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The ongoing coronavirus disease-19 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has drastically changed our way of life and continues to have an unmitigated socioeconomic impact across the globe. Research into potential vaccine design and production is focused on the spike (S) protein of the virus, which is critical for virus entry into host cells. Yet, whether the degree of glycosylation in the S protein is associated with vaccine efficacy remains unclear. Here, we first optimized the expression of the S protein in mammalian cells. While we found no significant discrepancy in purity, homogeneity, or receptor binding ability among S proteins derived from 293F cells (referred to as 293F S-2P), 293S GnTI- cells (defective in N-acetylglucosaminyl transferase I enzyme; 293S S-2P), or TN-5B1-4 insect cells (Bac S-2P), there was significant variation in the glycosylation patterns and thermal stability of the proteins. Compared with the partially glycosylated 293S S-2P or Bac S-2P, the fully glycosylated 293F S-2P exhibited higher binding reactivity to convalescent sera. In addition, 293F S-2P induced higher IgG and neutralizing antibody titres than 293S or Bac S-2P in mice. Furthermore, a prime-boost-boost regimen, using a combined immunization of S-2P proteins with various degrees of glycosylation, elicited a more robust neutralizing antibody response than a single S-2P alone. Collectively, this study provides insight into ways to design a more effective SARS-CoV-2 immunogen.

Titanium (Ti) and Ti-based alloy materials are ideal brackets that restore bone defect, and the mechanism of related genes inducing bone mesenchymal stem cells (BMSCs) to osteogenic differentiation is currently a hot research topic. In order to screen key genes of BMSCs during the osteogenic expression process, we acquired data sets (GSE37237 and GSE84500) which were in the database Gene Expression Omnibus (GEO). Investigations on differentially expressed genes (DEGs) and their enrichment of functions were conducted. We constructed relative protein-protein interaction (PPI) network by using Search Tool for the Retrieval of Interacting Genes (STRING) and visualized the expression of DEGs with Cytoscape. A total of 279 DEGs were discerned, which could be divided into 177 down regulated genes and 102 up regulated genes. In addition, the DEGs' enrichment and pathways included regulation of actin cytoskeleton, inflammatory mediator regulation of transient receptor potential (TRP) channels, peroxisome proliferator-activated receptors (PPAR) pathway, cell cycle, Rheumatoid arthritis, mitogen-activated protein kinases (MAPK) signaling pathway and Ras signaling pathway ect. It showed that 10 notable up regulated genes were mainly in AMP-activated protein kinase (AMPK) pathway. Then we used a technology named surface mechanical attrition treatment (SMAT) to prepare gradient nanostructured (GNS) surface Ti and seeded well-growing BMSCs on the surface of SMAT Ti and native pure Ti. Cell Counting Kits-8 (CCK-8), apoptosis experiment, immunofluorescence technology and staining experiments for alka-line phosphatase (ALP) and alizarin red staining (ARS) were used to research the proliferation, adhesion and differentiation ability of BMSCs seeded on SMAT Ti compared with native pure Ti. We used quantitative real-time PCR (qRT-PCR) technology so as to verify the expression of the most significant 5 genes. In summary, these results indicated novel point of views into candidate genes and potential mechanism for the further study of BMSCs' behaviors seeded on SMAT Ti.

Antibody therapeutics for the treatment of COVID-19 have been highly successful. However, the recent emergence of the Omicron variant has posed a challenge, as it evades detection by most existing SARS-CoV-2 neutralizing antibodies (nAbs). Here, we successfully generated a panel of SARS-CoV-2/SARS-CoV cross-neutralizing antibodies by sequential immunization of the two pseudoviruses. Of the potential candidates, we found that nAbs X01, X10, and X17 offer broad neutralizing potential against most variants of concern, with X17 further identified as a Class 5 nAb with undiminished neutralization against the Omicron variant. Cryo-electron microscopy structures of the three antibodies together in complex with each of the spike proteins of the prototypical SARS-CoV, SARS-CoV-2, and Delta and Omicron variants of SARS-CoV-2 defined three nonoverlapping conserved epitopes on the receptor-binding domain. The triple-antibody mixture exhibited enhanced resistance to viral evasion and effective protection against infection of the Beta variant in hamsters. Our findings will aid the development of antibody therapeutics and broad vaccines against SARS-CoV-2 and its emerging variants.

