Dependence of alignment direction on magnitude of strain in esophageal smooth muscle cells

• Published in: Biotechnology and bioengineering 2009-04, Vol.102 (6), p.1703-1711
• Main Authors: Ritchie, A.C, Wijaya, S, Ong, W.F, Zhong, S.P, Chian, K.S
• Format: Article
• Language: English
• Subjects: Actins - metabolism; alignment response; Animals; Biological and medical sciences; bioreactor; Bioreactors; Biotechnology; Cell culture; Cell Culture Techniques; Cell cycle; Cell Differentiation; Cell Proliferation; Cells, Cultured; Data Interpretation, Statistical; Desmin - metabolism; esophageal tissue engineering; Esophagus; Esophagus - cytology; Fundamental and applied biological sciences. Psychology; Gene Expression; mechanical stimulation; Membranes; Methods. Procedures. Technologies; Morphology; Myocytes, Smooth Muscle - cytology; Myocytes, Smooth Muscle - physiology; Proteins; smooth muscle cells; Stress, Mechanical; Swine; Tissue Engineering - methods; Tissue Scaffolds; Various methods and equipments; Vimentin - metabolism
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.22190

The response of cells in vitro to mechanical forces has been the subject of much research using devices to exert controlled mechanical stimulation on cultured cells or isolated tissue. In this study, esophageal smooth muscle cells were seeded on flexible polyurethane membranes to form a confluent cell layer. The cells were then subjected to uniform cyclic stretch of varying magnitudes at a frequency of approximately five cycles per minute in a custom made mechatronic bioreactor, providing similar strains experienced in the in vivo mechanical environment of the esophagus. The results show that the orientation response is dependent on the magnitude of cyclic stretch applied. Smooth muscle cells showed parallel alignment to the force direction at low cyclic strains (2%) compared to the hill-valley morphology of static controls. At higher strains (5% and 10% magnitude), the cells exhibited a consistent alignment perpendicular to the strain. To our knowledge, this is the first time that the alignment direction's dependence on strain magnitude has been demonstrated. MTS analysis indicated that cell metabolism was reduced when mechanical strain was applied, and proliferation was inhibited by mechanical strain. Protein expression indicates a decrease in smooth muscle α-actin, indicative of changes in cell phenotype, an increase in vimentin, which is associated with increased cell motility, and an increase in desmin, indicating differentiation in stimulated cells. Biotechnol. Bioeng. 2009;102: 1703-1711.