title: Genipin-crosslinked cartilage-derived matrix as a scaffold for human adipose-derived stem cell chondrogenesis.
abstract: Autologous cell-based tissue engineering using three-dimensional scaffolds holds much promise for the repair of cartilage defects. Previously, we reported on the development of a porous scaffold derived solely from native articular cartilage, which can induce human adipose-derived stem cells (ASCs) to differentiate into a chondrogenic phenotype without exogenous growth factors. However, this ASC-seeded cartilage-derived matrix (CDM) contracts over time in culture, which may limit certain clinical applications. The present study aimed to investigate the ability of chemical crosslinking using a natural biologic crosslinker, genipin, to prevent scaffold contraction while preserving the chondrogenic potential of CDM. CDM scaffolds were crosslinked in various genipin concentrations, seeded with ASCs, and then cultured for 4 weeks to evaluate the influence of chemical crosslinking on scaffold contraction and ASC chondrogenesis. At the highest crosslinking degree of 89%, most cells failed to attach to the scaffolds and resulted in poor formation of a new extracellular matrix. Scaffolds with a low crosslinking density of 4% experienced cell-mediated contraction similar to our original report on noncrosslinked CDM. Using a 0.05% genipin solution, a crosslinking degree of 50% was achieved, and the ASC-seeded constructs exhibited no significant contraction during the culture period. Moreover, expression of cartilage-specific genes, synthesis, and accumulation of cartilage-related macromolecules and the development of mechanical properties were comparable to the original CDM. These findings support the potential use of a moderately (i.e., approximately one-half of the available lysine or hydroxylysine residues being crosslinked) crosslinked CDM as a contraction-free biomaterial for cartilage tissue engineering.
authors.affiliation: Department of Orthopaedic Surgery, Duke University Medical Center, Durham, North Carolina 27710, USA.
authors.forename: Nai-Chen
authors.initials: NC
authors.lastname: Cheng
authors.forename: Bradley T
authors.initials: BT
authors.lastname: Estes
authors.forename: Tai-Horng
authors.initials: TH
authors.lastname: Young
authors.forename: Farshid
authors.initials: F
authors.lastname: Guilak
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000502:physiology*; D003432:Cross-Linking Reagents / Q000737:chemistry; D004343:Drug Implants / Q000008:administration & dosage / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005109:Extracellular Matrix / Q000737:chemistry*; D006801:Humans; D039823:Iridoids / Q000008:administration & dosage*; D050997:Nanomedicine; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D054457:Tissue Scaffolds
chemical_list: D003432:Cross-Linking Reagents; D004343:Drug Implants; D039823:Iridoids; C007834:genipin