Femtosecond pulsed laser photodynamic therapy activates melanin and eradicates malignant melanoma.

Photodynamic therapy (PDT) relies on a series of photophysical and photochemical reactions leading to cell death. While effective for various cancers, PDT has been less successful in treating pigmented melanoma due to high light absorption by melanin. Here, this limitation is addressed by 2-photon excitation of the photosensitizer (2p-PDT) using ~100 fs pulses of near-infrared laser light. A critical role of melanin in enabling rather than hindering 2p-PDT is elucidated using pigmented and non-pigmented murine melanoma clonal cell lines in vitro. The photocytotoxicities were compared between a clinical photosensitizer (Visudyne) and a porphyrin dimer (Oxdime) with ~600-fold higher σ2p value. Unexpectedly, while the 1p-PDT responses are similar in both cell lines, 2p activation is much more effective in killing pigmented than non-pigmented cells, suggesting a dominant role of melanin 2p-PDT. The potential for clinical translational is demonstrated in a conjunctival melanoma model in vivo, where complete eradication of small tumors was achieved. This work elucidates the melanin contribution in multi-photon PDT enabling significant advancement of light-based treatments that have previously been considered unsuitable in pigmented tumors.

Effects on Colonization Factors and Mechanisms Involved in Antimicrobial Sonophotodynamic Inactivation Mediated by Curcumin.

Photodynamic (PDI) and sonodynamic (SDI) inactivation have been successfully employed as antimicrobial treatments. Moreover, sonophotodynamic inactivation (SPDI), which is the simultaneous application of PDI and SDI, has demonstrated greater effects. This study assessed the effects of PDI (PDI group), SDI (SDI group) and SPDI (SPDI group) using curcumin as a sensitizer on the metabolism, adhesion capability, biofilm formation ability and structural effects in a Staphylococcus aureus biofilm. Moreover, the production of reactive oxygen species (ROS) and the degradation spectrum of curcumin under the irradiation sources were measured. SPDI was more effective in inactivating the biofilm than PDI and SDI. All treatments reduced the adhesion ability of the bacteria: 58 ± 2%, 58 ± 1% and 71 ± 1% of the bacterial cells adhered to the polystyrene plate after the SPDI, SDI and PDI, respectively, when compared to 79 ± 1% of the untreated cells (control group). This result is probably related to the metabolism cell reduction after treatments. The metabolism of cells from the PDI group was 89 ± 1% lower than the untreated cells, while the metabolic activity of SDI and SPDI groups were 82 ± 2% and 90 ± 1% lower, respectively. Regarding the biofilm formation ability, all treatments (SPDI, SDI and PDI) reduced the total biomass. The total biomass of the PDI, SDI and SPDI groups were 26 ± 2%, 31 ± 5% and 35 ± 6% lower than the untreated biofilm (control group), respectively. Additionally, all treatments produced ROS and caused significant structural changes, reducing cells and the extracellular matrix. The light caused a greater absorbance decay of the curcumin; however, the US did not expressively alter its spectrum. Finally, SPDI had improved antimicrobial effects, and all treatments exhibited similar effects in the colonization factors evaluated.

Recombinant Prevotella melaninogenica α-1,3 glucanase and Capnocytophaga ochracea α-1,6 glucanase as enzymatic tools for in vitro degradation of S. mutans biofilms.

