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Biological and Medical Applications of Materials and Interfaces

A Polysaccharide-Based Antibacterial Coating with
Improved Durability for Clear Overlay Appliances
Sohyeon Park, Hyun-hye Kim, Seok Bin Yang, Ji-Hoi Moon, Hyo-Won Ahn, and Jinkee Hong
ACS Appl. Mater. Interfaces, Just Accepted Manuscript • DOI: 10.1021/acsami.8b04433 • Publication Date (Web): 04 May 2018
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Article
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13 A Polysaccharide-Based Antibacterial Coating with
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17 Improved Durability for Clear Overlay Appliances
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22 Sohyeon Park†, Hyun-hye Kim‡, Seok Bin Yang§, Ji-Hoi Moon § , Hyo-Won Ahn* ⊥ , Jinkee
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25 Hong*†
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29 Department of Chemical and Biomolecular Engineering, Yonsei University, 50 Yonsei-ro,
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31 Seodaemun-gu, Seoul, Republic of Korea
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Graduate Student, Department of Dentistry, Graduate School, Kyung Hee University, Seoul,
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37 Korea, Republic of Korea
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40 Department of Maxillofacial Biomedical Engineering, School of Dentistry, Kyung Hee
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42 University, Seoul, Republic of Korea
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Department of Life and Nanopharmaceutical Sciences, Kyung Hee University, Seoul,
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Republic of Korea
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51 Department of Orthodontics, Kyung Hee University School of Dentistry, Seoul, Republic of
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3 ABSTRACT
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Clear overlay appliances (COAs) are widely used in orthodontic fields because they offer many
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9 advantages, such as cost-effectiveness, good formability, and good optical characteristics.
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11 However, it is necessary to frequently replace COAs because the thermoplastic polymers that
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13 are used to fabricate COAs have poor abrasion resistance and tend to induce bacterial
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16 accumulation. Here, we developed polysaccharide-based antibacterial multilayer films with
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18 enhanced durability, intended for COA applications. First, multilayer films composed of
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20 carboxymethylcellulose (CMC) and chitosan (CHI) were fabricated on polyethylene
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terephthalate glycol-modified (PETG), which was preferred material for COA fabrication, via
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25 a layer-by-layer (LbL) technique. Next, chemical crosslinking was introduced within the LbL-
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27 assembled multilayer films. The LbL-assembled CMC/CHI film, which was made porous and
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rough by the crosslinking, formed a super-hydrophilic surface to prevent the adhesion of
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32 bacteria and exhibited a bacterial reduction ratio of approximately 75%. Furthermore, the
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34 crosslinking of the multilayer film coated on the PETG also improved the chemical resistance
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36 and mechanical stability of the PETG under simulated intraoral conditions with artificial saliva,
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39 by increasing the bond strength between the polysaccharide chains. We attempted to
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41 accumulate datasets using our experimental design and to develop sophisticated methods to
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44 assess nanoscale changes through large-scale measurements. 소현
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51 KEYWORDS: clear overlay appliances, polysaccharide, layer by layer assembly, nano-film,
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53 antibacterial coating, durability
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6  INTRODUCTION
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8 Aesthetic factors are significant in the field of dentistry; this has led to the universalization
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10 of invisible removable appliances. For example, clear overlay appliances (COAs), such as clear
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aligners or retainers, are widely used in daily orthodontic clinics for dynamic orthodontic
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15 treatment and maintenance.1-4 The COAs are fabricated by imprinting thermoplastic sheets on
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17 the dental cast model through a heating cycle combined with either vacuum or pressure forming.
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Several thermoplastics, such as polyethylene, polycarbonate, polypropylene, and polyethylene
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22 terephthalate glycol-modified (PETG), are currently used to prepare COAs.5-7 However, these
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24 materials inevitably induce bacterial accumulation which is strong risk factor for the
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26 development of bacteria-related disease including dental caries and periodontal disease, and
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29 possess mechanical limitations, such as poor abrasion and corrosion resistance.8-9
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31 Consequently, it is necessary to frequently replace COAs, which causes the patient economic
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33 burden.7, 10
Thus, to overcome these limitations, several approaches have been attempted.
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Among them, increasing attention has recently focused on a coating technique that uses nano-
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38 films because it offers site-specific application and can be used to produce COAs with little
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40 increase in thickness. To maintain minimal thickness of COAs is critical for the function of
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temporomandibular joints and patients’ comfort.
