Characterization and in vivo study of decellularized aortic scaffolds using closed sonication system

Abstract

Extracellular matrix (ECM) based bioscaffolds prepared by decellularization has increasingly emerged in tissue engineering application because it has structural, biochemical, and biomechanical cues that have dramatic effects upon cell behaviors. Therefore, we developed a closed sonication decellularization system to prepare ideal bioscaffolds with minimal adverse effects on the ECM. The decellularization was achieved at 170 kHz of ultrasound frequency in 0.1% and 2% Sodium Dodecyl Sulphate (SDS) solution for 10 hours. The immersion treatment as control was performed to compare the decellularization efficiency with our system. Cell removal and ECM structure were determined by histological staining and biochemical assay. Biomechanical properties were investigated by the indentation testing to test the stiffness, a residual force and compression of bioscaffolds. Additionally, in vivo implantation was performed in rat to investigate host tissue response. Compared to native tissues, histological staining and biochemical assay confirm the absence of cellularity with preservation of ECM structure. Moreover, sonication treatment has not affected the stiffness [N/mm] and a residual force [N] of the aortic scaffolds except for compression [%] which 2% SDS significantly decreased compared to native tissues showing higher SDS has a detrimental effect on ECM structure. Finally, minimal inflammatory response was observed after 1 and 5 weeks of implantation. This study reported that the novelty of our developed closed sonication system to prepare ideal bioscaffolds for tissue engineering applications.

Xenogeneic Decellularized Extracellular Matrix-based Biomaterials For Peripheral Nerve Repair and Regeneration

“…The formation of micropores could destabilize cells and eventually lead to cell death. Although these physical decellularization methods alone have been used successfully in a few tissues, they are often used in conjunction with chemical or enzymatic methods to effectively clear genetic material from the tissue [ 45 , 46 ].…”

Vascular Tissue Engineering: Polymers and Methodologies for Small Caliber Vascular Grafts

“…The viscoelasticity properties can be analyzed by ball indentation. This test can provide information such as the stiffness, compressibility, and residual force of the scaffolds ( 32 ).…”

“…Various vessels have been decellularized in order to obtain a vascular graft, as for example, the carotid artery ( 30 ), aorta artery ( 32 , 60 ), internal mammary artery ( 61 ), umbilical artery ( 62 ), saphenous vein ( 63 ), coronary artery ( 64 ), femoral vein ( 58 ), and the vena cava ( 55 ). The protocols for vessel decellularization include a number of agents, such as surfactants, enzymes, solvents, and physical methods.…”

“…Some studies also combine physical methods for chemical or biological agents in order to increase the penetration of agents and increase the decellularization of vascular tissue ( 32 , 55 , 60 , 61 ). Simsa and colleagues evaluated the decellularization of vena cavas with TritonX-100, TnBP, and DNase under static, agitation, and perfusion conditions.…”

“…Non-effective decellularization can cause inflammatory response and antigenicity, elevating the risk of failure of a graft ( 32 ). It is therefore important to highlight that some criteria must be observed to ensure that the decellularization of tissue can be considered satisfactory.…”