Recent studies suggest that human adipose tissue contains pluripotent stem cells, which are similar to bone marrow-derived stem cells. The objective of the present study was to assess the effect in bone regenerating capability of human adipose-derived stem cells (ADSCs) cultured in osteogenic media layered over poly lactide-co-glycolic acid (PLGA) and implanted in a critical nude rat calvarial defect. Twenty-seven nude rats were randomized into 3 groups (n = 9): 1) PLGA alone (control), 2) PLGA with undifferentiated ADSCs, and 3) PLGA with differentiated ADSCs. These 3 groups were divided into 9 subgroups (n = 3) according to in vitro pre-cultured periods (day 1 pre-culture (Group1), day 7 pre-culture (Group2), and day 14 pre-culture (Group3)) before implantation. An 8 mm critical-size circular calvarial defect was made in each nude rat. Specimens were harvested at 12 weeks post-implantation and evaluated radiographically and histologically. Radiodensitometric analysis revealed significantly higher bone growth in implants pre-cultured in osteogenic media for 14 days for Group 3. Histomorphometric analysis demonstrated that Groups 2 and 3 had bone formation filling 35% to 72% of the area of the defect after transplantation with cells that had been pre-cultured for 14 days. Constructs with differentiated ADSCs (Group 3) had noticeably more maximal and robust bone tissue regeneration than constructs with undifferentiated ADSCs (Group 2). These data provide evidence that constructs or implants made of PLGA and osteogenically differentiated ADSCs pre-cultured for 14 days before transplantation have better, more-robust bone regeneration capability in critical-sized skeletal defects than constructs with undifferentiated ADSCs. Human adipose derived stem cells can therefore be used as seed cells to construct tissue-engineered bone.
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