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46 Recently, urushiol-based coatings have been developed in our laboratory, to enhance the
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48 mechanical strength and antibacterial effects of such materials. In addition, we fabricated
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50 organosilicate-based super-hydrophilic nano-films with enhanced durability for dental
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53 applications.11-12 However, coatings should ideally be prepared using easily obtainable
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55 materials and simple methods without employing a complicated synthesis process.
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57 Furthermore, rigorous testing and analysis with regard to the chemical and physical stabilities
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3 of such materials are required because the conditions in the oral cavity are very complicated
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6 owing to the various enzymes present, its variable pH and physical stimulation.
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9 Here, we developed biocompatible polysaccharide-based antibacterial films, optimized for
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11 oral conditions. Specifically, we fabricated multilayer films composed of chitosan (CHI) and
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carboxymethylcellulose (CMC) on PETG, which is the most commonly used material for
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16 COAs, using a layer-by-layer (LbL) technique; we also introduced a crosslinking technique to
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18 the LbL-assembled multilayer film. Both CHI and CMC are linear polysaccharides, which are
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highly compatible with each other owing to their similar chemical structures. CHI possesses
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23 rich amine groups that exert an antibacterial effect, and each molecule of CMC can form
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25 hydrogen bonds with six water molecules.13 Owing to these features, they are considered
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27 suitable materials for the production of super-hydrophilic coatings that exert an antibacterial
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30 effect.
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33 Bacteria often adhere to the surface of in-dwelling medical devices and form a complex
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35 microbial community called the "biofilm". Because biofilms have a strong tendency to adhere
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to surfaces and increase resistance to antibiotics and host immune systems, it is extremely
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40 significant to prevent their initial adhesion in medical devices associated with bacterial
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42 infection.14-15 The super-hydrophilic coating on medical devices inhibits biofilm adhesion by
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forming a thin water film which interferes with the interaction between the surface of the
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47 medical device and the biofilm.11, 16-19
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50 The LbL technique involves the alternating deposition of oppositely charged polyelectrolytes
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52 via electrostatic interaction; it can be applied to various substrates regardless of their shape and
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55 size.20-26 In addition, the crosslinking of such LbL-assembled multilayer films not only results
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57 in the production of a porous structure with a super-hydrophilic surface but also improves the
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3 chemical and physical stability of the coating by enhancing the strength of the bonds between
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6 the polysaccharide chains.27-28
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9 Therefore, the objective of this study is to investigate the antibacterial effect of the PETG
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11 coated with the crosslinked polysaccharide multilayer film, and assess the chemical resistance
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and mechanical properties of the coating under simulated intraoral conditions with artificial
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 RESULTS AND DISCUSSION
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24 Preparation of polysaccharide-based coating via LbL assembly and crosslinking.
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26 Both CMC and CHI are linear polysaccharides. Specifically, CMC is the sodium salt of a
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28 cellulose derivative, in which carboxymethyl groups (-CH2-COOH) are bound to some of the
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31 hydroxyl groups of the glucopyranose monomers that form the cellulose backbone. CMC is
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33 negatively charged in aqueous solutions because its -COOH groups are ionized to -COO-. CHI
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35 is composed of randomly distributed β-(1→4)-linked D-glucosamine (deacetylated unit) and
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N-acetyl-D-glucosamine (acetylated unit). CHI is positively charged in aqueous solutions
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40 because its NH2 groups are ionized to NH3+.28-29
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42 The CMC and CHI solutions were prepared with a pH of 4 because more than 80% of both
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these polysaccharides are ionized at this pH.13 Specifically, the driving force in this
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47 polysaccharide-based LbL assembly is considered to be owed to the electrostatic interaction
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49 between these materials. Prior to the LbL assembly, the oxygen plasma-treated substrates (Si
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51 wafer and PETG) were incubated in a branched polyethylene imine (BPEI) solution. BPEI is a
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54 polymer that is frequently used as a material for surface modification because it has inherently
