title: Effect of botulinum neurotoxin type A (BoNTA) on the morphology and viability of 3T3 murine fibroblasts.
abstract: OBJECTIVE
  BoNTA is used in the treatment of ophthalmological disorders, muscular hyperactivity and pain. In recent years it has been described that BoNTA reduces cellular viability and induces apoptosis in prostate cells lines. Studies about the effect of BoNTA are no well known. There have been studies about the effect of BoNTA on the expression levels of collagenase in fibroblasts, but not on its morphological impact on these cells. The aim of this study was to determine the effect of BoNTA on the morphology and viability of the 3T3 fibroblast cell line.
  
  
  METHODS
  The 3T3 fibroblast cell line was cultured and the experimental group received 10 U BoNTA added to a 0.9% sterile saline solution in a reconstituted vial. The control group received saline solution only. Cultured cells were observed and photographed at 5, 10, 15 and 20 h. Cell viability was evaluated by means of the trypan blue test, and cell proliferation with the Proliferation Assay kit (PROMEGA).
  
  
  RESULTS
  The application of BoNTA to 3T3 fibroblast cells induced morphological changes, such as a loss of normal fibroblast morphology. Additionally, we observed the cytoplasmic retraction and spread phenomena. The nuclei showed other important changes with Giemsa staining.
  
  
  CONCLUSIONS
  The results indicate that BoNTA induced a loss of spindle form, increase in cytoplasmic vesicles, and the presence of nuclear vesicles (compacted chromatin surrounded by a nuclear envelope). This suggests an apoptotic process and decreased cell viability. Further studies are needed to explore the mechanisms of these alterations.
authors.affiliation: Research Support Group, National Institute of Rehabilitation Mexico City, México ; Laboratory of Molecular Oncology and Oxidative Stress, Escuela Superior de Medicina, Instituto Politécnico Nacional Mexico City, México.
authors.forename: Cindy
authors.initials: C
authors.lastname: Bandala
authors.affiliation: Laboratory of Molecular Oncology and Oxidative Stress, Escuela Superior de Medicina, Instituto Politécnico Nacional Mexico City, México.
authors.forename: Juan Luis
authors.initials: JL
authors.lastname: Terán-Melo
authors.affiliation: Laboratory of Cell Biology, Centro de Investigación Biomédica de Oriente, Instituto Mexicano del Seguro Social Puebla, México.
authors.forename: Maricruz
authors.initials: M
authors.lastname: Anaya-Ruiz
authors.affiliation: Laboratory of Molecular Oncology and Oxidative Stress, Escuela Superior de Medicina, Instituto Politécnico Nacional Mexico City, México.
authors.forename: Cesar Miguel
authors.initials: CM
authors.lastname: Mejía-Barradas
authors.affiliation: Research Support Group, National Institute of Rehabilitation Mexico City, México.
authors.forename: Rene
authors.initials: R
authors.lastname: Domínguez-Rubio
authors.affiliation: Laboratory of Neurochemistry, National Institute of Rehabilitation Mexico City, México.
authors.forename: Paloma
authors.initials: P
authors.lastname: De la Garza-Montano
authors.affiliation: Laboratory of Neurochemistry, National Institute of Rehabilitation Mexico City, México.
authors.forename: Alfonso
authors.initials: A
authors.lastname: Alfaro-Rodríguez
authors.affiliation: Laboratory of Molecular Oncology and Oxidative Stress, Escuela Superior de Medicina, Instituto Politécnico Nacional Mexico City, México.
authors.forename: Eleazar
authors.initials: E
authors.lastname: Lara-Padilla
mesh_terms: D016475:3T3 Cells; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D019274:Botulinum Toxins, Type A / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D003593:Cytoplasm / Q000187:drug effects; D022162:Cytoplasmic Vesicles / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects*; D051379:Mice; D008941:Spindle Apparatus / Q000187:drug effects
chemical_list: D019274:Botulinum Toxins, Type A
keywords: Botulinum neurotoxin; apoptosis; cell viability; fibroblasts; genotoxicity; morphology