Decellularized tissue engineered pericardium as replacement for tricuspid valve in cardiac surgery

Abstract

Introduction: Tricuspid valve replacement is the last treatment choice in tricuspid valve pathology. The choice to insert mechanical or bioprosthetic valve remains controversial. Both prostheses have some limitations such as infection, risk of thromboembolism, need for life-long anticoagulation or limited durability. The following study aimed to develop a novel tissue-engineered tricuspid valve based on decellularized pericardium allograft. Materials and methods: Fresh ovine pericardium was harvested at the local slaughter house and decellularized using detergents. For disinfection all samples were treated for 24h with Phosphate Buffered Solution supplemented with 1% gentamicin and 1% streptomycin. The effectiveness of decellularization was evaluated by histological staining (hematoxylin-eosin, Movat’s Pentachrom and Van Gieson), Isolectin B4 staining (a-gal xenoantigen) and by DNA-quantification. Two valvular leaflets were manufactured out of decellularized pericardium and sutured ex-vivo into the tricuspid annulus of an ovine heart and suspended on papillary muscles. Hydraulic test were performed to prove valve competency. Discussion results: After detergent treatment pericardial tissue has been converted in a cell-free scaffold as proven by standard histological analysis. Immunofluorescent examinations revealed the absence of a-gal xenoantigens. DNA-quantification showed a substantial reduction in DNA content compared to the normal tissue. The alignment of collagenous fibers in decellularized scaffolds appeared well-preserved and was not affected by detergent decellularization procedure as proven by histological staining. Graft disinfection and storage in antibiotic solution after decellularization did not affect the texture of the scaffold. Furthermore, two leaflet structure created out of decellularized pericardium and surgically sutured in tricuspid position of ovine heart resulted in a competent valve prosthesis. Conclusion: The present results have shown successful decellularization of the ovine pericardium using detergents. Decellularized pericardial allograft can be used in cardiac surgery as a scaffold for valvular tissue engineering or for in-vivo guided tissue regeneration in tricuspid valve replacement.