scholarly journals Transcriptional analysis and target genes discovery of Pseudomonas aeruginosa biofilm developed ex vivo chronic wound model

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Xiaojuan Tan ◽  
Xi Cheng ◽  
Mei Hu ◽  
Yifan Zhang ◽  
Aiqun Jia ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Alkaline Phosphatase ◽  
Chronic Wound ◽  
Ex Vivo ◽  
Wound Infections ◽  
Porcine Skin ◽  
Expression Levels ◽  
Wound Model ◽  
Skin Explants ◽  
Chronic Wound Infections

AbstractBacterial biofilms formation is one of the major reasons for treatment failure in chronic wound infections. Therefore, diagnostic biomarkers remain the best option for prevention and treatment of chronic wound infections by biofilms. Herein, Pseudomonas aeruginosa PAO1 was used to mimic biofilm development in porcine skin explants wells as ex vivo wound model. The microscopic imaging showed that PAO1 in porcine skin explants wells formed micro-colonies at 24 h, developed mushroom-like structure at 48 h, and at 72 h mushroom-like structure disappeared, remaining a thin bacterial lawn. RNA-seq data analysis revealed that the expression levels of genes involved in the type II hxc secretion system were significantly higher in biofilms than in planktonic cells, especially the expression of lapA encoding alkaline phosphatase. However, the expression levels of genes associated with denitrification pathway were markedly decreased in biofilms, especially the transcription of nirS encoding nitrite reductase to produce nitric oxide (NO). Therefore, their expressions and products were further detected using RT-qPCR and biochemical assays, respectively. The results found that the expression of lapA and alkaline phosphatase activity were induced, but the expression of nirS and intracellular NO were reduced at the whole biofilms cycle. The study indicates that LapA and NO would play an important role for P. aeruginosa biofilm formation in chronic wound infections. LapA would serve as potential target to monitor chronic wound infections by P. aeruginosa biofilms. Inducing NO would be used to treat chronic wound infections due to P. aeruginosa biofilms.

scholarly journals Transcriptional Analysis and Target Genes Discovery of Pseudomonas Aeruginosa Biofilm Developed Ex Vivo Chronic Wound Model

2021 ◽  
Author(s):  
Xiaojuan Tan ◽  
Xi Cheng ◽  
Mei Hu ◽  
Yifan Zhang ◽  
Aiqun Jia ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Alkaline Phosphatase ◽  
Chronic Wound ◽  
Ex Vivo ◽  
Wound Infections ◽  
Porcine Skin ◽  
Expression Levels ◽  
Wound Model ◽  
Skin Explants ◽  
Chronic Wound Infections

Abstract Bacterial biofilms formation is one of the major reasons for treatment failure in chronic wound infections. Therefore, diagnostic biomarkers remain the best option for prevention and treatment of chronic wound infections by biofilms. Herein, Pseudomonas aeruginosa PAO1 was used to mimic biofilm development in porcine skin explants wells as ex vivo wound model. The microscopic imaging showed that PAO1 in porcine skin explants wells formed micro-colonies at 24 h, developed mushroom-like structure at 48 h, and at 72 h mushroom-like structure disappeared, remaining a thin bacterial lawn. RNA-seq data analysis revealed that the expression levels of genes involved in the type II hxc secretion system were significantly higher in biofilms than in planktonic cells, especially the expression of lapA encoding alkaline phosphatase. However, the expression levels of genes associated with denitrification pathway were markedly decreased in biofilms, especially the transcription of nirS encoding nitrite reductase to produce nitric oxide (NO). Therefore, their expressions and products were further detected using RT-qPCR and biochemical assays, respectively. The results found that the expression of lapA and alkaline phosphatase activity were induced, but the expression of nirS and intracellular NO were reduced at the whole biofilms cycle. The study indicates that LapA and NO would play an important role for P. aeruginosa biofilm formation in chronic wound infections. LapA would serve as potential target to monitor chronic wound infections by P. aeruginosa biofilms. Inducing NO would be used to treat chronic wound infections due to P. aeruginosa biofilms.

scholarly journals Hypertonic glucose inhibits growth and attenuates virulence factors of multidrug-resistant Pseudomonas aeruginosa

