Journal: Advanced Science

Article Title: Ultrafast Micrometric Laser Structuring of Polymer Surfaces for Highly Organized Skeletal Muscle Tissue Modeling

doi: 10.1002/advs.78129

Figure Lengend Snippet: Relative gene expression of C2C12 myoblasts cultured on the LS surface compared with the standard surface. Data are presented as mean ± SD with individual data points ( n = 3). Gene expression levels were normalized to the housekeeping gene and expressed as fold change relative to the standard group (Std). Panels correspond to: (a) d0, (b) d4, (c) d7, and (d) d11. Statistical significance was determined using Student's t ‐test: * p < 0.05; ** p < 0.01; *** p < 0.001 vs. Standard.

Article Snippet: Murine myoblasts C2C12 (ATCC) were cultured according to a previously published protocol [ ] in T‐75 Flasks (Sarstedt) at 37°C and 5% CO 2 in Dulbecco's modified Eagle medium GlutaMAX (DMEM, Gibco) supplemented with 10% heat‐inactivated fetal bovine serum (FBS, EuroClone), penicillin (100 IU mL −1 , Gibco), and streptomycin (100 mg mL −1 , Gibco).

Techniques: Gene Expression, Cell Culture

Journal: Advanced Science

Article Title: Ultrafast Micrometric Laser Structuring of Polymer Surfaces for Highly Organized Skeletal Muscle Tissue Modeling

doi: 10.1002/advs.78129

Figure Lengend Snippet: (a) Infrared spectra of the standard surface before (Std, red line) and after laser structuring (LS, blue line); (b) MTT assay. The C2C12 cell growth at three experimental time points (24 h, 48 h, and 72 h) is plotted for the cells seeded on a standard surface (Std, red line) and LS (ex. LS, blue line) surfaces. A theoretical curve was also plotted, taking into account the area available on the LS surface (th. LS, dashed dark blue line). (a.u. = arbitrary unit); (c) Brightfield pictures of the cells cultured on standard (top pictures) or LS (bottom pictures) surfaces at the three experimental time points (24 h, 48 h, and 72 h).

Article Snippet: Murine myoblasts C2C12 (ATCC) were cultured according to a previously published protocol [ ] in T‐75 Flasks (Sarstedt) at 37°C and 5% CO 2 in Dulbecco's modified Eagle medium GlutaMAX (DMEM, Gibco) supplemented with 10% heat‐inactivated fetal bovine serum (FBS, EuroClone), penicillin (100 IU mL −1 , Gibco), and streptomycin (100 mg mL −1 , Gibco).

Techniques: Infrared, MTT Assay, Brightfield, Cell Culture

Journal: Advanced Science

Article Title: Ultrafast Micrometric Laser Structuring of Polymer Surfaces for Highly Organized Skeletal Muscle Tissue Modeling

doi: 10.1002/advs.78129

Figure Lengend Snippet: Image analyses conducted on C2C12 after 7 days of differentiation. Pictures a, b, and c show representative immunofluorescence pictures superimposed on the top with their relative brightfield pictures. The myotubes positive for MHC (Red), are randomly oriented in the standard plates (a) while properly aligned on the LS surfaces (channel spacing of 75 µm (b) and 150 µm (c)). Orientation of muscle fibers. The distribution of orientation is plotted for unstructured standard plates (Std) and for all spacing (d, 75, 100, 125, and 150 µm) and depth (g, 3.5 – 5 – 8;5 – 11.5 – 14 – 15 ‐20 µm) values tested, revealing the presence of a prevalent peak for all conditions compared to their respective control (black plot); the quantification of the orientation through the coherency value (e, f) shows a significative increase for all the LS surfaces compared to the standard ( * ). Moreover, regarding the spacing analysis, the three higher values are also significative compared to 75 µm (e, $ ). Myotube lengths. The myotubes differentiated on LS surfaces with a spacing of 125 µm and a depth of 11.5 µm (h, red bar) are significantly longer than those grown on the standard plates (h, black bar). Sample size n = 3. p value list: spacing, **** p < 0.0001, *** p = 0.0001, $$$ p = 0.0006, $$ p = 0.0013 (75 µm vs 100 µm), $$ p = 0.0059 (75 µm vs 150 µm); depth, **** p < 0.0001, *** p = 0.0007, ** p = 0.001 (3.5 µm), ** p = 0.0025 (14 µm), ** p = 0.0085 (20 µm); length, **** p < 0.0001.

Article Snippet: Murine myoblasts C2C12 (ATCC) were cultured according to a previously published protocol [ ] in T‐75 Flasks (Sarstedt) at 37°C and 5% CO 2 in Dulbecco's modified Eagle medium GlutaMAX (DMEM, Gibco) supplemented with 10% heat‐inactivated fetal bovine serum (FBS, EuroClone), penicillin (100 IU mL −1 , Gibco), and streptomycin (100 mg mL −1 , Gibco).

Techniques: Immunofluorescence, Brightfield, Control, Analysis

Journal: Advanced Science

Article Title: Ultrafast Micrometric Laser Structuring of Polymer Surfaces for Highly Organized Skeletal Muscle Tissue Modeling

doi: 10.1002/advs.78129

Figure Lengend Snippet: Analysis of MHC and Desmin protein levels during C2C12 myoblast differentiation. Western blot analysis of C2C12 cells cultured on standard (Std) and LS surfaces at different stages of differentiation. Representative immunoblot for (a) MHC and Tubulin, and (c) Desmin and GAPDH. Densitometric analysis of (b) MHC and (d) Desmin protein levels. Data are presented as mean ± SD with individual data points ( n = 3). Statistical significance was assessed using Student's t ‐test ( * p < 0.05; ** p < 0.01; *** p < 0.001 vs. Standard at each time point).

Article Snippet: Murine myoblasts C2C12 (ATCC) were cultured according to a previously published protocol [ ] in T‐75 Flasks (Sarstedt) at 37°C and 5% CO 2 in Dulbecco's modified Eagle medium GlutaMAX (DMEM, Gibco) supplemented with 10% heat‐inactivated fetal bovine serum (FBS, EuroClone), penicillin (100 IU mL −1 , Gibco), and streptomycin (100 mg mL −1 , Gibco).

Techniques: Analysis, Western Blot, Cell Culture