Photodynamic Therapy for Colorectal Cancer: An Update and a Look to the Future.

This review provides an update on the current state of photodynamic therapy (PDT) for colorectal cancer (CRC) and explores potential future directions in this field. PDT has emerged as a promising minimally invasive treatment modality that utilizes photosensitizers and specific light wavelengths to induce cell death in targeted tumor tissues. In recent years, significant progress has been made in understanding the underlying mechanisms, optimizing treatment protocols, and improving the efficacy of PDT for CRC. This article highlights key advancements in PDT techniques, including novel photosensitizers, light sources, and delivery methods. Furthermore, it discusses ongoing research efforts and potential future directions, such as combination therapies and nanotechnology-based approaches. By elucidating the current landscape and providing insights into future directions, this review aims to guide researchers and clinicians in harnessing the full potential of PDT for the effective management of CRC.

Antibiotic Susceptibility Profiles and Frequency of Resistance Genes in Clinical Shiga Toxin-Producing Escherichia coli Isolates from Michigan over a 14-Year Period.

Shiga toxin-producing Escherichia coli (STEC) is an important foodborne pathogen that contributes to over 250,000 infections in the United States each year. Because antibiotics are not recommended for STEC infections, resistance in STEC has not been widely researched despite an increased likelihood for the transfer of resistance genes from STEC to opportunistic pathogens residing within the same microbial community. From 2001 to 2014, 969 STEC isolates were collected from Michigan patients. Antibiotic susceptibility profiles to clinically relevant antibiotics were determined using disc diffusion, while epidemiological data were used to identify factors associated with resistance. Whole-genome sequencing was used for serotyping, examining genetic relatedness, and identifying genetic determinants and mechanisms of resistance in the non-O157 isolates. Increasing frequencies of resistance to at least one antibiotic were observed over the 14 years (P = 0.01). While the non-O157 serogroups were more commonly resistant than O157 (odds ratio, 2.4; 95% confidence interval,1.43 to 4.05), the frequency of ampicillin resistance among O157 isolates was significantly higher in Michigan than the national average (P = 0.03). Genomic analysis of 321 non-O157 isolates uncovered 32 distinct antibiotic resistance genes (ARGs). Although mutations in genes encoding resistance to ciprofloxacin and ampicillin were detected in four isolates, most of the horizontally acquired ARGs conferred resistance to aminoglycosides, ß-lactams, sulfonamides, and/or tetracycline. This study provides insight into the mechanisms of resistance in a large collection of clinical non-O157 STEC isolates and demonstrates that antibiotic resistance among all STEC serogroups has increased over time, prompting the need for enhanced surveillance.

Adhesive/silane application effects on bond strength durability to a lithium disilicate ceramic.

OBJECTIVE: To test the effects of different adhesive protocols and silane application on the adhesive durability to a Lithium Disilicate reinforced glass ceramic. METHODS: Forty disks of 13 mm diameter (E.max Press) were used. After etching with 9.5% HF for 20 seconds, disks were randomly assigned into 4 groups according to the adhesive/silane protocol: silane application only (SIL); silane application followed by adhesive (SILXP-XP Bond); silane-containing adhesive (SBU-ScotchBond Universal); silane application followed by silane-containing adhesive (SILSBU). Four resin composite cylinders of 1-mm diameter and 3-mm height were made on each ceramic disk and tested in shear. Specimens were stored in water for 24 hours or 12 months prior to testing. Results were statistically analyzed by two-way ANOVA and Tukey test. RESULTS: After 24 hours, the highest SBS values were observed for SILXP and SILSBU. However, after 12 months, SILXP and SILSBU presented a significant reduction in SBS, while the highest SBS were observed for SIL. For SBU, no significant reduction in SBS was observed, however, it showed the lowest SBS after 12 months. CONCLUSIONS: Regardless of the presence of silane in the composition of SBU, previous silane application is still recommended prior to cementation of Lithium Disilicate. CLINICAL SIGNIFICANCE: The application of silane as a separate step is recommended prior to cementation of Lithium Disilicate reinforced glass-ceramic, independent of the presence of silane within the universal adhesive solution.

Isolation of Cells Specialized in Anticancer Alkaloid Metabolism by Fluorescence-Activated Cell Sorting.

