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. 2020 Dec 3;8(1):rbaa049.
doi: 10.1093/rb/rbaa049. eCollection 2021 Feb 1.

The study of dry biological valve crosslinked with a combination of carbodiimide and polyphenol

Li Yang  1 , Shuang Xie  1 , Kailei Ding  1 , Yang Lei  1 , Yunbing Wang  1
Affiliations

The study of dry biological valve crosslinked with a combination of carbodiimide and polyphenol

Li Yang et al. Regen Biomater. .

Abstract

The glutaraldehyde crosslinked pericardium has been used in bioprosthetic valves for about 50 years. However, problems such as glutaraldehyde residue and calcification still exist in current commercial products. Non-glutaraldehyde crosslinked dry valve is an important strategy to solve those problems. In this study, a non-glutaraldehyde crosslinked dry biological valve material was obtained by the combined crosslinking of carbodiimide (EDC) and polyphenol. The results showed that the comprehensive properties of EDC and curcumin crosslinked pericardium were superior to glutaraldehyde crosslinked pericardium, including unfolding property, anti-calcification, cytotoxicity, anticoagulant properties, mechanical properties, enzyme degradation resistance and thermal shrinkage temperature. EDC and curcumin crosslinked dry pericardium could flatten after being folded at 40°C for 3 days while glutaraldehyde crosslinked pericardium could not. The calcification of pericardium treated with EDC and curcumin was 1.21 ± 0.36 mg/g in rats after 60 days' subdermal implantation, much lower than that of glutaraldehyde treated control group (22.06 ± 3.17 mg/g).

Keywords: carbodiimide; dry biological valve; non-glutaraldehyde; polyphenol.

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Figures

Figure 1.
Figure 1.
Representative photos for the tissues (3 cm × 3 cm) pre-crimped in catheter after put into PBS are shown. Scale bar: 1 cm. The dry valve in the glutaraldehyde (GLUT) treatment group had obvious creases while the EDC/CC treatment group flattened without creases or damage.
Figure 2.
Figure 2.
Calcium contents of each explant were calculated. N = 6. The calcification content of EDC/CC and EDC/PC group was much lower than that of GLUT group.
Figure 3.
Figure 3.
A. Cell viability in each group. N = 3 (three independent experiments). B. Microscopic images (200×) of L929 cells are shown. Scale bar = 100. The cytotoxicity of polyphenols or polyphenols combined with EDC was significantly lower than that of glutaraldehyde.
Figure 4.
Figure 4.
Representative scanning electron microscope images of adherent platelets on the samples. Scale bar: 20 µm (2000×). There was more adhered platelet in the glutaraldehyde treatment group, and almost no adhered platelet was seen in the CC, EDC/CC, PC, or EDC/PCC treatment group.
Figure 5.
Figure 5.
Representative scanning electron microscope images of adherent whole blood on the samples. Scale bar: 20 µm (2000×). The results of whole blood adhesion tests were consistent with the trend of platelet adhesion.
Figure 6.
Figure 6.
Peak stress required to cause tearing (A) and ultimate tensile strength of pericardium samples (B) is shown. N = 4 (four independent experiments). Scale bar: 1 cm. The peak stress to cause tear of CC group was lower than that of glutaraldehyde group, while EDC/CC, PC, or EDC/PC treatment group was close to glutaraldehyde treatment group. The maximum tensile strength in CC group was lower than that in glutaraldehyde group, while EDC/CC, PC, EDC/PC treatment groups were close to glutaraldehyde treatment group.
Figure 7.
Figure 7.
Ultimate tensile strain of pericardium samples (A) and tangent modulus (B) are shown. N = 4 (four independent experiments). The maximum tensile strain of CC group was similar to that of glutaraldehyde group, while EDC/CC, PC, or EDC/PC treatment group was slightly lower than glutaraldehyde treatment group.
Figure 8.
Figure 8.
Collagenase and elastase enzymatic degradation in vitro. Change in mass post collagenase (A) and elastase (B) treatment. N = 4 (four independent experiments). The protective effect of the EDC/CC or EDC/PC combination treatment group on collagen was similar to that of the glutaraldehyde crosslinking group.
Figure 9.
Figure 9.
Thermal shrinkage temperature of each group. N = 4 (four independent experiments). The thermal shrinkage temperature of the EDC/CC or EDC/PC treatment group was close to that of the glutaraldehyde crosslinking group.
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