Osteogenic potency of dedifferentiated fat cells isolated from elderly people with osteoporosis.

Abstract

Mature adipocytes are the major cell type in adipose tissue. This study aimed to explore the osteogenic potency of dedifferentiated fat cells obtained from osteoporotic patients (opDFATs) in vitro and in vivo. Mature adipocytes and adipose-derived stem cells (opASCs) were harvested from subcutaneous adipose tissue. Mature adipocytes were dedifferentiated to produce opDFATs by the ceiling culture method. OpDFATs were osteogenically induced in vitro with opASCs as a control. Cell growth, alkaline phosphatase (ALPase) activity and cell mineralization were determined, and expression levels of osteogenesis-specific genes (collagen I, osteocalcin and bone sialoprotein) were analyzed using quantitative reverse transcription polymerase chain reaction. After 14 days, the opDFATs were combined with a poly(lactide-co-glycolide)-β-tricalcium phosphate porous scaffold after being suspended in collagen I gel and implanted into nude mice for 4 weeks prior to histological analysis. Unilocular lipid droplets in mature adipocytes gradually split into smaller droplets and disappeared from the cytoplasm. Mature adipocytes dedifferentiated to opDFATs and cell morphology changed from spherical to elongated. High levels of ALPase and cell mineralization were observed in opDFATs by staining. No significant differences were found between the growth curves, ALPase activity, cell mineralization and expression levels of osteogenesis-specific genes between opDFATS and opASCs. After implantation for 4 weeks, new bone tissue was observed histologically in the opDFATs-based biocomposite. OpDFATs are implicated as a novel type of seed cell for bone tissue engineering based on their osteogenic potency and higher abundance in adipose tissue compared with opASCs.