Plant specialized metabolism often presents a complex cell-specific compartmentation essential to accomplish the biosynthesis of valuable plant natural products. Hence, the disclosure and potential manipulation of such pathways may depend on the capacity to isolate and characterize specific cell types. Catharanthus roseus is the source of several medicinal terpenoid indole alkaloids, including the low-level anticancer vinblastine and vincristine, for which the late biosynthetic steps occur in specialized mesophyll cells called idioblasts. Here, the optical, fluorescence, and alkaloid-accumulating properties of C. roseus leaf idioblasts are characterized, and a methodology for the isolation of idioblast protoplasts by fluorescence-activated cell sorting is established, taking advantage of the distinctive autofluorescence of these cells. This achievement represents a crucial step for the development of differential omic strategies leading to the identification of candidate genes putatively involved in the biosynthesis, pathway regulation, and transmembrane transport leading to the anticancer alkaloids from C. roseus.

Gas-diffusion microextraction coupled with spectrophotometry for the determination of formaldehyde in cork agglomerates.

In this work, a simple methodology was developed for the extraction and determination of free formaldehyde content in cork agglomerate samples. For the first time, gas-diffusion microextraction was used for the extraction of volatile formaldehyde directly from samples, with simultaneous derivatization with acetylacetone (Hantzsch reaction). The absorbance of the coloured solution was read in a spectrophotometer at 412 nm. Different extraction parameters were studied and optimized (extraction temperature, sample mass, volume of acceptor solution, extraction time and concentration of derivatization reagent) by means of an asymmetric screening. The developed methodology proved to be a reliable tool for the determination of formaldehyde in cork agglomerates with the following suitable method features: low LOD (0.14 mg kg-1) and LOQ (0.47 mg kg-1), r 2 = 0.9994, and intraday and interday precision of 3.5 and 4.9%, respectively. The developed methodology was applied to the determination of formaldehyde in different cork agglomerate samples, and contents between 1.9 and 9.4 mg kg-1 were found. Furthermore, formaldehyde was also determined by the standard method EN 717-3 for comparison purposes; no significant differences between the results of both methods were observed. Graphical abstract Representation of the GDME system and its main components.

An Insight on Salting-out Assisted Liquid-Liquid Extraction for Phytoanalysis.

INTRODUCTION: Salting-out assisted liquid-liquid extraction (SALLE) is a technique that, although simple and not requiring any complex equipment, is very powerful and versatile. It has obtained growing interest in bioanalysis particularly when combined with chromatographic techniques. OBJECTIVES: Herein, fennel seeds (Foeniculum vulgare Mill.) were used as a case-study to show the application of SALLE in phytochemical analysis. MATERIAL AND METHODS: SALLE combined with HPLC-UV-MS/MS and GC-MS. RESULTS: By HPLC-UV-MS/MS analysis of the organic extract it was possible to identify various phenolic compounds, including quercetin derivatives, caffeic acid, p-coumaric acid and chlorogenic acid. The main compounds identified by GC-MS were estragole, fenchone, anisaldehyde, anethole, benzaldehyde, camphor and apiole. CONCLUSION: HPLC and GC analysis of the extracts showed that it is possible to isolate, in only one step, a wide range of compounds with distinct properties, allowing a detailed phytochemical analysis. Copyright © 2017 John Wiley & Sons, Ltd.

Influence of osteoporosis on the osteocyte density of human mandibular bone samples: a controlled histological human study.

OBJECTIVES: Osteocytes have emerged as key regulators of skeletal and mineral homeostasis. The number of these cells could be influenced by the presence of osteoporosis and osteopenia. Hence, the aim this study was to evaluate the osteocyte density in patients with osteopenia, and in patients with osteoporosis treated with bisphosphonates. MATERIALS AND METHODS: Thirty-nine patients were selected for this study and divided into three groups: (A) nine healthy patients (control), (B) 15 patients with osteopenia, and (C) 15 patients with osteoporosis. During the surgical insertion of dental implants in the lower jaw, bone samples were retrieved and processed for histological analysis of osteocyte density, measured as number of osteocytes/bone tissue area (µm(2) ). RESULTS: Patients with osteopenia showed statistically higher values of osteocyte density than patients with osteoporosis (P < 0.05) No significant differences were detected between osteopenia and osteoporosis subjects vs. healthy patients (P > 0.05). CONCLUSIONS: Bone metabolism diseases (osteoporosis and osteopenia) do not seem to influence the osteocyte density; this could be due to the administration of bisphosphonates in patients with osteoporosis. This information could play a fundamental role in the diagnosis and treatment of patients in a postmenopausal stage.

