Laser-induced modification of the patellar ligament tissue: comparative study of structural and optical changes

The effects of non-ablative infrared (IR) laser treatment of collagenous tissue have been commonly interpreted in terms of collagen denaturation spread over the laser-heated tissue area. In this work, the existing model is refined to account for the recently reported laser-treated tissue heterogeneity and complex collagen degradation pattern using comprehensive optical imaging and calorimetry toolkits. Patella ligament (PL) provided a simple model of type I collagen tissue containing its full structural content from triple-helix molecules to gross architecture. PL ex vivo was subjected to IR laser treatments (laser spot, 1.6 mm) of equal dose, where the tissue temperature reached the collagen denaturation temperature of 60 ± 2°C at the laser spot epicenterin the first regime, and was limited to 67 ± 2°C in the second regime. The collagen network was analyzed versus distance from the epicenter. Experimental characterization of the collagenous tissue at all structural levels included cross-polarization optical coherence tomography, nonlinear optical microscopy, light microscopy/histology, and differential scanning calorimetry. Regressive rearrangement of the PL collagen network was found to spread well outside the laser spot epicenter (>2 mm) and was accompanied by multilevel hierarchical reorganization of collagen. Four zones of distinct optical and morphological properties were identified, all elliptical in shape, and elongated in the direction perpendicular to the PL long axis. Although the collagen transformation into a random-coil molecular structure was occasionally observed, it was mechanical integrity of the supramolecular structures that was primarily compromised. We found that the structural rearrangement of the collagen network related primarily to the heat-induced thermo-mechanical effects rather than molecular unfolding. The current body of evidence supports the notion that the supramolecular collagen structure suffered degradation of various degrees, which gave rise to the observed zonal character of the laser-treated lesion

Norepinephrine Effects on Uropathogenic Strains Virulence

The degree of virulence correlates with adhesion, biofilm formation, motility and the capacity to quickly colonize biological surfaces. The virulence of the bacteria that have colonized the urinary tract may be modified by substances dissolved in urine. One such substance is the norepinephrine (NE) hormone, which may be present in human urine, especially in times of stress and under changes in the activity of the renin-angiotensin-aldesterone system. In this work, we study the influence of NE on the biomass, biofilm formation, matrix production, adhesion, motility and metabolism of uropathogenic strains of E. coli and S. aureus. We used Congo red and gentian violet staining for detection of matrix and biomass formation, respectively. The optical density was measured by a multichannel spectrophotometer. The motility of bacterial cells was measured on semi-solid agar at 24 h and 48 h. The metabolic activity was analyzed by MTT assay. It was shown that the metabolic activity of E. coli was stimulated by NE, which led to the increasing synthesis of virulence factors such as biofilm production, adhesion, and motility. At the same time, NE did not activate the S. aureus strain’s metabolism and did not change its adhesion and motility. Thus, the virulence activity of uropathogenic E. coli may be modified by NE in urine

Investigation of the Effect of Temperature Stabilization in Radiation–Heat Converters Based on a Strong Absorbing Coating

Light–heat converters are promising for further development of contact laser surgery. The simplest converter consists of an optical fiber with a strong absorbing layer at the tip. We studied the time dependence of the tip temperature at different CW laser powers and revealed that, in several seconds, the temperature evolution becomes almost power-independent. Mathematical modeling showed that laser ablation of the tip coating is the main reason for this phenomenon

Post-Operational Photodynamic Therapy of the Tumor Bed: Comparative Analysis for Cold Knife and Laser Scalpel Resection

In this paper, we report on a study regarding the efficiency of the post-operational phototherapy of the tumor bed after resection with both a cold knife and a laser scalpel in laboratory mice with CT-26 tumors. Post-operational processing included photodynamic therapy (PDT) with a topically applied chlorin-based photosensitizer (PS), performed at wavelengths of 405 or 660 nm, with a total dose of 150 J/cm2. The selected design of the tumor model yielded zero recurrence in the laser scalpel group and 92% recurrence in the cold knife group without post-processing, confirming the efficiency of the laser scalpel in oncology against the cold knife. The application of PDT after the cold knife resection decreased the recurrence rate to 70% and 42% for the 405 nm and 660 nm procedures, respectively. On the other hand, the application of PDT after the laser scalpel resection induced recurrence rates of 18% and 30%, respectively, for the considered PDT performance wavelengths. The control of the penetration of PS into the tumor bed by fluorescence confocal microscopy indicated the deeper penetration of PS in the case of the cold knife, which presumably provided deeper PDT action, while the low-dose light exposure of deeper tissues without PS, presumably, stimulated tumor recurrence, which was also confirmed by the differences in the recurrence rate in the 405 and 660 nm groups. Irradiation-only light exposures, in all cases, demonstrated higher recurrence rates compared to the corresponding PDT cases. Thus, the PDT processing of the tumor bed after resection could only be recommended for the cold knife treatment and not for the laser scalpel resection, where it could induce tumor recurrence

Efficacy of Photodynamic Inactivation against the Major Human Antibiotic-Resistant Uropathogens