Dental biofilms represent a serious oral health problem playing a key role in the development of caries and other oral diseases. In the present work, we cloned and expressed in E. coli two glucanases, Prevotella melaninogenica mutanase (PmGH87) and Capnocytophaga ochracea dextranase (CoGH66), and characterized them biochemically and biophysically. Their three-dimensional structures were elucidated and discussed. Furthermore, we tested the capacity of the enzymes to hydrolyze mutan and dextran to prevent formation of Streptococcus mutans biofilms, as well as to degrade pre- formed biofilms in low and abundant sugar conditions. The percentage of residual biofilm was calculated for each treatment group in relation to the control, as well as the degree of synergism. Our results suggest that both PmGH87 and CoGH66 are capable of inhibiting biofilm formation grown under limited or abundant sucrose conditions. Degradation of pre-formed biofilms experiments reveal a time-dependent effect for the treatment with each enzyme alone. In addition, a synergistic and dose-dependent effects of the combined enzymatic treatment with the enzymes were observed. For instance, the highest biomass degradation was 95.5% after 30 min treatment for the biofilm grown in low sucrose concentration, and 93.8% after 2 h treatment for the biofilm grown in sugar abundant condition. Strong synergistic effects were observed, with calculated degree of synergism of 5.54 and 3.18, respectively and their structural basis was discussed. Jointly, these data can pave the ground for the development of biomedical applications of the enzymes for controlling growth and promoting degradation of established oral biofilms.

Integrating Fluorescent Nanodiamonds into Polymeric Microstructures Fabricated by Two-Photon Polymerization.

Nitrogen-vacancy (NV) and other color centers in diamond have attracted much attention as non-photobleaching quantum emitters and quantum sensors. Since microfabrication in bulk diamonds is technically difficult, embedding nanodiamonds with color centers into designed structures is a way to integrate these quantum emitters into photonic devices. In this study, we demonstrate a method to incorporate fluorescent nanodiamonds into engineered microstructures using two-photon polymerization (2PP). We studied the optimal concentration of nanodiamonds in the photoresist to achieve structures with at least one fluorescent NV center and good structural and optical quality. Fluorescence and Raman spectroscopy measurements were used to confirm the presence and location of the nanodiamonds, while absorbance measurements assessed scattering losses at higher concentrations. Our results show the feasibility of fabricating microstructures embedded within fluorescent nanodiamonds via 2PP for photonics and quantum technology applications.

Sonophotodynamic inactivation of Pseudomonas aeruginosa biofilm mediated by curcumin.

The inactivation of Pseudomonas aeruginosa biofilm is a major challenge, as biofilms are less responsive to conventional treatments and responsible for persistent infections. This has led to the investigation of alternative approaches for biofilm control such as photodynamic (PDI) and sonodynamic (SDI) inactivation. The combination of them, known as Sonophotodynamic Inactivation (SPDI), has improved the effectiveness of the process. Curcumin, a well-established photosensitizer, has been identified as a potential sonosensitizer. This study evaluated the most effective combination for SPDI against P. aeruginosa biofilms in vitro, varying curcumin concentrations and ultrasound intensities. The results indicated that the inactivation was directly proportional to the curcumin concentration. Using curcumin 120 µM and 3.0 W.cm-2 of ultrasound intensity, SPDI demonstrated the highest and the best synergistic results, equivalent to 6.9 ± 2.1 logs of reduction. PDI reduced 0.7 ± 0.9 log and SDI had no effect. In conclusion, SPDI with curcumin is a promising approach for biofilm inactivation.

Staphylococcus aureus microbial biofilms degradation using cellobiose dehydrogenase from Thermothelomyces thermophilus M77.

This work reports biochemical characterization of Thermothelomyces thermophilus cellobiose dehydrogenase (TthCDHIIa) and its application as an antimicrobial and antibiofilm agent. We demonstrate that TthCDHIIa is thermostable in different ionic solutions and is capable of oxidizing multiple mono and oligosaccharide substrates and to continuously produce H2O2. Kinetics measurements depict the enzyme catalytic characteristics consistent with an Ascomycota class II CDH. Our structural analyses show that TthCDHIIa substrate binding pocket is spacious enough to accommodate larger cello and xylooligosaccharides. We also reveal that TthCDHIIa supplemented with cellobiose reduces the viability of S. aureus ATCC 25923 up to 32 % in a planktonic growth model and also inhibits its biofilm growth on 62.5 %. Furthermore, TthCDHIIa eradicates preformed S. aureus biofilms via H2O2 oxidative degradation of the biofilm matrix, making these bacteria considerably more susceptible to gentamicin and tetracycline.

