In VitroAssessment of the Differentiation Potential of Bone Marrow-Derived Mesenchymal Stem Cells on Genipin-Chitosan Conjugation Scaffold with Surface Hydroxyapatite Nanostructure for Bone Tissue Engineering

Abstract

Increasing evidence has revealed that the surface characteristics of biomaterials, such as chemical composition, stiffness, and topography, especially nanotopography, significantly influence cell growth and differentiation. In this study, we examined the effect of surface biomimetic apatite nanostructure of a new hydroxyapatite-coated genipin-chitosan conjugation scaffold (HGCCS) on cell shape, cytoskeleton organization, and osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells in vitro. Cell shape and cytoskeleton organization showed significant differences between cells cultured on genipin-cross-linked chitosan framework and those cultured on HGCCS with surface apatite network-like nanostructure after 7 days of incubation in the osteogenic medium. The result of specific alkaline phosphatase activity as an indicator of osteogenic differentiation showed that the alkaline phosphatase activity of rat bone marrow-derived mesenchymal stem cells was higher on HGCCS. Based on quantitative real-time polymerase chain reaction, HGCCS induced highest mRNA expression of osteogenic differentiation makers, runt-related transcription factor 2 by 7 days, osteopontin by 7 days, and osteocalcin by 14 days, respectively. The enhanced ability of cells on HGCCS to produce mineralized extracellular matrix and nodules was also assessed on day 14 with Alizarin red staining. The results of this study suggest that the surface biomimetic apatite nanostructure of HGCCS is a critical signal cue to promoting osteogenic differentiation in vitro. These findings open a new research avenue to controlling stem cell lineage commitment and provide a promising scaffold for bone tissue engineering.

Bone repair by periodontal ligament stem cell-seeded nanohydroxyapatite-chitosan scaffold

“…33 In this study, the alkaline phosphatase activity of PDLSCs on GCF and HGCCS was measured for up to 14 days. The results show that the alkaline phosphatase activity of the cells on both scaffolds peaked after 7 days of cultivation but decreased by day 14 ( Figure 5).…”

“…Moreover, the surface biomimetic apatite nanostructure of HGCCS can promote osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells in vitro. 33 In this present study, we isolated and cultured human PDLSCs and examined the effect of HGCCS on cell morphology, cell viability, and osteogenic differentiation of PDLSCs. Subsequently, a rat calvarial critical size defect model was used to evaluate the potential of PDLSC-seeded HGCCS constructs in vivo for bone tissue regeneration.…”

Graphene oxide-reinforced biodegradable genipin-cross-linked chitosan fluorescent biocomposite film and its cytocompatibility

“…24,25 In addition, CS-genipin conjugates possess a novel intrinsic fluorescent property, which broadens their use in biomedical applications, like in situ observation. 26,27 In our previous work, we fabricated a hydroxyapatite-coated genipin−CS conjugation scaffold with fluorescent properties, which could provide effective ways for imaging the scaffold−cell interface and for tracing the adhesion, migration, and proliferation of cells, by a confocal laser scanning microscope (CLSM). 26 Due to their individual excellent properties, both GO incorporated into CS and genipin-cross-linked CS (GCS) are expected to hold promising performance.…”

“…26,27 In our previous work, we fabricated a hydroxyapatite-coated genipin−CS conjugation scaffold with fluorescent properties, which could provide effective ways for imaging the scaffold−cell interface and for tracing the adhesion, migration, and proliferation of cells, by a confocal laser scanning microscope (CLSM). 26 Due to their individual excellent properties, both GO incorporated into CS and genipin-cross-linked CS (GCS) are expected to hold promising performance. Recently, Pan et al presented a simple approach to the fabrication of GO/CS nanocomposite films with enhanced mechanical properties.…”

Effects of hydroxyapatite nanostructure on channel surface of porcine acellular dermal matrix scaffold on cell viability and osteogenic differentiation of human periodontal ligament stem cells

“…9 Recently, the authors' group has successfully fabricated an HAp-PADM scaffold with a 3D two-level porous structure by assembling the HAp nanostructures on the channel surface of a PADM framework by a two-step biomimetic mineralization method. This HAp-PADM scaffold has both interconnected channels (∼100 µm in diameter) formed by collagen fiber bundles and nanopores (,100 nm in diameter).…”