Plastic Surgery Research Council

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Seeding Of ADSC-differentiated Chondrocytes Onto A Three-dimensional Allograft Matrix
Max Korzec, BS, Ashim Gupta, MS, PhD, Joel Reichensperger, BS, Andrew Klein, BS, Michael Neumeister, MD, Brian Mailey, MD.
Southern Illinois University School of Medicine, Springfield, IL, USA.

Seeding of ADSC-differentiated chondrocytes onto a 3-dimensional allograft matrix
Purpose: Degeneration of articular cartilage results from trauma and aging. No current available treatments target cartilage regeneration. Our ultimate aim is to utilize tissue engineering approaches to replace lost cartilage or perform autologous small joint replacements. For this study, our goal was to assess ability of rat ADSCs to differentiate into chondrocytes and seed them onto acellular dermal matrices (ADM) and three dimensional adipose allograft matrices (AAM).
Methods: We isolated ADSCs from green fluorescent protein (GFP) expressing Sprague Dawley rats (SD -Tg (UBC-EGFP) 2BalRrrc) and assessed their ability to differentiate into chondrocytes. These isolated cells were then seeded on ADMs (AlloDerm (LifeCell Corp., Branchburg, NJ, USA) and Adipose Allograft Matrix) and the biocompatibility was determined by performing Immunofluorescence staining to determine chondrogenic markers expression, and Scanning Electron Microscopy (SEM) to determine cell growth, adhesion, and morphology.
Results: GFP-ADSCs were successfully isolated and their multipotency confirmed via successful adipo-, chondro- and osteo-induction. Induced cells demonstrated expression of collagen-II indicating differentiation towards chondrocyte lineage. Approximately 1 million GFP-ADSCs
were successfully seeded onto each ADM and AAM, with each experiment requiring ~30 million isolated GFP-ADSCs. SEM imaging demonstrated cells adhered, grew, and maintained non-stressed morphology on ADMs and AAMs.
Conclusions: We confirmed ability of ADSCs to differentiate into chondrocytes and in-vitro biocompatibility of seeded ADMs and AAMs. Future studies will involve evaluating in-vivo biocompatibility and the potential of these seeded ADMs to form a 3-D cartilage structure.
Acknowledgments: The authors would like to acknowledge the Memorial Medical Center Foundation in Springfield, IL for funding.


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