A combined physicochemical approach towards human tenocyte phenotype maintenance

C N M Ryan; E Pugliese; N Shologu; D Gaspar; P Rooney; Md N Islam; A O'Riordan; M J Biggs; M D Griffin; D I Zeugolis

doi:10.1016/j.mtbio.2021.100130

A combined physicochemical approach towards human tenocyte phenotype maintenance

Mater Today Bio. 2021 Sep 10:12:100130. doi: 10.1016/j.mtbio.2021.100130. eCollection 2021 Sep.

Authors

C N M Ryan^{1

2}, E Pugliese^{1

2}, N Shologu^{1

2}, D Gaspar^{1

2}, P Rooney², Md N Islam^{2

3

4}, A O'Riordan⁵, M J Biggs², M D Griffin^{2

3}, D I Zeugolis^{1

2

6}

Affiliations

¹ Regenerative, Modular & Developmental Engineering Laboratory (REMODEL), Biomedical Sciences Building, National University of Ireland Galway (NUI Galway), Galway, Ireland.
² Science Foundation Ireland (SFI) Centre for Research in Medical Devices (CÚRAM), Biomedical Sciences Building, National University of Ireland Galway (NUI Galway), Galway, Ireland.
³ Regenerative Medicine Institute (REMEDI), School of Medicine, Biomedical Sciences Building, National University of Ireland Galway (NUI Galway), Galway, Ireland.
⁴ Discipline of Biochemistry, School of Natural Sciences, National University of Ireland Galway (NUI Galway), Galway, Ireland.
⁵ Tyndall National Institute, University College Cork (UCC), Cork, Ireland.
⁶ Regenerative, Modular & Developmental Engineering Laboratory (REMODEL), Charles Institute of Dermatology, Conway Institute of Biomolecular & Biomedical Research and School of Mechanical & Materials Engineering, University College Dublin (UCD), Dublin, Ireland.

Abstract

During in vitro culture, bereft of their optimal tissue context, tenocytes lose their phenotype and function. Considering that tenocytes in their native tissue milieu are exposed simultaneously to manifold signals, combination approaches (e.g. growth factor supplementation and mechanical stimulation) are continuously gaining pace to control cell fate during in vitro expansion, albeit with limited success due to the literally infinite number of possible permutations. In this work, we assessed the potential of scalable and potent physicochemical approaches that control cell fate (substrate stiffness, anisotropic surface topography, collagen type I coating) and enhance extracellular matrix deposition (macromolecular crowding) in maintaining human tenocyte phenotype in culture. Cell morphology was primarily responsive to surface topography. The tissue culture plastic induced the largest nuclei area, the lowest aspect ratio, and the highest focal adhesion kinase. Collagen type I coating increased cell number and metabolic activity. Cell viability was not affected by any of the variables assessed. Macromolecular crowding intensely enhanced and accelerated native extracellular matrix deposition, albeit not in an aligned fashion, even on the grooved substrates. Gene analysis at day 14 revealed that the 130 kPa grooved substrate without collagen type I coating and under macromolecular crowding conditions positively regulated human tenocyte phenotype. Collectively, this work illustrates the beneficial effects of combined physicochemical approaches in controlling cell fate during in vitro expansion.

Keywords: Cell phenotype maintenance; Collagen type I coating; In vitro microenvironment; Macromolecular crowding; Substrate elasticity; Surface topography.