Preparation of Ex Vivo–Based Biomaterials Using Convective Flow Decellularization

Abstract

With advantageous biomechanical properties, materials derived from ex vivo tissues are being actively investigated as scaffolds for tissue engineering applications. However, decellularization treatments are required before implantation to reduce the materials immune impact. The aim of these investigations was to assess a convective flow model as an enhanced methodology to decellularize ex vivo tissue. Isolated human umbilical veins were decellularized using two methods: rotary agitation at 100 rpm on orbital shaker plates, and convective flow run at 5, 50, and 150 mmHg within perfusion bioreactors. Extracted phospholipids and total soluble protein were assessed over time. Histology, SEM, and uniaxial tensile testing analysis were carried out to evaluate variation in the tissues. After 72 h, samples exposed to traditional rotary agitation showed retention of whole cells and cellular components, whereas pressure-based systems showed no visual sign of cells. The convective flow method was significantly more effective at removing phospholipid and total protein than the agitation model. High transmembrane pressure (150 mmHg) resulted in higher phospholipids extraction. However, a more efficient protein extraction occurred at 50 mmHg. Variation in extraction rates was dependent on tissue permeability, which varied as pressure increased. Collectively, these findings show significant improvements in decellularization efficiency that may lead to more immune compliant ex vivo-derived biomaterials.

Preparation of Cardiac Extracellular Matrix from an Intact Porcine Heart

“…13,14 Convincing arguments can be made for the advantages of tissue-specific ECM scaffolds for tissue-specific repair. [15][16][17][18][19][20][21][22] Logically, a biologic scaffold derived from the targeted tissue source would possess the ideal threedimensional (3D) architecture and biochemical composition to support tissue-specific cell phenotype, cell proliferation, and tissue biomechanical properties. If organs can be decellularized and still maintain their 3D integrity, the resulting scaffold would possibly represent the ideal scaffold for all components of the organ, including vascular and lymphatic structures, nerves, and the parenchymal cells.…”

Characterization and Implementation of a Decellularized Porcine Vessel as a Biologic Scaffold for a Blood Vessel Mimic

“…A lower elastic modulus implies the scaffold is more elastic, most likely due to a lack of cellular bonds that promote a firm structure. This trend was supported in the literature and is summarized in Table 8 (65,71,73,74). However, it is difficult to compare all tensile testing methods.…”

“…To evaluate scaffold mechanics, tensile and burst pressure evaluation were performed on both native and decellularized vessels. The mechanical testing methods utilized were modeled from the work done by Montoya et al in 2009 for the evaluation of a decellularized scaffold (65).…”

Arterial Decellularized Scaffolds Produced Using an Innovative Automatic System

“…These aspects represent major drawbacks when decellularization is being aimed at settings compliant with good manufacturing practice. In order to address these issues, we have already proposed a device for the automatic decellularization of biological tissues [Pellegata et al, 2012] and other devices have been proposed by other groups [Montoya and McFetridge, 2009;Sullivan et al, 2012;Fercana et al, 2014;Struecker et al, 2014;Price et al, 2015].…”