Understanding myoblast differentiation pathways when cultured on electroactive scaffolds through proteomic analysis

• Autor(a) principal: Ribeiro, Sylvie Oliveira
• Data de Publicação: 2022
• Outros Autores: Ribeiro, Clarisse, Martins, Vítor M., Honoré, Bent, Neves, Maria Teresa, Gomes, Andreia C, Lanceros-Méndez, S.
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: https://hdl.handle.net/1822/82060
• topic: Proteomic analysis; Active polymers; Poly(vinylidene fluoride) films; C2C12 myoblast cells; Myotube; Science & Technology

Electroactive materials allow to modulate cell-materials interactions and cell fate, leading to advanced tissue regeneration strategies. Nevertheless, their effect at the cellular level is still poorly understood. In this context, the proteome analysis of C2C12 cell differentiation cultured on piezoelectric polymer films with null average surface charge (non-poled), net positive surface charge (poled +) and net negative surface charge (poled -) has been addressed. Protein/pathway alterations for skeletal muscle development were identified comparing proteomic profiles of C2C12 cells differentiated on poly(vinylidene fluoride) with similar cells differentiated on polystyrene plate (control), using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS). Only significantly expressed proteins (P<0.01, analysis of variance) were used for bioinformatic analyses. A total of 37 significantly expressed proteins were detected on the C2C12 proteome with PVDF “poled –” at 24 h, whereas on the PVDF “poled +” a total of 105 significantly expressed proteins were considered. At 5 days of differentiation, the number of significantly expressed proteins decreased to 23 and 31 in cells grown on negative and positive surface charge, respectively, being the influence of surface charge more explicit in some proteins. In both cases, proteins such as Fbn1, Hspg2, Rcn3, Tgm2, Mylpf, Anxa2, Anxa6, involved in calcium-related signaling, were highly expressed during myoblast differentiation. Furthermore, some proteins involved in muscle contraction (Acta2, Anxa2, Anxa6) were detected in the PVDF “poled +” sample. Upregulation of several proteins that enhance skeletal muscle development was detected in the PVDF “poled –” sample, including Ckm (422%), Tmem14c (384%), Serpinb6a (460%), adh7 (199%), and Car3 (171%), while for the “poled +” samples these proteins were also upregulated at a smaller magnitude (254%, 317%, 253%, 123%, 72%, respectively). Other differentially expressed proteins such as Mylpf (189%), Mybph (168%), and Mbnl1 (168%) were upregulated only in PVDF “poled –” samples, while Hba-a1 levels (581%) were increased in PVDF “poled +” sample. On the other hand, cells cultured on non-poled samples have no differences with respect to the ones culured on the control, contrarily to the poled films, with overall surface charge, demonstrating the relevance of scaffolds surface charge on cell behavior. This study demonstrates that both positive and negative overall surface charge promote the differentiation of C2C12 cells through involvement of proteins related with the contraction of the skeletal muscle cells, with a more pronounced effect with the negative charged surfaces.