Extracellular matrix mechanics regulate transfection and SOX9-directed differentiation of mesenchymal stem cells
| Autor: | Kyle H. Vining, Marcos Garcia-Fuentes, Adriana M. Ledo, María J. Alonso, David J. Mooney |
|---|---|
| Přispěvatelé: | Universidade de Santiago de Compostela. Centro de Investigación en Medicina Molecular e Enfermidades Crónicas, Universidade de Santiago de Compostela. Departamento de Farmacoloxía, Farmacia e Tecnoloxía Farmacéutica |
| Rok vydání: | 2020 |
| Předmět: |
animal structures
0206 medical engineering Biomedical Engineering macromolecular substances 02 engineering and technology Gene delivery Transfection Biochemistry Regenerative medicine Article Biomaterials Directed differentiation Tissue engineering Humans GAMs Molecular Biology IPNs Chemistry Regeneration (biology) Mesenchymal stem cell technology industry and agriculture Cell Differentiation Hydrogels Mesenchymal Stem Cells SOX9 Transcription Factor General Medicine 021001 nanoscience & nanotechnology Chondrogenesis 020601 biomedical engineering Extracellular Matrix Cell biology embryonic structures Self-healing hydrogels 0210 nano-technology 3D transfection SOX9 Biotechnology |
| Zdroj: | Minerva: Repositorio Institucional de la Universidad de Santiago de Compostela Universidad de Santiago de Compostela (USC) Minerva. Repositorio Institucional de la Universidad de Santiago de Compostela instname Acta Biomater |
| Popis: | Gene delivery within hydrogel matrices can potentially direct mesenchymal stem cells (MSCs) towards a chondrogenic fate to promote regeneration of cartilage. Here, we investigated whether the mechanical properties of the hydrogel containing the gene delivery systems could enhance transfection and chondrogenic programming of primary human bone marrow-derived MSCs. We developed collagen-I-alginate interpenetrating polymer network hydrogels with tunable stiffness and adhesion properties. The hydrogels were activated with nanocomplexed SOX9 polynucleotides to direct chondrogenic differentiation of MSCs. MSCs transfected within the hydrogels showed higher expression of chondrogenic markers compared to MSCs transfected in 2D prior to encapsulation. The nanocomplex uptake and resulting expression of transfected SOX9 were jointly enhanced by increased stiffness and cell-adhesion ligand density in the hydrogels. Further, transfection of SOX9 effectively induced MSCs chondrogenesis and reduced markers of hypertrophy compared to control matrices. These findings highlight the importance of matrix stiffness and adhesion as design parameters in gene-activated matrices for regenerative medicine. STATEMENT OF SIGNIFICANCE: Gene-activated matrices (GAMs) are biodegradable polymer networks integrating gene therapies, and they are promising technologies for supporting tissue regeneration. Despite this interest, there is still limited information on how to rationally design these systems. Here, we provide a systematic study of the effect of matrix stiffness and cell adhesion ligands on gene transfer efficiency. We show that high stiffness and the presence of cell-binding sites promote transfection efficiency and that this result is related to more efficient internalization and trafficking of the gene therapies. GAMs with optimized mechanical properties can induce cartilage formation and result in tissues with better characteristics for articular cartilage tissue engineering as compared to previously described standard methods. |
| Databáze: | OpenAIRE |
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