Rose Bengal and Future Directions in Larynx Tumor Photodynamic Therapy.

Photodynamic Therapy (PDT) seems to be a promising method in the treatment of larynx tumor tissues. The aim of the present analysis was the study of photosensitizer penetration of larynx tissue associated with the application of PDT in vitro. This study is based on the use of photosensitive compounds Rose Bengal (RB) that selectively accumulate in larynx tissue. The selection of the study group of patients who will undergo surgery in accordance with medical principles was of key importance for the project. Histopathological examination of samples subjected to PDT revealed numerous changes in the morphology of the cancer cells and surrounding tissues. After PDT treatment, the number of tumor cells decreased compared with the cells number before PDT and the arrangement was relatively loose. After PDT with RB the nuclei morphology was incomplete and fragmented. The effects of the applied PDT of larynx in vitro were assessed under an optical microscope. The future directions in larynx tumor PDT with the use of upconversion nanoparticles (UPCNP) is also discussed.

The use of magnetic resonance imaging technique and 3D printing in order to develop a three-dimensional fistula model for patients with Crohn's disease: personalised medicine.

INTRODUCTION: Preoperative evaluation of magnetic resonance (MR) images may not be sufficient for the precise planning of anal fistula surgery or for stem cell therapy. Three-dimensional (3D) printing allows one to obtain spatial structures in a 1 : 1 scale with unprecedented precision. AIM: To combine magnetic resonance imaging (MRI) with 3D printing for more precise visualisation of perianal Crohn's disease. MATERIAL AND METHODS: MRI at 1.5T and a 3D printer were used. DICOM (Digital imaging and communications in medicine) images were imported into 3D Slicer v.4.8.0. Firstly, anal fistula was modelled on the basis of axial images. Fistula locations, the anus and anal canal, were marked with different coloured markers. The last step was to mark the skin that was connected to the anus and contact areas of the fistula with the skin. The prepared models were then exported to an STL format file. The anal fistula model was printed using a 3D printer. The development of the model, including printing, took approximately 6 h. RESULTS AND CONCLUSIONS: The accessibility of a rotatable 3D model before surgery allows for a more precise detection of the location and the degree of perianal disease. Moreover, this may also lower the inter-observer bias connected with interpretation of complex MR imaging before planned surgery. Development of MRI image transfer to 3D printing and the decreasing cost of 3D printers suggests a promising future of this technology in medical applications.

Alternative methods of photodynamic therapy and oxygen consumption measurements-A review.

Photooxidation generates reactive oxygen species (ROS) through the interaction of dyes or surfaces with light radiation of appropriate wavelength. The reaction is of wide utility and is highly effective in photodynamic therapy (PDT) of various types of cancer and skin disease. Understanding generation of singlet oxygen has contributed to the development of PDT and its subsequent use in vivo. However, this therapy has some limitations that prevent its use in the treatment of cancers located deep within the body. The limited depth of light penetration through biological tissue limits initiation of PDT action in deep tissue. Measurement of oxygen photo consumption is critical due to tumor hypoxia, and use of magnetic resonance imaging (MRI) is particularly attractive since it is non-invasive. This article presents bioluminescence (BL) and chemiluminescence (CL) phenomena based on publications from the last 20 years, and preliminary results from our lab in the use of MRI to measure oxygen concentration in water. Current work is aimed at improving the effectiveness of singlet oxygen delivery to deep tissue cancer.

A Neonatal Murine Escherichia coli Sepsis Model Demonstrates That Adjunctive Pentoxifylline Enhances the Ratio of Anti- vs. Pro-inflammatory Cytokines in Blood and Organ Tissues.

Introduction: Neonatal sepsis triggers an inflammatory response that contributes to mortality and multiple organ injury. Pentoxifylline (PTX), a phosphodiesterase inhibitor which suppresses pro-inflammatory cytokines, is a candidate adjunctive therapy for newborn sepsis. We hypothesized that administration of PTX in addition to antibiotics decreases live bacteria-induced pro-inflammatory and/or enhances anti-inflammatory cytokine production in septic neonatal mice without augmenting bacterial growth. Methods: Newborn C57BL/6J mice (< 24 h old) were injected intravenously with 105 colony forming units (CFUs)/g weight of a bioluminescent derivative of the encapsulated clinical isolate Escherichia coli O18:K1. Adequacy of intravenous injections was validated using in vivo bioluminescence imaging and Evans blue. Pups were treated with gentamicin (GENT), PTX, (GENT + PTX) or saline at 0, 1.5, or 4 h after sepsis initiation, and euthanized after an additional 4 h. CFUs and cytokines were measured from blood and homogenized organ tissues. Results: GENT alone inhibited bacterial growth, IL-1ß, and IL-6 production in blood and organs. Addition of PTX to GENT profoundly inhibited E. coli-induced TNF and enhanced IL-10 in blood of newborn mice at all timepoints, whereas it primarily upregulated IL-10 production in peripheral organs (lung, spleen, brain). PTX, whether alone or adjunctive to GENT, did not increase microbial colony counts in blood and organs. Conclusion: Addition of PTX to antibiotics in murine neonatal E. coli sepsis promoted an anti-inflammatory milieu through inhibition of plasma TNF and enhancement of IL-10 production in plasma and organs without increasing bacterial growth, supporting its utility as a potential adjunctive agent for newborn sepsis.

