Improving the biological function of decellularized heart valves through integration of protein tethering and three‐dimensional cell seeding in a bioreactor

Abstract

Decellularized xenogeneic heart valves (DHVs) are promising products for valve replacement. However, the widespread clinical application of such products is limited due to the risk of immune reaction, progressive degeneration, inflammation, and calcification. Here, we have developed an optimized decellularization protocol for a xenogeneic heart valve. We improved the biological function of DHVs by protein tethering onto DHV and three-dimensional (3D) cell seeding in a bioreactor. Our results showed that heart valves treated with a Triton X-100 and sodium deoxycholate-based protocol were completely cell-free, with preserved biochemical and biomechanical properties. The immobilization of stromal derived factor-1α (SDF-1α) and basic fibroblast growth factor on DHV significantly improved recellularization with endothelial progenitor cells under the 3D culture condition in the bioreactor compared to static culture conditions. Cell phenotype analysis showed higher fibroblast-like cells and less myofibroblast-like cells in both protein-tethered DHVs. However, SDF-DHV significantly enhanced recellularization both in vitro and in vivo compared to basic fibroblast growth factor DHV and demonstrated less inflammatory cell infiltration. SDF-DHV had less calcification and platelet adhesion. Altogether, integration of SDF-1α immobilization and 3D cell seeding in a bioreactor might provide a novel, promising approach for production of functional heart valves.

Human Heart Explant-Derived Extracellular Vesicles: Characterization and Effects on the In Vitro Recellularization of Decellularized Heart Valves

“…Although DHVs show promising results for valve replacement, the clinical application of these scaffolds remains a challenge because of the limited cell repopulation into the decellularized natural matrix [17]. One strategy to facilitate cell recruitment into the DHV is the use of bioactive molecules [42,43,44]. Complete recellularization of the acellular scaffold is highly desirable because it can modulate the host response to transplanted tissue [45] and inhibit tissue calcification [46].…”

Differentiation of human spermatogonial stem cells using a human decellularized testicular scaffold supplemented by platelet‐rich plasma

“…Effective decellularization of organs or tissues is essential for producing high-quality ECM scaffolds, and inefficient decellularization processes alter the ECM's composition. 18 Different factors, such as the organ's or tissue's structural properties and the periods of exposure to decellularization reagents, can influence the effectiveness of decellularization. 19 SDS (ionic surfactant) and Triton X-100 (nonionic surfactant) or a combination of both are widely employed for complete cell removal of various tissues and organs.…”

Next-generation tissue-engineered heart valves with repair, remodelling and regeneration capacity

“…However, this approach still presents several limitations: EC isolation and expansion, as well as EPC isolation and differentiation, are time consuming and costly procedures; cell seeding may be inefficient due to limited and/or inhomogeneous cell adhesion on the material 167 ; and cell retention and viability upon implantation via minimally-invasive transcatheter procedures is limited 168 . To overcome these limitations, a combination approach using SDF1a functionalization and EC pre-seeding has been evaluated, demonstrating enhanced recellularisation, less inflammation, calcification, and platelet adhesion 163 . In addition, recently developed bioreactors for 3D cell seeding should be considered to ensure homogeneous and controlled EC distribution 169 .…”

“…In addition, recently developed bioreactors for 3D cell seeding should be considered to ensure homogeneous and controlled EC distribution 169 . These results suggest that a more complex functionalization system, combining two or more bioactive components 161,170 , as well as surface topography 171 and EC pre-seeding 163 , should be further investigated to improve and control in-situ endothelialisation of regenerative materials.…”