Photodynamic inactivation (PDI) is considered to be an effective method of prevention of postoperative complications of urolithiasis. The present study shows a complex approach to assess the efficacy of PDI of drug resistant bacteria associated with renal calculi. Bacterial strains associated with renal calculi were isolated and identified using standard methods of bacteriological analysis and tested for drug resistance to 10 antibiotics by the disco-diffusion method. Uropathogenic bacterial strains present in 78.7 ± 5.2% of the infected samples from the total number of analyzed calculi. The most frequent representatives belonged to the genera Staphylococcus, Escherichia, and Enterococcus. All tested strains showed high antibiotic resistance. Representatives of the most common bacterial genera in the calculi were used as models for the selection of PD exposure modes. It was found that the maximum time of photosensitizer accumulation depends on the structure of the bacterial cell wall: 30 min for Gram-negative strains and 60 min for Gram-positive ones. Optimal modes of PD exposure to antibiotic-resistant uropathogenic microorganisms were selected: 50 µg/mL Fotoditazin and 150 mW laser power. The maximal bactericidal activity of PDI against uropathogenic microorganisms was shown for Enterococcus faecalis, and Staphylococcus aureus. The bacteriostatic effect was found against Escherichia coli and Proteus mirabilis

Efficacy of Photodynamic Inactivation against the Major Human Antibiotic-Resistant Uropathogens

Photodynamic inactivation (PDI) is considered to be an effective method of prevention of postoperative complications of urolithiasis. The present study shows a complex approach to assess the efficacy of PDI of drug resistant bacteria associated with renal calculi. Bacterial strains associated with renal calculi were isolated and identified using standard methods of bacteriological analysis and tested for drug resistance to 10 antibiotics by the disco-diffusion method. Uropathogenic bacterial strains present in 78.7 ± 5.2% of the infected samples from the total number of analyzed calculi. The most frequent representatives belonged to the genera Staphylococcus, Escherichia, and Enterococcus. All tested strains showed high antibiotic resistance. Representatives of the most common bacterial genera in the calculi were used as models for the selection of PD exposure modes. It was found that the maximum time of photosensitizer accumulation depends on the structure of the bacterial cell wall: 30 min for Gram-negative strains and 60 min for Gram-positive ones. Optimal modes of PD exposure to antibiotic-resistant uropathogenic microorganisms were selected: 50 µg/mL Fotoditazin and 150 mW laser power. The maximal bactericidal activity of PDI against uropathogenic microorganisms was shown for Enterococcus faecalis, and Staphylococcus aureus. The bacteriostatic effect was found against Escherichia coli and Proteus mirabilis.</i

Diode Laser Lithotription Technique Based on Optothermal Converter

Purpose: evaluation of the efficiency of the “hot spot” method for the fragmentation of urinary stones. Materials and methods: A retrospective analysis of clinical records of 1666 patients with urolithiasis who underwent percutaneous nephrolithoextraction/tripsy and contact ureterolithotripsy/extraction in the period from 2014 to 2017 at the urology clinic was performed to assess the incidence of postoperative infectious and inflammatory complications. The research objects were postoperative urinary stones (n-78). The X-ray density and linear dimensions of the stones were determined. Stone fragmentation was performed with a continuous-wave diode laser operating at wavelengths of 0.81 μm, 0.97 μm, and 1.47 μm. An absorbing coating of micro-size graphite powder was applied on the working tip of the optical fiber. In vitro fragmentation was carried out in liquid. Results: A group of patients (224/1666) (13.4 ± 0.86%) was identified, who developed infectious and inflammatory complications after: percutaneous nephrolithotripsy, 123/361 (34.1 ± 2.5%) cases; percutaneous nephrolithoextraction, 59/240 (24.6 ± 2.78%); contact ureterolithotripsy, 23/294 (7.8 ± 1.57%); and ureterolithoextraction, 19/771 (2.5 ± 0.56%). In liquid, the “hot spot” technique made it possible to fragment stones with an X-ray density of up to 1000 HU at a laser wavelength of 0.81 µm, up to 1400 HU at 0.97 µm, and up to 1400 HU at 1.47 µm

The bactericidal effect of continuous wave laser with strongly absorbing coating at the fiber tip

The bactericidal effect of laser radiation with a quartz fiber-based transmission system with a strong absorption coating converter against bacteria associated with urological stones has been studied. Gram-negative rod Escherichia coli and the Gram-positive coccus Staphylococcus epidermidis, Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium were used in this study. Each bacterial species was treated by continuous-wave near infrared laser coupled with bare fiber tip or strongly absorption coating fiber tip. After treatment, the temperature of bacterial suspension was measured. In addition, the temperature distribution was analyzed. It has been shown that using laser with a strongly absorption coating fiber tip results in significant bactericidal effect. The decrease of the amount of E. coli and S. epidermidis was 100% after treatment with an output power of 6W of radiation at a wavelength of 0.97μm for 40s. Number of S. aureus and Ent. faecium colony-forming unit was reduced to 70% after same exposure. The peak temperature of bacterial suspension was 86∘C after treatment by laser with a strongly absorption coating fiber tip. Laser with a strongly absorption coating fiber tip provides large-scale hydrodynamic flows directed away from the fiber tip. The laser with a strongly absorption coating fiber tip has bactericidal effect. The main role is associated with the effect of high temperature, which, in the form of flow in a liquid medium, affects bacteria