Comparative Study Between Triton X-100-SD Method And Detergent-Enzymatic Digestion Method For Decellularization Of Porcine Aortic Valves

Objective:Heart valve replacement is the primary treatment means of heart valve disease at the present. What’s more, Mechanical and biologic heart valve are not ideal substitute, both of which have apparent limitations in terms of durability, growth potential, compatibility as well as resistance to infection. However, the tissue engineering heart valve (TEHV), a living organ, is an ideal one which has many advantages in many ways. Especially it has the capabilities for growth, repair and remodeling in the same way that the native heart valve does. It avoid the shortcomings of ordinary replacements. Tissue engineered grafts based on biodegradable polymeric scaffolds or decellularized native biological scaffolds. It is a pity that there are still some problems crying out for solutions, such as control of degradation rate of biodegradable polymeric scaffolds and Mechanical properties of decellularized native biological scaffolds.In this study, to seek a better method of making biological scaffolds for tissue engineering heart valves, we tested out different decellularization procedures for their potential of cell removal and their ability of maintaining scaffolds.Methods:Aortic heart valve conduits (wall and leaflets) were obtained from healthy adult pigs slaughtered, then the valve leaflets were dissected from the conduits after washed repeatedly by physiological saline in aseptic conditions. Specimens of fresh porcine aortic valve leaflets were randomly divided into group Ⅰ (n=16), group Ⅱ (n=16) and group Ⅲ(n=16). The valve leaflets of groups I were not decellularized, the groups II were treated with Triton X-100-SD method, the groups Ⅲ were treated with detergent-Enzymatic digestion method. Having finished these procedures, we should inspect by making use of histological examination and analyze DNA content. The process should obey the followings:first, the surface characteristics of the color, the shape or something else shall be observed to the valve leaflets generally. Secondly, the tissue formation, the effect of decellularization and the row of decellularized scaffold also be observed under light and electron microscope.At last, the DNA content can be measured, meanwhile analyze the residual parts of nucleic acid in the comparison of untreated group.Results:Detergent-Enzymatic digestion method removed cells completely but caused the valve fiber structural alterations in GroupⅢ. Treatmenting with Triton X-100-SD method achieved both complete decellularization and preservation of the valve structure in GroupⅡ.Conclusions:It was a better technique of decellularization method that Triton X-100-SD achieved both complete decellularization and preservation of the valve scaffold structure in our study.