Antimicrobial Formulation of a Bacterial Nanocellulose/Propolis-Containing Photosensitizer for Biomedical Applications.

With the aim of contributing to the development of more efficient materials for wound care, new topical formulations based on bacterial nanocellulose (BNC) hydrogels containing propolis were produced. Characterizations confirmed the incorporation of propolis into the BNC matrix, maintaining its structure and properties. Rheological analysis confirmed that the hydrogels showed thixotropic behavior appropriate for topical application. Chromatographic profiles showed sustained release of propolis biomarkers for at least 20 h. The formulations did not present mutagenicity. For application in photodynamic inactivation (PDI), BNC/propolis hydrogels were prepared with the photosensitizers methylene blue (MB). Spectroscopy and confocal fluorescence microscopy confirmed the interaction of MB and propolis in BNC hydrogels, as well as the formation of a new composite material. In the antibacterial assays, formulations containing MB and propolis significantly reduced Staphylococcus aureus growth. In the presence of light, BNC/MB hydrogels completely inhibited the microorganism. Therefore, the results suggest potential materials for the prevention or treatment of Staphylococcus aureus infections in wounds.

Zinc-Based Nanoparticles Reduce Bacterial Biofilm Formation.

Biofilm formation is important for microbial survival in hostile environments and a phenotype that provides microorganisms with antimicrobial resistance. Zinc oxide (ZnO) and Zinc sulfide (ZnS) nanoparticles (NPs) present potential antimicrobial properties for biomedical and food industry applications. Here, we aimed to analyze, for the first time, the bactericidal and antibiofilm activity of ZnS NPs against Staphylococcus aureus, Klebsiella oxytoca, and Pseudomonas aeruginosa, all medically important bacteria in developed countries. We compared ZnS NPs antimicrobial activity to ZnO NPs, which have been extensively studied. Using the colorimetric XTT reduction assay to observe the metabolic activity of bacterial cells and the crystal violet assay to measure biofilm mass, we demonstrated that ZnS and ZnO had similar efficacy in killing planktonic bacterial cells and reducing biofilm formation, with S. aureus being more susceptible to both therapeutics than K. oxytoca and P. aeruginosa. Crystal violet staining and confocal microscopy validated that Zn NPs inhibit biofilm formation and cause architectural damage. Our findings provide proof of principle that ZnS NPs have antibiofilm activity, and can be potentially used in medical and food industry applications, such as treatment of wound infections or package coating for food preservation. IMPORTANCE Zinc (Zn)-based nanoparticles (NPs) can be potentially used in medical and food preservation applications. As proof of principle, we investigated the bactericidal and antibiofilm activity of zinc oxide (ZnO) and zinc sulfide (ZnS) NPs against medically important bacteria. Zn-based NPs were similarly effective in killing planktonic and biofilm-associated Staphylococcus aureus, Klebsiella oxytoca, and Pseudomonas aeruginosa cells. However, S. aureus was more susceptible to these investigational therapeutics. Although further studies are warranted, our findings suggest the possibility of future use of Zn-based NPs in the treatment of skin infections or preservation of food.

Photodynamic Inactivation of Microorganisms Using Semisynthetic Chlorophyll a Derivatives as Photosensitizers.

In this study, we describe the semisynthesis of cost-effective photosensitizers (PSs) derived from chlorophyll a containing different substituents and using previously described methods from the literature. We compared their structures when used in photodynamic inactivation (PDI) against Staphylococcus aureus, Escherichia coli, and Candida albicans under different conditions. The PSs containing carboxylic acids and butyl groups were highly effective against S. aureus and C. albicans following our PDI protocol. Overall, our results indicate that these nature-inspired PSs are a promising alternative to selectively inactivate microorganisms using PDI.

Effects of photobiomodulation on the redox state of healthy and cancer cells.

