Endothelial Progenitor Cell-Based in vitro Pre-Endothelialization of Human Cell-Derived Biomimetic Regenerative Matrices for Next-Generation Transcatheter Heart Valves Applications

Abstract

Hemocompatibility of cardiovascular implants represents a major clinical challenge and, to date, optimal antithrombotic properties are lacking. Next-generation tissue-engineered heart valves (TEHVs) made from human-cell-derived tissue-engineered extracellular matrices (hTEMs) demonstrated their recellularization capacity in vivo and may represent promising candidates to avoid antithrombotic therapy. To further enhance their hemocompatibility, we tested hTEMs pre-endothelialization potential using human-blood-derived endothelial-colony-forming cells (ECFCs) and umbilical vein cells (control), cultured under static and dynamic orbital conditions, with either FBS or hPL. ECFCs performance was assessed via scratch assay, thereby recapitulating the surface damages occurring in transcatheter valves during crimping procedures. Our study demonstrated: feasibility to form a confluent and functional endothelium on hTEMs with expression of endothelium-specific markers; ECFCs migration and confluency restoration after crimping tests; hPL-induced formation of neo-microvessel-like structures; feasibility to pre-endothelialize hTEMs-based TEHVs and ECFCs retention on their surface after crimping. Our findings may stimulate new avenues towards next-generation pre-endothelialized implants with enhanced hemocompatibility, being beneficial for selected high-risk patients.

Xenogeneic Serum‐Free Human Cell‐Derived Tissue Engineered Matrices for the Development of Clinical‐Grade Biomimetic Cardiovascular Devices

“…As reported by our mechanical tests, xenogeneic serum‐free hTEMs show similar soft material properties as control hTEMs, thereby potentially inducing a resolution of BMP‐2 expression over time, as the implants remodel in vivo. Moreover, such preliminary results together with the previously reported hTEMs soft material properties, [ 23 ] and their ability to rapidly endothelialize, [ 42 ] may warrant further in‐depth investigation and if successful may also hold promise for successful clinical translation.…”

“…[40,41] In this context, we have previously shown improved pre-endothelization potential of endothelial progenitor cells on hTEMs cultured with HPL, demonstrating the capacity to form microvessel-like structures. [42] However, while some studies reported HPL alone to successfully replace FBS in tissue culture, others have shown limited cellular proliferation rates, [43] which may in clinical setting limit the expansion of cells or ECM formation.…”

Combining Cell Technologies With Biomimetic Tissue Engineering Applications: A New Paradigm for Translational Cardiovascular Therapies

“… 15 B : Macroscopical and microscopic appearance of hTEMs before and after in vitro culture. 16 C : Design and fabrication of tubular, spatially heterogeneous, PCL MEW scaffolds (scale bar: 5 mm). 17 D : Focused rotary jet spinning for producing helical structures.…”

“…In parallel to hiPSC-based approaches, ECM-based technologies are also gaining attention for the design of suitable patient-specific adaptive biomaterials. Particularly, human cell-derived tissue-engineered matrices (hTEMs) being free of xenogeneic antigens could represent a promising candidate and holds great clinical translation potential 16 , 34-36 ( Fig. 1B ).…”

The path to a hemocompatible cardiovascular implant: Advances and challenges of current endothelialization strategies

“…Biomimetic acellular ECM-based TEHVs are manufactured from a polymer composite (PGA/P4HB) and an in vitro grown ECM, which are subsequently decellularized leaving a cell-free construct. Preclinical studies of such biomimetic valves have demonstrated their ability to undergo remodeling and recellularization, including endothelialization (102)(103)(104)(105)(106)(107)(108)(109). Biofunctionalization of decellularized scaffolds with antibodies (e.g., CD133) to attract and capture circulating EPCs was also performed, displaying rapid in situ endothelialization within the first month after implantation (110,111).…”