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The management of biofilm-related infections is a challenge in healthcare, and antimicrobial photodynamic therapy (aPDT) is a powerful tool that has demonstrated a broad-spectrum activity. Nanotechnology has been used to increase the aPDT effectiveness by improving the photosensitizer’s delivery properties. NewPS is a simple, versatile, and safe surfactant-free nanoemulsion with a porphyrin salt shell encapsulating a food-grade oil core with promising photodynamic action. This study evaluated the use of NewPS for aPDT against microorganisms in planktonic, biofilm, and in vivo models of infected wounds. First, the potential of NewPS-mediated aPDT to inactivate Streptococcus pneumoniae and Staphylococcus aureus suspensions was evaluated. Then, a series of protocols were assessed against S. aureus biofilms by means of cell viability and confocal microscopy. Finally, the best biofilm protocol was used for the treatment of S. aureus in a murine-infected wound model. A high NewPS-bacteria cell interaction was achieved since 0.5 nM and 30 J/cm 2 was able to kill S. pneumoniae suspension. In the S. aureus biofilm, enhanced efficacy of NewPS-aPDT was achieved when 100 µM of NewPS was applied with longer periods of incubation at the light dose of 60 J/cm 2 . The best single and double-session protocol reduced 5.56 logs and 6.03 logs, respectively, homogeneous NewPS distribution, resulting in a high number of dead cells after aPDT. The in vivo model showed that one aPDT session enabled a reduction of 6 logs and faster tissue healing than the other groups. In conclusion, NewPS-aPDT may be considered a safe and effective anti-biofilm antimicrobial photosensitizer.

Perspectives on this Article from Objective Detection and Delineation of Oral Neoplasia Using Autofluorescence Imaging

Organ transplantations have an increasing medical relevance. It is becoming a regular procedure with an increase in individuals waiting for organs.The increase in the number of discarded organs is mostly due to the donor bacterial and/or viral infection. In this article, we are demonstrating the feasibility of reduction of the bacterial load in kidney model by using Ultraviolet-C (UV-C) as a germicidal agent in circulating liquids. Using Staphylococcus aureus as a bacteria model, we were able to demonstrate that in less than 30 min of liquid circulation and associated to irradiation, the bacterial load of the perfusate Custodiol® HTK, histidine-tryptophan-ketoglutarate (solution with 5 log CFU mL

This is supportinginformation including additional data and further discussion on the manuscript "" by Vollet-Filho et al., accepted for publishing in Acta Cirurgica Brasileira in October, 2022. Purpose: Fluorescence spectroscopy techniques have been investigated aiming to reduce the invasiveness of methods for investigation of tissue. In transplantation procedures, it may offer the possibility of a complementary technique for the monitoring of liver grafts’ conditions prior to and during the transplantation procedure stages involving cold perfusion. The objective of this study was to evaluate fluorescence spectroscopy under violet light excitation (408 nm) for the monitoring of clinical hypothermic liver transplantation procedures. Methods: Organ grafts were monitored from before the removal of the donor’s body to 1 h after the implant into the receptor’s body. Fluorescence spectroscopy was assessed over five stages within these transplant stages. Results: The study provided evidence of a correlation between fluorescence information collected during liver grafts transplantation and the survival of patients. Conclusion: Fluorescence spectroscopy can become a tool to monitor transplantation grafts, providing objective information for the final decision of surgeons to use organs. Funded by: - Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) - Centro de Pesquisa, Inovação e Difusão/Centro de Pesquisas em Óptica e Fotônica (Grants No.s #2013/07276-1 and No. #98/14270-8) - Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) - Instituto Nacional de Ciência e Tecnologia/Instituto Nacional de Fotônica INCT/INOF program (Grant No. #573587/2008-6)

