Development of a biomimetic arch-like 3D bioprinted construct for cartilage regeneration using gelatin methacryloyl and silk fibroin-gelatin bioinks

Abstract

In recent years, engineering biomimetic cellular microenvironments have been a top priority for regenerative medicine. Collagen II, which is arranged in arches, forms the predominant fiber network in articular cartilage. Due to the shortage of suitable microfabrication techniques capable of producing 3D fibrous structures, in vitro replication of the arch-like cartilaginous tissue constitutes one of the major challenges. Hence, in the present study, we report a 3D bioprinting approach for fabricating arch-like constructs using two types of bioinks, gelatin methacryloyl(GelMa) and silk fibroin-gelatin(SF-G). The bioprinted SF-G constructs displayed increased proliferation of the encapsulated human bone marrow-derived mesenchymal stem cells compared to the GelMA constructs. Biochemical assays, gene, and protein expression exhibited the superior role of SF-G in forming the fibrous collagen network and chondrogenesis. Protein-protein interaction study using Metascape evaluated the function of the proteins involved. Further GeneMANIA and STRING analysis using Col2A1, SOX9, ACAN, and the genes upregulated on day 21 in RT-PCR, i.e., β-catenin, TGFβR1, Col1A1 in SF-G and PRG4, Col10A1, MMP13 in GelMA validated our in vitro results. These findings emphasized the role of SF-G in regulating the Wnt/β-catenin and TGF-β signaling pathways. Hence, the 3D bioprinted arch-like constructs possess a substantial potential for cartilage regeneration.


Covalent Conjugation of Small Molecule Inhibitors and Growth Factors to a Silk Fibroin-Derived Bioink to Develop Phenotypically Stable 3D Bioprinted Cartilage

“…GeneMANIA software was used to predict the correlation and interaction of the input genes with the pool of genes associated with articular cartilage homeostasis. Through this analysis, we can validate the importance of the chosen set of genes for qPCR . Furthermore, all the experimental groups were chosen from one single donor showing maximum chondrogenic characteristics (donor-2) and the respective 5 upregulated genes of each experimental group were selected for the analysis.…”

“…Through this analysis, we can validate the importance of the chosen set of genes for qPCR. 58 Furthermore, all the experimental groups were chosen from one single donor showing maximum chondrogenic characteristics (donor-2) and the respective 5 upregulated genes of each experimental group were selected for the analysis. The different colors represent the different forms of interactions between the respective genes.…”

Hierarchically structured biomaterials for tissue regeneration

“…In another study, the uniaxial arrangement of myoblasts in gelatin methacryloyl (GelMA) was accomplished by the in-situ crosslinking process during 3D bioprinting [39] . The related in vitro and in vivo experiments found that GelMA scaffolds loaded with cells with arranged cell structures are expected to become a promising platform for promoting muscle regeneration.…”

Enhanced chondrogenic potential in GelMA-based 3D cartilage model via Wnt3a surface immobilization

“…Gelatin methacryloyl (GelMA) is a commonly used hydrogel that exhibits excellent biocompatibility and can be easily functionalized to incorporate bioactive cues 4 , 7 , 8 . GelMA-based hydrogels have shown great potential for cartilage tissue engineering applications such as drug screening 9 – 11 . They provide a compatible and scaleable microenvironment for enhanced chondrogenic cell viability and functionality 12 .…”