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Cell-based therapies may hold significant promise for the treatment of early stage degeneration of the intervertebral disc (IVD). Given their propensity to proliferate and ability to form multiple tissue types, mesenchymal stem cells (MSCs) have been proposed as a potential cell source to promote repair of the nucleus pulposus (NP). However, for any successful cell-based therapy, a carrier biomaterial may be essential for targeted delivery providing key biophysical and biochemical cues to facilitate differentiation of MSCs. Two widely used biomaterials for NP regeneration are chitosan and alginate. The primary objective of this study was to assess the influence of alginate and chitosan hydrogels on bone marrow stem cells (BM) and NP cells in isolation or in coculture. A secondary objective of this study was to investigate coculture seeding density effects of BM and NP cells and simultaneously explore which cell type is responsible for matrix formation in a cocultured environment. Porcine NP and BM cells were encapsulated in alginate and chitosan hydrogels separately at two seeding densities (4×10(6) and 8×10(6) cells/mL) or in coculture (1:1, 8×10(6) cells/mL). Constructs (diameter=5 mm, height=3 mm) were maintained under IVD-like conditions [low-glucose, low (5%) oxygen] with or without transforming growth factor-β3 (TGF-β3) supplementation for 21 days. Results demonstrated differential viability depending on hydrogel type. NP cells remained viable in both biomaterial types whereas BM viability was diminished in chitosan. Further, hydrogel type was found to regulate sulfated glycosaminoglycan (sGAG) and collagen accumulation. Specifically, alginate better supports sGAG accumulation and collagen type II deposition for both NP and BM cell types compared with chitosan. Having identified that alginate more readily supports cell viability and matrix accumulation, we further explored additional effects of seeding density ratios (NP:BM--1:1, 1:2) for coculture studies. Interestingly, in coculture conditions, the BM cell population declined in number while NP cells increased, indicating that MSCs may in fact be signaling NP cells to proliferate rather than contributing to matrix formation. These findings provide exciting new insights on the potential of MSCs for NP tissue regeneration strategies.

Differential Response of Encapsulated Nucleus Pulposus and Bone Marrow Stem Cells in Isolation and Coculture in Alginate and Chitosan Hydrogels