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56 high adhesion and many amine groups. Thus, hydrogen bonding and electrostatic interaction
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58 are formed between the hydroxy groups of the oxygen plasma treated substrate and the amine
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3 groups of BPEI, thereby achieving surface modification for LbL deposition. BPEI also acts as
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6 a cushion layer by providing functional groups on both the substrate and the CMC due to the
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8 flexibility due to the branched structure.30-32 The surface modification allows the
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10 polysaccharide multilayer film to be robustly formed on the substrate surface. 33-34 In the case
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of the surface-amine modified substrates by BPEI, the LbL assembly commences with the
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15 deposition, via electrostatic interaction, of a negatively charged CMC layer, followed by a
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17 positively charged CHI layer. To demonstrate the successful assembly of (CMC/CHI)n
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nanofilms, the thicknesses of the LbL-assembled multilayered films on substrates (n = 5) were
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22 measured using a profilometer. Three sites were randomly selected for each substrate and the
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24 thickness was measured twice. By repeating these alternating depositions, the polysaccharide
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26 multilayer films are obtained. The thickness growth curve in figure 1 shows the successful
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29 deposition process of polysaccharides by the result that the multilayer film thickens with
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31 increasing the number of deposition. The multilayer films grow relatively linearly without any
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33 distinct irregularities, and the final thickness of the (CMC/CHI)20 film was approximately
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1,818 nm. According to our previous paper, the (CMC/CHI)10 films also had super-hydrophilic
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38 properties, and thus we believed that the (CMC/CHI)10 films also exhibit anti-biofilm adhesion
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40 effects. However, we concluded that thicker thickness than 10 bilayer films would be more
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effective, to improve the mechanical strength for practical use. Moreover, the film with a
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45 thickness more than 2μm is not suitable for coating because it can easily be peeled off from the
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47 substrate. Therefore, we used the (CMC/CHI)20 film with thickness of 1.5μm - 2μm in this
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22 Figure 1. Curve showing the increase in the thickness of the CMC/CHI multilayer film (a);
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24 FT-IR spectra of CMC (black), CHI (purple), non-crosslinked (CMC/CHI)20 film (green), and
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26 crosslinked (CMC/CHI)20 film (red) (b).
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31 Furthermore, Fourier transform-infrared (FT-IR) spectra shown in Figure 1b exhibited clear
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33 CMC/CHI film formation and crosslinking between multilayers. The spectra of CMC and CHI
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36 monotonous peaks due to the electrostatic interaction between CMC and CHI, rather than their
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38 respective singlets. Meanwhile, crosslinking of LbL assembled CMC/CHI film was performed
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using 1-Ethyl-3-(3-Dimethylaminopropyl) Carbodiimide hydrochloride (EDC)/N-
41 hydroxysulfosuccinimide (NHS) chemistry. First, EDC reacts with the carboxylate molecule,
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43 CMC, to form an unstable reactive o-acrylisourea ester. The formed ester reacts with NHS, and
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45 converts into a semi-stable amine-reactive NHS ester. Finally, the NHS ester promotes
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crosslinking between the polysaccharide chains by forming stable amide bonds with the amine
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48 groups of CHI. Glutaraldehyde was also used as a second crosslink agent, providing additional
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50 chemical bonds between the polysaccharide chains. The reactive end groups of glutaraldehyde
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52 allow the formation of covalent bonds between two polysaccharides containing hydroxyl
53 groups or primary amine groups.35-38 In the FT-IR spectra of the CMC/CHI film, there are
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55 several notable peaks, namely overlapping O–H and N–H peaks at around 3400 cm-1, a C–H
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57 bond peak at around 2800 cm-1, as well as C–O–H, C–O–C, and C–N peaks corresponding to
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the polysaccharide structure at around 1600 cm-1 and 1150 cm-1. CMC, CHI and CMC/CHI
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3 In the FT-IR spectra of the crosslinked CMC/CHI film, two distinctly separated peaks
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5 attributed to O=C, at around 1680 cm-1 and a N–H peak, at around 1645 cm-1 due to the amide
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7 bond formation, were observed.39-40 It is also possible to observe that the intensity of the C–N
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peak at around 1150 cm-1 was reduced. This corresponds to the formation of the C=N peak at
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12 also demonstrated successful chemical crosslinking using X-ray photoelectron spectroscopy
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14 (XPS) in our previous work.44
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Film morphology and wettability analysis.