In the context of the rapid increase of antibiotic-resistant infections, in particular of pneumonia, antimicrobial photodynamic therapy (aPDT), the microbiological application of photodynamic therapy (PDT), comes in as a promising treatment alternative since the induced damage and resultant death are not dependent on a specific biomolecule or cellular pathway. The applicability of aPDT using the photosensitizer indocyanine green with infrared light has been successfully demonstrated for different bacterial agents in vitro, and the combination of pulmonary delivery using nebulization and external light activation has been shown to be feasible. However, there has been little progress in obtaining sufficient in vivo efficacy results. This study reports the lung surfactant as a significant suppressor of aPDT in the lungs. In vitro, the clinical surfactant Survanta® reduced the aPDT effect of indocyanine green, Photodithazine®, bacteriochlorin-trizma, and protoporphyrin IX against Streptococcus pneumoniae . The absorbance and fluorescence spectra, as well as the photobleaching profile, suggested that the decrease in efficacy is not a result of singlet oxygen quenching, while a molecular dynamics simulation showed an affinity for the polar head groups of the surfactant phospholipids that likely impacts uptake of the photosensitizers by the bacteria. Methylene blue is the exception, likely because its high water solubility confers a higher mobility when interacting with the surfactant layer. We propose that the interaction between lung surfactant and photosensitizer must be taken into account when developing pulmonary aPDT protocols.

Summary Background A substantial proportion of organ donors test positive for hepatitis C virus (HCV) infection. To date, only a few studies have evaluated the safety of using lungs from these donors for transplantation, and no direct interventions to donor organs have been done with the aim of preventing HCV transmission via organ transplantation. We aimed to assess the safety and efficacy of lung transplantation in humans from HCV-positive donors to HCV-negative recipients after application of ex-vivo lung perfusion (EVLP) plus ultraviolet C (UVC) perfusate irradiation. Methods We did a single centre, prospective, open-label, non-randomised trial in which donor lungs from HCV-viraemic donors (HCV-positive) were transplanted into HCV-negative recipients at Toronto General Hospital, University Health Network (Toronto, ON, Canada). Donors were younger than 65 years old and tested positive for HCV by nucleic acid testing. Donors who tested positive for hepatitis B virus, HIV, human T-lymphotropic virus 1 or 2 were excluded. Recipients were on the lung transplant waiting list without significant liver disease (stage 2 fibrosis or higher were excluded) or active HCV infection. Before implantation, all HCV-positive donor lungs were treated with EVLP with or without UVC perfusate irradiation to reduce the concentration of HCV RNA and infectivity. For the first week after transplantation, patients' HCV RNA blood concentrations were measured once daily, then once per week for 12 weeks. All patients received 12 weeks of oral sofosbuvir 400 mg plus velpatasvir 100 mg, starting at least 2 weeks after transplantation. The primary endpoint was a composite of survival and HCV-free status at 6 months after transplantation in all patients who received HCV-positive lungs. Patient outcomes such as survival, time in hospital, and incidence of acute rejection were compared between those receiving HCV-positive lungs and all patients who received HCV-negative lung transplants during the study period. The study is registered with ClinicalTrials.gov, NCT03112044. Findings From Oct 1, 2017, to Nov 1, 2018, 209 patients had a transplantation; of 27 donors who were HCV-positive and initially considered, 22 were suitable for transplantation. The remaining 187 donors were HCV-negative. Before implantation, 11 of the HCV-positive donor lungs were treated with EVLP alone and the other 11 were treated with EVLP plus UVC. Lung disease, urgency status, and positive donor-recipient HLA crossmatch were similar between the patients who received HCV-positive and HCV-negative lungs. 20 (91%) patients in the HCV-positive group developed HCV viraemia within the first week after transplantation and had sofosbuvir plus velpatasvir treatment, starting at a median of 21 days after transplantation (IQR 16·76–24·75). Donor organ treatment with EVLP plus UVC was associated with significantly lower recipient viral loads in blood within the first week after transplantation than with EVLP alone (median of 167 IU/mL [IQR 20–12 000] vs 4390 IU/mL [1170–112 000] at day 7; p=0·048) and prevented transmission in two (18%) of 11 patients. All 20 infected patients achieved negative HCV PCR within 6 weeks of treatment initiation. The primary endpoint of survival and HCV-free status at 6 months after transplantation was achieved in 19 (86%) of 22 patients in the HCV-positive group. 6-month survival was 95% in recipients receiving lungs from HCV-viraemic donors versus 94% in recipients receiving lungs from HCV-negative donors. The most common grade 3–4 adverse events in the HCV-positive group were respiratory complications (five [23%]) and infections (four [18%]). Serious adverse events requiring admission to hospital occurred in ten (45%) patients. One (5%) patient who did not develop HCV infection died at day 31 from multiorgan failure related to pseudomonas sepsis. Two patients presented with HCV relapse within 3 months after sofosbuvir plus velpatasvir completion and required retreatment. Interpretation Early and intermediate clinical outcomes were not significantly different between patients receiving viraemic HCV donor lungs and HCV-negative donor lungs. Donor organ treatment with UVC perfusate irradiation during EVLP significantly decreased HCV viral loads within the first 7 days after transplantation and shows the proof-of-concept for a novel approach of minimising viral load ex vivo before transplantation, with intent of preventing donor–recipient transmission. Funding Canadian Institutes of Health Research.