Bone growth and metabolism are mainly regulated by a series of intracellular molecules and extracellular stimuli. Exosome, as a nanoscale substance secreted to the outside of the cells, plays an extensive role in intercellular communication. This review provides theoretical references and evidences for further exploration of exosomes as noncoding RNA carriers to regulate bone tissue recovery through the following aspects: (1) basic characteristics of exosomes, (2) research progress of exosomal noncoding RNA in bone tissue engineering, (3) current status and advantages of engineering exosomes as nanocarriers for noncoding RNA delivery, and (4) problems and application prospects of exosome therapy in the field of orthopedics.

Titanium (Ti) is widely used in oral implants. However, there is still a challenge to promote the osseointegration of bone tissue on the surface of Ti. To solve this problem, we prepared novel gradient nanostructured (GNS) Ti and studied the effect of GNS on the adhesion, proliferation, and apoptosis of MG63 cells in vitro. The results demonstrated that the GNS Ti promoted the adhesion effect and proliferation of MG63 cells better than the annealed Ti, while the ability of GNS Ti to inhibit cell apoptosis was better than that of the annealed Ti, the preliminary mechanism of which indicated by this study might be the enhanced mineralization capacity, protein adsorption ability, and hydrophilicity of the GNS Ti due to its specific nanostructure which improved the cell behaviours. The results in this study provide the theoretical and experimental foundations for the applications of GNS Ti in dental implants and joint replacements.

Antibody therapeutics for the treatment of COVID-19 has been highly successful while faces a challenge of the recent emergence of the Omicron variant which escapes the majority of existing SARS-CoV-2 neutralizing antibodies (nAbs). Here, we successfully generated a panel of SARS-CoV-2/SARS-CoV cross-neutralizing antibodies by sequential immunization of the two pseudoviruses. Of which, nAbs X01, X10 and X17 showed broadly neutralizing breadths against most variants of concern (VOCs) and X17 was further identified as a Class 5 nAb with undiminished neutralization against the Omicron variant. Cryo-EM structures of three-antibody in complex with the spike proteins of prototyped SARS-CoV-2, Delta, Omicron and SARS-CoV defined three non-overlapping conserved epitopes on the receptor-binding domain (RBD). The triple antibody cocktail exhibited enhanced resistance to viral escape and effective protection against the infection of Beta variant in hamsters. Our finding will aid the development of both antibody therapeutics and broad vaccines against SARS-CoV-2 and emerging variants.

Coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 has become disaster for human society. As the pandemic becomes more regular, we should develop more rapid and accurate detection methods to achieve early diagnosis and treatment. Antigen detection methods based on spike protein has great potential, however, it has not been effectively developed, probably due to the torturing conformational complexity. By utilizing cross-blocking data, we clustered SARS-CoV-2 receptor binding domain (RBD)-specific monoclonal antibodies (mAbs) into 6 clusters. Subsequently, the antigenic sites for representative mAbs were identified by RBDs with designed residue substitutions. The sensitivity and specificity of selected antibody pairs was demonstrated using serial diluted samples of SARS-CoV-2 S protein and SARS-CoV S protein. Furthermore, pseudovirus system was constructed to determine the detection capability against SARS-CoV-2 and SARS-CoV. 6 RBD-specific mAbs, recognizing different antigenic sites, were identified as potential candidates for optimal antibody pairs for detection of SARS-CoV-2 S protein. By considering relative spatial position, accessibility and conservation of corresponding antigenic sites, affinity and the presence of competitive antibodies in clinical samples, 6H7-6G3 was rationally identified as optimal antibody pair for detection of both SARS-CoV-2 and SARS-CoV. Furthermore, our results showed that 6H7 and 6G3 effectively bind to SARS-CoV-2 variants of concern (VOCs). Taken together, we identified 6H7-6G3 antibody pair as a promising rapid antigen diagnostic tool in containing COVID-19 pandemic caused by multiple VOCs. Moreover, our results also provide an important reference in screening of antibody pairs detecting antigens with complex conformation.

