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Tendon Regeneration with Tendon Hydrogel-Based Cell Delivery: A Comparison of Fibroblasts and Adipose Derived Stem Cells
Arhana Chattopadhyay, MD; Michael G. Galvez, MD; Anais Legrand, MD; Chris Crowe, MD; Rory McGoldrick, MD; Hung Pham, MD; James Chang, MD
Stanford University, Palo Alto, CA

Introduction: Tendon hydrogel is a promising injectable biomaterial for improving repair strength after tendon injury. This study compares the capacity of fibroblasts and adipose-derived stem cells (ASCs) to proliferate, survive, and acquire tenogenic properties when seeded into tendon hydrogel in vitro and in vivo.

Materials and Methods: To assess the effect of cell density on hydrogel contraction, cells were seeded in several concentrations in vitro and measured macroscopically. To assess tenogenic properties, RNA was isolated from cells seeded in hydrogel and tenocyte markers quantified. To assess proliferation and survival, cells were seeded in vitro and MTS and live-dead assays were performed respectively. Finally, to assess the in vivo survival of cells in hydrogel, subcutaneous injections were performed on rats and in vivo imaging system (IVIS) measurements acquired daily.

Results: At 0.5 million cells/mL, both the fibroblast and ASCs induced minimal hydrogel contraction compared to higher cellular concentrations. Fibroblasts and ASCs seeded at 0.5 million cells/mL in tendon hydrogel upregulated several specific tenocyte markers after one week. On MTS assay, fibroblasts and ASCs proliferate in hydrogel at similar rates in vitro. On live-dead assay, fibroblasts survive longer than ASCs in vitro. With IVIS imaging, fibroblasts survived longer than ASCs in hydrogel in vivo.

Conclusions: Tendon healing is mediated by the proliferation, survival, and tenogenic differentiation of cells at the site of injury. Tendon hydrogel delivering dermal fibroblasts may improve and stimulate this process compared to ASCs. Future studies will be needed to evaluate the effects of this hydrogel-based cell delivery on chronic tendon injuries.

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