Accurate early diagnosis of malignant skin lesions is critical in providing adequate and timely treatment; unfortunately, initial clinical evaluation of similar-looking benign and malignant skin lesions can result in missed diagnosis of malignant lesions and unnecessary biopsy of benign ones.To develop and validate a label-free and objective image-guided strategy for the clinical evaluation of suspicious pigmented skin lesions based on multispectral autofluorescence lifetime imaging (maFLIM) dermoscopy.We tested the hypothesis that maFLIM-derived autofluorescence global features can be used in machine-learning (ML) models to discriminate malignant from benign pigmented skin lesions. Clinical widefield maFLIM dermoscopy imaging of 41 benign and 19 malignant pigmented skin lesions from 30 patients were acquired prior to tissue biopsy sampling. Three different pools of global image-level maFLIM features were extracted: multispectral intensity, time-domain biexponential, and frequency-domain phasor features. The classification potential of each feature pool to discriminate benign versus malignant pigmented skin lesions was evaluated by training quadratic discriminant analysis (QDA) classification models and applying a leave-one-patient-out cross-validation strategy.Classification performance estimates obtained after unbiased feature selection were as follows: 68% sensitivity and 80% specificity with the phasor feature pool, 84% sensitivity, and 71% specificity with the biexponential feature pool, and 84% sensitivity and 32% specificity with the intensity feature pool. Ensemble combinations of QDA models trained with phasor and biexponential features yielded sensitivity of 84% and specificity of 90%, outperforming all other models considered.Simple classification ML models based on time-resolved (biexponential and phasor) autofluorescence global features extracted from maFLIM dermoscopy images have the potential to provide objective discrimination of malignant from benign pigmented lesions. ML-assisted maFLIM dermoscopy could potentially assist with the clinical evaluation of suspicious lesions and the identification of those patients benefiting the most from biopsy examination.

The aim of this study was to use microscopic and molecular techniques to evaluate the effects of a single session of antimicrobial photodynamic therapy (aPDT) on the alveolar repair process after tooth extraction in rats. The study sample included 84 rats divided into four groups, as follows: a) Control - untreated socket; b) Laser - socket treated using photobiomodulation; c) TBO - socket treated with topic application of the photosensitizer agent, toluidine blue O (TBO); and d) aPDT - socket treated with TBO and laser irradiation. An additional rat was used for thermal mapping during socket irradiation. The animals were euthanatized at 6, 15, and 28 days after unilateral extraction of the upper incisor. Quantitative and qualitative analyses of the connective and bone tissues, blood clot, blood vessel, and inflammatory infiltrate were performed, and real-time polymerase chain reaction was used to study the expression of genes (collagen type I, osteocalcin, alkaline phosphatase [ALP], runt-related transcription factor 2 [RUNX2], and vascular endothelial growth factor [VEGF]) involved in the bone healing process. No statistically significant differences in microscopic and molecular outcomes were observed between the groups (p > 0.05). A positive correlation was seen to exist between blood clot and VEGF (p = 0.000), and a negative correlation was observed between bone tissue and ALP (p = 0.028) and blood vessel and VEGF (p = 0.018). A single session of aPDT in the dental extraction site did not influence the alveolar repair process in rats.

