Plastic Surgery Research Council
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PSRC 60th Annual Meeting

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Dynamic Culture Enhances Osteogenic Performance Comparing Donor Matched Human Adipose and Bone Marrow Mesenchymal Stem Cells
Ajul Shah, MD, Miles Pfaff, BS, Wei Wu, MD, Laura Niklason, MD, Derek Steinbacher, DMD, DDS.
Yale University School of Medicine, New Haven, CT, USA.

PURPOSE:
Tissue engineered bone holds translational promise for myriad applications in reconstructive surgery. Both adipose-derived and bone marrow-derived stem cells (ADSCs and BMSCs) have been used for bone regeneration, and can be seeded on a variety of rigid scaffolds. This study aims to compare ADSCs and BMSCs from the same donor in three distinct bioreactor settings to create the most viable osseous engineered construct. We hypothesize that physiologic flow dynamics will optimize osteogenic cell viability and function, which are prerequisites to successful human tissue implantation.
METHODS:
Human ADSCs and BMSCs were isolated from the same donor, then cultured and seeded on decellularized porcine bone constructs. The constructs were then subjected to either static or dynamic (stirring or perfusion bioreactor) culture conditions for 7 to 21 days. Afterwards, the constructs were analyzed for cell adhesion and distribution using histology and electron scanning microscopy. Proliferation and osteogenic differentiation were further gauged using DNA quantification, alkaline phosphatase (ALP) assay, immunostaining for osteocalcin and real-time-PCR, and calcium deposition assay.
RESULTS:
hADSCs demonstrated higher seeding efficiency and proliferative potential in static culture than hBMSCs. However, dynamic culture, driven by stirring or perfusion flow, significantly increased BMSCs proliferation more than ADSCs proliferation. The highest cellularity was seen in the stirring bioreactor. In all conditions, BMSCs demonstrated stronger osteogenic activity compared to ADSCs, in ALP activity assay and gene expression for various bony markers. Conversely, ADSCs expressed more collagen I. In all constructs (ADSC and BMSC), dynamic conditions (stirring and perfusing bioreactors) enhance overall osteogenic gene expression. BMSCs in the stirring bioreactor exhibited the greatest calcium production, likely secondary to the greater cell proliferation and osteogenic function.
CONCLUSION:
Scaffolds seeded with BMSCs in dynamic conditions exhibit the greatest osteogenic proliferation and function. In particular, the stirring bioreactor optimizes the bone engineered construct, and may portend clinical success


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