title: Fixation-related autolysis and bioprosthetic aortic wall calcification.
abstract: OBJECTIVE
  It has been established previously that immediate fixation and increased glutaraldehyde (GA) concentrations are required to prevent severe autolytic tissue damage during bioprosthetic aortic root production. The study aim was to verify that structure-preserving fixation also reduces aortic wall calcification.
  
  
  METHODS
  Porcine aortic roots were fixed either instantly or after being kept on ice for 48 h (phosphate-buffered saline, PBS). Two concentrations of GA (0.2% and 3.0%) were chosen (4 degrees C, seven days, PBS). Discs of aortic wall tissue (1.2 cm diameter) were implanted subcutaneously in rats for 60 days (n = 10 per group), while aortic roots were implanted in the distal aortic arch of sheep for six weeks (n = 3 per group) and six months (n = 4 per group). Calcification was assessed by atomic absorption spectrophotometry and light microscopy. Fixation-related tissue damage was determined by transmission electron microscopy, and correlated with calcification.
  
  
  RESULTS
  No significant difference in calcification was found between immediate and delayed fixation if tissue was fixed with 0.2% GA. In the 3.0% GA group, both animal models showed a significantly lower level of calcification if tissue was immediately fixed. In the subcutaneous rat model, immediate fixation reduced calcification by 26% (p <0.0001). In the circulatory sheep model immediate fixation did not affect calcification in the short-term six-week implants, but markedly lowered it by 37% (p = 0.035) after six months. Ultrastructurally, there was a significant correlation between membrane damage, vacuolization and vesicle shedding on the one hand, and calcification on the other.
  
  
  CONCLUSIONS
  Coincidental fixation-related ultrastructural damage and increased calcification was demonstrated in bioprosthetic aortic wall tissue.
authors.affiliation: Department of Cardiothoracic Surgery, Cape Heart Centre, University of Cape Town Medical School, South Africa.
authors.forename: P
authors.initials: P
authors.lastname: Human
authors.forename: C
authors.initials: C
authors.lastname: Weissenstein
authors.forename: A
authors.initials: A
authors.lastname: Trantina
authors.forename: P
authors.initials: P
authors.lastname: Zilla
mesh_terms: D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000293:injuries / Q000648:ultrastructure; D001021:Aortic Valve / Q000187:drug effects / Q000293:injuries / Q000648:ultrastructure; D001329:Autolysis / Q000139:chemically induced*; D001705:Bioprosthesis; D002114:Calcinosis / Q000139:chemically induced*; D002118:Calcium / Q000032:analysis; D013504:Cardiovascular Surgical Procedures; D005404:Fixatives / Q000009:adverse effects*; D005976:Glutaral / Q000008:administration & dosage / Q000009:adverse effects; D006349:Heart Valve Diseases / Q000473:pathology*; D006350:Heart Valve Prosthesis; D008955:Models, Cardiovascular; D051381:Rats; D012720:Severity of Illness Index; D012756:Sheep; D013223:Statistics as Topic; D013552:Swine; D013997:Time Factors
chemical_list: D005404:Fixatives; D002118:Calcium; D005976:Glutaral