Effect of Cold Atmospheric Plasma (CAP) on Osteogenic Differentiation Potential of Human Osteoblasts.

Benedikt Eggers,Franz-Josef Kramer,Susanne Jung,Anna-Maria Wagenheim,Sonja Sielker,Johannes Kleinheinz,Marjan Nokhbehsaim

doi:10.3390/ijms23052503

Abstract

Bone regeneration after oral and maxillofacial surgery is a long-term process, which involves various mechanisms. Recently, cold atmospheric plasma (CAP) has become known to accelerate wound healing and have an antimicrobial effect. Since the use of CAP in dentistry is not yet established, the aim of the present study was to investigate the effect of CAP on human calvaria osteoblasts (HCO). HCO were treated with CAP for different durations of time and distances to the cells. Cell proliferation was determined by MTT assay and cell toxicity by LDH assay. Additionally, RT-qPCR was used to investigate effects on osteogenic markers, such as alkaline phosphatase (ALP), bone morphogenic protein (BMP)2, collagen (COL)1A1, osteonectin (SPARC), osteoprotegerin (OPG), osterix (OSX), receptor activator of NF-κB (RANK), RANK Ligand (RANKL), and Runt-related transcription factor (RUNX)2. There were small differences in cell proliferation and LDH release regarding treatment duration and distance to the cells. However, an increase in the expression of RANK and RANKL was observed at longer treatment times. Additionally, CAP caused a significant increase in mRNA expression of genes relevant to osteogenesis. In conclusion, CAP has a stimulating effect on osteoblasts and may thus represent a potential therapeutic approach in the regeneration of hard tissue defects.

Highlights

After oral and maxillofacial surgery, the healing of hard tissue is a much more complex and long-term process as compared to soft tissue
The process is conducted by genes such as bone morphogenic protein 2 (BMP2), Runt-related transcription factor 2 (RUNX2), osterix (OSX), and osteoprotegerin (OPG), and followed by markers that appeared later in osteogenic and mineralization cascade, as alkaline phosphatase (ALP), osteonectin (SPARC), and collagen 1 (COL1A1) [4,5,6,7,8,9,10,11]
Since possible clinical use requires precise understanding of its effect on the hard tissue cells of the skull and, must exclude damaging effects, the aim of the present study is to investigate the effects of cold atmospheric pressure plasma (CAP) on human calvaria osteoblasts (HCO)

Summary

Introduction

After oral and maxillofacial surgery, the healing of hard tissue is a much more complex and long-term process as compared to soft tissue. The regeneration of bone is extensive and involves many different mechanisms, such as the differentiation of osteoblasts or the formation of collagen fibres and its arrangement according to the mechanical forces [1,2,3]. Bone is highly vascularised and permanently remodels itself in human life [12,13]. Systemic factors such as increased age, malnutrition, or metabolic syndrome can negatively influence bone regeneration [14,15,16]. Cold atmospheric pressure plasma (CAP) has become known to improve wound healing [17,18]. Different technologies are known for medical applications, such as dielectric barrier discharge (DBD) devices, plasma jets, or hybrid devices [21]

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