Feasibility Study On The Preparation Of Artificial Small Blood Vessel By Fluorinated Decellularized Rabbit Aorta

Objective:To investigate the feasibility of fluorinated acellular rabbit aorta as a small artificial vessel for tissue engineering.Methods:The aorta was separated and removed from the midline of the abdomen and the left chest of the rabbit.The aorta was stored in PBS solution and stored in a refrigerator at 4℃ until subsequent processing within a week.The descending aorta was randomly divided into six groups(A-F),in which A-E was decellularized experimental group and F was control group.Decellularized fluid and biological enzyme preparations were prepared at different concentrations.The decellularized fluid consisted of polyethylene glycol octyl phenyl ether(Triton-X),sodium dodecyl sulfate(SDS),and sodium deoxycholate(SD),and the bioenzyme preparation consisted of dnase and ribonuclease.Group A: 0.625%Triton-X,0.625%SDS,0.625%SD,25ug/ml rnase,250ug/ml dnase;Group B: 0.5%Triton-X,0.5%SDS,0.5%SD,20ug/ml rnase,200ug/ml dnase;Group C: 0.375%Triton-X,0.375%SDS,0.375%SD,15ug/ml rnase,100ug/ml dnase;Group D: 0.25%Triton-X,0.25%SDS,0.25%SD,10ug/ml rnase,100ug/ml dnase;Group E: 0.125%Triton-X,0.125%SDS,0.125%SD,5ug/ml rnase,50ug/ml dnase.The obtained rabbit aorta was decellularized to remove endothelial cells as much as possible while retaining its relatively intact tissue structure.The decellularization effect was identified by HE staining,Masson staining,Verhoff-von Gieson staining and scanning electron microscopy,and the morphological changes of decellularized vessels were observed.Pressure blasting was performed to observe its mechanical properties.After obtaining the ideal concentration of decellularized solution,the rabbit aorta was decellularized by this decellularization method.The decellularized rabbit aorta was used as the arterialized artificial small vessel scaffold,and the decellularized small vessel lumen was modified by liquid perfluorocarbon coating to make a new type of artificial small vessel.The artificial small vessel was observed by light microscope and electron microscope,and its hydrophobicity analysis,anti-platelet aggregation property in vitro and fluoride coating identification were studied.To determine the feasibility of fluorinated decellularized rabbit aorta as a small blood vessel prosthesis.Results:The combination of 0.375% Triton-X,0.375%SDS,0.375%SD,15ug/ml DNAse and 15ug/ml RNAse could effectively remove cells and maintain good tissue structure.The fluorinated coating can make the acellular blood vessels have very low signifying energy and anti-platelet aggregation without affecting their mechanical properties.Conclusion:The fluorinated acellular rabbit aorta is a kind of tissue engineered small diameter artificial blood vessel with good mechanical properties,physical stability,anticoagulation and bioblood compatibility.