In light of the growing use of natural products in medicinal therapy, this study evaluated the proliferative effects of clove oil (CO) in two forms—native and loaded onto silica nanoparticles (CO@SNPs)—on Human Gingival Fibroblasts (HGFs), both singly and in combination with low-dose gamma irradiation (0.25 Gy). Silica nanoparticles were chemically synthesized and loaded with clove oil, with thorough characterization conducted using dynamic light scattering (DLS) for particle size, zeta-potential measurements, Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). The EC50 values of CO and CO@SNPs were determined to be 1.88 µmol/mL and 0.172 µmol/mL, respectively, demonstrating a superior proliferative efficacy of the CO@SNPs nanoformulation. The biological effects were assessed using the MTT assay for cell viability, Trypan blue exclusion for cell count, qPCR for gene expression of c-Myc, MAPK, and NF-κB, and antioxidant capacity assays (MDA and catalase activity). Moreover, the in vitro wound-healing scratch assay showed enhanced fibroblast migration and proliferation, with relative wound density reaching 48% and 62% for CO@SNPs alone and combined with low-dose gamma irradiation, respectively. Further analysis through immunofluorescence staining of TGF-β and hematoxylin staining confirmed cell proliferation. Our results showed that loading CO on silica nanoparticles (CO@SNPs) were significantly more efficient than native clove oil in promoting cell proliferation and wound healing, with the combined treatment of CO@SNPs and low-dose gamma irradiation producing the most pronounced regenerative effect. These findings suggest that CO@SNPs, especially when combined with low-dose gamma irradiation, hold significant promise for enhancing oral tissue regeneration. Graphical abstract <UnorderedList Mark="Bullet"> <ItemContent> Nanotechnology-enhanced clove oil delivery: The present study successfully encapsulated clove oil (CO) in silica nanoparticles (CO@SNPs), which improved its stability and controlled release and provided its therapeutic efficacy. </ItemContent> <ItemContent> Antioxidant and wound-healing properties: CO@SNPs significantly reduced oxidative stress markers (e.g., malondialdehyde levels decreased by 93%) and enhanced antioxidant enzyme activity (catalase activity increased by 83%) in human gingival fibroblasts (HGFs) under low-dose radiation conditions. </ItemContent> <ItemContent> Improved cellular proliferation: CO@SNPs promoted HGF proliferation and wound closure in vitro, demonstrating a synergistic effect when combined with low-dose gamma irradiation, achieving optimal healing outcomes. </ItemContent> <ItemContent> Biocompatibility and safety: CO and CO@SNPs exhibited a high cell viability with low cytotoxicity at effective concentrations, supporting their safe application in clinical settings. </ItemContent> <ItemContent> Implications for dentistry and beyond: These findings highlight the potential of CO@SNPs in nanodentistry and other biomedical applications for managing oral and tissue wounds efficiently. </ItemContent> </UnorderedList>

Synergistic effect of low-dose gamma radiation and/or clove oil loaded on silica nanoparticles on regeneration of human gingival fibroblast cell line: an in vitro study

期刊论文
出版模式：

开放获取
发表日期：

2025年9月18日

查看全文

Asmaa M. Abdeen,
Mustafa Gharib,
Ibrahim Y. Abdelrahman

摘要

In light of the growing use of natural products in medicinal therapy, this study evaluated the proliferative effects of clove oil (CO) in two forms—native and loaded onto silica nanoparticles (CO@SNPs)—on Human Gingival Fibroblasts (HGFs), both singly and in combination with low-dose gamma irradiation (0.25 Gy). Silica nanoparticles were chemically synthesized and loaded with clove oil, with thorough characterization conducted using dynamic light scattering (DLS) for particle size, zeta-potential measurements, Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). The EC50 values of CO and CO@SNPs were determined to be 1.88 µmol/mL and 0.172 µmol/mL, respectively, demonstrating a superior proliferative efficacy of the CO@SNPs nanoformulation. The biological effects were assessed using the MTT assay for cell viability, Trypan blue exclusion for cell count, qPCR for gene expression of c-Myc, MAPK, and NF-κB, and antioxidant capacity assays (MDA and catalase activity). Moreover, the in vitro wound-healing scratch assay showed enhanced fibroblast migration and proliferation, with relative wound density reaching 48% and 62% for CO@SNPs alone and combined with low-dose gamma irradiation, respectively. Further analysis through immunofluorescence staining of TGF-β and hematoxylin staining confirmed cell proliferation. Our results showed that loading CO on silica nanoparticles (CO@SNPs) were significantly more efficient than native clove oil in promoting cell proliferation and wound healing, with the combined treatment of CO@SNPs and low-dose gamma irradiation producing the most pronounced regenerative effect. These findings suggest that CO@SNPs, especially when combined with low-dose gamma irradiation, hold significant promise for enhancing oral tissue regeneration.

Graphical abstract

Nanotechnology-enhanced clove oil delivery: The present study successfully encapsulated clove oil (CO) in silica nanoparticles (CO@SNPs), which improved its stability and controlled release and provided its therapeutic efficacy.

Antioxidant and wound-healing properties: CO@SNPs significantly reduced oxidative stress markers (e.g., malondialdehyde levels decreased by 93%) and enhanced antioxidant enzyme activity (catalase activity increased by 83%) in human gingival fibroblasts (HGFs) under low-dose radiation conditions.

Improved cellular proliferation: CO@SNPs promoted HGF proliferation and wound closure in vitro, demonstrating a synergistic effect when combined with low-dose gamma irradiation, achieving optimal healing outcomes.

Biocompatibility and safety: CO and CO@SNPs exhibited a high cell viability with low cytotoxicity at effective concentrations, supporting their safe application in clinical settings.

Implications for dentistry and beyond: These findings highlight the potential of CO@SNPs in nanodentistry and other biomedical applications for managing oral and tissue wounds efficiently.