title: Crosslinkable hydrogels derived from cartilage, meniscus, and tendon tissue.
abstract: Decellularized tissues have proven to be versatile matrices for the engineering of tissues and organs. These matrices usually consist of collagens, matrix-specific proteins, and a set of largely undefined growth factors and signaling molecules. Although several decellularized tissues have found their way to clinical applications, their use in the engineering of cartilage tissue has only been explored to a limited extent. We set out to generate hydrogels from several tissue-derived matrices, as hydrogels are the current preferred cell carriers for cartilage repair. Equine cartilage, meniscus, and tendon tissue was harvested, decellularized, enzymatically digested, and functionalized with methacrylamide groups. After photo-cross-linking, these tissue digests were mechanically characterized. Next, gelatin methacrylamide (GelMA) hydrogel was functionalized with these methacrylated tissue digests. Equine chondrocytes and mesenchymal stromal cells (MSCs) (both from three donors) were encapsulated and cultured in vitro up to 6 weeks. Gene expression (COL1A1, COL2A1, ACAN, MMP-3, MMP-13, and MMP-14), cartilage-specific matrix formation, and hydrogel stiffness were analyzed after culture. The cartilage, meniscus, and tendon digests were successfully photo-cross-linked into hydrogels. The addition of the tissue-derived matrices to GelMA affected chondrogenic differentiation of MSCs, although no consequent improvement was demonstrated. For chondrocytes, the tissue-derived matrix gels performed worse compared to GelMA alone. This work demonstrates for the first time that native tissues can be processed into crosslinkable hydrogels for the engineering of tissues. Moreover, the differentiation of encapsulated cells can be influenced in these stable, decellularized matrix hydrogels.
authors.affiliation: 1 Department of Orthopaedics, University Medical Center Utrecht , Utrecht, The Netherlands .
authors.forename: Jetze
authors.initials: J
authors.lastname: Visser
authors.forename: Peter A
authors.initials: PA
authors.lastname: Levett
authors.forename: Nikae C R
authors.initials: NC
authors.lastname: te Moller
authors.forename: Jeremy
authors.initials: J
authors.lastname: Besems
authors.forename: Kristel W M
authors.initials: KW
authors.lastname: Boere
authors.forename: Mattie H P
authors.initials: MH
authors.lastname: van Rijen
authors.forename: Janny C
authors.initials: JC
authors.lastname: de Grauw
authors.forename: Wouter J A
authors.initials: WJ
authors.lastname: Dhert
authors.forename: P René
authors.initials: PR
authors.lastname: van Weeren
authors.forename: Jos
authors.initials: J
authors.lastname: Malda
mesh_terms: D000818:Animals; D002356:Cartilage / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects; D019245:Compressive Strength / Q000187:drug effects; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism; D055119:Elastic Modulus / Q000187:drug effects; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006025:Glycosaminoglycans / Q000378:metabolism; D006736:Horses; D020100:Hydrogels / Q000494:pharmacology*; D008592:Menisci, Tibial / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000166:cytology; D013710:Tendons / Q000166:cytology*
chemical_list: D003432:Cross-Linking Reagents; D006025:Glycosaminoglycans; D020100:Hydrogels; D004247:DNA