Photobiomodulation therapy (PBMT) uses light to stimulate cells. The molecular basis of the effects of PBMT is being unveiled, but it is stated that the cytochrome-c oxidase enzyme in mitochondria, a photon acceptor of PBMT, contributes to an increase in ATP production and modulates the reduction and oxidation of electron carriers NADH and FAD. Since its effects are not fully understood, PBMT is not used on tumors. Thus, it is interesting to investigate if its effects correlate to mitochondrial metabolism and if so, how it could be linked to the optical redox ratio (ORR), defined as the ratio of FAD/(NADH + FAD) fluorescences. To that end, fibroblasts (HDFn cell line) and oral squamous cell carcinoma (SCC-25 cell line) were irradiated with a light source of 780 nm and a total dose of 5 J/cm2, and imaged by optical microscopy. PBMT down-regulated the SCC-25 ORR by 10%. Furthermore, PBMT led to an increase in ROS and ATP production in carcinoma cells after 4 h, while fibroblasts only had a modest ATP increase 6 h after irradiation. Cell lines did not show distinct cell cycle profiles, as both had an increase in G2/M cells. This study indicates that PBMT decreases the redox state of oral cancer by possibly increasing glycolysis and affects normal and tumor cells through distinct pathways. To our knowledge, this is the first study that investigated the effects of PBMT on mitochondrial metabolism from the initiation of the cascade to DNA replication. This is an essential step in the investigation of the mechanism of action of PBMT in an effort to avoid misinterpretations of a variety of combined protocols.

Use of wide-field optical fluorescence for visualization of oral biofilm in a patient with peri-implant mucositis: a new approach.

Peri-implant diseases, caused by bacteria from biofilm related to dental implants, are one of the main causes of late loss of implants. In this sense, peri-implant diseases are divided into peri-implant mucositis, when it affects only the soft tissues, and peri-implantitis, when there is a bone involvement, which can lead to the failure of dental implant therapy. Thus, biofilm removal is essential for peri-implant health, allowing long-term success in implant therapy. To improve the visualization of oral biofilm, which is usually transparent or colorless, disclosing agents have been routinely used. However, disclosing agents have allergenic potential and can cause staining extrinsically in restorative and prosthetic materials, leading to aesthetic impairment. Thus, the use of fluorescence has been studied as an alternative for visualization of oral biofilm. Therefore, this report describes the use of wide-field optical fluorescence for visualization of oral biofilm associated with implants and teeth, in a routine appointment and follow-up of a partially edentulous patient with peri-implant mucositis. In addition, this report showed wide-field optical fluorescence can be used in a clinical routine of care of patients with dental implants. In this sense, wide-field optical fluorescence allowed easy and immediate visualization of the mature oral biofilm for its adequate removal, evaluation of the quality of restoration to sealing of screw access-hole of implant and identification of cariogenic lesions, without risk of allergic reactions or staining of prostheses and restorations.

Photodynamic therapy with a new bacteriochlorin derivative: Characterization and in vitro studies.

Photodynamic therapy presents a therapeutic choice that can be utilized to treat diverse neoplasms. In this technique, the critical element is a photosensitive molecule that absorbs light energy and transfers it to molecular oxygen or biological molecules to form reactive oxygen species, thus inducing irreversible damage to target cells and ultimately leading to cell death. Bacteriochlorin derivatives are employed as photosensitizers (PSs), possessing light-absorbing capacity in the near-infrared region. The objective of this study was to prepare a semi-synthetic bacteriochlorin from Rhodopseudomonas faecalis and adding Trizma® to improve solubility. Cell viability tests, flow cytometry (apoptotic and necrotic cells were identified by Annexin V and propidium iodide), and confocal microscopy were used to evaluate the photoactivity of bacteriochlorin-Trizma (Bchl-T) in fibroblast (HFF-1-control cells) and breast cancer (MCF-7 cells-target cells) cells. At concentrations above 0.5 µM, Bchl-T demonstrated 80 % cell death, presenting the highest PS interaction (via fluorescence microscopy) with lysosomes, mitochondria, and the endoplasmic reticulum; the cell death type was revealed as apoptosis (via cytometry). Our findings indicated the suitability of Bchl-T for future application in photodynamic therapy against cancer cells by inducing apoptosis.