Photodynamic therapy (PDT) has been reported as an excellent option for the treatment of small nodular basal cell carcinomas (nBCC). The standard protocol consists of two sessions, one week apart. Sometimes, returning to the hospital after one week can be impractical for elderly patients, due to comorbidities and mobility issues. Therefore, a new technique performed in one day could be superior for those patients.Evaluate the effectiveness of a PDT Single-visit protocol comparing to the standard protocol, as well as pain and long-term recurrence-free follow-up for nBCC.A total of 120 nBCC were treated through a Standard PDT protocol(two sessions, one week apart), and 120 nBCC were treated through a Single-visit PDT(two sessions in one day). A 30-day-after biopsy was performed in order to evaluate the results after the treatment. The lesions that had successful treatment were clinically and dermoscopically evaluated every 6 months up to 60 months. The pain score was compared between the groups(assessed every 3 min during PDT).A complete response at 30-days-after PDT biopsy was observed in 85% of Standard PDT and in 93.3% of Single-visit PDT. Regarding the pain during the illumination, less pain was observed during the second session of the Single-visit PDT. The recurrence-free follow up showed, after 60 months, an 69.0% cumulative probability of recurrence-free for Standard PDT and 80.6% for Single-visit PDT.The suggested Single-visit PDT protocol resulted in better outcomes at 30-day-after PDT biopsy and in lower recurrence rates than the Standard PDT protocol. A more comfortable and more efficient treatment was offered for the patients, with lower pain.

Made available in DSpace on 2022-04-29T08:36:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-03-01 Background: Photodynamic therapy with photosensitizers can reduce the microbial load. However, few studies have evaluated the effects of photosensitizers on the color stability of endodontically treated intraradicular dentin. This in vitro study investigated the effects of methylene blue and curcumin photosensitizers used in photodynamic therapy on the color stability of intraradicular dentin. Methods: Sixty human incisors were divided into three experimental groups according to the photosensitizer solutions used and their concentrations: methylene blue (25 mg/L), curcumin (1000 mg/L), and curcumin (1500 mg/L). The color stability of endodontically treated intraradicular dentin was evaluated using a portable reflectance spectrophotometer before and after the samples been storage in 2 mL of the photosensitizer solutions during 5 min (n = 20). Color stability data were subjected to a normality test, and statistical analysis was performed using the one-way analysis of variance and Tukey least significant difference test (α = 0.05). Results: Samples treated with 25 mg/L methylene blue photosensitizer showed a higher level of color alteration than those stored in 1000 mg/L curcumin (p = .03322). However, there was no significant difference in the color alteration profiles between the samples treated with 25 mg/L methylene blue and 1500 mg/L curcumin (p = .36428). Furthermore, there was no difference in the color alteration profiles between the dentin samples immersed in 1000 mg/L and 1500 mg/L curcumin photosensitizer solutions (p = .45321). Conclusions: Methylene blue and curcumin photosensitizers influenced the color stability of endodontically treated intraradicular dentin, and this color alteration exceeded the clinical acceptability threshold. Samples treated with 25 mg/L methylene blue showed the highest level of color alteration. Department of Preventive and Restorative Dentistry Discipline of Endodontics Araçatuba School of Dentistry São Paulo State University - UNESP Department of Dental Materials and Prosthodontics Araçatuba School of Dentistry São Paulo State University - UNESP Optic Group Physics Institute of São Carlos - IFSC University of São Paulo – USP Department of Preventive and Restorative Dentistry Discipline of Endodontics Araçatuba School of Dentistry São Paulo State University - UNESP Department of Dental Materials and Prosthodontics Araçatuba School of Dentistry São Paulo State University - UNESP

