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. 2024 Apr 30;17(5):575.
doi: 10.3390/ph17050575.

Characterization, Biocompatibility and Antioxidant Activity of Hydrogels Containing Propolis Extract as an Alternative Treatment in Wound Healing

Lindalva Maria de Meneses Costa Ferreira  1 , Yuri Yoshioka Modesto  1 , Poliana Dimsan Queiroz de Souza  2 , Fabiana Cristina de Araújo Nascimento  3 , Rayanne Rocha Pereira  4 , Attilio Converti  5 , Desireé Gyles Lynch  6 , Davi do Socorro Barros Brasil  3 , Edilene Oliveira da Silva  2 , José Otávio Carréra Silva-Júnior  1 , Roseane Maria Ribeiro-Costa  1
Affiliations

Characterization, Biocompatibility and Antioxidant Activity of Hydrogels Containing Propolis Extract as an Alternative Treatment in Wound Healing

Lindalva Maria de Meneses Costa Ferreira et al. Pharmaceuticals (Basel). .

Abstract

Hydrogels consist of a network of highly porous polymeric chains with the potential for use as a wound dressing. Propolis is a natural product with several biological properties including anti-inflammatory, antibacterial and antioxidant activities. This study was aimed at synthesizing and characterizing a polyacrylamide/methylcellulose hydrogel containing propolis as an active ingredient, to serve as a wound dressing alternative, for the treatment of skin lesions. The hydrogels were prepared using free radical polymerization, and were characterized using scanning electron microscopy, infrared spectroscopy, thermogravimetry, differential scanning calorimetry, swelling capacity, mechanical and rheological properties, UV-Vis spectroscopy, antioxidant activity by the DPPH, ABTS and FRAP assays and biocompatibility determined in Vero cells and J774 macrophages by the MTT assay. Hydrogels showed a porous and foliaceous structure with a well-defined network, a good ability to absorb water and aqueous solutions simulating body fluids as well as desirable mechanical properties and pseudoplastic behavior. In hydrogels containing 1.0 and 2.5% propolis, the contents of total polyphenols were 24.74 ± 1.71 mg GAE/g and 32.10 ± 1.01 mg GAE/g and those of total flavonoids 8.01 ± 0.99 mg QE/g and 13.81 ± 0.71 mg QE/g, respectively, in addition to good antioxidant activity determined with all three methods used. Therefore, hydrogels containing propolis extract, may serve as a promising alternative wound dressing for the treatment of skin lesions, due to their anti-oxidant properties, low cost and availability.

Keywords: Apis mellifera; cytotoxicity; dressing; hydrogel; natural product; propolis.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Macroscopic characteristics of hydrogels. (A) Blank hydrogel. (B) Hydrogel containing 1.0% propolis. (C) Hydrogel containing 2.5% propolis.
Figure 2
Figure 2
Hydrogel SEM micrographs: (A) Blank hydrogel. (B) Hydrogel containing 1.0% propolis. (C) Hydrogel containing 2.5% propolis. Magnitude: 500×, 2000× and 9000×, respectively.
Figure 3
Figure 3
SEM Micrographs of hydrogels at 9000x magnitude with indication of pore size. (A) Blank hydrogel. (B) Hydrogel containing 2.5% propolis.
Figure 4
Figure 4
Swelling capacity of blank hydrogel and of hydrogels containing 1.0 and 2.5% propolis in phosphate-buffered saline, pH 7.4, and distilled water.
Figure 5
Figure 5
Results of hydrogel Texture Profile Analysis: (A) Blank hydrogel. (B) Hydrogel containing 1.0% propolis. (C) Hydrogel containing 2.5% propolis. F1: strength 1 (1st cycle), hardness. A1: area 1. D1: distance 1. F2: strength 2 (2nd cycle), hardness. A2: area 2. A3: adhesiveness. D2: distance 2. Cohesivity is represented by A1/A2. Elasticity is represented by D2/D1.
Figure 6
Figure 6
Rheological properties of the blank hydrogel and hydrogels containing 1.0 and 2.5% propolis. (A) Viscosity versus shear rate; (B) Viscosity versus time.
Figure 7
Figure 7
TG/DTG curves of non-lyophilized hydrogels. Conditions: nitrogen atmosphere, flow rate of 50 mL/min, heating rate of 10 °C/min.
Figure 8
Figure 8
TG/DTG curves of lyophilized hydrogels and propolis extract. Conditions: nitrogen atmosphere, flow rate of 50 mL/min, heating rate of 10 °C/min.
Figure 9
Figure 9
DSC curves of non-lyophilized hydrogels: (A) Hydrogel containing 2.5% propolis. (B) Hydrogel containing 1.0% propolis. (C) Blank hydrogel. Conditions: nitrogen atmosphere, flow rate of 50 mL/min, heating rate of 10 °C/min.
Figure 10
Figure 10
DSC curves of lyophilized hydrogels: (A) Hydrogel containing 2.5% propolis. (B) Hydrogel containing 1.0% propolis. (C) Blank hydrogel. (D) Propolis extract. Conditions: nitrogen atmosphere, flow rate of 50 mL/min, heating rate of 10 °C/min.
Figure 11
Figure 11
FTIR spectra of lyophilized hydrogels: (A) Hydrogel containing 2.5% propolis. (B) Hydrogel containing 1.0% propolis. (C) Blank hydrogel. (D) Propolis extract. The analyses were performed in the wavenumber range of 4000 to 400 cm−1, with 32 scans and resolution of 4 cm−1.
Figure 12
Figure 12
Values of the antioxidant activity of propolis extract, by the ABTS, DPPH and FRAP assays. (A) Expressed in µM Trolox equivalent (TE)/g. (B) Percentage of inhibition.
Figure 13
Figure 13
Values of the antioxidant activity of blank and propolis-containing hydrogels determined by the ABTS, DPPH and FRAP assays. Different letters in the same column indicate significant difference. a** (p < 0.01) and b* (p < 0.05).
Figure 14
Figure 14
Viability of (A) Vero fibroblast-like kidney cells and (B) macrophages (J774) after 24 h of treatment with different extract and hydrogel concentrations (2.5, 5, 10, 20, 50, 100, 250 and 500 μg/mL). Control: Without treatment. The results were analyzed by the analysis of variance (ANOVA) followed by the Tukey test, b**** p < 0.0001, a*** p < 0.001, c** p < 0.01.
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