Médecine

Paramédical

Autres domaines


S'abonner

Micropatterning and Alignment of Skeletal Muscle Myoblasts Using Microflowed Plasma Process - 10/09/19

Doi : 10.1016/j.irbm.2019.08.003 
K.Y. Vajanthri a, 1, R.K. Sidu a, 1, S.K. Mahto a, b,
a Tissue Engineering and Biomicrofluidics Laboratory, School of Biomedical Engineering, Indian Institute of Technology (Banaras Hindu University), Varanasi 221005, Uttar Pradesh, India 
b Center for Advanced Biomaterials and Tissue Engineering, Indian Institute of Technology (Banaras Hindu University), Varanasi 221005, Uttar Pradesh, India 

Corresponding author at: Tissue Engineering and Biomicrofluidics Laboratory, School of Biomedical Engineering, Indian Institute of Technology (Banaras Hindu University), Varanasi 221005, Uttar Pradesh, India.Tissue Engineering and Biomicrofluidics LaboratorySchool of Biomedical EngineeringIndian Institute of Technology (Banaras Hindu University)VaranasiUttar Pradesh221005India
Sous presse. Épreuves corrigées par l'auteur. Disponible en ligne depuis le mardi 10 septembre 2019

Abstract

Objectives

The primary objective of the study was to optimize micropatterning environments using the microchannel flowed plasma process for controlling the orientation and behaviour of skeletal muscle cells. We have studied the cellular patterning and alignment of skeletal myoblast cells on the various micropattern widths developed on glass substrates.

Materials and Methods

In this method, we have utilized the microchannel flowed plasma process to create micropatterned self-assembled monolayers of octadecyltrichlorosilane and 3-aminopropyltrichlorosilane for creating cell adhesive widths of 20, 200 and 1000 microns on the glass substrates. The micropatterned substrates were characterized by using fluorescein 5(6)-isothiocyanate. Thereafter, the substrates were used to culture and pattern C2C12 and primary rat skeletal muscle cells. Further, we have studied the spatiotemporal variation in the orientation of the cells by using bright field and fluorescence microscopy. The microscopic images were analysed by using orientation order parameter and orientation distribution analysis.

Results

FITC based characterization of micropatterns reveals that the adopted process for micropatterning can effectively create cell adhesive widths with dimensions comparable to the diameter of myofiber. Microscopic observations and the orientation order parameter analysis reveal the precise alignment and specific orientation of myoblasts along the designated cell adhesive widths that closely mimics the physiological scenario. Both the cells showed immediate alignment within smaller cell adhesive widths of 20 and 200 μm. Actin cytoskeletal staining and its orientation distribution analysis of micropattrned C2C12 cells emphasises the influence of micropatterned environment on cytoskeletal actin orientation.

Conclusion

This study corroborates the alignment of the myoblasts using surface cues facilitated by changing surface chemistry of the glass substrates. The study promotes the application of a simple micropatterning technique as a useful tool to regulate the orientation and behaviour of skeletal muscle cells. Also, the study emphasizes the role of spatial topography created by surface modification and its effect on cell adhesion and communication of alignment information across the micropatterns. The microchannel flowed plasma process could be applied to selectively pattern different adherent cell types, which could prove to be a useful platform for the exploration of various cellular processes.

Le texte complet de cet article est disponible en PDF.

Graphical abstract

Le texte complet de cet article est disponible en PDF.

Highlights

Micro flowed plasma process enables cell micropatterning.
Micropatterning of primary and secondary myoblasts leads to gradual alignment of the cells.
Orientation order parameter (OOP) values nearing 1 confirm the alignment of myoblasts under micropatterned conditions.
The micropatterning process influences the orientation of actin cytoskeleton in myoblasts.
The myoblast alignment information propagates from boundaries of micropatterns to the centre.

Le texte complet de cet article est disponible en PDF.

Keywords : Micropatterning, Alkylsilanes, Primary skeletal muscle myoblasts, C2C12 mouse myoblasts, Alignment and orientation analysis


Plan


© 2019  AGBM. Publié par Elsevier Masson SAS. Tous droits réservés.
Ajouter à ma bibliothèque Retirer de ma bibliothèque Imprimer
Export

    Export citations

  • Fichier

  • Contenu

Bienvenue sur EM-consulte, la référence des professionnels de santé.
L’accès au texte intégral de cet article nécessite un abonnement ou un achat à l’unité.

Déjà abonné à cette revue ?

;

Mon compte


Plateformes Elsevier Masson

Déclaration CNIL

EM-CONSULTE.COM est déclaré à la CNIL, déclaration n° 1286925.

En application de la loi nº78-17 du 6 janvier 1978 relative à l'informatique, aux fichiers et aux libertés, vous disposez des droits d'opposition (art.26 de la loi), d'accès (art.34 à 38 de la loi), et de rectification (art.36 de la loi) des données vous concernant. Ainsi, vous pouvez exiger que soient rectifiées, complétées, clarifiées, mises à jour ou effacées les informations vous concernant qui sont inexactes, incomplètes, équivoques, périmées ou dont la collecte ou l'utilisation ou la conservation est interdite.
Les informations personnelles concernant les visiteurs de notre site, y compris leur identité, sont confidentielles.
Le responsable du site s'engage sur l'honneur à respecter les conditions légales de confidentialité applicables en France et à ne pas divulguer ces informations à des tiers.