Made available in DSpace on 2020-12-11T05:11:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Background: Blood can be the target of microbial cells in the human body. Erythrocytes, platelets, and plasma concentrates in blood bags used in hemotherapy for blood transfusion are contamination targets, which can trigger serious diseases in blood. These infections can cause septicemia that can lead to death if not recognized rapidly and treated adequately. The aim of this study was to evaluate the photodynamic inactivation in the in vitro decontamination of Staphylococcus aureus in whole blood, erythrocytes and platelet-rich plasma. Methods: Photodynamic inactivation using light doses of 10, 15 and 30 J/cm(2) at 630 nm and an hematoporphyrin-derivative photosensitizer (Photogem (R)) solutions at 25 and 50 mu g/mL were evaluated. Toxicity of treatment was determined by hemolysis and cell viability assays. Results: The S. aureus reduction in phosphate buffered saline (PBS), whole blood, erythrocytes and platelet-rich plasma at 15 J/cm(2) and 50 mu g/mL were 7.2, 1.0, 1.3 and 0.4 log CFU/mL, respectively. Quantitative and qualitative analyses were performed in whole blood samples, and Photogem (R) showed a low risk of hemolysis (10.7%) in whole blood. However, 100% of erythrocytes suffered hemolysis in the absence of plasma. The cell viability assay showed 13.9% of apoptosis in erythrocytes, but normal platelet viability. Conclusion: S. aureus inactivation of whole blood samples using 50 mu g/mL Photogem (R) and 15 J/cm(2) resulted in better outcomes, providing promising indications for treatment of bacterial contamination of blood, and in this work, alternative possibilities to apply the technique for blood decontamination are discussed. Univ Fed Sao Carlos, PPG Biotec, BR-13565905 Sao Carlos, SP, Brazil Univ Sao Paulo, Sao Carlos Inst Phys, POB 369,Av Trabalhador Sao Carlense, BR-13565905 Sao Carlos, SP, Brazil Sao Paulo State Univ, Botucatu Med Sch, BR-18618687 Botucatu, SP, Brazil Sao Paulo State Univ, Botucatu Med Sch, BR-18618687 Botucatu, SP, Brazil FAPESP: 2013/07276-1 CAPES: 1500213

Availability of organs is a limiting factor for lung transplantation, leading to substantial mortality rates on the wait list. Use of organs from donors with transmissible viral infections, such as hepatitis C virus (HCV), would increase organ donation, but these organs are generally not offered for transplantation due to a high risk of transmission. Here, we develop a method for treatment of HCV-infected human donor lungs that prevents HCV transmission. Physical viral clearance in combination with germicidal light-based therapies during normothermic ex-vivo Lung Perfusion (EVLP), a method for assessment and treatment of injured donor lungs, inactivates HCV virus in a short period of time. Such treatment is shown to be safe using a large animal EVLP-to-lung transplantation model. This strategy of treating viral infection in a donor organ during preservation could significantly increase the availability of organs for transplantation and encourages further clinical development., Organs from donors with transmissible viral infections, such as hepatitis C virus (HCV), are not offered for transplantation due to a high risk of transmission. Here, Galasso et al. develop a method for treatment of HCV-infected human donor lungs that is safe and prevents HCV transmission in the pig model.

