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. 2023 Nov 22;15(23):4484.
doi: 10.3390/polym15234484.

Salvia officinalis-Hydroxyapatite Nanocomposites with Antibacterial Properties

Steluta Carmen Ciobanu  1 , Daniela Predoi  1 , Mariana Carmen Chifiriuc  2   3   4 , Simona Liliana Iconaru  1 , Mihai Valentin Predoi  5 , Marcela Popa  3   4 , Krzysztof Rokosz  6 , Steinar Raaen  7 , Ioana Cristina Marinas  3
Affiliations

Salvia officinalis-Hydroxyapatite Nanocomposites with Antibacterial Properties

Steluta Carmen Ciobanu et al. Polymers (Basel). .

Abstract

In the present study, sage-coated zinc-doped hydroxyapatite was incorporated into a dextran matrix (7ZnHAp-SD), and its physico-chemical and antimicrobial activities were investigated. A 7ZnHAp-SD nanocomposite suspension was obtained using the co-precipitation method. The stability of the nanocomposite suspension was evaluated using ultrasound measurements. The stability parameter calculated relative to double-distilled water as a reference fluid highlights the very good stability of the 7ZnHAp-SD suspension. X-ray diffraction (XRD) experiments were performed to evaluate the characteristic diffraction peak of the hydroxyapatite phase. Valuable information regarding the morphology and chemical composition of 7ZnHAp-SD was obtained via scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and X-ray photoelectron spectroscopy (XPS) studies. Fourier-transform infrared spectroscopy (FTIR) measurements were performed on the 7ZnHAp-SD suspensions in order to evaluate the functional groups present in the sample. Preliminary studies on the antimicrobial activity of 7ZnHAp-SD suspensions against the standard strains of Staphylococcus aureus 25923 ATCC, Enterococcus faecalis 29212 ATCC, Escherichia coli 25922 ATCC, and Pseudomonas aeruginosa 27853 ATCC were conducted. More than that, preliminary studies on the biocompatibility of 7ZnHAp-SD were conducted using human cervical adenocarcinoma (HeLa) cells, and their results emphasized that the 7ZnHAp-SD sample did not exhibit a toxic effect and did not induce any noticeable changes in the morphological characteristics of HeLa cells. These preliminary results showed that these nanoparticles could be possible candidates for biomedical/antimicrobial applications.

Keywords: antimicrobial applications; dextran; morphology; sage; zinc doped hydroxyapatite.

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Conflict of interest statement

The authors declare no conflict of interest; The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Figure 1
Figure 1
Evolution in time of the ultrasonic signal relative amplitudes through the 7ZnHAp-SD suspension (a) and the frequency spectra of all recorded signals and the reference liquid and the 7ZnHAp-SD suspension (b).
Figure 2
Figure 2
Attenuation of ultrasonic signals vs. frequency for (a) and attenuation of ultrasonic signals vs. time for 7ZnHAp-SD suspension (b).
Figure 3
Figure 3
XRD patterns of 7ZnHAp-SD and the lines of the ICDD-PDF#9-432 reference file for hexagonal hydroxyapatite (in orange).
Figure 4
Figure 4
Low (a) and high (b) magnification micrographs of 7ZnHAp-SD particles in suspension; size distribution (c) and EDS spectrum (d) of the 7ZnHAp-SD suspension.
Figure 5
Figure 5
Elemental EDS mapping analysis of the 7ZnHAp-SD suspension.
Figure 6
Figure 6
Absorbance (a) and second-derivative (b) FTIR spectra of the 7ZnHAp-SD sample.
Figure 7
Figure 7
XPS general spectra of the 7ZnHAp-SD composite (a); and high-resolution spectra of calcium (b), phosphorus (c), oxygen, (d), and zinc (e).
Figure 8
Figure 8
The diameters of the inhibitory zones of 7ZnHAp-SD nanoparticles obtained after 24 h (a), 48 h (b), and 72 h (c) against E. coli and the graphical representation of the values obtained for the DZI evaluated at various time intervals after incubation (d) (comparisons across time intervals were not statistically significant, p > 0.05 for up to 72 h) in the case of 7ZnHAp-SD nanoparticles against E. coli. Statistical analysis revealed that the changes in diameter at different time intervals are not statistically significant (p > 0.05).
Figure 9
Figure 9
Graphical representation of the values obtained for HeLa cell viability after 24, 48, and 72 h of incubation in the presence of 7ZnHAp-SD. The results of the experiments are graphically represented as the mean ± SD. The data were statistically analyzed using paired and two-sample t-tests for means, with p ≤ 0.05 accepted as statistically significant.
Figure 10
Figure 10
Two-dimensional fluorescence microscopy images of the morphology of HeLa cells treated with 7ZnHAp-SD for 24 h, 48 h, and 72 h (d–f) relative to the untreated Hela cell culture used as a control (a–c).
Figure 11
Figure 11
Three-dimensional representation of the fluorescence micrographs of the morphology of HeLa cells treated with 7ZnHAp-SD for 24 h, 48 h, and 72 h (d–f) compared to the untreated Hela cell culture used as a control (a–c).
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