In Vitro Behavior of Osteoblasts on Zirconia After Different Intensities of Erbium, Chromium-Doped: Yttrium, Scandium, Gallium, and Garnet-Laser Irradiation.

OBJECTIVE: This study investigated the osteoblastic behavior to yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) surface after different intensities of erbium, chromium-doped:yttrium, scandium, gallium, and garnet (Er,Cr:YSGG) laser irradiation treatment. MATERIALS AND METHODS: Y-TZP blocks were randomized divided into 4 groups to Er,Cr:YSGG laser irradiation: group G1 = control (no laser irradiation); group G2 = 1.5 W; group G3 = 3.0 W; group G4 = 5.0 W, with air-water irrigation. The irradiated surface roughness (Sa and Ra) topography was evaluated. MC3T3-E1 osteoblast cells were cultured and seeded onto the treated surfaces at a plate density of 1 × 10 cells/mL. At the third and seventh days cell proliferation was measured by the colorimetric assay methods. The specimens were prepared to scanning electron microscope to observe adhesion and morphology of cells. RESULTS: MTT assay showed adhesion and proliferation of MC3T3-E1 cells in all groups. Compared with group G1 (control), the group G2 had a proliferation 1.4% higher (P > 0.05), the group G3 had a proliferation 3.1% higher (P < 0.05) and group G4 had a proliferation 4.5% higher (P < 0.01). After 7 days there was no difference among the groups (ANOVA, Newman-Keuls). CONCLUSIONS: Er,Cr:YSGG laser irradiation may have contributed to the greater and earlier cell spreading that was observed with the Y-TZP without alterations on surface topography.

Effect of lateral static load on immediately restored implants: histologic and radiographic evaluation in dogs.

OBJECTIVES: The aim of this study was to evaluate the effect of lateral static load in immediately restored implants in the canine mandible. MATERIAL AND METHODS: In 7 mongrel dogs, all premolars and the first molars were extracted bilaterally in the mandible. Two months after the extractions, 6 implants with sandblasted acid-etched surfaces were placed in each animal, 3 on each side. Randomly, two implants per side were immediately restored with an orthodontic expansion device that promoted lateral excessive static load (test group) or load (control) while the third implant of each side, remained submerged (unloaded group). These loaded devices were cleaned daily with chlorhexidine 0.12%, during the 4 months of study, when the animals were sacrificed and biopsies removed for histometric study. Intra-oral periapicals were made using positioners at the beginning and at the end of the study. RESULTS: Five implants from test group were lost in 3 dogs. Radiographically, there was a marginal bone loss of 3.68 ± 0.74 mm for the test group, 1.63 ± 0.2 mm for the control group and 0.45 ± 0.5 mm for the unloaded group. Histometrically, the percentage of bone-to-implant contact (BIC%) was 35.52 ± 7.32%, for the test group, 63.16 ± 5.16% for the control group and 42.33 ± 2.14% for the unloaded group. CONCLUSION: In conclusion, the excessive lateral static load negatively affected the behavior of peri-implant bone around immediate restored implants.

Effects of tobacco smoking on the survival rate of dental implants placed in areas of maxillary sinus floor augmentation: a systematic review.

OBJECTIVES: The objective of this systematic review was to evaluate the effects of smoking on the survival rate of dental implants placed in areas of maxillary sinus floor augmentation. MATERIAL AND METHODS: The Cochrane Oral Health Group's Trials Register (CENTRAL), MEDLINE and EMBASE were searched in duplicate up to, and including, October 2012 without language restrictions. Studies were considered eligible for inclusion if they involved the treatment of smokers and non-smokers with titanium implants and sinus floor elevation procedures. The Newcastle-Ottawa Scale and the Cochrane Collaboration's quality assessment tool were used for the assessment of the risk of bias in included studies. Random effects meta-analyses were used to assess the number of implants lost in smokers vs. number of implants lost in non-smokers. RESULTS: Of 3360 potentially eligible papers, eight studies were included. More than half (62.5%) of the studies found that smoking adversely affects implant survival in sites of sinus floor augmentation. Similarly, the pooled analysis indicated a statistically significantly increased risk of implant failure in smokers when the outcomes of all studies available to be included into meta-analysis were evaluated [RR: 1.87 (95% CI: 1.35, 2.58), P = 0.0001]. Conversely, a subgroup analysis including only prospective studies (3 studies) did not reveal significant differences in implant failure between smokers and non-smokers [RR: 1.55 (95% CI: 0.91, 2.65), P = 0.11]. CONCLUSIONS: Although smoking was associated with implant failure in most of individual studies and in the overall meta-analysis, the detrimental effect of smoking was not confirmed when only prospective data were assessed.

