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. 2023 Apr 12:10:rbad028.
doi: 10.1093/rb/rbad028. eCollection 2023.

A CS-based composite scaffold with excellent photothermal effect and its application in full-thickness skin wound healing

Jing Wang  1 , Shijia Fu  1 , Huishan Li  1 , Yue Wu  1
Affiliations

A CS-based composite scaffold with excellent photothermal effect and its application in full-thickness skin wound healing

Jing Wang et al. Regen Biomater. .

Abstract

The development of natural polymer-based scaffolds with excellent biocompatibility, antibacterial activity, and blood compatibility, able to facilitate full-thickness skin wound healing, remains challenging. In this study, we have developed three chitosan (CS)-based porous scaffolds, including CS, CS/CNT (carbon nanotubes) and CS/CNT/HA (nano-hydroxyapatite, n-HA) using a freeze-drying method. All three scaffolds have a high swelling ratio, excellent antibacterial activity, outstanding cytocompatibility and blood compatibility in vitro. The introduction of CNTs exhibited an obvious increase in mechanical properties and exerts excellent photothermal response, which displays excellent healing performance as a wound dressing in mouse full-thickness skin wound model when compared to CS scaffolds. CS/CNT/HA composite scaffolds present the strongest ability to promote full-thickness cutaneous wound closure and skin regeneration, which might be ascribed to the synergistic effect of photothermal response from CNT and excellent bioactivity from n-HA. Overall, the present study indicated that CNT and n-HA can be engineered as effective constituents in wound dressings to facilitate full-thickness skin regeneration.

Keywords: carbon nanotubes; chitosan; hydroxyapatite nanoparticles; photothermal effect; skin regeneration.

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Figures

None
Graphical abstract
Figure 1.
Figure 1.
SEM images of CS, CS/CNT and CS/CNT/HA scaffolds (black arrow indicated the n-HA in scaffolds) at low and high resolution (a); TEM image of n-HA particles (b); FTIR spectra of CS, CS/CNT and CS/CNT/HA scaffolds (c); XRD patterns of CS, CS/CNT and CS/CNT/HA scaffolds (d).
Figure 2.
Figure 2.
Full spectrum of XPS survey scan for CS, CS/CNT and CS/CNT/HA scaffolds (a); high-resolution XPS spectra of signal C1s (b), Ca2p (c), O1s (d), N1s (e) and P2p (f) element for CS, CS/CNT and CS/CNT/HA scaffolds.
Figure 3.
Figure 3.
The schematic diagram and results of the compressive test (a and c) and extension test (b and d). *P < 0.05.
Figure 4.
Figure 4.
The swelling properties (a) and the degradation behaviors (b) of CS, CS/CNT and CS/CNT/HA scaffolds. *P < 0.05.
Figure 5.
Figure 5.
The photothermal properties of CS, CS/CNT and CS/CNT/HA scaffolds. Photothermal images (a); time–temperature curve of CS, CS/CNT and CS/CNT/HA scaffolds exposed to 808 nm NIR irradiation at a 1.0 W cm−2 output power intensity for different times (b); the heating and cooling cycle curve of CS/CNT (c) and CS/CNT/HA (d) scaffolds exposed to 808 nm NIR irradiation at a 1.0 W cm−2 output power intensity for four cycles.
Figure 6.
Figure 6.
The biocompatibility of CS-based scaffolds. SEM images of L929 morphology on CS, CS/CNT and CS/CNT/HA scaffolds after incubation for 2 days (a); the cell cytoskeleton and nucleus images of L929 co-cultured with control and the extracts from CS, CS/CNT and CS/CNT/HA scaffolds (b and c); proliferation results of the L929 (d) and HUVECs (e) co-cultured with control, CS, CS/CNT, CS/CNT/HA scaffolds, respectively. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.
Figure 7.
Figure 7.
SEM images of blood cells(black arrow in CS group; white circle region in CS/CNT and CS/CNT/HA groups) and platelet adsorption on CS, CS/CNT and CS/CNT/HA scaffolds.
Figure 8.
Figure 8.
Antibacterial properties of CS, CS/CNT and CS/CNT/HA scaffolds. Images showing the growth status of E.coli (a) and S.aureus (d) after co-incubated with control, CS, CS/CNT and CS/CNT/HA scaffolds, respectively. The OD value at 600 nm of E.coli (b) and S.aureus (e) after co-incubated with control, CS, CS/CNT and CS/CNT/HA scaffolds, respectively. The SEM images and bacteriostasis circle of E.coli (c) and S.aureus (f) after contact with control, CS, CS/CNT and CS/CNT/HA scaffolds. ****P < 0.0001.
Figure 9.
Figure 9.
Biocompatibility of CS, CS/CNT and CS/CNT/HA scaffolds in vivo after implantation for 3, 7 and 14 days. Histological analysis of tissue growth into the scaffolds and the bonding state (H&E staining, yellow * = scaffolds, blue * = tissue growth into the pores of scaffolds, yellow # = scaffolds site, blue # = host tissue site, yellow dotted line = the interface between host tissue and scaffolds).
Figure 10.
Figure 10.
Wound healing properties of CS, CS/CNT and CS/CNT/HA scaffolds in vivo. Photothermal images of a mouse wound treated with or without scaffolds and exposed to 808 nm NIR light at 1.0 W cm−2 output power (a). H&E staining images of wound skin after 13 days of treatment with or without NIR radiation on 808 nm at 1.0 W cm−2 output power (b). The quantification analysis of the thickness for newly formed granulation tissue and epidermis at the wound sites (c). The images of the wound healing process (0, 3, 7 and 14 days) treated by different scaffolds with or without NIR radiation on 808 nm at 1.0 W cm−2 output power (d). Quantification analysis of wound area at Day 3, 7 and 14 in different groups (e) (*difference of scaffolds compared with control, ****P < 0.0001; #differences between the same scaffold with or without NIR radiation on 808 nm at 1.0 W cm−2 output power, ####P < 0.0001).
Figure 11.
Figure 11.
Histological analysis of CS, CS/CNT and CS/CNT/HA scaffolds in full-thickness skin wound healing using Masson trichrome staining (a) and Sirius red staining (c). The quantitative analysis of collagen deposition (b and d) in different groups according to the images of Masson trichrome staining and Sirius red staining (*difference of scaffolds compared with control, *P < 0.05, **P < 0.01 and ****P < 0.0001).
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