Dental implants are commonly used to repair missing teeth. The implant surface plays a critical role in promoting osseointegration and implant success. However, little information is available about which implant surface treatment technology best promotes osseointegration and implant stability. The aim of this network meta-analysis was to evaluate the osseointegration and stability of four commonly used dental implants (SLA, SLActive, TiUnite, and Osseotite). The protocol of the current meta-analysis is registered in PROSPERO (International Prospective Register of Systematic Reviews) under the code CRD42020190907 (https://www.crd.york.ac.uk). We conducted a systematic review following PRISMA and Cochrane Recommendations. Medline (PubMed), Cochrane Library, Embase, and the Web of Science databases were searched. Only randomized controlled trials were considered. Twelve studies were included in the current network meta-analysis, eleven studies were included concerning the osseointegration effect and five studies were included for stability analysis (four studies were used to assess both stability and osseointegration). Rank possibility shows that the SLActive surface best promoted bone formation at an early healing stage and TiUnite seemed to be the best surface for overall osseointegration. For stability, TiUnite seemed to be the best surface. The present network meta-analysis showed that the SLActive surface has the potential to promote osseointegration at an early stage. The TiUnite surface had the best effect on osseointegration regarding the overall healing period. The TiUnite surface also had the best effect in stability.

Background Though titanium (Ti) is widely used as dental materials in the clinic, effective methods to treat Ti for higher surface biological activity still lack. Through Surface mechanical attrition treatment (SMAT) technology we could endow Ti with gradient nanostructured surface (GNS Ti). To investigate the biocompatibility of GNS Ti for its further application in dental implant field, we study the effects of GNS Ti on cell responses in vitro and osseointegration of the implant with surrounding bone tissues in vivo. Methods In this study, GNS Ti was fabricated by SMAT. In vitro experiment, we co-cultured GNS Ti with bone mesenchymal stem cells (BMSCs), surface characterization was detected by transmission electron microscope (TEM). Adhesion, proliferation and differentiation of BMSCs were evaluated by scanning electron microscope (SEM), MTT, flow cytometry (FCM), alkaline phosphatase (ALP) and osteocalcin (OCN) tests. In vivo experiment, the GNS Ti was implanted into the rabbit mandible. Osteogenesis and osseointegration were evaluated by Micro CT, toluidine blue staining, and immunohistochemical staining at 4, 8, and 12 weeks postoperatively. Results Both results showed that compared with the coarse grained (CG) Ti, the GNS Ti stimulated the adhesion, proliferation, and differentiation of BMSCs and improved osteogenesis and osseointegration. Conclusions This study indicates that gradient nanostructured Ti is a promising material for dental implant application.

Respiratory diseases, including pulmonary fibrosis, silicosis, and allergic pneumonia, can be caused by long-term exposure to dental prosthesis grinding dust. The extent of the toxicity and pathogenicity of exposure to PMMA dust, Vitallium dust, and dentin porcelain dust differs. The dust from grinding dental prosthesis made of these three materials was characterized in terms of morphology, particle size, and elemental composition. The adverse effects of different concentrations of grinding dust (50, 150, 300, 450, and 600 μg ml−l) on RAW264.7 macrophages were evaluated, including changes in cell morphology and the production of lactate dehydrogenase (LDH) and reactive oxygen species (ROS). The dust particles released by grinding dental prosthesis made of these materials had different morphologies, particle sizes, and elemental compositions. They also induced varying degrees of cytotoxicity in RAW264.7 macrophages. A possible cytotoxicity mechanism is the induction of lipid peroxidation and plasma membrane damage as the dust particles penetrate cells. Therefore, clinicians who regularly work with these materials should wear the appropriate personal protection equipment to minimize exposure and reduce the health risks caused by these particulates.

Background To analyze and identify the circular RNAs (circRNAs) involved in promoting the osteogenic differentiation of human bone mesenchymal stem cells (hBMSCs) on titanium by surface mechanical attrition treatment (SMAT). Methods The experimental material was SMAT titanium and the control material was annealed titanium. Cell Counting Kits-8 (CCK-8) was used to detect the proliferation of hBMSCs, and alkaline phosphatase (ALP) activity and alizarin red staining were used to detect the osteogenic differentiation of hBMSCs on the sample surfaces. The bioinformatics prediction software miwalk3.0 was used to construct competing endogenous RNA (ceRNA) networks by predicting circRNAs with osteogenesis-related messenger RNAs (mRNAs) and microRNAs (miRNAs). The circRNAs located at the key positions in the networks were selected and analyzed by quantitative real-time PCR (QRT-PCR). Results Compared with annealed titanium, SMAT titanium could promote the proliferation and osteogenic differentiation of hBMSCs. The total number of predicted circRNAs was 51. Among these, 30 circRNAs and 8 miRNAs constituted 6 ceRNA networks. Circ-LTBP2 was selected for verification. QRT-PCR results showed that the expression levels of hsa_circ_0032599, hsa_circ_0032600 and hsa_circ_0032601 were upregulated in the experimental group compared with those in the control group; the differential expression of hsa_circ_0032600 was the most obvious and statistically significant, with a fold change (FC) = 4.25 ± 1.60, p-values (p) < 0.05.

