The beneficial effect of cold atmospheric plasma on parameters of molecules and cell function involved in wound healing in human osteoblast-like cells in vitro

Abstract

The aim of this study was to analyse the effect of cold atmospheric plasma (CAP) on human osteoblast-like cells in vitro. Additionally, underlying intracellular mechanisms were to be studied. Human osteoblast-like (MG63) cells were exposed to CAP for 60 s. The effects of CAP on key molecules essential for the wound healing response were studied using real-time PCR, ELISA and immunocytochemistry. For studying intracellular signalling pathways, MAP kinase MEK 1/2 was blocked. Cell viability was analysed by an XTT assay and with an EVE automated cell counter. Cell migration was examined by an in vitro wound healing assay. CAP exposition on osteoblast-like cells caused a significant upregulation of interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor (TNF)α, cyclooxygenase (COX)2, collagen (COL) 1α, matrix metalloproteinase (MMP)1, Ki67, proliferating-cellnuclear-antigen (PCNA) and chemokine ligand (CCL)2 mRNA expression at 1 day. Interestingly, after blocking of MAP kinase, CAP-induced upregulation of Ki67 was inhibited by 57%. Moreover, CAP treatment improved significantly osteoblastlike cell viability as compared to untreated cells at 1 day. Beneficial effect of CAP treatment was shown by an in vitro wound healing assay, displaying a significant faster wound closure. Our findings provide evidence that CAP exposure effects gene and protein regulation in human osteoblast-like cells. Furthermore, CAP treatment has a positive impact on wound closure in an in vitro setting and might improve existing concepts of hard tissue regeneration in the future.

Inhibiting proliferation and metastasis of osteosarcoma cells by downregulation of long non‑coding RNA colon cancer‑associated transcript 2 targeting microRNA‑143

“…The cells were cultured with RPMI-1640 medium with 10% FBS ( 14 , 15 ). A straight line was then scratched on the monolayer surface (confluence of 70–80%) using a pipette tip ( 16 ). After treatment, the wells were washed twice to remove the dead cells from the medium.…”

“…Osteosarcoma is a malignant bone tumor that accounts for 1-2% of all cancer cases 012-2016) (1). Although surgical and chemotherapy approaches for osteosarcoma have been greatly improved, the 5-year survival rate (2009-2015) was only 67% (birth to age 14) and ~ 69% (ages [15][16][17][18][19] (1). Hence, it is essential to determine novel and effective therapeutic targets and prognostic biomarkers for osteosarcoma.…”

Anticancer Effects of Cold Atmospheric Plasma in Canine Osteosarcoma Cells

“…Applying CAP on the surgical site after removing the tumor could not only kill the residual cancer cells in situ, but also suppress potential metastasis, and could activate an immune response to the residual cancer cells, considering that tumor recurrence is reported in 15–25% of the patients [ 34 ]. Furthermore, CAP has a beneficial effect on wound healing [ 42 , 43 , 44 ]. We assume that applying CAP on surgical sites could accelerate wound closure.…”

Fighting Mixed-Species Microbial Biofilms With Cold Atmospheric Plasma

“…The "cocktail" constituents of plasma, including molecules and neutral atoms, charged particles, metastable radicals, and photons, play synergistic functions in microbial inactivation (Guo et al, 2018). In recent years, cold atmospheric plasma (CAP) has been extensively investigated for its potential as an alternative treatment in wound healing, dental cure, oncological therapy, and food decontamination (Scholtz et al, 2015;Jiao et al, 2019;Eggers et al, 2020). Significant progress has been made in controlling mixed-species biofilms with CAP, which is considered a rapid, environmentally friendly, energy saving, and versatile antimicrobial technology (Šimončicová et al, 2019).…”