Simultaneous determination of carbonyl compounds related to thermal treatment and oxidative stability of infant formulas by gas-diffusion microextraction and high-performance liquid chromatography with ultraviolet detection.

Infant formulae are the only possible alternative to breastfeeding during the first year of life, so it is crucial to assure their innocuousness. Infant formula undergoes heat treatments to ensure safety and shelf life. However, such processes impact health as they lead to the formation of malondialdehyde, acrolein, and α-dicarbonyl compounds, related to Maillard reaction. Thus, there is a need for improved analytical methods to ensure the safety, quality, and nutritional value of infant formulae, and also exploring the potential of specific compounds as indicators for quality control and monitoring purposes. We developed and validated a novel, efficient, and cost-effective method using gas-diffusion microextraction for the simultaneous quantification of carbonyl compounds in infant formula. Malondialdehyde, acrolein, glyoxal, methylglyoxal, and diacetyl were detected as o-phenylenediamine derivatives using HPLC with UV detection. Parameters influencing extraction efficiency were studied using an asymmetric screening design. The validated method has shown excellent linearity, sensitivity, accuracy, and precision. It was applied to analyze 26 infant formula samples, including starter, follow-up, and special formulated powdered infant formula. Methylglyoxal was found in all samples (0.201-3.153 µg mL-1), while malondialdehyde was present only in certain starter formulas (1.033-1.802 µg mL-1). Acrolein (0.510-3.246 µg mL-1), glyoxal (0.109-1.253 µg mL-1), and diacetyl (0.119-2.001 µg mL-1) were detected in various sample types. Principal components and hierarchical cluster analyses have showcased distinct sample clustering based on analyte contents. This study presents a novel methodology for the analysis of markers of thermal treatment and oxidative stability in infant formula. It contributes to the characterization of the products' composition and quality control of infant formulae, thereby enhancing their safety and nutritional adequacy. This study also presents the first reported quantification of acrolein in infant formula and introduces the application of the acrolein-o-phenylenediamine derivative for food analysis.

Chromatographic analysis of methylglyoxal and other α-dicarbonyls using gas-diffusion microextraction.

Many α-dicarbonyl compounds such as methylglyoxal, diacetyl and pentane-2,3-dione are important quality markers of processed foods. They are produced by enzymatic and chemical processes, the Maillard reaction is the most known chemical route for α-dicarbonyl formation. In the case of methylglyoxal, there are obstacles to be overcome when analysing this compound due to its high reactivity, low volatility and low concentration. The use of extraction techniques based on the volatilization of methylglyoxal (like solid-phase microextraction) showed to be ineffective for the methylglyoxal extraction from aqueous solutions. Therefore, derivatization is typically applied to increase analyte's volatility. In this work a new methodology for the extraction and analysis of methylglyoxal and also diacetyl and pentane-2,3-dione from selected food matrices is presented. It is based on a gas-diffusion microextraction step followed by high performance liquid chromatographic analysis. It was successfully applied to port wines, black tea and soy sauce. Methylglyoxal, diacetyl and pentane-2,3-dione were quantified in the following concentration ranges: 0.24-1.74 mg L(-1), 0.1-1.85 mg L(-1) and 0.023-0.15 mg L(-1), respectively. The main advantages over existing methodologies are its simplicity in terms of sample handling, not requiring any chemical modification of the α-dicarbonyls prior to the extraction, low reagent consumption and short time of analysis.

Xanthohumol modulates inflammation, oxidative stress, and angiogenesis in type 1 diabetic rat skin wound healing.

Type 1 diabetes mellitus is responsible for metabolic dysfunction, accompanied by chronic inflammation, oxidative stress, and endothelium dysfunction, and is often associated with impaired wound healing. Phenol-rich food improves vascular function, contributing to diabetes prevention. This study has evaluated the effect of phenol-rich beverage consumption in diabetic rats on wound healing, through angiogenesis, inflammation, and oxidative stress modulation. A wound-healing assay was performed in streptozotocin-induced diabetic Wistar rats drinking water, 5% ethanol, and stout beer with and without 10 mg/L xanthohumol (1), for a five-week period. Wounded skin microvessel density was reduced to normal values upon consumption of 1 in diabetic rats, being accompanied by decreased serum VEGF-A and inflammatory markers (IL-1ß, NO, N-acetylglucosaminidase). Systemic glutathione and kidney and liver H2O2, 3-nitrotyrosine, and protein carbonylation also decreased to healthy levels after treatment with 1, implying an improvement in oxidative stress status. These findings suggest that consumption of xanthohumol (1) by diabetic animals consistently decreases inflammation and oxidative stress, allowing neovascularization control and improving diabetic wound healing.