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21 The crosslinking induces additional bonds and reassembly between the polysaccharide chains
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23 that form the multilayer film, transforming the dense structure of the multilayer film into a
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porous structure. The effect of crosslinking on the morphologies of the multilayer thin films
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28 was investigated via field-emission scanning electron microscope (FE-SEM) and atomic force
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30 microscopy (AFM) observation. First, the roughness of the films was measured by AFM.
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32 According to the results of AFM measurement in figure 2, the Rq values of non-crosslinked
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35 (CMC/CHI)20 film and crosslinked (CMC/CHI film)20 were 20.32 nm and 57.71 nm,
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37 respectively. The SEM images in Figure 3 also show the increased roughness and porosity of
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3 Figure 2. AFM images of the non-crosslinked (CMC/CHI)20 film (a) and the crosslinked
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6 (CMC/CHI)20 film (b), and roughness of the films measured using AFM.
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9 The Brunauer−Emmett−Teller (BET) analysis results included in our previous study,
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11 demonstrate that the total porosity increased after crosslinking.44 This surface morphology
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13 change is the result of the effect of PBS used in post-crosslinking washes as well as additional
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16 bond formation due to crosslinking. The salts of PBS used for washing of the EDC/NHS
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18 crosslinking agent were sandwiched between the multilayered films which have already been
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20 swollen by the wet cross-linking, thereby ensuring the area where the pores can be formed. A
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porous structure was then formed in the area of the PBS salt removed after several washes with
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25 distilled water. Such a change in the morphology also influences the wettability of the
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27 multilayer film. According to the Wenzel and Cassie-Baxter theory, rough surfaces with a high
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surface energy are super-hydrophilic, while rough surfaces with a low surface energy are super-
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32 hydrophobic.45-47 To compare the wettability of the films with changes in the surface
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34 morphology prior to and following crosslinking, statics water contact angles of non-crosslinked
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36 (CMC/CHI)10 film and crosslinked (CMC/CHI film)10 were measured. As expected, prior to
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39 crosslinking, the surface of the multilayer film exhibited hydrophilicity, with a contact angle
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41 of 35°; this was owed to the hydrophilic functional groups, namely –COOH and –NH2 of the
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43 CMC and CHI. However, following crosslinking, the multilayer film exhibited super-
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hydrophilicity, with a contact angle of less than 5° (Figure 3). It is unlikely that the surface
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48 energy has increased rapidly because of the crosslinking, since surface energy is a material-
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50 specific property. Therefore, the change in the contact angle signifies the increased surface
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roughness, which is owed to crosslinking.47
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27 of the (CMC/CHI)10 films (a) and crosslinked (CMC/CHI)10 films (b). Static water-contact-
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angle images of the (CMC/CHI)10 films (c), and crosslinked (CMC/CHI)10 films (d).
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35 Cell viability and antibacterial function test.
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37 Using the polysaccharides as coating material contributed to high cell viability of the coating.
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Figure 4 shows the viability of the C2C12 cells for the coating, which was normalized to the
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42 control cell group. The coating composed of biocompatible polysaccharides exhibited 98% cell
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44 viability and this result demonstrated the biocompatibility of the coating. Meanwhile, the
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46 antibacterial effect of the coating against S. mutans is shown in Figure 5. S. mutans is the most
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49 prominent acidogenic oral bacteria, which can cause white spot lesions (WSLs) or serious
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51 demineralization of the tooth enamel.15 The SEM images show that there are enormous
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53 biofilms in the bare PETG, whereas there are few biofilms in the coated PETG. In addition, the
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56 ATP luminescent signal of the coated PETG is approximately four times lower than that of the
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58 bare PETG, which indicates that the coated PETG exhibits low bacterial viability. The
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3 physiological conditions, super-hydrophilic surface forms a thin water film, which interferes
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Figure 4. Cell viability graph with regard to the crosslinked (CMC/CHI)20 films; data was
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3 Analysis of the chemical stability of the coatings.
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6 Figure 6 shows the normalized-thickness graphs of crosslinked (CMC/CHI)20 films (Group 1)
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8 and non-crosslinked (CMC/CHI)20 films (Group 2) under each condition. The thickness of the
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10 crosslinked (CMC/CHI)20 films (Group 1) was almost maintained, without significant change,
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under the artificial saliva, regardless of the pH level and enzymes present. In contrast, the
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15 thickness of the non-crosslinked (CMC/CHI)20 films (Group 2) significantly reduced with time.