In the business scenario, the level of product pricing or customer preferences is not only affected by the individual attributes of the product or customer, but also by the interdependency of other products or customers. For example, a customer’s preference may be more similar to its closer neighbor. This research introduces a complete system of models to study the preference interdependency among individual products or customers, so that the matrix for describing interdependence can be introduced into this model. This paper used the general Spatial Autoregressive Regression (SAR) to study these preference interdependency, which cannot be modeled with other standard machine learning methods. Moreover, an improved iterative lasso regression method is introduced to perform variable selection in the presence of interdependence. These models were illustrated by studying factors affecting Washington D. C. housing prices, where the prices are proved to be highly related to geographic networks.

Background: The intricate mechanisms of articular chondrogenesis are largely unknown. Gradually, with the help of high-throughput platforms, microarrays have become an important and useful method to testify hub genes in desease. Today, advanced bioinformatic analysis of available microarray data can provide more reliable and accurate screening results by duplicating related data sets. Results: Microarray datasets GSE9451 and GSE104113 were downloaded from Gene Expression Omnibus (GEO) database. The differentially expressed genes (DEGs) were performed, and function enrichment analyses were demonstrated. The protein-protein interaction network (PPI) was constructed and the module analysis was performed by using STRING and Cytoscape. Quantitative PCR was used to confirm the results of bioinformatics analysis. Conclusion: Compared to individual studies, this study can provide extra reliable and accurate screening results by duplicating relevant records. Additional molecular experiments are required to confirm the discovery of candidate genes identified by chondrogenesis. S100A4 is predicted to integrate with miR-325-3p to promote osteogenesis.

Soil nitrogen (N) and phosphorus (P) contents, and soil acidification have greatly increased in grassland ecosystems due to increased industrial and agricultural activities. As major environmental and economic concerns worldwide, nutrient enrichment and soil acidification can lead to substantial changes in the diversity and structure of plant and soil communities. Although the separate effects of N and P enrichment on soil food webs have been assessed across different ecosystems, the combined effects of N and P enrichment on multiple trophic levels in soil food webs have not been studied in semiarid grasslands experiencing soil acidification. Here we conducted a short-term N and P enrichment experiment in non-acidified and acidified soil in a semiarid grassland on the Mongolian Plateau. We found that net primary productivity was not affected by N or P enrichment alone in either non-acidified or acidified soil, but was increased by combined N and P enrichment in both non-acidified and acidified soil. Nutrient enrichment decreased the biomass of most microbial groups in non-acidified soil (the decrease tended to be greatest with combined N and P enrichment) but not in acidified soil, and did not affect most soil nematode variables in non-acidified or acidified soil. Nutrient enrichment also changed plant and microbial community structure in non-acidified but not in acidified soil, and had no effect on nematode community structure in non-acidified or acidified soil. These results indicate that the responses to short-term nutrient enrichment were weaker for higher trophic groups (nematodes) than for lower trophic groups (microorganisms) and primary producers (plants). The findings increase our understanding of the effects of nutrient enrichment on multiple trophic levels of soil food webs, and highlight that soil acidification, as an anthropogenic stressor, reduced the responses of plants and soil food webs to nutrient enrichment and weakened plant-soil interactions.

Multi-wall carbon nanotubes/hydroxyapatite (MWCNTs/HA) composites with 20%, 40% and 60% HA were successfully fabricated at 1200 °C and 120 MPa by spark plasma sintering (SPS). Scanning electron microscope (SEM) observations showed that MWCNTs/HA composites kept the nanostructure of MWCNTs after SPS treatment. The bulk density of the MWCNTs/HA composites was increased by the addition of HA. However, the Vickers hardness, porosity, Young's modulus, flexure strength and compression strength of the MWCNTs/HA composites were decreased with increasing content of HA. To test their biocompatibility, the MWCNTs/40% HA composites were inserted into rat femur. The in vivo reaction of four weeks showed that the composites possessed better osseointegration compared to pure MWCNTs. The results indicated that the MWCNTs/HA composites have potential for a wide variety of bonegraft applications in the future.