2020 ◽  
Author(s):  
Tao Chen ◽  
Ye Xu ◽  
Wenya Xu ◽  
Wenli Liao ◽  
Chunquan Xu ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Virulence Factors ◽  
Diabetic Foot ◽  
Biofilm Formation ◽  
Chronic Wound ◽  
Multidrug Resistant ◽  
Wound Infections ◽  
Hypertonic Glucose ◽  
Diabetic Foot Infections ◽  
Chronic Wound Infections

Abstract Background: Pseudomonas aeruginosa is the most common Gram-negative pathogen responsible for chronic wound infections, such as diabetic foot infections, and further exacerbates the treatment options and cost of such conditions. Hypertonic glucose, a commonly used prolotherapy solution, can accelerate the proliferation of granulation tissue and improve microcirculation in wounds. However, the action of hypertonic glucose on bacterial pathogens that infect wounds is unclear. In this study, we investigated the inhibitory effects of hypertonic glucose on multidrug-resistant P. aeruginosa strains isolated from diabetic foot infections. Hypertonic glucose represents a novel approach to control chronic wound infections caused by P. aeruginosa. Results: Four multidrug-resistant P. aeruginosa clinical strains isolated from diabetic foot ulcers from a tertiary hospital in China and the reference P. aeruginosa PAO1 strain were studied. Hypertonic glucose significantly inhibited the growth, biofilm formation, and swimming motility of P. aeruginosa clinical strains and PAO1. Furthermore, hypertonic glucose significantly reduced the production of pyocyanin and elastase virulence factors in P. aeruginosa. The expression of major quorum sensing genes (lasI, lasR, rhlI, and rhlR) in P. aeruginosa were all downregulated in response to hypertonic glucose treatment. In a Galleria mellonella larvae infection model, the administration of hypertonic glucose was shown to increase the survival rates of larvae infected by P. aeruginosa strains (3/5).Conclusions: Hypertonic glucose inhibited the growth, biofilm formation, and swimming motility of P. aeruginosa, as well as reduced the production of virulence factors and quorum sensing gene expression. Further studies that investigate hypertonic glucose therapy should be considered in treating chronic wound infections.

scholarly journals Hypertonic glucose inhibits growth and attenuates virulence factors of multidrug-resistant Pseudomonas aeruginosa by regulating quorum sensing

2020 ◽  
Author(s):  
Tao Chen ◽  
Ye Xu ◽  
Wenya Xu ◽  
Wenli Liao ◽  
Chunquan Xu ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Virulence Factors ◽  
Diabetic Foot ◽  
Chronic Wound ◽  
Multidrug Resistant ◽  
Infection Model ◽  
Wound Infections ◽  
Hypertonic Glucose ◽  
Chronic Wound Infections

Abstract Background: Pseudomonas aeruginosa is the most common Gram-negative pathogen responsible for chronic wound infections such as diabetic foot infections, which make chronic wound treatment become increasingly difficult and costly, further exacerbated. Hypertonic glucose, a commonly used prolotherapy solution, can accelerate the proliferation of granulation tissue and improve microcirculation in the wound, but the knowledge of the impacts that hypertonic glucose has on wound infecting pathogens has lagged. In this study, we aim to investigate the inhibitory effects of hypertonic glucose on diabetic foot infecting multidrug-resistant P. aeruginosa, to offer a novel approach to control the chronic wound infections caused by P. aeruginosa. Results: Four multidrug-resistant P. aeruginosa clinical strains isolated from diabetic foot ulcers from a tertiary hospital in China and P. aeruginosa PAO1 were studied. Hypertonic glucose had a remarkable inhibitory effect on growth of PAO1 and multidrug-resistant P. aeruginosa strains. Furthermore, hypertonic glucose significantly inhibited the formation of biofilm, the ability of swimming, and the production of virulence factors including pyocyanin and elastase in all P. aeruginosa strains. We examined the expressions of major quorum sensing related genes (lasI, lasR, rhlI, and rhlR) in P.aeruginosa and found these genes were all downregulated after hypertonic glucose treatment. In infection model of Galleria mellonella larvae, groups inoculated with P. aeruginosa strains which were treated with hypertonic glucose showed no increased survival rate than non-hypertonic glucose-treated groups. Conclusions: Our research suggested that hypertonic glucose displayed the ability to inhibit growth, biofilm formation and swimming motility as well as attenuate virulence factors of multidrug-resistant P. aeruginosa via regulating quorum sensing system. Further clinical studies combining hypertonic glucose with traditional antibiotics should be considered in chronic wound infections.