Allosteric site variants affect GTP hydrolysis on Ras.

RAS GTPases are proto-oncoproteins that regulate cell growth, proliferation, and differentiation in response to extracellular signals. The signaling functions of RAS, and other small GTPases, are dependent on their ability to cycle between GDP-bound and GTP-bound states. Structural analyses suggest that GTP hydrolysis catalyzed by HRAS can be regulated by an allosteric site located between helices 3, 4, and loop 7. Here we explore the relationship between intrinsic GTP hydrolysis on HRAS and the position of helix 3 and loop 7 through manipulation of the allosteric site, showing that the two sites are functionally connected. We generated several hydrophobic mutations in the allosteric site of HRAS to promote shifts in helix 3 relative to helix 4. By combining crystallography and enzymology to study these mutants, we show that closure of the allosteric site correlates with increased hydrolysis of GTP on HRAS in solution. Interestingly, binding to the RAS binding domain of RAF kinase (RAF-RBD) inhibits GTP hydrolysis in the mutants. This behavior may be representative of a cluster of mutations found in human tumors, which potentially cooperate with RAF complex formation to stabilize the GTP-bound state of RAS.

Pangenomic analyses of antibiotic-resistant Campylobacter jejuni reveal unique lineage distributions and epidemiological associations.

Application of whole-genome sequencing (WGS) to characterize foodborne pathogens has advanced our understanding of circulating genotypes and evolutionary relationships. Herein, we used WGS to investigate the genomic epidemiology of Campylobacter jejuni, a leading cause of foodborne disease. Among the 214 strains recovered from patients with gastroenteritis in Michigan, USA, 85 multilocus sequence types (STs) were represented and 135 (63.1 %) were phenotypically resistant to at least one antibiotic. Horizontally acquired antibiotic resistance genes were detected in 128 (59.8 %) strains and the genotypic resistance profiles were mostly consistent with the phenotypes. Core-gene phylogenetic reconstruction identified three sequence clusters that varied in frequency, while a neighbour-net tree detected significant recombination among the genotypes (pairwise homoplasy index P<0.01). Epidemiological analyses revealed that travel was a significant contributor to pangenomic and ST diversity of C. jejuni, while some lineages were unique to rural counties and more commonly possessed clinically important resistance determinants. Variation was also observed in the frequency of lineages over the 4 year period with chicken and cattle specialists predominating. Altogether, these findings highlight the importance of geographically specific factors, recombination and horizontal gene transfer in shaping the population structure of C. jejuni. They also illustrate the usefulness of WGS data for predicting antibiotic susceptibilities and surveillance, which are important for guiding treatment and prevention strategies.

Application of gas-diffusion microextraction to the analysis of free and bound acetaldehyde in wines by HPLC-UV and characterization of the extracted compounds by MS/MS detection.

In wines, the presence of high levels of acetaldehyde (AA) not only is responsible for undesirable characteristic odours but can also cause health adverse effects. Such sensorial activity of AA can be overcome by adding sulphites during winemaking, due to the formation of adducts between AA and sulphites, which lower the sensorial impact of AA. Nevertheless, bound AA can be released during wine storage; therefore, the knowledge of its total amount can be important to estimate the long-term wine quality. The proposed methodology is based on the extraction of AA from wines using gas-diffusion microextraction and determination by liquid chromatography. Free and bound forms of AA could be differentiated and determined using an alkaline hydrolysis step to dissociate the sulphites-AA adducts. This methodology was successfully applied to different wine types, with free AA values ranging between 5 and 26 mg L(-1) and total form between 154 and 906 mg L(-1). Bound AA was above 90% of the total content determined for all samples analysed, and higher amounts were obtained for white wines (around 98%). Other carbonyl compounds were also identified in the extracts using mass spectrometry.

Analysis of cardamonin by square wave voltammetry.