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17 Besides, the stability of the coatings under the artificial saliva with the enzymes could not be
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measured after 96 h because the group 2 peeled off (Figure S3). Table 1 exhibits the changes
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22 in the film thickness for Groups 1 and 2 owing to crosslinking by repeated measures ANOVA
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24 for each artificial saliva condition. Significant differences were observed between Groups 1
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26 and 2, except for the case involving artificial saliva at a pH of 4.7 (Table 1). In the case of both
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29 Group 1 and 2 under the artificial saliva with a pH of 4.7, there was no significant reduction in
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31 thickness. This was because the pH 4.7 was similar to that of the environment in which the
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33 multilayer film was prepared. However, it could be observed that the non-crosslinked film
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(Group 2) degrades more severely under conditions where the pH is higher than 4.7. This is
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38 owed to the aggregation of CHI, which occurs as the pH increases, and the poor stability of the
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40 multilayer film prior to crosslinking.50 Table 2 shows the results of examining the effects of
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artificial saliva conditions on each group according to the cross-linking. The change in
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45 thickness was not significant in Group 1 with crosslinks in both time, acid, enzyme, or
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47 combined conditions. These results demonstrate that crosslinking of the films improved
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49 chemical stability under the simulated intraoral conditions. A Bonferroni post hoc test was
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52 performed for Group 2, which revealed significant differences in the film thickness changes
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54 according to the pH conditions (Table 3). The non-crosslinked (CMC/CHI)20 films (Group 2)
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significantly as the pH increased (Table 1, Figures 6a to c).
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29 Figure 6. Chemical resistance test results obtained for the nanofilms under the simulated
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31 intraoral conditions with artificial saliva. The normalized thickness (%) of the crosslinked
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33 (CMC/CHI)20 films (red line, group 1) and that of the non-crosslinked films (black line, group
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2) are indicated in each graph. (a) Artificial saliva with a pH of 4.7. (b) Artificial saliva with a
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38 pH of 6.7. (c) Artificial saliva with a pH of 8.7. (d) Artificial saliva with a pH 6.7, containing
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40 α-amylase. (e) Artificial saliva with a pH of 6.7, containing lysozyme.
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(* indicates a P-value of < 0.05, which represents statistical significance in a repeated measures
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3 Table 1. Change in the film thickness for Groups 1 and 2, with respect to the crosslinking,
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6 under each artificial saliva condition
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9 Time Crosslinking Time * crosslinking
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pH 4.7 0.1049 0.4958 0.2374
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14 pH pH 6.7 0.0002*** 0.0005*** 0.0012**
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16 pH 8.7 <0.0001*** <0.0001*** 0.0003***
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19 α-amylase <0.0001*** 0.0002*** <0.0001***
20 Enzymes
21 Lysozyme <0.0001*** 0.0015** 0.0005***
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The Repeated measures ANOVA was performed.
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*P-value <0.05, **P-value <0.01, ***P-value <0.001 for significance.
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32 Table 2. Comparison of the change of the film thickness with respect to the artificial saliva
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34 condition for each group.
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37 Time *
38 Time pH Time * pH Enzymes
enzymes
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40 Group 1
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42 (crosslinked 0.0836 0.0557 0.1397 0.3746 0.3938
43 films)
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45 Group 2
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47 (non- <0.0001*** <0.0001*** <0.0001*** 0.0561 0.0042**
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The repeated measures ANOVA was performed.
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53 *P-value < 0.05, **P-value < 0.01, ***P-value < 0.001 for significance.
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3 Table 3. Post hoc test for group 2, performed with respect to the pH.
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7 Time pH Time * pH
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9 pH 4.7 –
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Group 2 6.7
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(non- pH 4.7 - 8.7 <0.0001*** <0.0001*** <0.0001***
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14 crosslinked
15 films)
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pH 6.7 - 8.7 <0.0001*** 0.0015** 0.0333*
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18 Bonferroni post hoc test was performed.
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20 *P-value < 0.05, **P-value < 0.01, ***P-value < 0.001 for significance.
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25 2.5. Analysis of the mechanical properties of the coatings.
