Improving the osteogenesis of human bone marrow mesenchymal stem cell sheets by microRNA-21-loaded chitosan/hyaluronic acid nanoparticles via reverse transfection

Abstract

Cell sheet engineering has emerged as a novel approach to effectively deliver seeding cells for tissue regeneration, and developing human bone marrow mesenchymal stem cell (hBMMSC) sheets with high osteogenic ability is a constant requirement from clinics for faster and higher-quality bone formation. In this work, we fabricated biocompatible and safe chitosan (CS)/hyaluronic acid (HA) nanoparticles (NPs) to deliver microRNA-21 (miR-21), which has been proved to accelerate osteogenesis in hBMMSCs; then, the CS/HA/miR-21 NPs were cross-linked onto the surfaces of culture plates with 0.2% gel solution to fabricate miR-21-functionalized culture plates for reverse transfection. hBMMSC sheets were induced continuously for 14 days using a vitamin C-rich method on the miR-21-functionalized culture plates. For the characterization of CS/HA/miR-21 NPs, the particle size, zeta potential, surface morphology, and gel retardation were sequentially investigated. Then, the biological effects of hBMMSC sheets on the miR-21-functionalized culture plates were evaluated. The assay results demonstrated that the hBMMSC sheets could be successfully induced via the novel reverse transfection approach, and miR-21 delivery significantly enhanced the in vitro osteogenic differentiation of hBMMSC sheets in terms of upregulating calcification-related gene expression and enhancing alkaline phosphatase production, collagen secretion, and mineralized nodule formation. The enhanced osteogenic activity of hBMMSC sheets might promisingly lead to more rapid and more robust bone regeneration for clinical use.

Role of Epigenomics in Bone and Cartilage Disease

“…A number of miRNAs enhance the osteogenic differentiation of MSCs; hence, they have been pointed out as potential therapies to promote bone regeneration. For example, scaffolds and particles loaded with analogs of miR‐21 or miR‐148 potentiate bone regeneration in experimental models, at least in part, by interacting with the RUNX2 pathway . On the other hand, anti‐sense oligonucleotides and “sponges” inhibiting miR‐22, miR‐29, miR‐31, miR‐133, miR‐138, or miR‐214 have a stimulatory effect on osteogenesis and may improve fracture healing .…”

A Concise Review on the Use of Mesenchymal Stem Cells in Cell Sheet-Based Tissue Engineering with Special Emphasis on Bone Tissue Regeneration

“…There are ongoing investigations to obtain the best results in bone regeneration with MSC sheets. Some of the biomaterials and/or biological agents reported to be combined with MSC sheets to enhance osteogenesis include coral particles [ 47 ], ceramics [ 48 ], surface-modified titanium and zirconia [ 82 ], simvastatin [ 83 ], β -tricalcium phosphate [ 84 ], coumarin-like derivative osthole [ 85 ], CD34+ peripheral blood cells [ 86 ], a complex of polyethylenimine-alginate nanocomposites plus BMP2 gene [ 87 ], nonviral oligonucleotide antimiR-138 delivery to MSC sheets [ 88 ], platelet-rich fibrin [ 89 , 90 ], vitamin C [ 91 ], poly(dimethylsiloxane) surface silanization [ 92 ], stromal cell-derived factor-1 [ 93 ], microRNA-21-loaded chitosan/hyaluronic acid nanoparticles [ 94 ], hydroxyapatite particles [ 90 , 95 ], and Notch activation by Jagged1 in MSC sheet cultures [ 96 ].…”

Signaling pathways of nucleic acids for bone healing: A review

“…used a microarc-oxidized titanium surface with miRNA-21loaded hyaluronic acid nanoparticles, and they confirmed that this scaffold promotes osteogenic differentiation on human bone marrow mesenchymal stem cells. [44][45][46] There are no clinical trials entries using miRNAs or antagomirs as bone diseases therapy in clinicaltrial.gov to date. However, there is study from Moscow State University started in 2017, that evaluates the safety and efficacy of a gene-activated bone substitute consisting of octacalcium phosphate and plasmid DNA encoding vascular endothelial growth factor for maxillofacial bone regeneration.…”