Abstract:We investigated the proliferation and osteogenic differentiation of mesenchymal stem cells cultured on fibroin microcarriers. Effective cell proliferation on the surface of the microcarriers, determined by the large surface area, and the contribution of microcarrier mineralization to the stimulation of the osteogenic differentiation of mesenchymal stem cells was revealed.
“…Osteosarcoma cell lines (MG‐63) and MSCs cell adhesion occurred for all types of MCs. Cell proliferation was found to be significatively lower for mineralised MCs, probably since substrate mineralisation contributed to the induction of osteogenic differentiation in the absence of inductors, reducing the rate of proliferation [81]. While gelatin can be added to the material in order to promote the cellular response [76], calcium phosphate surface increases the MCs roughness.…”
Section: Silk Fibroin‐based Mcsmentioning
confidence: 99%
“…Fibroin MCs modified by calcium phosphate mineralisation have also been studied. In the work of Kotliarova et al [29] and Goncharenko et al [81], porous MCs with the size ranging from 100 to 250 μm were obtained by cryodestruction of the matrices to generate fibroin, fibroin/gelatin, fibroin/calcium phosphate and fibroin/gelatin/calcium phosphate materials. Osteosarcoma cell lines (MG‐63) and MSCs cell adhesion occurred for all types of MCs.…”
“…Osteosarcoma cell lines (MG‐63) and MSCs cell adhesion occurred for all types of MCs. Cell proliferation was found to be significatively lower for mineralised MCs, probably since substrate mineralisation contributed to the induction of osteogenic differentiation in the absence of inductors, reducing the rate of proliferation [81]. While gelatin can be added to the material in order to promote the cellular response [76], calcium phosphate surface increases the MCs roughness.…”
Section: Silk Fibroin‐based Mcsmentioning
confidence: 99%
“…Fibroin MCs modified by calcium phosphate mineralisation have also been studied. In the work of Kotliarova et al [29] and Goncharenko et al [81], porous MCs with the size ranging from 100 to 250 μm were obtained by cryodestruction of the matrices to generate fibroin, fibroin/gelatin, fibroin/calcium phosphate and fibroin/gelatin/calcium phosphate materials. Osteosarcoma cell lines (MG‐63) and MSCs cell adhesion occurred for all types of MCs.…”
“…Particle size and size distribution can be modulated to some degree by varying the viscosity of the continuous phase, the rate of emulsification, and the impeller geometry, as well as by the addition of surfactants and post‐production filtration 23,24 . In contrast, microfluidic or flow‐focusing techniques are capable of generating particles with a narrow size distribution; however, the rate of production is comparatively low 25,26 …”
Section: Introductionmentioning
confidence: 99%
“…23,24 In contrast, microfluidic or flow-focusing techniques are capable of generating particles with a narrow size distribution; however, the rate of production is comparatively low. 25,26 Gelatin and other collagen-derived materials have been used extensively as base materials for microcarriers due to their broad availability, ease of processing, and generally excellent cytocompatibility.…”
mentioning
confidence: 99%
“…The fabrication of collagen 23,30 and gelatin [31][32][33][34][35][36] microparticles has been demonstrated using bulk emulsification, 37,38 microfluidics, 39 coacervation, 40 and solgel synthesis. 12 In some cases these microcarriers are augmented with other macromolecules such as chitosan 41 or silk fibroin, 25,39 or with a mineral phase such as tricalcium phosphate, 15,30 calcium-deficient hydroxyapatite, 42,43 or hydroxyapatite. 38,44 Collagen and collagen-peptides foster the attachment and reorganization by many cell types, including a variety of progenitor cells known to be involved in bone formation.…”
Biomaterial‐based bone regeneration strategies often include a cellular component to accelerate healing. Modular approaches have the potential for minimally‐invasive delivery and the ability to conformally fill complex defects. In this study, spherical gelatin microparticles were fabricated via water‐in‐oil emulsification and were subsequently crosslinked with genipin. Microparticle diameter depended on impeller geometry, and increased stirring rates consistently produced smaller particles with narrower size distributions. Increasing the concentration of gelatin resulted in larger particles with a broader size distribution. Viscoelastic characterization showed that increased gelatin concentration produced stiffer matrices, though the mechanical properties at lower gelatin concentration were more stable across strain rate. Microparticles of 6.0% wt/vol gelatin were then applied as microcarriers for packed‐bed culture of human mesenchymal stromal cells (MSC) at seeding densities of 5.0 × 103, 2.5 × 104, or 5.0 × 104 cells/cm2 of surface area, in either control or osteogenic medium. Cell viability was uniformly high (>90%) across seeding densities over 22 days in culture. MSC number stayed approximately constant in the 5.0 × 103 and 2.5 × 104 cells/cm2 samples, while it dropped over time at 5.0 × 104 cells/cm2. Alkaline phosphatase activity was significantly upregulated in osteogenic conditions relative to controls at day 15, and absolute calcium deposition was strongly induced by days 15 and 22. However, calcium deposition per cell was highest in the lowest cell density, suggesting an inhibitory effect of high cell numbers. These results show that genipin‐crosslinked gelatin microcarriers can be reproducibly fabricated and used as microcarriers for progenitor cells, which may have utility in treating large and complex bone defects.
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