Reveal crucial subtype of natural chondroitin sulfate on the functionalized coatings for cardiovascular implants

doi:10.1016/j.jmst.2021.02.038

Abstract

Abstract:

Surface functionalization is generally accepted as one of the most useful methods for endowing better biocompatibility of the cardiovascular implants. Chondroitin sulfate (CS) is a commercial drug applied for preventing cardiovascular diseases in clinics, suggesting a potential application for functionalized coatings. Our previous work demonstrated that the CS conjugated onto the poly-dopamine (PDA)/hexanediamine (HD) co-deposited 316L stainless steel (316L SS) could endow the surface stronger multi-functions for cardiovascular implants. However, the natural CS has two subtypes, CS A and CS C, while it is still unclear which subtype makes more contribution to the surface modification of cardiovascular implants. Thus, the present study focus on comparing CS A and CS C via preparing functionalized coatings on the cardiovascular implants. Our data suggested PDA/HD-CS C coating possessed better blood compatibility and pro-endothelialization function through controlling the phenotype and distribution of macrophages, smooth muscle cells and endothelial cells in time sequence and space sequence.

Key words: Cardiovascular implant, Surface functionalization, Chondroitin sulfate A, Chondroitin sulfate C

Dan Zou, Jingan Li, Fang Kou, Xiao Luo, Ping Yang. Reveal crucial subtype of natural chondroitin sulfate on the functionalized coatings for cardiovascular implants[J]. J. Mater. Sci. Technol., 2021, 91: 67-77.

Figures/Tables 11

Fig. 1. Scheme of preparing PDA/HD-CS A and PDA/HD-CS C coatings on the 316L SS discs respectively.

Fig. 2. (A) FTIR spectra of 316L SS, PDA/HD, PDA/HD-CS A and PDA/HD-CS C; (B) Surface scanning X-ray photoelectron full spectrum of PDA/HD, PDA/HD-CS A and PDA/HD-CS C; (C) Water contact angles of 316L SS, PDA/HD, PDA/HD-CS A and PDA/HD-CS C (mean ± SD, n=6).

Table 1 Elemental composition of PDA/HD, PDA/HD-CS A and PDA/HD-CS C obtained by XPS.

	PDA/HD	PDA/HD-CS A	PDA/HD-CS C
C1s	74.82	66.98	68.05
N1s	12.73	13.69	10.74
O1s	12.21	18.9	20.05
S2p	0.23	0.43	1.16

Fig. 3. (A) Fluorescent images and SEM images of the adherent platelets on 316L SS, PDA/HD, PDA/HD-CS A and PDA/HD-CS C; (B) Quantitative analysis of the total platelets on PDA/HD, PDA/HD-CS A and PDA/HD-CS C by the LDH method (*p < 0.05 compared with other samples, mean ± SD, n=3); (C) Quantitative analysis of the activated platelets on PDA/HD, PDA/HD-CS A and PDA/HD-CS C by the GMP140 method (*p < 0.05 compared with other samples, **p < 0.05 compared with 316L SS and PDA/HD, mean ± SD, n=3).

Fig. 4. (A) Sectional view and surface view of the cavity with the samples of PDA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS after the ex vivo experiment (B) SEM images of the whole blood composition adherent on the PDA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS surfaces after the ex vivo experiment; (C) Blood flow rate and (D) thrombosis weight of the PDA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS samples calculated from the ex vivo experiment (*p < 0.05 compared with 316L SS, PDA/HD and PDA/HD-CSA, mean ± SD, n=3).

Fig. 5. (A) Immunofluorescence staining images of CD34 and DAPI on the PDA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS samples after the ex vivo experiment; (B) Quantity of EPC ratios of the adherent cells on the DA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS samples after the ex vivo experiment (?p < 0.05 compared with all the other samples, ??p < 0.05 compared with 316L SS and PDA/HD, mean ± SD, n=3).

Fig. 6. Dead/living staining of EC on the PDA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS surfaces. Green: living cells, blue: dead cells.

Fig. 7. The total and dead EC grown on the PDA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS surfaces ((???p < 0.05 compared with all the other samples, ??p < 0.05 compared with 316L SS and PDA/HD, mean ± SD, n=3).

Fig. 8. Release rates of NO generated by EC cultured on PDA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS after incubation for 1 day and 3 days.

Fig. 9. (A) Fluorescence images of SMC stained with rhodamine on the PDA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS after incubation for 1 day and 3 days; (B) SMC numbers of PDA/HD-CS A, PDA/HD-CS C, PDA/HD and 316L SS samples after incubation for 1 day and 3 days (*p < 0.05, ***p < 0.001, mean ± SD, n=3).

Fig. 10. (A) HE stained optical photographs of the PDA/HD-CS C-coated 316L SS stents implanted into New Zealand white rabbit's iliac artery; (B) Immunofluorescence analysis of the stented arterial segments: immunofluorescence staining for CD31, α-SMA, OPN, CD32 and CD206 of tissues treated with the PDA/HD-CS C-coated 316L SS stents implanted into New Zealand white rabbit's iliac artery (the white star marks the location of the stents’ wires implanted).

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