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Copper Doped Carbon Dots for Addressing

Bacterial Biofilm Formation, Wound Infection,
and Tooth Staining
Meng Liu, Ling Huang, Xingyi Xu, Xiaoming Wei, Xianfeng Yang, Xiaolei Li, Bingnan Wang, Yue Xu,*
Lihua Li,* and Zhongmin Yang*
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ABSTRACT: Oral infectious diseases and tooth staining, the main challenges of
dental healthcare, are inextricably linked to microbial colonization and the formation
of pathogenic biofilms. However, dentistry has so far still lacked simple, safe, and
universal prophylactic options and therapy. Here, we report copper-doped carbon
dots (Cu-CDs) that display enhanced catalytic (catalase-like, peroxidase-like) activity
in the oral environment for inhibiting initial bacteria (Streptococcus mutans) adhesion
and for subsequent biofilm eradication without impacting the surrounding oral tissues
Figure 9. Analysis of nondestructive teeth whitening. (A) SEM pictures of tooth surface enamel and cross-sectional dentin treated with CDs/
via oxygen (O2) and reactive oxygen species (ROS) generation. Especially, Cu-CDs Cu-CDs + 3% H2O2 and 30% H2O2 after 3 h of bleaching. Origin group without any treatment. Scale bar = 500 μm. (B) Vickers hardness of
exhibit strong affinity for lipopolysaccharides (LPS) and peptidoglycans (PGN), thus tooth samples before and after 3 h of bleaching. (C) Microhardness loss of tooth samples after bleaching for 3 h. (D) Residual pH values of
conferring them with excellent antibacterial ability against Gram-positive bacteria the used bleaching solutions after 1 h of bleaching. (E) Variation of the pH value of bleaching solutions composed of different H2O2
(Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli), such that they concentrations and CDs/Cu-CDs after bleaching for 1 h. Significance analysis was carried out by t test. *p < 0.05, **p < 0.01, ***p < 0.001,
can prevent wound purulent infection and promoting rapid wound healing. n = 5. All groups were compared to the H2O2 group.
Additionally, the Cu-CDs/H2O2 system shows a better performance in tooth
whitening, compared with results obtained with other alternatives, e.g., CDs and clinically used H2O2, particularly its 0.05) than the H2O2 group and CD + H2O2 group, indicating a we observed that the Cu-CD + H2O2 group achieved the best
negligible enamel and dentin destruction. It is anticipated that the biocompatible Cu-CDs presented in this work are a better external teeth whitening effect (Figure S23C,D). whiteness at 5 h, which was significantly stronger than the
promising nano-mouthwash for eliminating oral pathogenic biofilms, prompting wound healing as well as tooth whitening, Intrinsic staining mainly manifested as dentin staining, which H2O2 group and the CD + H2O2 group (p < 0.001). The final
highlighting their significance in oral health management. cannot be removed by external brushing and needs to be ΔE of the crowns in the Cu-CD + H2O2 group was calculated
KEYWORDS: oral diseases, copper doped carbon dots (Cu-CDs), enzymatic activity, antibacterial, tooth whitening removed by chemical bleaching. The tooth internal bleaching to be about 48.396, whereas that of the H2O2 group was only
effect of Cu-CDs was tested with rhodamine B (RhB) stained 30.222 (Figure S24C). The above experimental results indicate
tooth models.36 The whitening procedure is shown in Figure that the Cu-CD + H2O2 system can enable powerful tooth
INTRODUCTION and are highly acidogenic or acid-proof,16 making them hard to 8A. According to the photographs, it is found that the fading whitening. Moreover, the degradation of RhB in vitro using
remove, thereby turning into dental plaque.17 The glyco- degree of the Cu-CD + H2O2 group in the first hour was different concentrations of Cu-CDs and CDs (30−240 μg·
Oral diseases are major global public health problems affecting
metabolism activity of such a high density of acid-producing
clearly much stronger than that of the H2O2 group and CD + mL−1) was studied. As expected, the maximum absorption
more than 3.5 billion people, and greatly reduced quality of H2O2 group (Figure 8B). Under the influence of Cu-CD + peak of RhB (degraded by Cu-CDs + H2O2) at 550 nm
life, especially in many low- and middle-income countries.1−5 bacteria clustered within the microbial ecological network of
H2O2 bleaching solution, the inner color of teeth continued to continued to decrease with concentration increased after 1 h,
The most prevalent oral threats are infectious diseases caused plaque produces a highly acidic microenvironment (pH < 5.5),
lighten with the increase of bleaching time, and a satisfactory while the CD + H2O2 group exhibited negligible effects on
by biofilms, such as tooth decay (dental caries), pulp infection, which ensures continuous demineralization of adjacent tooth tooth color could be achieved by the fifth hour. Because of the RhB degradation (Figure S25A). Particularly, as the reaction
periodontitis, peri-implantitis, induced wound infection, and enamel and periodontal tissue damage.8,18,19 Meanwhile, structural differences of the tooth profile resulting in different temperature increased from 25 to 37 °C, the faster degradation
tooth staining.6−8 Therefore, it is urgently needed to develop dormant bacteria within the biofilm can cause the recurrence degrees of discoloration, the ΔE of the outermost enamel, rate of RhB in each group resulted in less residual RhB content
efficient and inexpensive multifunctional preventive and of localized chronic infections.20 Moreover, exogenous tooth outer dentin, and inner dentin were counted separately. Figure been observed (Figure S25B), standing for the oral environ-
treatment methods. staining is usually attributed to stains in various drinks and 8C shows that the ΔE of the outermost enamel of Cu-CDs + ment being more suitable for Cu-CD’s performing its
The oral cavity of humans, as a bacterial environment, is foods that are absorbed on the sticky substances on the surface H2O2 within 1 h was about 4 times that of the other two whitening function.
perched by hundreds of different bacterial species,9,10 some of of the biofilms to form plaque and tartar, and then the groups and, at subsequent time points, the gap gradually The most commonly used peroxide bleaching agent in
which form oral pathogenic biofilms through irreversible nonenzymatic browning between carbohydrates and proteins narrowed, evidencing the short-term bleaching function of Cu- clinical practice is mainly 30% H2O2, and the strong oxidizing
attachment, reproduction, differentiation and secretion of CDs + H2O2 on slight tooth enamel staining. For deeper property can easily cause irreversible irritation to the oral soft
highly structured water-soluble polysaccharides, such as Received: March 13, 2022 dentin staining, the ΔE brought by Cu-CDs + H2O2 was and hard tissues.87 The corroded tooth surface is also more
exopolysaccharides (EPS).11−14 The microanaerobic environ- Accepted: June 10, 2022 significantly higher than the other groups at every point in time likely to cause biofilm growth and recoloring because of
ment is more beneficial to the growth and reproduction of (p < 0.05), whereas the CD + H2O2 and H2O2 groups almost roughness. As a result, to verify the oral adaptability and
bacteria upon biofilm.15 The embedded resident microbial maintained a low level steadily (Figure 8D,E). To evaluate the nondestructive of the enzyme-powered whitening of Cu-CDs,
communities, e.g., Streptococcus mutans (S. mutans), encased changes in crown appearance, a simulation model of SEM was first applied to reflect the bleached tooth tissue
intramedullary bleaching was developed that was consistent (represented by tooth enamel and dentin) (Figure 9A). It was
and protected in the matrix are resistant to general antibiotics
with clinical practice (Figure S24A). As shown in Figure S24B, worth noting that the enamel surface after 3 h bleaching with
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