title: High mesenchymal stem cell seeding densities in hyaluronic acid hydrogels produce engineered cartilage with native tissue properties.
abstract: Engineered cartilage based on adult mesenchymal stem cells (MSCs) is an alluring goal for the repair of articular defects. However, efforts to date have failed to generate constructs with sufficient mechanical properties to function in the demanding environment of the joint. Our findings with a novel photocrosslinked hyaluronic acid (HA) hydrogel suggest that stiff gels (high HA concentration, 5% w/v) foster chondrogenic differentiation and matrix production, but limit overall functional maturation due to the inability of the formed matrix to diffuse away from the point of production and form a contiguous network. In the current study, we hypothesized that increasing the MSC seeding density would decrease the required diffusional distance, and so expedite the development of functional properties. To test this hypothesis bovine MSCs were encapsulated at seeding densities of either 20,000,000 or 60,000,000 cells ml(-1) in 1%, 3%, and 5% (w/v) HA hydrogels. Counter to our hypothesis the higher concentration HA gels (3% and 5%) did not develop more rapidly with increased MSC seeding density. However, the biomechanical properties of the low concentration (1%) HA constructs increased markedly (nearly 3-fold with a 3-fold increase in seeding density). To ensure that optimal nutrient access was delivered, we next cultured these constructs under dynamic culture conditions (with orbital shaking) for 9 weeks. Under these conditions 1% HA seeded at 60,000,000 MSCs ml(-1) reached a compressive modulus in excess of 1 MPa (compared with 0.3-0.4 MPa for free swelling constructs). This is the highest level we have reported to date in this HA hydrogel system, and represents a significant advance towards functional stem cell-based tissue engineered cartilage.
authors.affiliation: McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, University of Pennsylvania, 424 Stemmler Hall, 36th Street and Hamilton Walk, Philadelphia, PA 19104, USA.
authors.forename: Isaac E
authors.initials: IE
authors.lastname: Erickson
authors.forename: Sydney R
authors.initials: SR
authors.lastname: Kestle
authors.forename: Kilief H
authors.initials: KH
authors.lastname: Zellars
authors.forename: Megan J
authors.initials: MJ
authors.lastname: Farrell
authors.forename: Minwook
authors.initials: M
authors.lastname: Kim
authors.forename: Jason A
authors.initials: JA
authors.lastname: Burdick
authors.forename: Robert L
authors.initials: RL
authors.lastname: Mauck
mesh_terms: D000818:Animals; D002356:Cartilage / Q000187:drug effects / Q000502:physiology*; D002417:Cattle; D002452:Cell Count; D018914:Cells, Immobilized / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D020219:Chondrogenesis / Q000187:drug effects*; D003094:Collagen / Q000378:metabolism; D004058:Diffusion / Q000187:drug effects; D055119:Elastic Modulus / Q000187:drug effects; D005452:Fluoresceins / Q000378:metabolism; D005453:Fluorescence; D006025:Glycosaminoglycans / Q000378:metabolism; D006820:Hyaluronic Acid / Q000494:pharmacology*; D020100:Hydrogels / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D005452:Fluoresceins; D006025:Glycosaminoglycans; D020100:Hydrogels; C085925:calcein AM; C013786:A73025; D006820:Hyaluronic Acid; D003094:Collagen