Tissue‐engineering acellular scaffolds—The significant influence of physical and procedural decellularization factors

Abstract

The importance of decellularized medical products has significantly increased during the last years. In this paper, we evaluated the effects of selected physical and procedural decellularization (DC) factors with the aim to systematically assess their influence on DC results. 72 porcine aortic walls (AW) were divided into three groups and exposed to a DC solution for 4 h and 8 h, either continuously or in repeated cycles. The AW were rocked (90bpm), whirled (10 l/min), sonicated (120W, 45 kHz) or exposed to a combination of these treatments, followed by 10 washing cycles. Defining successful DC as removal of nuclei while keeping an intact extracellular matrix (ECM), we equalized the efficiency to the penetration depth (PD), obtained by DAPI fluorescence and H&E staining. Additionally, we performed scanning electron microscopy (SEM), Pentachrome and Picrosirius-Red staining. Results showed that significantly higher DC depths are achieved on outer compared to inner surfaces (61 ± 7%; p < 0.001). Furthermore, the PD showed a high time dependency for all samples. Compared to continuous rocking, we achieved a significant increase in the DC efficiency through cyclic treatments ( ∼ 43%), whirling ( ∼ 19%) and sonication ( ∼ 49%). The combined treatment supported these results. In all procedures, a skeletonized but intact Collagen fibrous network was obtained as confirmed by SEM analysis. In conclusion, we systematically identified essential factors to significantly enhance DC procedures. We highly recommend considering these factors in future DC protocols. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 153-162, 2018.

Preparation and characterization of small-diameter decellularized scaffolds for vascular tissue engineering in an animal model

“…Apart from what has been discussed above, other factors can influence the decellularization process [ 54 ], such as physical factors (rocking/whirling/sonicating) or device procedural factors. We developed a simple combination for decellularization, but three-dimensional printing technology is growing fast [ 55 ].…”

Tissue-Specific Decellularization Methods: Rationale and Strategies to Achieve Regenerative Compounds

“…Mechanical agitation and sonication are useful in combination with a chemical treatment to assist in cell lysis and the removal of cellular debris [40,41]. Mechanical agitation can be applied by using a magnetic stir plate, an orbital shaker, or a low-profile roller.…”

Successful decellularization of thick-walled tissue: Highlighting pitfalls and the need for a multifactorial approach

“…As published previously, no negative effects were induced by sonication treatment. 21,34 The slight loosening (higher transparency) of the tissue could be explained by the removal of cellular components and GAGs. It is known that DC normally affects and reduces the GAG content of tissue.…”

“…20 In a previous publication, we could show that DC efficiency can be significantly enhanced by procedural and physical modifications of established protocols. 21 Based on these findings, we developed a stable, safe, and reproducible protocol for successful aortic DC with a multifactorial approach.…”