The Influence of κ-Carrageenan-R-Phycoerythrin Hydrogel on In Vitro Wound Healing and Biological Function

Abstract

Wound healing is widely recognized as a critical issue impacting the healthcare sector in numerous countries. The application of wound dressings multiple times in such instances can result in tissue damage, thereby increasing the complexity of wound healing. With the aim of tackling this necessity, in the present study, we have formulated a hydrogel using natural polysaccharide κ-carrageenan and phycobiliprotein R-phycoerythrin from Pyropia yezoensis. The formulated hydrogel κ-Carrageenan-R-Phycoerythrin (κ-CRG-R-PE) was analyzed for its antioxidant and antimicrobial activity. The wound healing potential of the κ-CRG-R-PE was evaluated in Hs27 cells by the wound scratch assay method. The hydrogel showed dose-dependent antioxidant activity and significant antimicrobial activity at 100 μg/mL concentration. κ-CRG-R-PE hydrogels promoted more rapid and complete wound closure than κ-Carrageenan (κ-CRG) hydrogel at 24 and 48 h. κ-CRG-R-PE hydrogels also filled the wound within 48 h of incubation, indicating that they positively affect fibroblast migration and wound healing.

Keywords: R-phycoerythrin; antimicrobial activity; antioxidant activity; hydrogel; wound healing; κ-carrageenan.

Figure 1

Synthesized hydrogel: (A) κ-carrageenan (κ-CRG) hydrogel, (B) κ-carrageenan-R-phycoerythrin (κ-CRG-R-PE) hydrogel, (C) Gelation property of the κ-carrageenan-R-phycoerythrin (κ-CRG-R-PE) hydrogel (D) Swelling analysis of hydrogels. Values are expressed as mean ± SD (n = 3). Different letters of the alphabet denote significant differences (p < 0.05), while the same letters of the alphabet indicate no significant difference (p > 0.05). (E) SEM image of κ-carrageenan (κ-CRG) hydrogel, (F) κ-carrageenan-R-phycoerythrin (κ-CRG-R-PE) hydrogel, (G) pore size of the hydrogels (H) FTIR spectra of κ-CRG hydrogel and κ-CRG-R-PE hydrogel (scale bar represents 100 μm), (I) Degradation analysis of hydrogels. Values are expressed as mean ± SD (n = 3). Different letters of the alphabet denote significant differences (p < 0.05), while the same letters of the alphabet indicate no significant difference (p > 0.05).

Figure 2

(A) Linear viscoelastic region (LVE) during an amplitude sweep test, (B) Complex viscosity (η*), (C) Storage modulus (G′), (D) Loss modulus (G″) during an angular frequency sweep test for hydrogels κ-CRG and κ-CRG-R-PE.

Figure 3

Antioxidant activity of κ-carrageenan (κ-CRG) hydrogel, κ-carrageenan-R-phycoerythrin (κ-CRG-R-PE) hydrogel: (A) ABTS radical scavenging activity of κ-CRG, (B) ABTS radical scavenging activity of κ-CRG-R-PE, (C) DPPH radical scavenging activity κ-CRG, (D) DPPH radical scavenging activity κ-CRG-R-PE. Values are expressed as mean ± SD (n = 3). Different letters of the alphabet denote significant differences (p < 0.05), while the same letters of the alphabet indicate no significant difference (p > 0.05).

Figure 4

Antibacterial activity of κ-carrageenan (κ-CRG) hydrogel and κ-carrageenan-R-phycoerythrin (κ-CRG-R-PE) hydrogel: (A) Antibacterial activity against P. aeruginosa, (B) Antibacterial activity against S. aureus. Kanamycin was used as a positive control (50 μg/mL) and κ-carrageenan (κ-CRG) hydrogel (100 μg/mL) was used as a negative control.

Figure 5

Cell viability assay and live–dead cell assay. (A) Cytotoxicity of κ-carrageenan (κ-CRG) hydrogel and κ-carrageenan-R-phycoerythrin (κ-CRG-R-PE) hydrogel analyzed in Hs27 cells. Values are expressed as mean ± SD (n = 3), ns (no significance). (B) Live–dead cell assay of Hs27 cells treated with different concentrations of hydrogels using propidium iodide and acridine orange staining. The scale bar represents 100 μm.

Figure 6

(A) Wound scratch assay showing gap closure at 0 h, 24 h, 48 h of control, κ-CRG hydrogel, κ-CRG-R-PE hydrogel, (B) Quantitative estimation of wound healing percentage in control, κ-CRG hydrogel, κ-CRG-R-PE hydrogel. The scale bar represents 100 μm. Values are expressed as mean ± SD (n = 3).