scholarly journals Hypertonic glucose inhibits growth and attenuates virulence factors of multidrug-resistant Pseudomonas aeruginosa

2020 ◽  
Author(s):  
Tao Chen ◽  
Ye Xu ◽  
Wenya Xu ◽  
Wenli Liao ◽  
Chunquan Xu ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Virulence Factors ◽  
Diabetic Foot ◽  
Biofilm Formation ◽  
Chronic Wound ◽  
Multidrug Resistant ◽  
Wound Infections ◽  
Hypertonic Glucose ◽  
Diabetic Foot Infections ◽  
Chronic Wound Infections

Abstract Background: Pseudomonas aeruginosa is the most common Gram-negative pathogen responsible for chronic wound infections, such as diabetic foot infections, and further exacerbates the treatment options and cost of such conditions. Hypertonic glucose, a commonly used prolotherapy solution, can accelerate the proliferation of granulation tissue and improve microcirculation in wounds. However, the action of hypertonic glucose on bacterial pathogens that infect wounds is unclear. In this study, we investigated the inhibitory effects of hypertonic glucose on multidrug-resistant P. aeruginosa strains isolated from diabetic foot infections. Hypertonic glucose represents a novel approach to control chronic wound infections caused by P. aeruginosa. Results: Four multidrug-resistant P. aeruginosa clinical strains isolated from diabetic foot ulcers from a tertiary hospital in China and the model P. aeruginosa PAO1 strain were studied. Hypertonic glucose significantly inhibited the growth, biofilm formation, and swimming motility of P. aeruginosa clinical strains and PAO1. Furthermore, hypertonic glucose significantly reduced the production of pyocyanin and elastase virulence factors in P. aeruginosa. The expression of major quorum sensing genes (lasI, lasR, rhlI, and rhlR) in P. aeruginosa were all downregulated in response to hypertonic glucose treatment. In a Galleria mellonella larvae infection model, the administration of hypertonic glucose was shown to increase the survival rates of larvae infected by P. aeruginosa strains (3/5).Conclusions: Hypertonic glucose inhibited the growth, biofilm formation, and swimming motility of P. aeruginosa, as well as reduced the production of virulence factors and quorum sensing gene expression. Further studies that investigate hypertonic glucose therapy should be considered in treating chronic wound infections.

scholarly journals Biofilm Formation Reducing Properties of Manuka Honey and Propolis in Proteus mirabilis Rods Isolated from Chronic Wounds

2020 ◽  
Vol 8 (11) ◽  
pp. 1823
Author(s):  
Joanna Kwiecińska-Piróg ◽  
Jana Przekwas ◽  
Michał Majkut ◽  
Krzysztof Skowron ◽  
Eugenia Gospodarek-Komkowska
Keyword(s):  
Proteus Mirabilis ◽  
Biofilm Formation ◽  
Chronic Wounds ◽  
Chronic Wound ◽  
Ethanol Extract ◽  
Minimal Bactericidal Concentration ◽  
Wound Infections ◽  
Manuka Honey ◽  
The Impact ◽  
Chronic Wound Infections

Chronic wound infections are difficult to manage because of the biofilm formation in the wound environment. New measures for eliminating infections are necessary to increase the chance of wound healing. Apitherapy may be the new solution. The aim of this study was to assess the prevalence of wound infection factors and to examine the impact of Manuka honey and ethanol extract of propolis on biofilm formation of Proteus mirabilis isolated from chronic wound infections. According to the findings, the most frequent factors of infection are Staphylococcus aureus (46.1%), Pseudomonas aeruginosa (35.0%), and Proteus mirabilis (10.6%). Minimal inhibitory concentration and minimal bactericidal concentration values were assigned using the microbroth dilution test according to the Clinical and Laboratory Standards Institute. Biofilm of Proteus mirabilis isolates was formed in 96-well polystyrene plates and treated with Manuka honey (concentrations from 1.88% to 30.0%) and ethanol extract of propolis (1.0% to 40.0%). After 24 h, the biofilm viability was expressed by formazan absorbance (λ = 470 nm). Manuka honey reduced the biofilm viability in all, and ethanol extract of propolis in most, of the concentrations tested. Ethanol extract of propolis at the concentrations of 20.0% and 40.0%, reduced biofilm viability stronger than ethanol itself. With these results comes the conclusion that these substances can reduce biofilm formation.

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