This study evaluated curcumin incubation time in photodynamic inactivation with bacteria of different wall compositions: Staphylococcus aureus and Pseudomonas aeruginosa. Bacteria samples were cultured for 24h (37°C). Curcumin was dissolved in dimethylsulphoxide and subsequently diluted for final concentrations. Fluence used was 30J/cm2 (at 460nm excitation). After incubation with photosensitizer and irradiation, ten-fold serial dilutions were plated and incubated at 37°C for 48h for CFU counting. Treatment reduced approximately 7 logs of 1µM S. aureus after 30min incubation, whereas 2 logs only for P. aeruginosa at 50µM maximum concentration. Photosensitizer removed from the medium before irradiation resulted in lower reductions.

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 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.

Melanoma is the most aggressive type of skin cancer and a relevant health problem due to its poor treatment response with high morbidity and mortality rates. This study, aimed to investigate the tissue changes of an improved photodynamic therapy (PDT) response when combined with optical clearing agent (OCA) in the treatment of cutaneous melanoma in mice. Photodithazine (PDZ) was administered intraperitoneally and a solution of OCA was topically applied before PDT irradiation. Due to a resultant refractive index matching, OCA-treated tumors are more optically homogenous, improving the PDT response. Raman analysis revealed, when combined with OCA, the PDT response was more homogenous down to 725 µm-depth in thickness.

Cancer remains a main public health issue and the second cause of mortality worldwide. Photodynamic therapy is a clinically approved therapeutic option. Effective photodynamic therapy induces cancer damage and death through a multifactorial manner including reactive oxygen species-mediated damage and killing, vasculature damage, and immune defense activation. Anticancer efficiency depends on the improvement of photosensitizers drugs used in photodynamic therapy, their selectivity, enhanced photoproduction of reactive species, absorption at near-infrared spectrum, and drug-delivery strategies. Both experimental and clinical studies using first- and second-generation photosensitizers had pointed out the need for developing improved photosensitizers for photodynamic applications and achieving better therapeutic outcomes. Bioconjugation and encapsulation with targeting moieties appear as a main strategies for the development of photosensitizers from their precursors. Factors influencing cellular biodistribution and uptake are briefly discussed, as well as their roles as cancer diagnostic and therapeutic (theranostics) agents. The two-photon photodynamic approach using third-generation photosensitizers is present as an attempt in treating deeper tumors. Although significant advances had been made over the last decade, the development of next-generation photosensitizers is still mainly in the developmental stage.

Photodynamic therapy (PDT) has been used to treat certain types of non-melanoma skin cancer with promising results. However, some skin lesions have not fully responded to this treatment, suggesting a potential PDT-resistant phenotype. Therefore, novel therapeutic alternatives must be identified that improve PDT in resistant skin cancer. In this study, we analyzed the cell viability, intracellular protoporphyrin IX (PpIX) content and subcellular localization, proliferation profile, cell death, reactive oxygen species (ROS) detection and relative gene expression in PDT-resistant HSC-1 cells. PDT-resistant HSC-1 cells show a low quantity of protoporphyrin IX and low levels of ROS, and thus a low rate of death cell. Furthermore, the resistant phenotype showed a downregulation of HSPB1, SLC15A2, FECH, SOD2 and an upregulation of HMBS and BIRC5 genes. On the other hand, epigallocatechin gallate catechin enhanced the MAL-PDT effect, increasing levels of protoporphyrin IX and ROS, and killing 100% of resistant cells. The resistant MAL-PDT model of skin cancer squamous cells (HSC-1) is a reliable and useful tool to understand PDT cytotoxicity and cellular response. These resistant cells were successfully sensitized with epigallocatechin gallate catechin. The in vitro epigallocatechin gallate catechin effect as an enhancer of MAL-PDT in resistant cells is promising in the treatment of difficult skin cancer lesions.

