteogenesis in AT-MSCs. This method could be applicable for gene-cell therapy of bone defects.