Response of human trabecular meshwork cells to topographic cues on the nanoscale level.
purpose To determine how primary human trabecular meshwork (HTM) cells are influenced by their interaction with nanopatterned substrates.


METHODS
HTM cells from several individuals were grown on planar or anisotropically ordered nanopatterned surfaces. Microscopy was used to measure cellular elongation and alignment. Cells were also incubated with 10(-7) M dexamethasone for comparison to control cells. Quantitative PCR for myocilin and versican isoforms was performed in addition to Western blots of myocilin and alphaB-crystallin.


RESULTS
Cells on anisotropically ordered nanopatterned substrates aligned with the surface nanopatterns and displayed actin filaments that were parallel to the patterned ridges and grooves. The cells became more elongated on the nanogrooved surfaces compared with the planar control cells. Myocilin mRNA and protein levels increased when HTM cells were plated onto 400-nm pitch surfaces. With some HTM cells, myocilin increased to a greater extent when untreated cells were plated on nanosurfaces compared with the cells grown on planar surfaces with dexamethasone. The V0 and V1 isoforms of versican had increased expression on patterned surfaces.


CONCLUSIONS
Nanopatterned surfaces containing biomimetic length scale features clearly influenced cellular behavior of HTM cells. Increased mRNA and protein levels of myocilin were observed when cells were grown on 400-nm pitch surfaces, suggesting that the reduction of myocilin mRNA when cells are plated onto flat tissue culture plastic is an artifact of a nonphysiologic culture environment that lacks appropriate topographic cues.
Russell
Paul
P
Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53703, USA. russellp@svm.vetmed.wisc.edu
Gasiorowski
Joshua Z
JZ
Nealy
Paul F
PF
Murphy
Christopher J
CJ
D000199:Actins / Q000378:metabolism; D015153:Blotting, Western; D048430:Cell Shape; D002478:Cells, Cultured; D003463:Cues; D003598:Cytoskeletal Proteins / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology; D005136:Eye Proteins / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics; D006801:Humans; D007429:Intraocular Pressure; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014129:Trabecular Meshwork / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D053675:Versicans / Q000235:genetics; D038203:alpha-Crystallin B Chain / Q000378:metabolism; D000099179:Myocilin
D000199:Actins; D003598:Cytoskeletal Proteins; D003907:Dexamethasone; D005136:Eye Proteins; D006023:Glycoproteins; D012333:RNA, Messenger; D053675:Versicans; D038203:alpha-Crystallin B Chain; D000099179:Myocilin; C508322:VCAN protein, human