Tight Regulation of Mechanotransducer Proteins Distinguishes the Response of Adult Multipotent Mesenchymal Cells on PBCE-Derivative Polymer Films with Different Hydrophilicity and Stiffness

Abstract

Mechanotransduction is a molecular process by which cells translate physical stimuli exerted by the external environment into biochemical pathways to orchestrate the cellular shape and function. Even with the advancements in the field, the molecular events leading to the signal cascade are still unclear. The current biotechnology of tissue engineering offers the opportunity to study in vitro the effect of the physical stimuli exerted by biomaterial on stem cells and the mechanotransduction pathway involved in the process. Here, we cultured multipotent human mesenchymal/stromal cells (hMSCs) isolated from bone marrow (hBM-MSCs) and adipose tissue (hASCs) on films of poly(butylene 1,4-cyclohexane dicarboxylate) (PBCE) and a PBCE-based copolymer containing 50 mol% of butylene diglycolate co-units (BDG50), to intentionally tune the surface hydrophilicity and the stiffness (PBCE = 560 Mpa; BDG50 = 94 MPa). We demonstrated the activated distinctive mechanotransduction pathways, resulting in the acquisition of an elongated shape in hBM-MSCs on the BDG50 film and in maintaining the canonical morphology on the PBCE film. Notably, hASCs acquired a new, elongated morphology on both the PBCE and BDG50 films. We found that these events were mainly due to the differences in the expression of Cofilin1, Vimentin, Filamin A, and Talin, which established highly sensitive machinery by which, rather than hASCs, hBM-MSCs distinguished PBCE from BDG50 films.

Novel Nanostructured Scaffolds of Poly(butylene trans-1,4-cyclohexanedicarboxylate)-Based Copolymers with Tailored Hydrophilicity and Stiffness: Implication for Tissue Engineering Modeling

“…No cytotoxicity was observed in cells on the PBCE and BDG50 scaffolds, as shown by the similar mitochondrial dehydrogenase activity curves to those of the control systems ( Figure 11 c). Confirming previous work [ 14 ], no cytotoxicity was observed in cells on the PBCE and BDG50 films ( Figure 11 c). These findings indicated the biocompatibility of the PBCE and BDG50 scaffolds for hBM-MSCs long-term cultures and further validate our previous results on the biocompatibility of PBCE and BDG50 polymers for hBM-MSCs [ 14 ].…”

“…Confirming previous work [ 14 ], no cytotoxicity was observed in cells on the PBCE and BDG50 films ( Figure 11 c). These findings indicated the biocompatibility of the PBCE and BDG50 scaffolds for hBM-MSCs long-term cultures and further validate our previous results on the biocompatibility of PBCE and BDG50 polymers for hBM-MSCs [ 14 ].…”

“…We compared the properties of PBCE and BD50 scaffolds with those of PBCE and BDG50 films obtained through compression molding techniques and already used by our group [ 10 , 12 , 14 ].…”

“…Next, we evaluated the biocompatibility of the PBCE and BDG50 scaffolds toward hBM-MSCs. In comparison, experiments were conducted by seeding hBM-MSCs on PBCE and BDG50 films [ 14 ].…”

“…Poly(butylene trans -1,4-cyclohexane dicarboxylate) (PBCE) is a biocompatible and easily processable cycloaliphatic polyester, previously investigated for tissue engineering applications by some of the authors of the present paper [ 10 , 11 , 12 , 13 , 14 ]. Although PBCE films have high crystallinity and rigidity characteristics, copolymerization with different amounts of butylene diglycolate (BDG) co-units permits a tunable modulation of the properties (i.e., hydrophilicity, stiffness) without alteration of the biocompatibility of the parent homopolymer [ 10 , 15 ], thus making both the polymer and copolymer attractive for the generation of innovative tunable supports for biological applications.…”