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. 2020 Apr 3:8:281.
doi: 10.3389/fbioe.2020.00281. eCollection 2020.

Polylysine Enriched Matrices: A Promising Approach for Vascular Grafts

Luca Fusaro  1   2 , Marta Calvo Catoira  2   3 , Martina Ramella  1   2 , Federico Sacco Botto  4 , Maria Talmon  1 , Luigia Grazia Fresu  1 , Araida Hidalgo-Bastida  5   6   7 , Francesca Boccafoschi  1   2
Affiliations

Polylysine Enriched Matrices: A Promising Approach for Vascular Grafts

Luca Fusaro et al. Front Bioeng Biotechnol. .

Abstract

Cardiovascular diseases represent the leading cause of death in developed countries. Modern surgical methods show poor efficiency in the substitution of small-diameter arteries (<6 mm). Due to the difference in mechanical properties between the native artery and the substitute, the behavior of the vessel wall is a major cause of inefficient substitutions. The use of decellularized scaffolds has shown optimal prospects in different applications for regenerative medicine. The purpose of this work was to obtain polylysine-enriched vascular substitutes, derived from decellularized porcine femoral and carotid arteries. Polylysine acts as a matrix cross-linker, increasing the mechanical resistance of the scaffold with respect to decellularized vessels, without altering the native biocompatibility and hemocompatibility properties. The biological characterization showed an excellent biocompatibility, while mechanical tests displayed that the Young's modulus of the polylysine-enriched matrix was comparable to native vessel. Burst pressure test demonstrated strengthening of the polylysine-enriched matrix, which can resist to higher pressures with respect to native vessel. Mechanical analyses also show that polylysine-enriched vessels presented minimal degradation compared to native. Concerning hemocompatibility, the performed analyses show that polylysine-enriched matrices increase coagulation time, with respect to commercial Dacron vascular substitutes. Based on these findings, polylysine-enriched decellularized vessels resulted in a promising approach for vascular substitution.

Keywords: decellularized vessels; matrix degradation; polylysine; surface grafting; vascular substitutes.

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Figures

FIGURE 1
FIGURE 1
Decellularization of porcine artery showing (A) DAPI and HandE histology before and after decellularization process. (B) Quantitative determination of DNA in a native and decellularized matrix. Data are expressed as mean values ± SD (n = 3, *p ≤ 0.05). (C) Agarose gel electrophoresis of DNA derived from native and decellularized matrices. The image is representative of three different analyses.
FIGURE 2
FIGURE 2
X-ray photoelectron spectroscopy (XPS) spectra of (A) decellularized and (B) polylysine enriched matrices. (C) AT-IR spectra of decellularized (red), polylysine enriched matrices (blue), and Polylysine as a control (green).
FIGURE 3
FIGURE 3
(A) Confocal microscopy of decellularized and polylysine-enriched vessels. (B) Micro-CT analysis of native vessel, decellularized and polylysine-enriched matrices.
FIGURE 4
FIGURE 4
Mechanical testing results under static conditions: (A, left panel) Young’s modulus and tensile strength representation for native, decellularized and polylysine grafting. Data are expressed normalized with a native vessel. The data origin is the stress–strain curves. Data are expressed as mean values ± SD (n = 3, ∗p ≤ 0.05). (B) Burst pressure data for native, decellularized and polylysine grafting. Data are expressed as mean values ± SD (n = 3, ∗p ≤ 0.05). Mechanical testing results under dynamic conditions: (C) Burst pressure data for native, decellularized, and polylysine grafting for static and dynamic conditions. Data are expressed as mean values ± SD (n = 5, ∗p ≤ 0.05). (D) Degradation assays in terms of weight loss. Data are expressed as mean values ± SD (n = 5, ∗p ≤ 0.05). (E) Degradation assays in terms of tensile strength loss. Data are expressed as mean values ± SD (n = 3, ∗p ≤ 0.05).
FIGURE 5
FIGURE 5
(A) Endothelial cells viability assays. MTS test was performed after 1, 3, and 7 days. Data are expressed as mean values ± SD (n = 3, ∗p ≤ 0.05). (B) Hematoxylin-Eosin staining of decellularized and polylysine enriched matrices with cells cultured for 3 and 7 days. (C) DAPI staining of decellularized and polylysine enriched matrices after dynamic conditions.
FIGURE 6
FIGURE 6
(A) Monocyte viability assay. MTT assay was performed after 24 h. (B) ELISA assay on TNF-α concentration after 7 days of culture. Data are expressed as mean values ± SD (n = 3, *p ≤ 0.05).
FIGURE 7
FIGURE 7
Haemocompatibility test. Coagulation parameters (A: reaction time; B: coagulation time; C: platelet aggregation; D: fibrinogenic activity) were evaluated on synthetic vascular substitutes (DACRON) and treated matrices. Data are expressed as mean values ± SD (n = 3 from different healthy donors, ∗p ≤ 0.05).
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