Monitoring photodynamic oxygen consumption by endogenous oxygen contrast MRI.

Photodynamic oxygen consumption was measured by changes in spin-lattice relaxation time (T1) in aqueous solution in a clinical GE scanner at 1.5 T. Similar measurements were attempted in excised laryngeal and thyroid tissues that were infused with Rose Bengal. First, T1 was measured as a function of dissolved oxygen in argon and in oxygen pre-saturated water samples that were opened to the atmosphere in a series of steps allowing air to diffuse into or out of solution; for both argon and oxygen saturated water solutions, stepwise air re-equilibration resulted in a return to air-saturated water T1. Secondly, T1 was measured as a function of time under type II photooxidative conditions in aqueous solution. Under type II photooxidative conditions, a 492 ± 53 ms increase in T1 was measured following 300 s of visible light illumination of aqueous solutions containing the photosensitizer Rose Bengal (2.5 × 10-6 M) and the singlet oxygen trap methionine (0.0012 M). The 492 ± 53 ms increase in T1 corresponded to consumption of all the measurable dissolved oxygen (˜ 0.1 mg O2 in 15.0 mL of H2O) during photooxidation of methionine in air saturated water. This rapid oxygen consumption, indicated by an increase in T1, is due to irreversible trapping of photogenerated singlet oxygen by methionine. Thirdly, an increase in T1 was observed in Rose Bengal infused normal laryngeal tissue, and in normal and cancerous thyroid tissue samples following 20 min of exposure to visible light. An increase in T1 was not observed after 40 min of illumination which suggests that the increases in T1 observed after 20 min were not due to water uptake, but rather to photoconsumption of interstitial dissolved oxygen.

Pentoxifylline Alone or in Combination with Gentamicin or Vancomycin Inhibits Live Microbe-Induced Proinflammatory Cytokine Production in Human Cord Blood and Cord Blood Monocytes In Vitro.

Neonatal sepsis and its accompanying inflammatory response contribute to substantial morbidity and mortality. Pentoxifylline (PTX), a phosphodiesterase inhibitor which suppresses transcription and production of proinflammatory cytokines, is a candidate adjunctive therapy for newborn sepsis. We hypothesized that PTX decreases live microbe-induced inflammatory cytokine production in newborn blood. Cord blood was stimulated with live microorganisms commonly encountered in newborn sepsis (Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, or Candida albicans) and simultaneously treated with antimicrobial agents (gentamicin, vancomycin, or amphotericin B) and/or clinically relevant concentrations of PTX. Microbial colony counts were enumerated by plating, supernatant cytokines were measured by multiplex assay, intracellular cytokines and signaling molecules were measured by flow cytometry, and mRNA levels were measured by quantitative reverse transcription-PCR. PTX inhibited concentration-dependent E. coli-, S. aureus-, S. epidermidis-, and C. albicans-induced tumor necrosis factor (TNF) and E. coli-induced interleukin-1ß (IL-1ß) production in whole blood, with greater suppression of proinflammatory cytokines in combination with antimicrobial agents. Likewise, PTX suppressed E. coli-induced monocytic TNF and IL-1ß, whereby combined PTX and gentamicin led to significantly greater reduction of TNF and IL-1ß. The anti-inflammatory effect of PTX on microbe-induced proinflammatory cytokine production was accompanied by inhibition of TNF mRNA expression and was achieved without suppressing the production of the anti-inflammatory IL-10. Of note, microbial colony counts in newborn blood were not increased by PTX. Our findings demonstrated that PTX inhibited microbe-induced proinflammatory cytokine production, especially when combined with antimicrobial agents, without enhancing microbial proliferation in human cord blood in vitro, thus supporting its utility as candidate adjunctive agent for newborn sepsis.