A carbon nanotube (CNT) monolith without any binders was obtained by spark plasma sintering (SPS) treatment at 1100 °C under 40 MPa pressure. Transmission electron microscope results confirmed that this material maintained the nanosized tube microstructure of raw CNT powder after SPS treatment. The density and mechanical properties of this material were similar to cancellous bone. The material was implanted in subcutaneous tissue and femurs of rats and tissue samples were analyzed by histopathology at 1 and 4 weeks after surgery. Although some foreign body giant cells were seen around the CNTs, no severe inflammatory response such as necrosis was observed, and CNT implants were surrounded by newly formed bone in the femur. The study provides the first in vivo testing evidence that pure bulk carbon nanotubes are not a strongly inflammatory substance and have no toxicity for bone regeneration. Our study is the first successful experiment to consolidate CNTs without binders, and may provide an effective method for CNT monolith synthesis, as well as demonstrating that a binderless carbon nanotube material with a strength matching that of bone could be a candidate bone substitute material and a bone tissue engineering scaffold material.

Objective The purpose of this paper was to investigate the in vitro biodegradation of a guided tissue regeneration composite membrane, nano-carbonated hydroxyapatite/collagen/poly(lactic-co-glycolic acid) (nCHAC/PLGA). Especially for periodontal therapy, the functional graded material (FGM) nCHAC/PLGA membrane was prepared that consisted of three layers with 8 wt% nCHAC + PLGA/4 wt% nCHAC + PLGA/PLGA, where one face of the membrane is porous, thereby allowing cell growth thereon and the opposite face of the membrane smooth, thereby inhibiting cell adhesion. Methods For evaluation, in vitro degradation specimens of nCHAC/PLGA were immersed into artificial saliva solution at 37 °C for 1, 2, 4, 8 and 12 weeks to detect the weight loss over the period, and set pure PLGA membrane as control to compare the degraded behaviors. pH value and calcium concentration of the residual solution were measured, and morphology change was investigated by scanning electron microscopy (SEM). Results During the experimental period in vitro, the whole shape of the membrane could be kept for 4 weeks, after that it became powder at between 8 and 12 weeks. The results demonstrated that weight loss increased continuously with a reduction in mass of 23.1% after 4 weeks and 88% after 12 week for the nCHAC/PLGA three FGM layers composite membrane. The calcium concentration in the residual solution showed a significant increase after 4 weeks, which referred to the nano-carbonated hydroxyapatite degradation. Moreover, the pH value in the solution of the nCHAC/PLGA membrane was a little higher than that of the pure PLGA membrane, which demonstrated the possible neutralization effect from nCHAC composite for the acid outcome of PLGA in the solution. The pore structure of 8 wt% nCHAC + PLGA was enlarged on the porous surface, while the nonporous surface of pure PLGA also showed a small porous structure after increased time. Significance Degradation of the composite membrane is appropriate for practical periodontal repair. Moreover, the new mineral formation on the surface of the composite membrane referred to the possible positive effect in vivo for new bone tissue regeneration.

Multiwalled carbon nanotube/polycarbosilane (MWCNT/PCS) composites were fabricated by the spark plasma sintering (SPS) method. The MWCNT/PCS composites consisted of MWCNTs and nanosized SiC particles pyrolyzed from PCS and possessing good mechanical properties for bone tissue repair or dental implantation. The MWCNT/PCS composites were implanted in the subcutaneous tissue and femur of rats at 1 and 4 weeks after implantation. Histological investigations showed that there was little inflammatory response in the subcutaneous tissue, and newly formed bone tissue was observed in the femur. These results indicated that the MWCNT/PCS composite had little prophlogistic effect and good osteoconductivity. The study suggested the possibility that the MWCNT/PCS composite could be a candidate bone-substitute and dental-implant material in the future.