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28 1) Tensile strength test
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31 A significant difference in the mean maximum tensile load (P < 0.05, Table 4) was observed
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33 between coated PETG without thermoforming (Groups A-1) and bare PETG (Group A-2). The
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35 mean maximum tensile load was significantly higher for Group A-1 (292.47 N) than for Group
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38 A-2 (258.18 N). On the other hand, there was no significant difference between coated PETG
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40 with thermoforming (Groups B-1) and bare PETG with thermoforming (Group B-2) in the case
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42 of the final thermoforming process using a thermoforming machine. The two-way ANOVA
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45 revealed the interaction between the thermoforming process and the (CMC/CHI)20 films with
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47 regard to the maximum tensile load (P < 0.05, Table 5). It suggested that the thermoforming
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49 process offset the effect of increasing the tensile strength of the (CMC/CHI)20-coated PETG.
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3 Table 4. Mean maximum tensile loads of the PETG sheets of the four groups
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6 Group A Group B
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(Without thermoforming) (With thermoforming)
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9 Group A-1 Group B-1
Group A-2 Group B-2
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11 (Coated PETG†) (Bare PETG) (Coated PETG†) (Bare PETG)
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13 Maximum 292.47 ± 5.58 258.18 ± 24.02 100.01 ± 28.07 113.33 ± 9.61
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15 tensile load (N) (1.76) (7.60) (8.88) (3.04)
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18 P-value* 0.0145* 0.0685
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20 †,
21 Crosslinked (CMC/CHI)20-coated PETG.
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23 Values are presented as mean ± standard deviation (standard error).
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25 Mann Whitney U test* was performed.
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27 P-value <0.05 for significance.
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31 Table 5. Results of two-way ANOVA of the maximum tensile load
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34 Source F Value Pr>F
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36 Thermoforming 764.42 <.0001***
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38 (CMC/CHI)20 films 2.95 0.0942
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40 Thermoforming * (CMC/CHI)20 films 15.24 0.0004***
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Two-way ANOVA was performed.
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*P-value <0.05, **P-value <0.01, ***P-value <0.001 for significance.
45
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49 2) Fracture resistance test
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52 Figure S4 shows the superimposed load−elongation curves for a series of specimens with
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54 varying notch depths for Group A-1. Each area under the load−elongation curves represents
55
56
the normalized Wf and was plotted as Wf vs. L.
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Figure 7. Results of fracture-resistance test for the non-heat-treated samples (a) and heat-
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23 treated samples (b).
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26 Figure 7 indicates the linearity of Wf vs. L for the four groups. According to the curve-fitted
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28 equations, without the thermoforming process, the EWF value for group A-1 was 492.1 KJ/m2,
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31
which was greater than that for group A-2 (4.1 KJ/m2) (Figure 7a). Similarly, in case of the
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33 final thermoforming process using a thermoforming machine, the EWF value for group B-1
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35 was 570 KJ/m2, which was greater than that for group B-2 (101.6 KJ/m2) (Figure 7b). This
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37 means that the (CMC/CHI)20 nanofilms contributed to the increased resistance to crack
38
39
40 initiation and fracture, which is a critical factor for the long-term use of COAs.
41
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43 In this analysis, it was assumed that the increased mechanical properties of the crosslinked
44
45 (CMC/CHI)20 nanofilms were due to the following reasons. First, CMC and CHI are both
46
47
polysaccharides with a high molecular weight and good mechanical strength. And also, the
48
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50 crosslinked multilayer film has a porous structure that differs from that of the non-crosslinked
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52 multilayer film because the crosslinking process of the multilayer film results in additional
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54 bond formations and reassembly between the polysaccharide chains. Porous structure due to
55
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57 additional bond formation is more flexible than stiff conventional coatings because they swell
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3 in physiological condition. Accordingly, (CMC/CHI)20 nanofilms with a thickness of only
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6 1,818 nm can exhibit significantly increased mechanical strength.
7
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9 Thermoforming is a necessary process for manufacturing COAs. The COAs are fabricated by
10
11 imprinting thermoplastic sheets on the dental cast model through a heating cycle combined
12
13 under pressure or vacuum. However, the enhanced mechanical properties of (CMC/CHI)20-
14
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16 coated PETG were diminished after the thermoforming process. The thermal or pressure
17
18 treatment usually reinforces the bonding strength between film layers.12 However, the
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20 imprinting process on the model causes significant morphological changes in the sheets, which
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23
could have a detrimental effect on the mechanical and structural properties of the nanofilms.