Strategies to Improve the Antimicrobial Efficacy of Photodynamic, Sonodynamic, and Sonophotodynamic Therapies.

BACKGROUND AND OBJECTIVES: This work evaluated antimicrobial photodynamic therapy (PDT), sonodynamic therapy (SDT), and the association of both therapies (sonophotodynamic therapy [SPDT]), mediated by curcumin (Cur) against Staphylococcus aureus biofilm. Next, additional strategies for these treatments were assessed. MATERIALS AND METHODS: S. aureus biofilms received PDT, SDT, and SPDT, mediated by Cur (80 µM), LED light (450 nm), and 1 MHz ultrasound. The same treatments were also performed adding a strategy: Cur with sodium dodecyl sulfate (SDS), Cur with potassium iodide (KI) or a pre-treatment with ultrasound. Cell viability was determined and biofilm architecture was evaluated under confocal microscopy. RESULTS: SPDT was more effective to inactivate the bacteria than PDT and SDT. SDS achieved the greatest viability reductions, followed by KI and ultrasound pre-treatment. Confocal images revealed biofilm disruption and a reduced number of cells in all treatments. However, SPDT exhibited a pronounced effect and it was greater using SDS. CONCLUSION: SPDT was more effective and additional strategies potentiated its effectiveness. Lasers Surg. Med. © 2021 Wiley Periodicals LLC.

Photodynamic and Sonodynamic Therapy with Protoporphyrin IX: In Vitro and In Vivo Studies.

Sono-photodynamic therapy is a promising anticancer technique based on the combination of sonodynamic and photodynamic therapy to improve the cancer treatment effectiveness. This study was aimed at analyzing the effects of the sono-photodynamic (SPD) activity on protoporphyrin IX (PpIX) solution and PpIX-loaded rat liver. In vitro, PpIX 5 µM solutions were irradiated with light (635 nm, 30-50 mW/cm2), ultrasound (1 MHz, 1-2 W/cm2) and both. The PpIX absorption spectra recorded over exposure time revealed that the PpIX decay rate induced by SPD activity (combined irradiation) was approximately the sum of those induced by photodynamic and sonodynamic activity. In vivo, rats were intraperitoneally injected with 5-aminolevulinic acid at the dose of 500 mg/kg weight. After 3 h of injection, the PpIX-loaded livers were irradiated with light (635 nm, 180 ± 9 J/cm2), ultrasound (1.0 MHz, 770 ± 40 J/cm2) and both using a single probe capable of illuminating and sonicating the liver simultaneously. After 30 h, the liver damage induced by each protocol was analyzed histologically. It was found that a greater necrosis depth was induced by the SPD activity. These results suggest that the SPD activity could improve the PpIX decay rate and have greater scope than photodynamic or sonodynamic activity. Further studies should be performed to gain a better understanding of this protocol.

Dissolving microneedles containing aminolevulinic acid improves protoporphyrin IX distribution.

One important limitation of topical photodynamic therapy (PDT) is the limited tissue penetration of precursors. Microneedles (MNs) are minimally invasive devices used to promote intradermal drug delivery. Dissolving MNs contain drug-associated to polymer blends, dissolving after insertion into skin, allowing drug release. This study comprises development and characterization of a pyramidal model of dissolving MNs (500 µm) prepared with 5% wt/wt aminolevulinic acid and 20% wt/wt Gantrez AN-139 in aqueous blend. Protoporphyrin IX formation and distribution were evaluated in tumor mice model by using fluorescence widefield imaging, spectroscopy, and confocal microscopy. MNs demonstrated excellent mechanical resistance penetrating about 250 µm with minor size alteration in vitro, and fluorescence intensity was 5-times higher at 0.5 mm on average compared to cream in vivo (being 10 ± 5 a.u. for MNs and 2.4 ± 0.8 a.u. for cream). Dissolving MNs have overcome topical cream application, being extremely promising especially for thicker skin lesions treatment using PDT.

