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  • 73.3% of people 20 or older have had acne in their lifetimes (1)
  • Biofilm: a collection of microorganisms that form on different types of surfaces (2)
  •  Biofilms are naturally tolerant to antibiotics when they are clustered together on the skin, causing acne to be more difficult to treat
  • Acne vulgaris: a skin disease caused by the inflammation and lesions from excessive oil secretions and the formation of biofilms within our skin (3)
  • Cutibacterium acnes and Staphylococcus epidermidis are linked to acne biofilms (4)
  • Currently, there is no effective way to treat acne completely
  • Goal: Find a way to bypass the natural tolerance to antibiotics in biofilms that makes acne difficult to treat and the most effective combination treatment for biofilms, specifically a two-step treatment process

INTRODUCTION

HYPOTHESIS

CONCLUSIONS & FUTURE WORK

Figure 2. Two-Step Treatment Process on Cutibacterium acnes Biofilm Growth. (a) Represents a control: C. acnes are treated with a general antibiotic. (b) The biofilm is treated with a combination of DNase I enzyme and Nicotinamide, destroying the extracellular matrix of the biofilm (6). Then, the chemical treatment Chitosan-Caffeic Acid (CCA) is added. (c) Similar to (b), both Nicotinamide and DNase I are introduced to the biofilm, destroying the extracellular matrix. Then, the natural oregano essential oil is added.

HYPOTHESIZED TWO-STEP TREATMENT PROCESS

REFERENCES

If C. acnes biofilms are first disturbed with a biofilm disruptor, then treated in combination with an acne treatment, they can more effectively be destroyed.

CHEMICAL VS NATURAL

Figure 3. A comparison of the effectiveness of chemical vs. natural treatments for two strains of bacteria, S. epidermidis and C. acnes, that cause acne. MIC (minimum inhibition concentration) represents the lowest concentration needed of an antimicrobial agent to hinder visible growth of microorganisms in mg/mL. The smaller the MIC value, the more effective the treatment. Even though oregano was the most effective natural treatment (4), it is still less effective than the least effective chemical treatment (7). The difference between the highest and lowest MIC values of natural treatments (4) is much greater than the difference between the highest and lowest MIC values of chemical treatments (7).

* Ranges the MIC of 2 strains of C. acnes (C. acne isolate 2874 - C. acne isolate 2875)

Investigating the Effectiveness of the Biofilm Disrupting Agents in Combination with Acne Treatments 

Shannon Kelleher, Francesca Mendejar, Carleigh Newman, Adrianna Scalo, Teresa Scotto, & Sarina Shield

Microbial Biofilms in Human Health Stream, First-year Research Immersion Program

METHODS

DISPERSION METHODS

Figure 1. Two-Step Treatment Process on Cutibacterium acnes Biofilm Growth. A comparison of using a typical antibiotic acne treatment (a) to using the DNase I and Nicotinamide model (b), in order to find the most effective biofilm combination treatment (c). (a) Represents a control: C. acnes are treated with a general antibiotic. (b) The biofilm is treated with a combination of DNase I enzyme, which destroys the extracellular matrix, and Nicotinamide (NAM), reducing the amount of acne left (5). (c) An unknown dispersion agent is added along with an unknown drug, which has the potential to destroy the biofilm more effectively than DNase I and NAM.

  • Test each dispersion method mentioned above on C. acnes biofilms
  • Compare the effectiveness of different types of biofilm disruptors and different types of antibiotic treatments
    • Biofilm disruptors must be tested on C. acnes and compared to the DNase I and NAM model
    • The most effective dispersion method, Nitric oxide, and the most effective chemical treatment, CCA, would be tested as a combination treatment 
    • Test chemical vs. natural antibiotic treatments to find which works most effectively with a biofilm disruptor
  • Apply research to different acne-causing strains, such as S. epidermidis

Figure 2. Method for Testing Combined Treatments. The biofilms were first grown in test tubes. After 1 minute, the tubes were emptied out by a pipette and were flipped outside without undergoing a wash step. Next, the tubes were left overnight at room temperature. After growing the biofilms overnight, they were placed into their specific wells. Well (a) represents our negative control. Well (b) represents the positive control. Well (c) represents the biofilm and the addition of NAM. Well (d) represents the biofilms with the addition of DNase I, and well (e) represents the biofilms being submerged with the combination of DNase I and NAM. Overall, this shows how the biofilms were grown, and that when a dispersion agent and treatment are added together, the treatment is more effective. (6)

Figure 4. A Comparison of Dispersion Efficacy. Six different dispersion methods were tested for their ability to disperse Pseudomonas aeruginosa biofilms. Dispersion methods were tested in their most efficient concentrations: 310nM chloroform-extracted spent medium (CSM), 3.5mg/L Iron, 18mM Glutamate, 500μM Nitric oxide, 10mM Ammonium chloride, and 2mM Mercury chloride. Dispersion efficacy was measured through optical density (OD 600); the higher the optical density, the more effective the dispersion method. Nitric Oxide possessed the highest dispersion efficacy; therefore, it would be the most effective dispersion method out of the six tested methods. Dispersion methods have been commonly studied using P. aeruginosa; however, this data can be applied to other bacteria, such as C. acnes. In this way, one can compare these proven dispersion methods with the two-step treatment model proposed earlier. From here, it can be understood that using Nitric Oxide in conjunction with a natural or chemical treatment has potential to be an effective treatment for acne.

3.Syal S, Pandit V, Ashawat MS. 2020. Traditional herbs to treat acne vulgaris. Asian J Pharm Research 10:195–201

4.Taleb MH, Abdeltawab NF, Shamma RN, Abdelgayed SS, Mohamed SS, Farag MA, Ramadan MA. 2018. Origanum vulgare L. Essential oil as a potential anti-acne topical nanoemulsion—in vitro and in vivo study. Molecules 23.

5.Shih Y-H, Liu D, Chen Y-C, Liao M-H, Lee W-R, Shen S-C. 2021. pharmaceutics Activation of Deoxyribonuclease I by Nicotinamide as a New Strategy to Attenuate Tetracycline-Resistant Biofilms of Cutibacterium acnes https://doi.org/10.3390/pharmaceutics.

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  • If biofilms are first disrupted, then they can be treated more effectively
    • A biofilm disruptor interferes with the structure of the biofilm, making it more susceptible to treatment
    • An antibiotic destroys the bacteria in the biofilm
  • It was hypothesized that this model two-step combination treatment would more effectively eradicate tolerant biofilms that cause acne vulgaris compared to traditional antibiotic acne treatments
  • Nitric Oxide is the most effective dispersion method because it has the greatest measured dispersion efficacy