Nicolas G. Loebel

Reduction of Endotracheal Tube Biofilms Using Antimicrobial Photodynamic Therapy.

Ventilator‐associated pneumonia (VAP) is reported to occur in 12–25% of patients who require mechanical ventilation with a mortality rate of 24–71%. The endotracheal tube (ETT) has long been recognized as a major factor in the development of VAP since biofilm harbored within the ETT become dislodged during mechanical ventilation and have direct access to the lungs. The objective of this study was to demonstrate the safety and effectiveness of a non‐invasive antimicrobial photodynamic therapy (aPDT) treatment method of eradicating antibiotic resistant biofilms from ETT in an in vitro model.

In vitro photodynamic eradication of Pseudomonas aeruginosa in planktonic and biofilm culture.

Photodynamic disinfection (PDD) is a nonantibiotic approach to treating drug‐resistant bacterial infections. Pseudomonas aeruginosa, an opportunistic pathogen, is problematic because of its propensity to develop antibiotic resistance and its ability to secrete a protective biofilm matrix. This study examined the ability of PDD to eradicate planktonic and biofilm cultures of P. aeruginosa in vitro. Planktonic P. aeruginosa cultures were briefly exposed to a methylene blue‐based photosensitizer formulation and subjected to energy doses ranging from 1.7 to 20.6 J cm−2 using a 670 nm nonthermal diode laser. Biofilms were grown for 24 and 48 h and exposed to photosensitizer for 30 s before illumination with 13.2 or 26.4 J of energy. A single exposure of planktonic P. aeruginosa to photosensitizer at >15.5 J cm−2 resulted in 100% eradication (>7 log10 reduction from control), an effect that could be decreased significantly in the presence of the singlet oxygen quenchers l‐tryptophan and sodium azide. Decreasing the energy dose below this threshold by varying both power density and illumination duration resulted in a dose‐dependent decrease in bacterial kill. In addition, 24 h biofilm viability was reduced by 99% with single exposure and 99.9% with double exposure, while 48 h biofilm viability was reduced by >99.999% with both single and double exposures. This study shows that PDD is effective in eradicating planktonic and biofilm cultures of P. aeruginosa, supporting the concept that translation into clinical practice for indications such as otitis externa and wound disinfection is a viable option.

Photodynamic therapy of antibiotic‐resistant biofilms in a maxillary sinus model

Chronic rhinosinusitis (CRS) is one of the most common chronic conditions in the United States. There is a significant subpopulation of CRS patients who remain resistant to cure despite rigorous treatment regimens including surgery, allergy therapy, and prolonged antibiotic therapy. Antimicrobial photodynamic therapy (aPDT) is a noninvasive nonantibiotic broad spectrum antimicrobial treatment. Our previous in vitro studies demonstrated that aPDT reduced CRS polymicrobial planktonic bacteria and fungi by >99.9% after a single treatment. However, prior to human treatment, the effectiveness of aPDT to eradicate polymicrobial biofilms in a maxillary sinus cavity must be demonstrated. The objective of this study was to demonstrate the effectiveness of a noninvasive aPDT treatment of antibiotic resistant biofilms known to cause CRS in a novel anatomically correct maxillary sinus in vitro model using an enhanced photosensitizer solution.

Energy Dose Parameters Affect Antimicrobial Photodynamic Therapy–Mediated Eradication of Periopathogenic Biofilm and Planktonic Cultures

OBJECTIVE This study evaluated the in vitro efficacy of a commercially available aPDT system in eradication of the periopathogens Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans in both planktonic and biofilm cultures. BACKGROUND DATA Antimicrobial photodynamic therapy (aPDT) is an effective antibacterial approach in vitro; however, few data are available regarding effective light-energy parameters. MATERIALS AND METHODS Planktonic and biofilm cultures of periopathogens were exposed to a methylene blue-based formulation and irradiated with a 670-nm nonthermal diode laser. Energy doses were varied from 2.3 to 9.4 J/cm(2) through adjustments in illumination time and a constant power density. Controls consisted of no treatment, light only, and photosensitizer only. Temperature changes were recorded in experimental samples before and after illumination. RESULTS aPDT with an energy dose of 9.4 J/cm(2) was effective in eradicating P. gingivalis, F. nucleatum, and A. actinomycetemcomitans in biofilm and planktonic form. Reductions from control in planktonic cultures at this energy dose were 6.8 +/- 0.7, 5.2 +/- 0.6, and 1.9 +/- 0.6 log(10), respectively, whereas biofilm reductions were 4.5 +/- 1.2, 3.4 +/- 1.1, and 4.9 +/- 1.4 log(10). Decreasing the treatment time produced an energy dose-dependent killing effect in both models. Changes in sample temperature did not exceed 3 degrees C under these exposure parameters. CONCLUSION This study demonstrated that three important periopathogens are susceptible to aPDT-mediated killing, regardless of whether they are present in planktonic or biofilm form. Furthermore, a clear energy dose-dependence exists with this treatment that should to be taken into account when determining optimal treatment times in clinical application.