Basal cell carcinoma (BCC) is the most common type of skin cancer. Diagnosis and edge assessment of BCC lesions are based on clinical and dermoscopy evaluation, which are strongly dependent on the expertise and training of the physician. There is a high rate of underdiagnosis because BCC is frequently confused with certain common benign lesions and is often indistinguishable from the surrounding healthy tissue. In the present study, a multispectral fluorescence lifetime imaging (FLIm) dermoscopy system, designed for imaging and analyzing the autofluorescence emission of skin tissue, was used to image thirty-eight patients with diagnosed nodular BCC (nBCC) lesions, using clinically acceptable levels of excitation light exposure. With this system, skin autofluorescence was imaged simultaneously using three emission bands: 390 ± 20 nm, 452 ± 22 nm, and >496 nm, preferentially targeting collagen, NADH, and FAD autofluorescence, respectively. Statistical classifiers based on FLIm features developed to discriminate BCC from healthy tissue showed promising performance (ROC area-under-the-curve of 0.82). This study demonstrates the feasibility of clinically performing multispectral endogenous FLIm dermoscopy providing baseline results indicating the potential of this technology as an image-guided tool to improve the delineation of nBCC during surgical lesion resection.

Every year more than 5.4 million new cases of skin cancer are reported in the US. Melanoma is the most lethal type with only 5% occurrence rate, but accounts for over 75% of all skin cancer deaths. Non-melanoma skin cancer, especially basal cell carcinoma (BCC) is the most commonly occurring and often curable type that affects more than 3 million people and causes about 2000 deaths in the US annually. The current diagnosis involves visual inspection, followed by biopsy of the lesions. The major drawbacks of this practice include difficulty in border detection causing incomplete treatment and, the inability to distinguish between clinically similar lesions. Melanoma is often mistaken for the benign lesion pigmented seborrheic keratosis (pSK), making it extremely important to differentiate benign and malignant lesions. In this work, a novel feature extraction algorithm based on phasors was performed on the Fluorescence Lifetime Imaging (FLIM) images of the skin to reliably distinguish between benign and malignant lesions. This approach, unlike the standard FLIM data processing method that requires time-deconvolution of the instrument response from the measured time-resolved fluorescence signal, is computationally much simpler and provides a unique set of features for classification. Subsequently, FLIM derived features were selected using a double step cross validation approach that assesses the reliability and the performance of the resultant trained classifier. Promising FLIM-based classification performance was attained for detecting benign from malignant pigmented (sensitivity: ~80%, specificity: 79%, overall accuracy: ~79%) and nonpigmented (sensitivity: ~88%, specificity: 83%, overall accuracy: ~87%) lesions.

Treatment using light-activated photosensitizers (photodynamic therapy, PDT) has shown limited efficacy in pigmented melanoma, mainly due to the poor penetration of light in this tissue. Here, an optical clearing agent (OCA) was applied topically to a cutaneous melanoma model in mice shortly before PDT to increase the effective treatment depth by reducing the light scattering. This was used together with cellular and vascular-PDT, or a combination of both. The effect on tumor growth was measured by longitudinal ultrasound/photoacoustic imaging in vivo and by immunohistology after sacrifice. In a separate dorsal window chamber tumor model, angiographic optical coherence tomography (OCT) generated 3D tissue microvascular images, enabling direct in vivo assessment of treatment response. The optical clearing had minimal therapeutic effect on the in control, non-pigmented cutaneous melanomas but a statistically significant effect (p &lt, 0.05) in pigmented lesions for both single- and dual-photosensitizer treatment regimes. The latter enabled full-depth eradication of tumor tissue, demonstrated by the absence of S100 and Ki67 immunostaining. These studies are the first to demonstrate complete melanoma response to PDT in an immunocompromised model in vivo, with quantitative assessment of tumor volume and thickness, confirmed by (immuno) histological analyses, and with non-pigmented melanomas used as controls to clarify the critical role of melanin in the PDT response. The results indicate the potential of OCA-enhanced PDT for the treatment of pigmented lesions, including melanoma.

In this chapter, we introduce the fundamentals of photodynamic reactions and applications for the treatment of oral-facial lesions, and microbiological control are presented.