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Understanding Myoblast Differentiation Pathways When Cultured on Electroactive Scaffolds through Proteomic Analysis.
Sylvie Ribeiro1,2,3, Clarisse Ribeiro1,2, Vítor M Martins3,4
1Physics Centre of Minho and Porto Universities (CF-UM-UP), University of Minho, 4710-057 Braga, Portugal.
ACS Applied Materials & Interfaces
|May 31, 2022
Summary
This study investigated how charged piezoelectric polymer films affect C2C12 cell differentiation for skeletal muscle regeneration. Negatively charged surfaces showed a more pronounced positive effect on cell differentiation and protein expression compared to positively charged surfaces.
Area of Science:
- Biomaterials Science
- Cell Biology
- Proteomics
Background:
- Electroactive materials offer tunable control over cell-material interactions for tissue regeneration.
- Understanding the cellular-level impact of these materials, particularly their surface charge, is crucial but remains limited.
- Piezoelectric polymers, like poly(vinylidene fluoride) (PVDF), are promising for modulating cell behavior.
Purpose of the Study:
- To analyze the proteome of C2C12 myoblasts differentiating on PVDF films with varying surface charges (non-poled, poled +, poled -).
- To identify specific protein and pathway alterations related to skeletal muscle development influenced by surface charge.
- To compare the effects of charged PVDF films against a control (polystyrene) to elucidate the role of surface charge.
Main Methods:
- Proteomic analysis using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS).
- Culturing C2C12 cells on non-poled, poled +, and poled - PVDF films, alongside a polystyrene control.
- Bioinformatic analysis of significantly expressed proteins (P < 0.01, ANOVA).
Main Results:
- PVDF films with overall surface charge (poled + and poled -) significantly altered C2C12 cell proteomes compared to non-poled films and controls.
- Both positive and negative charges enhanced skeletal muscle development proteins, with a more pronounced effect observed on negatively charged surfaces.
- Specific proteins involved in calcium signaling and muscle contraction showed differential expression patterns based on surface charge, e.g., Ckm, Tmem14c, Serpinb6a, Mylpf, Mybph, Mbnl1, and Hba-a1.
Conclusions:
- Scaffold surface charge is a critical factor influencing C2C12 cell behavior and differentiation.
- Both positive and negative surface charges promote skeletal muscle cell differentiation via proteins involved in muscle contraction.
- Negatively charged piezoelectric polymer surfaces demonstrate a more significant impact on promoting skeletal muscle development compared to positively charged surfaces.
