Human Corneal Fibroblast Pattern Evolution and Matrix Synthesis on Mechanically Biased Substrates.
In a fibroblast colony model of corneal stromal development, we asked how physiological tension influences the patterning dynamics of fibroblasts and the orientation of deposited extracellular matrix (ECM). Using long-term live-cell microscopy, enabled by an optically accessible mechanobioreactor, a primary human corneal fibroblast colony was cultured on three types of substrates: a mechanically biased, loaded, dense, disorganized collagen substrate (LDDCS), a glass coverslip, and an unloaded, dense, disorganized collagen substrate (UDDCS). On LDDCS, fibroblast orientation and migration along a preferred angle developed early, cell orientation was correlated over long distances, and the colony pattern was stable. On glass, fibroblast orientation was poorly correlated, developed more slowly, and colony patterns were metastable. On UDDCS, cell orientation was correlated over shorter distances compared with LDDCS specimens. On all substrates, the ECM pattern reflected the cell pattern. In summary, mechanically biasing the collagen substrate altered the early migration behavior of individual cells, leading to stable emergent cell patterning, which set the template for newly synthesized ECM.
1 Department of Bioengineering, Northeastern University , Boston, Massachusetts.
Ramin
R
Zareian
1 Department of Bioengineering, Northeastern University , Boston, Massachusetts.
Monica E
ME
Susilo
1 Department of Bioengineering, Northeastern University , Boston, Massachusetts.
Jeffrey A
JA
Paten
2 Department of Electrical and Computer Engineering, Northeastern University , Boston, Massachusetts.
James P
JP
McLean
3 The Institute of Photonic Sciences , Castelldefels (Barcelona), Spain .
Joseph
J
Hollmann
4 Department of Ophthalmology, Dean McGee Eye Institute, Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma.
Dimitrios
D
Karamichos
1 Department of Bioengineering, Northeastern University , Boston, Massachusetts.
Conor S
CS
Messer
5 Departments of Mechanical Engineering and Department of Pharmacology and Center for Lung Biology, University of South Alabama , Mobile, Alabama.
Dhananjay T
DT
Tambe
6 Department of Surgery, Harvard Medical School , Boston, Massachusetts.
Nima
N
Saeidi
7 Schepens Eye Research Institute , Boston, Massachusetts.
James D
JD
Zieske
1 Department of Bioengineering, Northeastern University , Boston, Massachusetts.
Jeffrey W
JW
Ruberti
D002465:Cell Movement; D003094:Collagen / Q000096:biosynthesis*; D003315:Cornea / Q000166:cytology / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D006801:Humans
D003094:Collagen