A Suitable Protocol to Prepare Decellularized Porcine Vessel for Cardiovascular

Abstract

Although autologous and artificial grafts are commonly used to solve cardiovascular disease or trauma, decellularized vessels scaffolds are recent promising materials for vascular transplantation. The purpose of this study was to introduce a simple and effective protocol to prepare decellularized porcine vessels. Porcine carotid arteries (pCA) were decellularized by 4 methods including 3 commonly protocols: SDS 0.5% for 24 h or distilled water for 24 h or Triton X100 0.1% for 24 h and our protocol, combination of SDS 0.5% 24 h and distilled water for 24 h. After removing the cells, structure and collagen were determined by Heamatoxylin/Eosin (HE) and Trichrome staining. In vitro cytotoxicity test on fibroblasts was performed according to ISO 10993-5. Additionally, mesenchymal stem cells (MSCs) were seeded on the acellular porcine carotid arteries (aCA) pieces to examine their attachment and growth for 11 days. To investigate in vivo behaviors, aCA were implanted subcutaneously into mouse dorsal skin (Dacron grafts were used as control) and grafts were then isolated and stained with HE after 7, 14, 28 days. The results showed that our protocol removed all cells and maximally preserve extracellular matrix (ECM) comparing to other basis methods. aCA are non-toxic to fibroblast (level 0) and evoked weak inflammation in vivo which was absent in week 4 (similar to Dacron grafts). MSCs attached and grow successfully on the lumen of aCA. Conclusion: Our research may provide a promising protocol to prepare a potential vascular graft with its quality is suitable for cardiovascular surgery.