Carbide layer was formed on the surface of Ti by heating in hydrocarbon atmosphere (benzene C6H6) at 1000-1400 degrees C using a high frequency induction heating method. Physical and mechanical properties of carbide-coated Ti were investigated to examine its potential as an abrasion-resistant implant material. Scanning electron microscopy (SEM) showed that the surface of Ti was covered with fine grains of 1-4 microm diameter, depending on heating conditions. In addition, carbide layer of about 1-25 microm thickness was observed on the cross-section of specimens by SEM and energy dispersive spectroscopy. Vickers hardness of surface carbide was found to be more than 2000. Further, Martens scratch test and ultrasonic scaler abrasion test showed that the indentation depth and width of carbide-coated Ti were much smaller than pure Ti, thereby confirming its high abrasion resistance. These results showed that for Ti implant materials that require high abrasion resistance, such as the abutment for dental implants, surface carbide coatings would be an effective means to improve their wear properties.

In recent years, lots of researches have been launched on nanomaterials for biomedical applications. It has been shown that the performances of many biomaterials used in prosthodontics have been significantly enhanced after their scales were reduced by nanotechnology, from micron-size into nanosize. On the other hand, many nanocomposites composed of nanomaterials and traditional metals, ceramics, resin, or other matrix materials have been widely used in prosthodontics because their properties, such as modulus elasticity, surface hardness, polymerization shrinkage, and filler loading, were significantly increased after the addition of the nanomaterials. In this paper, the latest research progress on the applications of nanometals, nanoceramic materials, nanoresin materials, and other nanomaterials in prosthodontics was reviewed, which not only gives a detailed description of the new related investigations, but also hopefully provides important elicitation for future researches in this field. Published version

The dose verification methods in advanced radiotherapy are elaborated in this paper. The usage and application results for various dosimeters in dose verification are explained. As a theoretical method, Monte Carlo simulation, which has been developed greatly in recent years based on the technical progress in computer science, can be also used in dose verification with unique advantages. On the other hand, the principle of dose verification on proton and heavy-ion therapy is discussed briefly. Finally, the evaluation criteria for verification and the future development for dose verification are presented.

This review paper reported carbon nanotubes reinforced composites for biomedical applications. Several studies have found enhancement in the mechanical properties of CNTs-based reinforced composites by the addition of CNTs. CNTs reinforced composites have been intensively investigated for many aspects of life, especially being made for biomedical applications. The review introduced fabrication of CNTs reinforced composites (CNTs reinforced metal matrix composites, CNTs reinforced polymer matrix composites, and CNTs reinforced ceramic matrix composites), their mechanical properties, cell experimentsin vitro, and biocompatibility testsin vivo.

To determine the effect of different denture cleaning methods on surface roughness in resin denture base.We established 20 wafer samples of fuser base resin (14 mm×14 mm×3 mm), and then randomly divided them into 4 groups: group A was the control group, which were placed in water, while group B, C and D were the experimental groups, whose cleaning methods were toothbrush and water, toothbrush and toothpaste, denture cleaning tablets, respectively. Each procedure in group B and C lasted for 3 seconds, but group D lasted 5 min and repeated for 1080 times. We cleaned twice a day to simulate the method of cleaning dentures. Surface roughness was checked after different procedures by laser scanning confocal microscopy.Significant difference on surface roughness was found between group B, C and A (P0.05), while no significant difference in the surface roughness between group A and D (P0.05).Significant surface roughness alterations have been observed in toothbrush and toothpaste group, but no change has been found in denture cleaning tablets group, which does not produce scratches on the surface of resin denture base.

This study investigated the efficacy of cold light bleaching using different bleaching times and the effects thereof on tooth enamel. Before and after bleaching, stained tooth specimens were subjected to visual and instrumental colorimetric assessments using Vita Shade Guide and spectrophotometric shade matching. Enamel surface alterations were examined using scanning electron microscopy (SEM) to analyze surface morphology, surface microhardness (SMH) measurement to determine changes in mechanical properties, and X-ray diffraction (XRD) to characterize post-bleaching enamel composition. Cold light bleaching successfully improved tooth color, with optimal efficacy when bleaching time was beyond 10 min. Significant differences in surface morphology were observed among the different bleaching times, but no significant differences were observed for enamel composition and surface microhardness among the different bleaching times. Results of this study revealed an association between the bleaching time of cold light bleaching and its whitening efficacy. Together with the results on enamel surface changes, this study provided positive evidence to support cold light bleaching as an in-office bleaching treatment.