Pentoxifylline, dexamethasone and azithromycin demonstrate distinct age-dependent and synergistic inhibition of TLR- and inflammasome-mediated cytokine production in human newborn and adult blood in vitro.

INTRODUCTION: Neonatal inflammation, mediated in part through Toll-like receptor (TLR) and inflammasome signaling, contributes to adverse outcomes including organ injury. Pentoxifylline (PTX), a phosphodiesterase inhibitor which potently suppresses cytokine production in newborn cord blood, is a candidate neonatal anti-inflammatory agent. We hypothesized that combinations of PTX with other anti-inflammatory agents, the steroid dexamethasone (DEX) or the macrolide azithromycin (AZI), may exert broader, more profound and/or synergistic anti-inflammatory activity towards neonatal TLR- and inflammasome-mediated cytokine production. METHODS: Whole newborn and adult blood was treated with PTX (50-200 µM), DEX (10-10-10-7 M), or AZI (2.5-20 µM), alone or combined, and cultured with lipopolysaccharide (LPS) (TLR4 agonist), R848 (TLR7/8 agonist) or LPS/adenosine triphosphate (ATP) (inflammasome induction). Supernatant and intracellular cytokines, signaling molecules and mRNA were measured by multiplex assay, flow cytometry and real-time PCR. Drug interactions were assessed based on Loewe's additivity. RESULTS: PTX, DEX and AZI inhibited TLR- and/or inflammasome-mediated cytokine production in newborn and adult blood, whether added before, simultaneously or after TLR stimulation. PTX preferentially inhibited pro-inflammatory cytokines especially TNF. DEX inhibited IL-10 in newborn, and TNF, IL-1ß, IL-6 and interferon-α in newborn and adult blood. AZI inhibited R848-induced TNF, IL-1ß, IL-6 and IL-10, and LPS-induced IL-1ß and IL-10. (PTX+DEX) synergistically decreased LPS- and LPS/ATP-induced TNF, IL-1ß, and IL-6, and R848-induced IL-1ß and interferon-α, while (PTX+AZI) synergistically decreased induction of TNF, IL-1ß, and IL-6. Synergistic inhibition of TNF production by (PTX+DEX) was especially pronounced in newborn vs. adult blood and was accompanied by reduction of TNF mRNA and enhancement of IL10 mRNA. CONCLUSIONS: Age, agent, and specific drug-drug combinations exert distinct anti-inflammatory effects towards TLR- and/or inflammasome-mediated cytokine production in human newborn blood in vitro. Synergistic combinations of PTX, DEX and AZI may offer benefit for prevention and/or treatment of neonatal inflammatory conditions while potentially limiting drug exposure and toxicity.

Pentoxifylline inhibits TLR- and inflammasome-mediated in vitro inflammatory cytokine production in human blood with greater efficacy and potency in newborns.

BACKGROUND: Toll-like receptor (TLR)-mediated inflammation may contribute to neonatal sepsis, for which pentoxifylline (PTX), a phosphodiesterase inhibitor that raises intracellular cAMP, is a candidate adjunctive therapy. We characterized the anti-inflammatory effects of PTX toward TLR-mediated production of inflammatory (tumor necrosis factor (TNF) and interleukin (IL)-1ß) and proresolution (IL-6 and IL-10) cytokines in human newborn and adult blood. METHODS: Newborn cord and adult blood were treated with PTX (50-400 µmol/l) before, during or after stimulation with LPS (TLR4 agonist), R848 (TLR7/8 agonist) or LPS/ATP (inflammasome activation). Cytokines were measured by multiplex assay (supernatants), intracellular cytokines and signaling molecules by flow cytometry, and mRNA by quantitative real-time PCR. RESULTS: Whether added 2 h pre-, simultaneously to, or 2 h post-TLR stimulation, PTX inhibited TLR-mediated cytokine production in a concentration-dependent manner, with greater efficacy and potency in newborn blood, decreasing intracellular TNF and IL-1ß with relative preservation of IL-10 and IL-6. PTX decreased TLR-mediated TNF mRNA while increasing IL-10 mRNA. Neonatal plasma factors contributed to the anti-inflammatory effects of PTX in newborn blood that were independent of soluble TNF receptor concentrations, p38 MAPK phosphorylation and IĸB degradation. CONCLUSION: PTX is a potent and efficacious inhibitor of TLR-mediated inflammatory cytokines in newborn cord blood and a promising neonatal anti-inflammatory agent.