Use of wide-field optical fluorescence for visualization of oral biofilm in a patient with peri-implant mucositis: a new approach / Uso da fluorescência óptica de campo amplo para visualização de biofilme oral em paciente com mucosite peri-implantar: uma nova abordagem

ABSTRACT Peri-implant diseases, caused by bacteria from biofilm related to dental implants, are one of the main causes of late loss of implants. In this sense, peri-implant diseases are divided into peri-implant mucositis, when it affects only the soft tissues, and peri-implantitis, when there is a bone involvement, which can lead to the failure of dental implant therapy. Thus, biofilm removal is essential for peri-implant health, allowing long-term success in implant therapy. To improve the visualization of oral biofilm, which is usually transparent or colorless, disclosing agents have been routinely used. However, disclosing agents have allergenic potential and can cause staining extrinsically in restorative and prosthetic materials, leading to aesthetic impairment. Thus, the use of fluorescence has been studied as an alternative for visualization of oral biofilm. Therefore, this report describes the use of wide-field optical fluorescence for visualization of oral biofilm associated with implants and teeth, in a routine appointment and follow-up of a partially edentulous patient with peri-implant mucositis. In addition, this report showed wide-field optical fluorescence can be used in a clinical routine of care of patients with dental implants. In this sense, wide-field optical fluorescence allowed easy and immediate visualization of the mature oral biofilm for its adequate removal, evaluation of the quality of restoration to sealing of screw access-hole of implant and identification of cariogenic lesions, without risk of allergic reactions or staining of prostheses and restorations.

RESUMO Doenças peri-implantares, causadas por bactérias de biofilme relacionadas a implantes dentários, são uma das principais causas de perda tardia de implantes. Nesse sentido, as doenças peri-implantares são divididas em mucosite peri-implantar, quando afeta apenas tecidos moles, e peri-implantite, quando há comprometimento ósseo, o que pode levar ao fracasso da terapia com implantes dentários. Assim, a remoção do biofilme é essencial para a saúde peri-implantar, permitindo sucesso a longo prazo na terapia com implantes. A fim de melhorar a visualização do biofilme oral, que geralmente é transparente ou incolor, agentes reveladores têm sido rotineiramente utilizados. No entanto, esses agentes têm potencial alergênico e podem causar manchas extrinsecamente em materiais restauradores e protéticos, levando a prejuízo estético. Assim, o uso da fluorescência tem sido estudado como alternativa para visualização do biofilme oral. Este relato descreve o uso da fluorescência óptica de campo amplo para visualização do biofilme oral associado a implantes e dentes em uma consulta de acompanhamento de rotina de uma paciente parcialmente edêntula com mucosite peri-implantar. Além disso, este relato evidenciou que a fluorescência óptica de campo amplo pode ser utilizada dentro da rotina clínica de atendimento de pacientes com implantes dentários. Nesse sentido, a fluorescência óptica de campo amplo permitiu a visualização fácil e imediata do biofilme oral maduro para sua remoção adequada, a avaliação da qualidade da restauração do selamento do orifício de acesso do parafuso do implante e a identificação de lesões cariogênicas, sem risco de reações alérgicas ou manchamento de próteses e restaurações.

Theoretical and Experimental Analysis of Protoporphyrin IX Photodegradation Using Multi-Wavelength Light Sources.

Photodynamic procedures have been used in many applications, ranging from cancer treatment to microorganism inactivation. Photodynamic reactions start with the activation of a photosensitizing molecule with light, leading to the production of cytotoxic molecules that promote cell death. However, establishing the correct light and photosensitizer dosimetry for a broadband light source remains challenging. In this study, we proposed a theoretical mathematical model for the photodegradation of protoporphyrin IX (PpIX), when irradiated by multi-wavelength light sources. The theoretical model predicts the experimental photobleaching (temporal change in PpIX concentration) of PpIX for different light sources. We showed that photobleaching occurs independently of the light source wavelengths but instead depends only on the number of absorbed photons. The model presented here can be used as an important mathematical approach to better understand current photodynamic therapy protocols and help achieve optimization of the doses delivered.