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25 To evaluate the effect of thermoforming on the surface morphology of the crosslinked
26
27 (CMC/CHI)20-coated PETG, FE-SEM images were obtained at 226× and 5,000× magnification
28
29
before and after thermoforming at an acceleration voltage of 5 kV (Figure S5). As shown in
30
31
32 Figures S4A and B, the crosslinked (CMC/CHI)20 multilayer film had an equally porous
33
34 structure because the crosslinking of the multilayer film resulted in additional bond formation
35
36 and reassembly between the polysaccharide chains. On the other hand, Figures S4C and D
37
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39 show that, after the thermoforming process, the (CMC/CHI)20 film had a collapsed porous
40
41 structure with several cracks.
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43 In a previous study, the thermoforming process did not cause significant surface changes in
44
45
46
the thermoplastic COAs. The thermoforming only led to a slightly smoother surface; no
47
48 crystallization of the PETG sheets occurred.7 Accordingly, the thermoforming process had a
49
50 considerable effect not on the thermoplastic polymer sheet but on the surface of the
51
52
(CMC/CHI)20 films. Therefore, we concluded that nanofilm fabrication should be preceded by
53
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55 the thermoforming process in clinical applications.
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4 ■ CONCLUSION
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7 In this study, we developed a durable antibacterial coating, which was optimized for clear
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9 overlay appliances (COAs) intended for intraoral applications. This coating was produced by
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11 fabricating a multilayer film composed of CMC and CHI on the surface of the COA material,
12
13
14 via LbL assembly and the introduction of crosslinking techniques. The polysaccharide, as a
15
16 multilayer film material, enhanced the biocompatibility of the coating, and the introduction of
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18 crosslinking contributed to the formation of a super-hydrophilic coating, which inhibited
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20
21 bacterial adhesion by altering the morphology of the multilayer film. In addition, the coating
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23 that was strengthened via crosslinking remained very stable without any degradation or
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25 detachment occurring under the various simulated intraoral conditions, which involved
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27
28
artificial saliva, various enzymes, and a range of acidity conditions. Furthermore, the coated
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30 COA material exhibited improved tensile strength and fracture resistance compared with those
31
32 of the bare COA material. The effect regarding the improvement in the mechanical properties
33
34 decreased when the material underwent a thermoforming process. Therefore, it is
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37 recommended that during clinical applications, the nanofilm is introduced following the
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39 thermoforming process. In conclusion, this study offers a practical approach for the
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41 development of multifunctional nano-coatings intended for dental materials.
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49
■ ASSOCIATED CONTENT
50
51
52
53 Supporting information
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55
56 Chemical structure of CMC and chitosan, Experimental section, Appearance of each film
57
58 following the stability tests, Design of the PETG sheets used for mechanical testing,
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3 load−elongation curve, FE-SEM images of crosslinked (CMC/CHI)20-coated PETG before and
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6 after thermoforming, Pictures of Layer-by-layer assembly process using PETG as substrate and
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8 completed coated PETG. (PDF)
9
10
11
12 ■ AUTHOR INFORMATION
13
14
15 Corresponding Authors
16
17 *E-mail: hyowon@khu.ac.kr
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19 *E-mail: jinkee.hong@yonsei.ac.kr
20
21 ORCID
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23 Jinkee Hong: 0000-0003-3243-8536
24
25 Notes
26
27 The authors declare no competing financial interest.
28
29
30
31 ■ ACKNOWLEDGMENT
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33
34 This research was supported by a grant of the Korea Health Technology R&D Project through
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36 the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health
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38
39 & Welfare, Republic of Korea (Grants HI14C3266) and also supported by the Basic Science
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41 Research Program through the National Research Foundation of Korea (NRF) funded by the
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43 Ministry of Science and ICT (NRF-2017R1E1A1A01074343).
44
45
46
47 ■ ABBREVIATIONS
48
49 COA, clear overlay appliance
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51 PETG, polyethylene terephthalate glycol
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53 LbL, layer-by-layer assembly
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55 CMC, carboxymethyl cellulose
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57 CHI, chitosan
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59 FE-SEM, field-emission scanning electron microscope
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3 Wf, work of fracture
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5 EWF, essential work of fracture
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10 ■ REFERENCES
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