Three-dimensional bioprinting of multilayered constructs containing human mesenchymal stromal cells for osteochondral tissue regeneration in the rabbit knee joint

Abstract

The use of cell-rich hydrogels for three-dimensional (3D) cell culture has shown great potential for a variety of biomedical applications. However, the fabrication of appropriate constructs has been challenging. In this study, we describe a 3D printing process for the preparation of a multilayered 3D construct containing human mesenchymal stromal cells with a hydrogel comprised of atelocollagen and supramolecular hyaluronic acid (HA). This construct showed outstanding regenerative ability for the reconstruction of an osteochondral tissue in the knee joints of rabbits. We found that the use of a mechanically stable, host-guest chemistry-based hydrogel was essential and allowed two different types of extracellular matrix (ECM) hydrogels to be easily printed and stacked into one multilayered construct without requiring the use of potentially harmful chemical reagents or physical stimuli for post-crosslinking. To the best of our knowledge, this is the first study to validate the potential of a 3D printed multilayered construct consisting of two different ECM materials (atelocollagen and HA) for heterogeneous tissue regeneration using an in vivo animal model. We believe that this 3D printing-based platform technology can be effectively exploited for regeneration of various heterogeneous tissues as well as osteochondral tissue.

Three‐dimensional direct cell bioprinting for tissue engineering

“…Therefore, new studies focus on using alternative hydrogels such as decellularized extracellular matrix bioink to print cell‐laden constructs, which provides an optimized microenvironment for the growth of 3D tissue . Furthermore, researchers focus on fabricate 3D multilayered constructs consisting live cells and different ECM materials for heterogenous tissue regeneration …”

“…21 Furthermore, researchers focus on fabricate 3D multilayered constructs consisting live cells and different ECM materials for heterogenous tissue regeneration. 22 Extrusion-based bioprinting process can also be used for direct printing of cellular-aggregates. In extrusion-based bioprinting processes, biomaterial concentration, loaded cell viscosity, nozzle pressure, and nozzle diameter are determinant parameters for continuous deposition of cylindrical cell aggregates and hydrogel biomaterials.…”

From intricate to integrated: Biofabrication of articulating joints

“…Biofabricated osteochondral constructs have been already adopted in an in vivo study involving a MSC‐laden collagen and hyaluronic acid hydrogel construct reinforced by PCL. This artificial osteochondral plug was implanted in a rabbit knee and appeared to be mechanically stable and to integrate well with the native cartilage and the underlying bone . The success of this study was, at least in part, due to a perfectly fitting design of the prosthesis, as well as the good integration in both bone and cartilage region, achieved by stack crosslinking with the same cell‐friendly chemistry.…”

“…A wide range of smaller osteochondral constructs have been successfully generated using AM alone or in combination with conventional techniques, including casting, freeze‐drying and solvent casting/particle leaching . Even though the generation of a long‐term functional solution in osteochondral tissue engineering remains challenging, in vivo approaches with 3D printed osteochondral plugs have already been reported …”

A Dual-Bonded Approach for Improving Hydrogel Implant Stability in Cartilage Defects

“…Without blood vessels, nerves, and lymphoid tissues, once there is injury to the articular cartilage, it has almost no effective ability to repair and rebuild itself [1]. Hydrogels are a kind of three-dimensional porous network with similar mechanical characteristics to those of articular cartilage [2,3,4]. As an ideal material substitute to repair articular cartilage, the physical and chemical properties of hydrogels such as their excellent biocompatibility have received extensive attention from researchers [5,6].…”