title: Substratum compliance regulates human trabecular meshwork cell behaviors and response to latrunculin B.
abstract: OBJECTIVE
  To determine the impact of substratum compliance and latrunculin-B (Lat-B), both alone and together, on fundamental human trabecular meshwork (HTM) cell behavior. Lat-B is a reversible actin cytoskeleton disruptor that decreases resistance to aqueous humor outflow and decreases intraocular pressure.
  
  
  METHODS
  HTM cells were cultured on polyacrylamide hydrogels possessing values for compliance that mimic those reported for normal and glaucomatous HTM, or tissue culture plastic (TCP). Cells were treated with 0.2 μM or 2.0 μM Lat-B in dimethyl sulfoxide (DMSO) or DMSO alone. The impact of substratum compliance and/or Lat-B treatment on cell attachment, proliferation, surface area, aspect ratio, and migration were investigated.
  
  
  RESULTS
  HTM cells had profoundly decreased attachment and proliferation rates when cultured on hydrogels possessing compliance values that mimic those found for healthy HTM. The effect of Lat-B treatment on HTM cell surface area was less for cells cultured on more compliant hydrogels compared with TCP. HTM cell migration was increased on stiffer hydrogels that mimic the compliance of glaucomatous HTM and on TCP in comparison with more compliant hydrogels. Lat-B treatment decreased cellular migration on all surfaces for at least 7 hours after treatment.
  
  
  CONCLUSIONS
  Substratum compliance profoundly influenced HTM cell behaviors and modulated the response of HTM cells to Lat-B. The inclusion of substratum compliance that reflects healthy or glaucomatous HTM results in cell behaviors and responses to therapeutic agents in vitro that may more accurately reflect in vivo conditions.
authors: Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, Davis, CA 95616, USA.
authors: Joshua A
authors: JA
authors: Wood
authors: Clayton T
authors: CT
authors: McKee
authors: Sara M
authors: SM
authors: Thomasy
authors: Marion E
authors: ME
authors: Fischer
authors: Nihar M
authors: NM
authors: Shah
authors: Christopher J
authors: CJ
authors: Murphy
authors: Paul
authors: P
authors: Russell
mesh_terms: D000180:Acrylic Resins; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects; D016326:Extracellular Matrix Proteins; D005901:Glaucoma; D006801:Humans; D020100:Hydrogels; D008387:Marine Toxins / Q000494:pharmacology; D018625:Microscopy, Atomic Force; D053778:Thiazolidines / Q000494:pharmacology*; D014129:Trabecular Meshwork / Q000187:drug effects* / Q000473:pathology*; D000099166:Polyacrylamides
chemical_list: D000180:Acrylic Resins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D016326:Extracellular Matrix Proteins; D020100:Hydrogels; D008387:Marine Toxins; D053778:Thiazolidines; D000099166:Polyacrylamides; C037068:latrunculin B