INTRODUCTION: Several biochemical studies have already shown that cardamonin has health promoting properties, such is in agreement with typical characteristics of chalcones. Although being a very promising compound for the nutraceutical field there is a lack of studies concerning its electroanalytical properties. OBJECTIVE: To develop an electroanalytical methodology for the quantification of cardamonin in cardamom. METHODOLOGY: Cardamonin was analysed electrochemically by means of a hanging mercury drop electrode (HMDE) using square wave voltammetry (SWV). It was extracted from cardamom spice and quantified thereafter using the standard additions method to overcome matrix effects. RESULTS: A limit of detection (LOD) of 0.15 mg/L and good linearity (r² = 0.9998) were obtained. Decoction using ethanol as the extraction solvent appears to be the simplest extraction technique. Spectrophotometric analysis (maximum absorbance peak was found in ethanol at 344 nm with a value of molar extinction coefficient of (2.8 ± 0.1) × 104 L mol⁻¹ cm⁻¹) and mass spectrometry analysis by electrospray in the positive ion mode were also performed. CONCLUSION: Cardamonin was detected voltammetrically. The LOD and limit of quantification (LOQ) of the proposed voltammetric methodology are adequate for trace analysis of this compound in several phytochemical matrices.

Enhanced Photodynamic Therapy: A Review of Combined Energy Sources.

Photodynamic therapy (PDT) has been used in recent years as a non-invasive treatment for cancer, due to the side effects of traditional treatments such as surgery, radiotherapy, and chemotherapy. This therapeutic technique requires a photosensitizer, light energy, and oxygen to produce reactive oxygen species (ROS) which mediate cellular toxicity. PDT is a useful non-invasive therapy for cancer treatment, but it has some limitations that need to be overcome, such as low-light-penetration depths, non-targeting photosensitizers, and tumor hypoxia. This review focuses on the latest innovative strategies based on the synergistic use of other energy sources, such as non-visible radiation of the electromagnetic spectrum (microwaves, infrared, and X-rays), ultrasound, and electric/magnetic fields, to overcome PDT limitations and enhance the therapeutic effect of PDT. The main principles, mechanisms, and crucial elements of PDT are also addressed.

A Novel Approach for Monitoring the Volatile Metabolome in Biological Samples from Ruminants through Miniaturized Liquid-Liquid Extraction and Multiclass Gas Chromatography Analysis.

A straightforward and versatile methodology for the extraction of volatile metabolites in biological samples from ruminants for gas chromatography analysis is proposed. The methodology was applied in the determination of multiclass metabolites (short-chain fatty acids, aldehydes, alcohols, ketones, esters, phenols, and sulfides) in different analytical matrices (rumen fluid, urine, and feces) collected from Holstein cows. The 24 multiclass volatile metabolites reported in the different biological samples and their respective concentrations were critically discussed in the context of digestive physiology. Most detected compounds are derived from the rumen and lower gut fermentation of carbohydrates, proteins, and lipids or their metabolism, being consistent with the prior state of the art. The proposed method also takes advantage of the already existing tools in animal nutrition laboratories, providing a novel methodological ground that can generate relevant bioanalytical information with a significant impact on ruminant's nutritional studies.

Effect of different concentrations of carbamide peroxide on microhardness of dental ceramics.

PURPOSE: To evaluate the effect of 10% and 16% carbamide peroxide bleaching agents on microhardness of dental ceramics after different periods of bleaching treatment. METHODS: 15 specimens with 5 x 3 x 1 mm3 were created with four dental ceramics following manufacturers' instructions: IPS Classic (Ivoclar-Vivadent); IPS d.Sign (Ivoclar-Vivadent); EX3 (Noritake); VMK-95 (Vita). A microhardness tester with a Knoop diamond with a 100 g load was used to evaluate the baseline microhardness values of all ceramics. Afterwards, the specimens were submitted to 6-hour daily bleaching treatments with 10% or 16% carbamide peroxide (Whiteness Perfect-FGM) for 21 days, while control groups from each ceramic system were maintained in artificial saliva. The microhardness of all groups was evaluated at 18, 42, 84, and 126 hours of bleaching treatment. The mean value of five indentations performed at each specimen in each time was obtained and all data were submitted to two-way repeated measures ANOVA and Tukey's post-hoc test (alpha = 0.05). RESULTS: No significant differences in ceramic microhardness were observed among either bleaching intervals or bleaching treatments. Ceramic restorations are not affected by carbamide peroxide 10% or 16% gel during bleaching treatment.