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Behavior of Muscle-Derived Stem Cells on Silica Nanostructured Substrates.

ABSTRACT: The aim of the present work was to evaluate the responses of rat muscle-derived stem cells (rMDSCs) to growth on silica nanostructured substrates (SN) with nanoscale topographic surfaces. SN of different sizes (SN-60, SN-150, SN-300, SN-500, and SN-700) were prepared using silica nanoparticles with sizes of 60-700 nm. The prepared SN showed roughness at the nanoscale level. The total number of adherent cells on SN increased with increasing nanoscale level and incubation time. The rMDSCs attached to SN-500 and SN-700 were extensively flattened, whereas those grown on SN-60, SN-150, and SN-300 were more rounded. The rank order of the cell length and height of attached rMDSCs at 5 d on different surfaces was SN-60 ? SN-150 >> SN-300 > SN-500 > SN-700 > glass. Compared with rMDSCs grown on SN-60, SN-150, or SN-300, those attached to SN-500 and SN-700 exhibited a distinct morphology with filopodial extensions and stronger expression of focal adhesion, integrin, and actin. An evaluation of the gene expression of adhered rMDSCs showed that rMDSCs grown on SN-300 exhibited a higher environmental stress response than those grown on glass or SN-700. Collectively, our data provide fundamental insight into the cellular response and gene expression of rMDSCs grown on nanostructured substrates.

SUBMITTER: Kim HS

PROVIDER: S-EPMC7558836 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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Behavior of Muscle-Derived Stem Cells on Silica Nanostructured Substrates.

Kim Hyo-Sop HS Lee Bit Na BN Choi Sangdun S Kim Moon Suk MS Kim Jae-Ho JH

Nanomaterials (Basel, Switzerland) 20200822 9

The aim of the present work was to evaluate the responses of rat muscle-derived stem cells (rMDSCs) to growth on silica nanostructured substrates (SN) with nanoscale topographic surfaces. SN of different sizes (SN-60, SN-150, SN-300, SN-500, and SN-700) were prepared using silica nanoparticles with sizes of 60-700 nm. The prepared SN showed roughness at the nanoscale level. The total number of adherent cells on SN increased with increasing nanoscale level and incubation time. The rMDSCs attached ...[more]

PMID: 32842628

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Project description:Silica nonwoven fabrics (SNFs) with enough mechanical strength are candidates as implantable scaffolds. Culture of cells therein is expected to affect the proliferation and differentiation of the cells through cell-cell and cell-SNF interactions. In this study, we examined three-dimensional (3D) SNFs as a scaffold of mesenchymal stem cells (MSCs) for bone tissue engineering applications. The interconnected highly porous microstructure of 3D SNFs is expected to allow omnidirectional cell-cell interactions, and the morphological similarity of a silica nanofiber to that of a fibrous extracellular matrix can contribute to the promotion of cell functions. 3D SNFs were prepared by the sol-gel process, and their mechanical properties were characterized by the compression test and rheological analysis. In the compression test, SNFs showed a compressive elastic modulus of over 1 MPa and a compressive strength of about 200 kPa. These values are higher than those of porous polystyrene disks used for in vitro 3D cell culture. In rheological analysis, the elastic modulus and fracture stress were 3.27 ± 0.54 kPa and 25.9 ± 8.3 Pa, respectively. Then, human bone marrow-derived MSCs were cultured on SNFs, and the effects on proliferation and osteogenic differentiation were evaluated. The MSCs seeded on SNF proliferated, and the thickness of the cell layer became over 80 μm after 14 days of culture. The osteogenic differentiation of MSCs on SNFs was induced by the culture in the commercial osteogenic differentiation medium. The alkaline phosphatase activity of MSCs on SNFs increased rapidly and remained high up to 14 days and was much higher than that on two-dimensional tissue culture-treated polystyrene. The high expression of RUNX2 and intense staining by alizarin red s after differentiation supported that SNFs enhanced the osteogenic differentiation of MSCs. Furthermore, permeation analysis of SNFs using fluorescein isothiocyanate-dextran suggested a sufficient permeability of SNFs for oxygen, minerals, nutrients, and secretions, which is important for maintaining the cell viability and vitality. These results suggested that SNFs are promising scaffolds for the regeneration of bone defects using MSCs, originated from highly porous and elastic SNF characters.

Project description:BACKGROUND:The unmet medical needs in repairing large muscle defects promote the development of tissue regeneration strategy. The use of bioactive molecules in combination with biomaterial scaffold has become an area of great interest. SW033291, a small-molecule inhibitor targeting 15-hydroxyprostaglandin dehydrogenase (15-PDGH) and subsequently elevating the production of prostaglandin E2 (PGE2), has been proved to accelerate the recovery and potentiate the regeneration of multiple tissues including the bone, liver, and colon. The limited understanding of the potential therapeutic effects on myogenesis motivated us to investigate the role of SW033291 in regulating muscle-derived stem cell (MDSC) myogenic differentiation and MDSC-mediated muscle regeneration. METHODS:The characteristics of rat MDSCs, including cell-specific markers and myogenic differentiation potential, were determined. MDSCs were incubated with SW033291 to evaluate PGE2 production and cytotoxicity. The effects of SW033291 on MDSC myogenic differentiation were assessed by quantitative real-time polymerase chain reaction (qPCR), western blot, and immunocytochemistry. The fibrin gel containing MDSCs and SW033291 was used for muscle regeneration in a tibialis anterior muscle defect model. RESULTS:Our data demonstrated that MDSCs were well-tolerated to SW033291 and treatment with SW033291 significantly promoted the production of PGE2 by MDSCs. In vitro analysis showed that SW033291 enhanced the myogenic differentiation and myotube formation by upregulating a series of myogenic markers. Additionally, the activation of PI3K/Akt pathway was involved in the mechanism underlying these promotive effects. Then, in situ casting of fibrin gel containing MDSCs and SW033291 was used to repair the tibialis anterior muscle defect; the addition of SW033291 significantly promoted myofiber formation within the defect region with mild immune response, less fibrosis, and sufficient vascularization. CONCLUSION:SW033291 acted as a positive regulator of MDSC myogenic differentiation, and incorporating the compound with MDSCs in fibrin gel could serve as an effective method to repair large skeletal muscle defects.

Dataset Information

Behavior of Muscle-Derived Stem Cells on Silica Nanostructured Substrates.

Publications

Behavior of Muscle-Derived Stem Cells on Silica Nanostructured Substrates.

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