title: Heat shock attenuates VEGF expression in three-dimensional myoblast sheets deteriorating therapeutic efficacy in heart failure.
abstract: BACKGROUND
  Myoblast sheet transplantation is a promising novel treatment for ischemic heart failure. The aim of this study was to test the hypothesis that heat shock (HS) pre-treatment affects the angiogenic properties of myoblast sheets in vivo and in vitro.
  
  
  METHODS
  We studied HS preconditioning of L6 myoblast sheets in relation to their apoptosis, proliferation, and vascular endothelial growth factor (VEGF)-associated responses under normoxia and under hypoxia in vitro. In vivo evaluation of their therapeutic effect was performed with 60 male Wistar rats divided into 3 groups (20 each): sole left anterior descending (LAD) ligation (control); LAD ligation and non-conditioned sheet transplantation (L6 No-Shock); and LAD ligation and L6-heat shock conditioned sheet transplantation (L6 Heat-Shock). Left ventricular function was evaluated by echocardiography after 3, 10, and 28 days.
  
  
  RESULTS
  Expression of HSP70/72 was strongly induced 24 hours after HS, and thereafter it decreased notably during 72 hours in hypoxia. Under normal growth conditions, HSP70/72 expression remained stable. HS delayed apoptosis-associated caspase-3 expression during 24-hour hypoxia compared to non-treated controls. However, VEGF expression reduced significantly in the heat shock pretreated sheets. Ejection fraction of the L6-myoblast HS pre-treatment group (L6 Heat-Shock) decreased gradually during follow-up, in the same pattern as the controls. However, these functional parameters improved in the L6-myoblast normal sheet group (L6 No-Shock) at the tenth day and remained significantly better.
  
  
  CONCLUSIONS
  HS protects myoblast sheets from hypoxia-associated apoptosis in vitro, but reduces VEGF expression of the sheet, leading to lower therapeutic effect in heart failure.
authors: Department of Cardiothoracic Surgery, Helsinki Universtity Meilahti Hospital and Cell Therapy Research Consortium, Institute of Biomedicine, Pharmacology, University of Helsinki, Helsinki, Finland.
authors: Mona
authors: M
authors: Augustin
authors: Muhammad
authors: M
authors: Ali Asim Mahar
authors: Päivi
authors: P
authors: Lakkisto
authors: Ilkka
authors: I
authors: Tikkanen
authors: Antti
authors: A
authors: Vento
authors: Tommi
authors: T
authors: Patila
authors: Ari
authors: A
authors: Harjula
mesh_terms: D000818:Animals; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D015687:Cell Hypoxia; D002460:Cell Line; D049109:Cell Proliferation; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D050884:HSP72 Heat-Shock Proteins / Q000378:metabolism; D006333:Heart Failure / Q000150:complications / Q000000981:diagnostic imaging / Q000503:physiopathology / Q000628:therapy*; D018869:Heat-Shock Response; D008297:Male; D032446:Myoblasts / Q000378:metabolism* / Q000473:pathology / Q000637:transplantation*; D009203:Myocardial Infarction / Q000150:complications / Q000000981:diagnostic imaging / Q000503:physiopathology; D018384:Oxidative Stress; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013318:Stroke Volume; D016896:Treatment Outcome; D014463:Ultrasonography; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D018840:HSP70 Heat-Shock Proteins; D050884:HSP72 Heat-Shock Proteins; D018809:Proliferating Cell Nuclear Antigen; D042461:Vascular Endothelial Growth Factor A; D053148:Caspase 3