A method for investigating the cellular response to cyclic tension or compression in three-dimensional culture

Babak Sarrafpour; Philip Boughton; Ramin M Farahani; Stephen C Cox; Gareth Denyer; Elizabeth Kelly; Hans Zoellner

doi:10.1016/j.jmbbm.2018.08.006

A method for investigating the cellular response to cyclic tension or compression in three-dimensional culture

J Mech Behav Biomed Mater. 2018 Dec:88:11-17. doi: 10.1016/j.jmbbm.2018.08.006. Epub 2018 Aug 8.

Authors

Babak Sarrafpour¹, Philip Boughton², Ramin M Farahani³, Stephen C Cox⁴, Gareth Denyer⁵, Elizabeth Kelly⁶, Hans Zoellner⁷

Affiliations

¹ The University of Sydney, Faculty of Dentistry, Department of Oral Pathology and Oral Medicine, Cellular and Molecular Pathology Research Unit, Westmead Centre for Oral Health, Westmead Hospital, NSW 2145, Australia. Electronic address: babak.sarrafpour@sydney.edu.au.
² The University of Sydney, The Institute of Biomedical Engineering and Technology, Sydney, NSW 2006, Australia. Electronic address: philip.boughton@sydney.edu.au.
³ The University of Sydney, Faculty of Dentistry, Institute of Dental Research, Westmead Hospital, NSW 2145, Australia. Electronic address: ramin.mostofizadehfarahani@sydney.edu.au.
⁴ The University of Sydney, Department of Oral Surgery, Westmead Centre for Oral Health, Westmead Hospital, Westmead, NSW, Australia. Electronic address: stephen.cox@sydney.edu.au.
⁵ The University of Sydney, School of Molecular Bioscience, NSW 2006, Australia. Electronic address: gareth.denyer@sydney.edu.au.
⁶ The University of Sydney, Faculty of Dentistry, Department of Oral Pathology and Oral Medicine, Cellular and Molecular Pathology Research Unit, Westmead Centre for Oral Health, Westmead Hospital, NSW 2145, Australia. Electronic address: elizabeth.kelly@sydney.edu.au.
⁷ The University of Sydney, Faculty of Dentistry, Department of Oral Pathology and Oral Medicine, Cellular and Molecular Pathology Research Unit, Westmead Centre for Oral Health, Westmead Hospital, NSW 2145, Australia. Electronic address: hans.zoellner@sydney.edu.au.

PMID: 30118920
DOI: 10.1016/j.jmbbm.2018.08.006

Abstract

We have an interest in the cellular response to mechanical stimuli, and here describe an in-vitro method to examine the response of cells cultured in a three-dimensional matrix to mechanical compressive and tensile stress. Synthetic aliphatic polyester scaffolds coated with 45S5 bioactive glass were seeded with human dental follicular cells (HDFC), and attached to well inserts and magnetic endplates in six well palates. Scaffolds were subjected to either cyclic 10% tensile deformation, or 8% compression, at 1 Hz and 2 Hz respectively for 6, 24 or 48 h, by uniaxial motion of magnetically-coupled endplates. It was possible to isolate high quality mRNA from cells in these scaffolds, as demonstrated by high RNA integrity numbers scores, and ability to perform meaningful cRNA microarray analysis, in which 669 and 727 genes were consistently upregulated, and 662 and 518 genes down regulated at all times studied under tensile and compressive loading conditions respectively. MetaCore analysis revealed the most regulated gene ontogenies under both loading conditions to be for: cytoskeletal remodelling; cell adhesion-chemokines and adhesion; cytoskeleton remodelling-TGF WNT and cytoskeletal remodelling pathways. We believe the method here described will be of value for analysis of the cellular response to cyclic loading.

Keywords: C-RNA Microarray; Cyclic tension and compression; Cytoskeleton Remodelling; Dental follicle; In vitro biomechanical actuation device.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biomechanical Phenomena
Compressive Strength*
Dental Sac / cytology*
Dental Sac / metabolism
Gene Expression Regulation
Humans
Stress, Mechanical*