Non-Thermal Atmospheric Pressure Plasma-Conditioned Root Dentin Promotes Attraction and Attachment of Primary Human Dental Pulp Stem Cells in Real-Time Ex Vivo

Yeon-Jee Yoo,Seung-Ho Baek,Min-Ji Kang,Hiran Perinpanayagam,Kee-Yeon Kum,Joo-Cheol Park

doi:10.3390/app11156836

Abstract

This study investigated if non-thermal atmospheric pressure plasma (NTAPP) treatment of root dentin surfaces promotes human dental pulp stem cell (hDPSCs) adhesion. Freshly extracted human single-rooted teeth (n = 36) were decoronated and cut (first vertically, then horizontally) into root dentin slices (3 mm thick). Primary hDPSCs cultures were seeded onto slices randomly assigned to pretreatment groups (n = 9/group): NaOCl (1.5%), EDTA (17%) then NTAPP (Group I); NaOCl then NTAPP (Group II); NaOCl then EDTA (Group III); and NaOCl alone (Group IV). Cell viability and proliferation were measured using MTT assay with log-linear statistical analysis. Cell attachment and spreading morphologies on dentin slices (n = 3/group) were examined through scanning electron microscopy. Early cell adhesion events and subcellular activities were observed in real time by live-cell imaging through holotomographic microscopy. Cell viability and proliferation were significantly higher on NTAPP-treated dentin (p < 0.05), without interactions with EDTA (p > 0.05). The attachment, spreading, extensions and multiple layers of hDPSCs were heightened on NTAPP-treated dentin. Cell adhesion, spreading, and dentinal tubule penetration were hastened on NTAPP-treated dentin surfaces in real-time, with elevated subcellular activities and intracellular lipid droplet formation. NTAPP-treated root dentin surfaces support enhanced cellular responses, potentially promoting pulp-dentin regeneration.

Highlights

The purpose of this study was to investigate the effects of Non-Thermal Atmospheric Pressure Plasma (NTAPP) treatment of human root dentin surfaces in the recruitment, attachment and growth of human dental pulp stem cells (hDPSCs)
Dentin slices were randomly assigned (n = 9/group) for conditioning as follows: Group I, NaOCl (1.5%, 10 mL/5 min), ethylenediaminetetraacetic acid (EDTA) (17%, 10 mL/5 min), followed by NTAPP; Group II, NaOCl, NTAPP; Group III, NaOCl
Dentin slices were randomly assigned (n = 9/group) for conditioning as follows: Group I, NaOCl (1.5%, 10 mL/5 min), EDTA (17%, 10 mL/5 min), followed by NTAPP; Group II, NaOCl, NTAPP; Group III, NaOCl, EDTA; Group IV, NaOCl alone

Summary

Introduction

Non-thermal atmospheric pressure plasma (NTAPP) involves partially ionized gas containing electrically charged particles at atmospheric pressure. These include reactive oxygen and nitrogen species that induce various physiological effects, with promising clinical and preclinical applications in plasma medicine. Recent developments include direct and indirect applications of NTAPP for microbial biofilm eradication, tissue disinfection, wound healing, tissue rejuvenation, and surface modification of biological scaffolds. NTAPP has been studied for periodontal pocket decontamination, adhesion, caries treatment, root canal disinfection, implant surface treatment, and tooth bleaching [1,2]

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