载外泌体银纳米颗粒抗菌水凝胶的构建及其对人表皮HaCaT细胞增殖的影响

doi:10.13481/j.1671-587X.20240206

Abstract

Abstract:

Objective To construct the antibacterial hydrogel containing mesenchymal stem cell exosomes （MSC-Exo） and silver nanoparticles （AgNPs）， and to preliminarily clarify its antibacterial activity and proliferative effect on the human epidermal HaCaT cells， and to clarify its application potential in treatment of skin wound. Methods The basic hydrogel scaffold was fabricated using oxidized dextran （ODex） and modified hyaluronic acid （HA-ADH）. The AgNPs were synthesized with cetyltrimethylammonium bromide （CTAB） as the stabilizer， while the MSC-Exo was isolated from the cultured mesenchymal stem cells （MSC）. The AgNP/HA-ADH/ODex hydrogel and MSC-Exo@AgNP/HA-ADH/ODex hydrogel were constructed. The loading rate of the Msc-Exo was evaluated through BCA assay； dynamic light scattering （DLS）， Zeta potential measurement， and ultraviolet-visible spectroscope were used to analyze the morphology， the charged status of the AgNPs； fourier transform infrared spectra was used to detect the compositions of the HA-ADH/ODex hydrogel framework； transmission electron microscope （TEM）， nanoparticle tracking analysis， and Western blotting methods were used to identify the presence of MSC-Exo；the cell survival rates of the HA-ADH/ODex hydrogel and the AgNP/HA-ADH/ODex hydrogel， and the effect of MSC-Exo@AgNP/HA-ADH/ODex hydrogel on the proliferation rate of HaCaT cells were detected by CCK-8 assay；the antibacterial efficacy of the AgNP/HA-ADH/ODex hydrogel was detected by spread plate method and turbidity measurement. Results The DLS， Zeta potential， and ultraviolet-visible spectrum results showed the successful synthesis of uniform and positively charged AgNPs. The fourier transform infrared spectrometry results showed the HA-ADH/ODex hydrogel was properly formulated. The identification of MSC-Exo was verified by TEM， nanoparticle tracking， and Western blotting methods. The BCA assay results showed that the loading rate of the MSC-Exo on the MSC-Exo@AgNP/HA-ADH/ODex hydrogel was 77.8%.The CCK-8 assay results showed that the survival rates of the HaCaT cells were close to 100% when treated with the dilution ratios of 0，1， 2， 4， 8， 16， 32， and 64 of culture medium-extract of HA-ADH/ODex hydrogel. For the AgNP/HA-ADH/ODex hydrogel extracted with dilution ratio of 8 and above， the survival rates were greater than 85%. The spread plate method results showed that the antibe terial rates of AgNP/HA-ADH/ODex and MSC-Exo@AgNP/HA-ADH/ODex hydrogels against Staphylococcus aureus （S. aureus） and Escherichia coli （E. coli）， and the antibacterial rates against S.aureus and E. coli were （96.44±0.16）%，（96.62±0.16）% and （95.73±0.28）%，（95.58±0.14）%， respectively. The CCK-8 assay results showed that the MSC-Exo@AgNP/HA-ADH/ODex hydrogel promoted the proliferation of the HaCaT cells，and the proliferation rate was （115.00 ± 7.42）%. Conclusion The MSC-Exo@AgNP/HA-ADH/ODex hydrogel exhibits the excellent antibacterial properties and good ability to promote the proliferation of human epidermal HaCaT cells； it is a safe and effective bio-dressing for the treatment of skin wounds.

Key words: Hyaluronic acid, Dextran, Silver nanoparticles, Mesenchymal stem cell exosome, Skin wound, Drug delivery

CLC Number:

R751

Tao LIU,Jiacai FU,Chengming ZHONG,Xu TENG,Ling QI. Construction of exosome and AgNPs-loaded antibacterial hydrogel and its effect on proliferation of human epidermal HaCaT cells[J].Journal of Jilin University(Medicine Edition), 2024, 50(2): 336-345.

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Construction of exosome and AgNPs-loaded antibacterial hydrogel and its effect on proliferation of human epidermal HaCaT cells

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