Antimicrobial efficacy of irradiation with visible light on oral bacteria in vitro: a systematic review.

AIM: Resistances to antibiotics employed for treatment of infectious diseases have increased to alarming numbers making it more and more difficult to treat diseases caused by microorganisms resistant to common antibiotics. Consequently, novel methods for successful inactivation of pathogens are required. In this instance, one alternative could be application of light for treatment of topical infections. Antimicrobial properties of UV light are well documented, but due to its DNA-damaging properties use for medical purposes is limited. In contrast, irradiation with visible light may be more promising. METHODS: Literature was systematically screened for research concerning inactivation of main oral bacterial species by means of visible light. RESULTS: Inactivation of bacterial species, especially pigmented ones, in planktonic state showed promising results. There is a lack of research examining the situation when organized as biofilms. CONCLUSION: More research concerning situation in a biofilm state is required.

Lethal photosensitisation of Prevotellaceae under anaerobic conditions by their endogenous porphyrins.

BACKGROUND: Oxygen is generally considered essential for lethal photosensitisation by photodynamic processes. The oral anaerobes, Prevotella intermedia and P. nigrescens are known to be photosensitive, but are also extremely sensitive to the cytotoxic effects of oxygen. METHODS: The Prevotellaceae were exposed to two 405 nm light sources for different exposure times in an anaerobic chamber. Viable counts of the light exposed samples were compared to light-free controls to determine the proportion of bacteria killed. RESULTS: Lethal photosensitivity was demonstrated against P. intermedia and P. nigrescens. The proportions of bacteria killed by either the light-emitting diode or laser pointer were similar at a given energy density (J/cm(2)). CONCLUSIONS: Lethal photosensitivity was demonstrated in two species of Prevotella under anaerobic conditions.

Characterization of oral bacterial diversity of irradiated patients by high-throughput sequencing.

The objective of this study was to investigate the compositional profiles and microbial shifts of oral microbiota during head-and-neck radiotherapy. Bioinformatic analysis based on 16S rRNA gene pyrosequencing was performed to assess the diversity and variation of oral microbiota of irradiated patients. Eight patients with head and neck cancers were involved in this study. For each patient, supragingival plaque samples were collected at seven time points before and during radiotherapy. A total of 147,232 qualified sequences were obtained through pyrosequencing and bioinformatic analysis, representing 3,460 species level operational taxonomic units (OTUs) and 140 genus level taxa. Temporal variations were observed across different time points and supported by cluster analysis based on weighted UniFrac metrics. Moreover, the low evenness of oral microbial communities in relative abundance was revealed by Lorenz curves. This study contributed to a better understanding of the detailed characterization of oral bacterial diversity of irradiated patients.

Phototargeting oral black-pigmented bacteria.

We have found that broadband light (380 to 520 nm) rapidly and selectively kills oral black-pigmented bacteria (BPB) in pure cultures and in dental plaque samples obtained from human subjects with chronic periodontitis. We hypothesize that this killing effect is a result of light excitation of their endogenous porphyrins. Cultures of Prevotella intermedia and P. nigrescens were killed by 4.2 J/cm2, whereas P. melaninogenica required 21 J/cm2. Exposure to light with a fluence of 42 J/cm2 produced 99% killing of P. gingivalis. High-performance liquid chromatography demonstrated the presence of various amounts of different porphyrin molecules in BPB. The amounts of endogenous porphyrin in BPB were 267 (P. intermedia), 47 (P. nigrescens), 41 (P. melaninogenica), and 2.2 (P. gingivalis) ng/mg. Analysis of bacteria in dental plaque samples by DNA-DNA hybridization for 40 taxa before and after phototherapy showed that the growth of the four BPB was decreased by 2 and 3 times after irradiation at energy fluences of 4.2 and 21 J/cm2, respectively, whereas the growth of the remaining 36 microorganisms was decreased by 1.5 times at both energy fluences. The present study suggests that intraoral light exposure may be used to control BPB growth and possibly benefit patients with periodontal disease.

Cloning, sequencing and complementation analysis of the recA gene from Prevotella ruminicola.

Degenerate PCR primers based on conserved RecA protein regions were used to amplify a portion of recA [corrected] from Prevotella ruminicola strain 23, which was used as a probe to isolate the full-length recA gene from the P. ruminicola genomic library. The P. ruminicola recA gene encoded a protein of 340 amino acids with a molecular mass of 36.81 kDa, P. ruminicola RecA was highly similar to other RecA proteins and most closely resembled that of Bacteroides fragilis (75% identity). It alleviated the methyl methanesulfonate and mitomycin C sensitivities of Escherichia coli recA mutants, but did not restore the resistance to UV-light irradiation. Mitomycin C treatment of otherwise isogenic E. coli strains showed a higher level of prophage induction in a recA harboring lysogen.

Phototoxicity of argon laser irradiation on biofilms of Porphyromonas and Prevotella species.

Species of Prevotella (Pr.) and Porphyromonas (Po.) and other microorganisms were cultivated as biofilms on agar medium and examined for their susceptibility to argon laser irradiation (continuous mode; wavelengths, 488-514 nm; fluences, 20-200 J cm(-2)). Fluences of 35 to 80 J cm(-2) inhibited biofilm growth in Po. endodontalis, Po. gingivalis, Pr. denticola, Pr. intermedia, Pr. melaninogenica and Pr. nigrescens. A fluence of 70 J cm(-2) did not affect biofilm growth in species of Bacillus, Candida, Enterobacter, Proteus, Pseudomonas, Staphylococcus and Streptococcus. The phototoxic effects of argon laser irradiation against Prevotella and Porphyromonas species were: (1) caused by the radiation alone; (2) modified by biofilm age; (3) dependent on the presence of atmospheric oxygen; (4) influenced by medium supplements of hemin, hemoglobin and blood; (5) greater when compared with other microbial species; (6) demonstrated without augmentation with an exogenous photosensitizer; and (7) apparently unrelated to the protoporphyrin content of the cells. Overall, these in vitro findings suggest that low doses of argon laser radiation may be effective in the treatment and/or prevention of clinical infections caused by biofilm-associated species of Prevotella or Porphyromonas.

Sensitivity of Porphyromonas and Prevotella species in liquid media to argon laser.

The phototoxicity of argon laser irradiation was studied in aqueous suspensions of Porphyromonas endodontalis (American Type Culture Collection [ATCC] 35406), Porphyromonas gingivalis (ATCC 33277), Prevotella denticola (ATCC 33184) and two strains of Prevotella intermedia (ATCC 15033 and 49046), all "black-pigmented bacteria," BPB, that accumulate cellular porphyrins. Several of these species have been implicated in the etiology of periodontal disease. Non-black-pigmented bacteria were also studied to test the specificity of irradiation as a potential photodynamic treatment for periodontal infections. Cell suspensions were irradiated with an argon laser at fluences of 20-200 J/cm2. When cultured in hemin-supplemented media, ATCC 15033 was the most sensitive to irradiation. However, a second strain of the same species (ATCC 49046) was resistant. The photosensitivity of other species ranked ATCC 33277 > 35406 = 33184 = 35496. When hemin was replaced in media by hemoglobin, ATCC 33277 became resistant to irradiation. Protoporphyrin IX content in BPB cells was shown not to be a major factor determining photosensitivity. Oxygen was required during irradiation for BPB species to be affected. Non-black-pigmented bacteria were much less sensitive to irradiation than BPB.