Heparin Modification Improves Biocompatibility Of Decellularized Porcine Liver Scaffolds

Objective: This article researched a method of ameliorating thebiocompatibility of the decellularized porcine liver as scaffold for liver tissueengineering research through the modification of heparin. Methods: We manufactureddifferent kinds of heparinized and decellularized porcine liver scaffolds thoughdifferent methods. The t-LBL scaffolds were made by decellularized porcine liverscaffolds immobilized with heparin by ionic binding via layer-by-layer self-assemblytechnique (LBL). The t-EPA scaffolds were made by decellularized porcine liverscaffolds immobilized with heparin by covalent binding via multi-point attachment(MPA). The t-EBL scaffolds were made by decellularized porcine liver scaffoldsimmobilized with heparin by covalent binding via end-point attachment (EPA). Weevaluated the effect of heparinization, anticoagulant ability, and cytocompatibility inthis article via Toluidine blue (TB) staining, toluidine-blue colorimetric method, HEstaining, coagulation time tests in vitro, Platelet adhesion, Plasma recalcification time,Cell culture of HepG2, MTT assay, Haemolysis ratio measurement, scanning electronmicroscopy (SEM). Results: the contents of heparin of t-LBL, t-MPA and t-EPA weredifferent. The heparinized and decellularized scaffolds displayed better performanceof the anticoagulant activity than the nonheparinized and decellularized scaffolds. Thet-EPA possessed more heparin, released heparin slower, and retained heparin morethan other heparinized counterparts. Compared with t-LBL, t-MPA, the thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (APTT) oft-EPA scaffold all exceeded the range of measurement. What’s more, therecalcification time of the t-EPA is longest. In addition, matched with control, t-LBLand t-MPA, the t-EPA scaffold further promoted the proliferation, distribution,aggregate and extension of HepG2cells since a more complex, dynamic environmentwas formed to accelerate the cell growth. Conclusion: the decellularized porcine liverscaffold immobilized with heparin by covalent binding via end-point attachment (EPA)obtained a good ability of anticoagulation and cytocompatibility. Seeing that the mildaqueous preparation condition and the good anticoagulant ability in the liver tissueengineering, our study has also opened a door to prepare various scaffolds with goodanticoagulant activity.