The effect of antimicrobial photodynamic therapy on human ciliated respiratory mucosa

Chronic recurrent sinusitis (CRS) is one of the most common chronic conditions in the United States. There is a significant subpopulation of CRS patients who remain resistant to cure despite rigorous treatment regimens including surgery, allergy therapy, and prolonged antibiotic therapy. Antimicrobial photodynamic therapy (aPDT) is a noninvasive nonantibiotic broad spectrum antimicrobial treatment. Our previous in vitro studies demonstrated that aPDT reduced CRS polymicrobial biofilm and planktonic bacteria and fungi by > 99.9% after a single treatment. Prior to human treatment however, aPDT treatment must be demonstrated to not result in histologic damage to the sinus ciliated respiratory epithelium. The objective of this study was to demonstrate the safety of aPDT treatment on a living human ciliated respiratory mucosal model (EpiAirway™).

Antimicrobial photodynamic therapy for the decolonization of methicillin-resistant Staphylococcus aureus from the anterior nares

The nosocomial infection rate has increased dramatically due to emergence of antibiotic resistant bacterial strains such as methicillin resistant Staphylococcus aureus (MRSA). The primary anatomical site of MRSA colonization is the anterior nares, and this reservoir represents a primary vector of transmission from non-infected carriers to susceptible individuals. Antimicrobial photodynamic therapy (aPDT) has been used successfully for topical disinfection in the oral cavity. The aim of this study was to evaluate the utility of aPDT for nasal MRSA decolonization at the preclinical and clinical level. The nasal aPDT system consists of a 670 nm diode laser fibre-optically coupled to a disposable light diffusing tip, used to activate a methylene blue based photosensitizer formulation. Preclinical testing was done both in a custom nasal reservoir model and on human skin cultures colonized on the epithelial surface with MRSA. Human clinical testing was performed by clinicians in regions in which the system is approved by the regulatory authority. In vitro testing demonstrated that aPDT eradicated planktonic MRSA in an energy and photosensitizer concentration dependent manner. Furthermore, aPDT eliminated sustained colonization of MRSA on cultured human epithelial surfaces, an effect that was sustained over multiple days post-treatment. In preliminary human testing, aPDT eradicated MRSA completely from the nose with total treatment times <10 minutes. aPDT is effective against MRSA when used topically in the nose. Energy dose and photosensitizer parameters have been optimized for the nasal environment. Controlled clinical studies are currently underway to further evaluate safety and efficacy.

Periowave demonstrates bactericidal activity against periopathogens and leads to improved clinical outcomes in the treatment of adult periodontitis

Periodontitis affects half of the U.S. population over 50, and is the leading cause of tooth loss after 35. It is believed to be caused by growth of complex bacterial biofilms on the tooth surface below the gumline. Photodynamic therapy, a technology used commonly in antitumor applications, has more recently been shown to exhibit antimicrobial efficacy. We have demonstrated eradication of the periopathogens Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans in vitro using PeriowaveTM; a commercial photodisinfection system. In addition, several clinical studies have now demonstrated the efficacy of this treatment. A pilot study in the U.S. showed that 68% of patients treated with PeriowaveTM adjunctively to scaling and root planing (SRP) showed clinical attachment level increase of >1 mm, as opposed to 30% with SRP alone. In a subsequent larger study, a second PeriowaveTM treatment 6 weeks after initial treatment led to pocket depth improvements of >1.5 mm in 89% of patients. Finally, in the most recent multicenter, randomized, examiner-blinded study conducted on 121 subjects in Canada, PeriowaveTM treatment produced highly significant gains in attachment level (0.88 mm vs. 0.57 mm; p=0.003) and pocket depth (0.87 mm vs. 0.63 mm; p=0.01) as compared to SRP alone. In summary, PeriowaveTM demonstrated strong bactericidal activity against known periopathogens, and treatment of periodontitis using this system produced significantly better clinical outcomes than SRP alone. This, along with the absence of any adverse events in patients treated to date demonstrates that PDT is a safe and effective treatment for adult chronic periodontitis.

Meta-analysis of five photodisinfection clinical trials for periodontitis

Photodynamic therapy(PDT) has been demonstrated to effectively kill human periopathogens in vitro. To evaluate the efficacy of PDT in vivo a series of clinical trials was carried out in multiple centers and populations. Clinical parameters including clinical attachment level, pocket probing depth and bleeding on probing were all evaluated. All groups received the standard of care, scaling and root planing, and the treatment group additionally received a single treatment of PDT. Of the total 309 patients and over 40,000 pockets treated in these 5 trials it was determined that photodynamic therapy provided a statistically significant improvement in clinical parameters over scaling and root planing alone.

Photodynamic dosimetry in the treatment of periodontitis

Photodynamic therapy has been demonstrated to effectively kill human periopathogens in vitro. However, the translation of in vitro work to in vivo clinical efficacy has been difficult due to the number of variables present in any given patient. Parameters such as photosensitizer concentration, duration of light therapy and amount of light delivered to the target tissue all play a role in the dose response of PDT in vivo. In this 121 patient study we kept all parameters the same except for light dose which was delivered at either 150 mW or 220 mW. This clearly demonstrated the clinical benefits of a higher light dose in the treatment of periodontitis.