Skeletal muscle regeneration is one of the major areas of interest in sport medicine as well as trauma centers. Three-dimensional (3D) bioprinting (BioP) is nowadays widely adopted to manufacture 3D constructs for regenerative medicine but a comparison between the available biomaterial-based inks (bioinks) is missing. The present study aims to assess the impact of different hydrogels on the viability, proliferation, and differentiation of murine myoblasts (C2C12) encapsulated in 3D bioprinted constructs aided to muscle regeneration. We tested three different commercially available hydrogels bioinks based on: (1) gelatin methacrylate and alginate crosslinked by UV light; (2) gelatin methacrylate, xanthan gum, and alginate-fibrinogen; (3) nanofibrillated cellulose (NFC)/alginate-fibrinogen crosslinked with calcium chloride and thrombin. Constructs embedding the cells were manufactured by extrusion-based BioP and C2C12 viability, proliferation, and differentiation were assessed after 24 h, 7, 14, 21, and 28 days in culture. Although viability, proliferation, and differentiation were observed in all the constructs, among the investigated bioinks, the best results were obtained by using NFC/alginate-fibrinogen-based hydrogel from 7 to 14 days in culture, when the embedded myoblasts started fusing, forming at day 21 and day 28 multinucleated myotubes within the 3D bioprinted structures. The results revealed an extensive myotube alignment all over the linear structure of the hydrogel, demonstrating cell maturation, and enhanced myogenesis. The bioprinting strategies that we describe here denote a strong and endorsed approach for the creation of in vitro artificial muscle to improve skeletal muscle tissue engineering for future therapeutic applications.

Myoblast 3D bioprinting to burst in vitro skeletal muscle differentiation / Ronzoni, Flavio L; Aliberti, Flaminia; Scocozza, Franca; Benedetti, Laura; Auricchio, Ferdinando; Sampaolesi, Maurilio; Cusella, Gabriella; Redwan, Itedale Namro; Ceccarelli, Gabriele; Conti, Michele. - In: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE. - ISSN 1932-7005. - 16:5(2022), pp. 484-495. [10.1002/term.3293]

Myoblast 3D bioprinting to burst in vitro skeletal muscle differentiation

Sampaolesi, Maurilio;
2022

Abstract

Skeletal muscle regeneration is one of the major areas of interest in sport medicine as well as trauma centers. Three-dimensional (3D) bioprinting (BioP) is nowadays widely adopted to manufacture 3D constructs for regenerative medicine but a comparison between the available biomaterial-based inks (bioinks) is missing. The present study aims to assess the impact of different hydrogels on the viability, proliferation, and differentiation of murine myoblasts (C2C12) encapsulated in 3D bioprinted constructs aided to muscle regeneration. We tested three different commercially available hydrogels bioinks based on: (1) gelatin methacrylate and alginate crosslinked by UV light; (2) gelatin methacrylate, xanthan gum, and alginate-fibrinogen; (3) nanofibrillated cellulose (NFC)/alginate-fibrinogen crosslinked with calcium chloride and thrombin. Constructs embedding the cells were manufactured by extrusion-based BioP and C2C12 viability, proliferation, and differentiation were assessed after 24 h, 7, 14, 21, and 28 days in culture. Although viability, proliferation, and differentiation were observed in all the constructs, among the investigated bioinks, the best results were obtained by using NFC/alginate-fibrinogen-based hydrogel from 7 to 14 days in culture, when the embedded myoblasts started fusing, forming at day 21 and day 28 multinucleated myotubes within the 3D bioprinted structures. The results revealed an extensive myotube alignment all over the linear structure of the hydrogel, demonstrating cell maturation, and enhanced myogenesis. The bioprinting strategies that we describe here denote a strong and endorsed approach for the creation of in vitro artificial muscle to improve skeletal muscle tissue engineering for future therapeutic applications.
2022
commercially hydrogel bioinks; murine myoblasts (C2C12); muscle differentiation; three-dimensional (3D) bioprinting
01 Pubblicazione su rivista::01a Articolo in rivista
Myoblast 3D bioprinting to burst in vitro skeletal muscle differentiation / Ronzoni, Flavio L; Aliberti, Flaminia; Scocozza, Franca; Benedetti, Laura; Auricchio, Ferdinando; Sampaolesi, Maurilio; Cusella, Gabriella; Redwan, Itedale Namro; Ceccarelli, Gabriele; Conti, Michele. - In: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE. - ISSN 1932-7005. - 16:5(2022), pp. 484-495. [10.1002/term.3293]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1709667
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