Development of a system to treat and online monitor photodynamic therapy of skin cancer using PpIX near-infrared fluorescence.

The limited adoption of photodynamic therapy (PDT) around the medical field may be tied to the unpredicted treatment response that an unmonitored therapy could deliver. Given the high variability in the lesions optical and physiological parameters, it is of fundamental importance to monitor PDT, since different lesions require different therapeutic parameters. We developed a system to treat and online monitor PDT of skin cancer, using protoporphyrin-IX (PpIX) near-infrared fluorescence imaging. The system can be operated up to 150 mW/cm2 at 633 nm, with real-time fluorescence monitoring around 700 nm, using the treatment light itself for fluorescence excitation. This technology allows system portability, simplicity, and low cost. This study describes the system development and its comparison with a 400-450 nm commercial system to detect the PpIX fluorescence during a PDT in murine skin cancer model. The developed device was able to acquire considerably more fluorescence signal from deeper regions when compared to the violet excitation device.

Use of dermograph for improvement of PpIX precursor's delivery in photodynamic therapy: Experimental and clinical pilot studies.

BACKGROUND: Photodynamic Therapy (PDT) is a treatment for non-melanoma skin cancer. One of the main challenges of topical PDT is to increase the precursor penetration when applied on the lesion. Protoporphyrin IX (PpIX) is an endogenous photosensitizer (PS) widely used, obtained by the administration of precursors such as aminolevulinic acid and methyl aminolevulinate. Aiming for the technique improvement by providing greater PS penetration in skin lesions, we tested a new approach for drug delivery with a minimally invasive technique. A dermograph is a device currently used in aesthetic procedures to promote skin rejuvenation or to micropigmentation. The use of dermograph for drug delivery has not been particularly explored for PDT so far, and the present study explores that approach as its main goal. METHODS: This study evaluated the PpIX distribution and PDT damage in normal rat skin model; the response of dermograph application in a pilot clinical study was also investigated. RESULTS: The animal tests showed that more homogeneous PpIX distribution and greater penetration in the tissue was observed with dermograph when compared to the topical application. Six nodular basal cell carcinoma lesions were treated with PDT using intradermal delivery by dermograph, and no recurrence was observed after 28 months of follow-up. CONCLUSIONS: The precursor's penetration improvement and the consequent increase in PpIX distribution in-depth both favor PDT response, providing upgrades concerning problems that hinder the clinical practice acceptance.

Quantitative image of fluorescence of ceramic and resin-cement veneers.

The main of the study was quantify the effect of two ceramics with two underlying resin cements on apparent fluorescence levels. Buccal surfaces of two bovine incisors were ground flat producing one enamel and one dentin substrate. The veneers were fabricated (0.5 and 1.0 mm thickness) using two ceramics (IPSe.max Press and IPSe.max Zirpress, Ivoclar Vivadent). Veneers were cemented using either light-cured (Variolink II, Ivoclar Vivadent) or self-adhesive dual (Rely X U200, 3M ESPE) cement. The layered Control group materials had no cement application. Semi-quantitative fluorescence image analysis (Matlabs software, Matworks) involved processing the images as captured under each daylight (DL, Gretagmacbeth) and ultraviolet illuminants (UVA, Sylvania) within a neutral-gray lightbox (Macbeth Spectral Light). Statistical analysis of the quantitative fluorescence values was performed using two-way ANOVA and Tukey's test (p < 0.05). The e.max Zirpress on the dentin substrate produced greater fluorescence (p < 0.05) when subjected to UV illumination and more fluorescence (p < 0.05) than e.max Press in both cement groups. Light-cured cement produced higher (p < 0.05) fluorescence than the dual-cement with e.max Press on enamel under UV illumination. The fluorescence for e.max Press on the dentin substrate was greater (p < 0.05) than for e.max Zirpress using dual self-adhesive cement subjected to daylight illumination. Thus, it is possible to conclude that the combination of ceramic and cement produce definite, significant effects on the apparent fluorescence, vital quality for restorative dentistry.