ABSTRACT
In cartilage regeneration, the biomimetic functionalization of hydrogels with growth factors is a promising approach to improve the in vivo performance and furthermore the clinical potential of these materials. In order to achieve this without compromising network properties, multifunctional linear poly(glycidol) acrylate (PG-Acr) is synthesized and utilized as crosslinker for hydrogel formation with thiol-functionalized hyaluronic acid via Michael-type addition. As proof-of-principle for a bioactivation, transforming growth factor-beta 1 (TGF-ß1) is covalently bound to PG-Acr via Traut's reagent which does not compromise the hydrogel gelation and swelling behavior. Human mesenchymal stromal cells (MSCs) embedded within these bioactive hydrogels show a distinct dose-dependent chondrogenesis. Covalent incorporation of TGF-ß1 significantly enhances the chondrogenic differentiation of MSCs compared to hydrogels with supplemented noncovalently bound TGF-ß1. The observed chondrogenic response is similar to standard cell culture with TGF-ß1 addition with each medium change. In general, multifunctional PG-Acr offers the opportunity to introduce a range of biomimetic modifications (peptides, growth factors) into hydrogels and, thus, appears as an attractive potential material for various applications in regenerative medicine.
Subject(s)
Cell Differentiation/drug effects , Chondrocytes/drug effects , Hyaluronic Acid/chemistry , Hydrogels/chemistry , Mesenchymal Stem Cells/drug effects , Propylene Glycols/chemistry , Transforming Growth Factor beta1/pharmacology , Acrylates/chemistry , Cells, Immobilized/cytology , Cells, Immobilized/drug effects , Cells, Immobilized/metabolism , Chondrocytes/cytology , Chondrocytes/metabolism , Chondrogenesis/drug effects , Glycoconjugates/chemistry , Humans , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Primary Cell Culture , Protein Binding , Tissue Engineering/methods , Tissue Scaffolds , Transforming Growth Factor beta1/chemistryABSTRACT
This study investigates the use of allyl-functionalized poly(glycidol)s (P(AGE-co-G)) as a cytocompatible cross-linker for thiol-functionalized hyaluronic acid (HA-SH) and the optimization of this hybrid hydrogel as bioink for 3D bioprinting. The chemical cross-linking of gels with 10 wt.% overall polymer concentration was achieved by a UV-induced radical thiol-ene coupling between the thiol and allyl groups. The addition of unmodified high molecular weight HA (1.36 MDa) enabled the rheology to be tuned for extrusion-based bioprinting. The incorporation of additional HA resulted in hydrogels with a lower Young's modulus and a higher swelling ratio, especially in the first 24 h, but a comparable equilibrium swelling for all gels after 24 h. Embedding of human and equine mesenchymal stem cells (MSCs) in the gels and subsequent in vitro culture showed promising chondrogenic differentiation after 21 d for cells from both origins. Moreover, cells could be printed with these gels, and embedded hMSCs showed good cell survival for at least 21 d in culture. To achieve mechanically stable and robust constructs for the envisioned application in articular cartilage, the formulations were adjusted for double printing with thermoplastic poly(ε-caprolactone) (PCL).
