With the surge of myopia, contact lenses are used by more and more people for their advantages of convenience and beauty. However, the incidence of corneal diseases also increases year by year because they are not worn, cleaned, disinfected and stored as instructed. Approximately 99% of wearers reported at least one contact lens hygiene risk behavior [1]. It can cause some damage to the eyes, secondly, lens cleaning and improper handling will breed a variety of bacteria, contact lens related complications are very common, 1 in 2 of the wearer surveyed in the United States have red or painful eyes, need emergency eye care [2].
Most contact lens-associated infections, about 80 – 95% by bacteria, cause [3,4] and the rest by pathogens such as Acanthamoeba and filamentous fungi. The acute red eye response induced by the contact lens is an inflammatory response in the conjunctiva and cornea associated with prolonged wear and eye closure; reported predisposing factors include endotoxin released from Gram-negative bacteria, such as Pseudomonas aeruginosa [5].
Repeated soft glasses are mainly Pseudomonas aeruginosa [6], which is the main cause of lens-associated microbial keratitis. The US CDC added Pseudomonas aeruginosa to its most serious threat list, [7], and the microbial keratitis caused by it has become one of the causes of blindness worldwide, while contact lens wearing is a major trigger [8]. The contact lens storage box contains the environmental bacteria [9], which also puts forward high requirements for the contact lens care solution. This is where we focus on it, and we hope to make corresponding efforts to improve this situation and improve the safety and comfort of contact lenses.
For the existing contact lenses caused by eye problems and the shortage of nursing fluid composition, we designed a to inhibit pseudomonas aeruginosa guide poor inflammation of the engineering bacteria as the main body of the integration of contact lens box, in vitro from the simulated tear components and reduce daily life after contact lenses wearing eye infection, remove contact lenses users trouble back at home.
Tear DMBT 1 and DMBT 1 purified from saliva play a protective role against mucosal tissues by inhibiting the twitching motility of P. aeruginosa. Tear DMBT 1 causes bacterial chain formation and clumping, can interfere with the formation of contact lens associated biofilm [7], and can lead to loss of bacterial motility, for swimming in fluid and for traveling on the surface; we identified DMBT 1 as a component of tears responsible for inhibiting the twitching motility function of P. aeruginosa. Importantly, we found that DMBT 1 prevents P. aeruginosa from penetrating multilayer cultures of human corneal epithelial cells, [11].
The design we expect is innovative and unique; it is an external contact lens box that mimics the unique design of engineered bacteria for DMBT 1 antibacterial, sterilization, contact lens bioactivity and comfort.
To achieve the function of engineered bacteria to produce DMBT 1 protein to inhibit Pseudomonas aeruginosa, our goal was to construct four-part systems separately for the regulation of engineered bacteria. Plasmids capable of producing DMBT 1 protein, systems with a toxin antitoxin system, control of population density by quorum sensing, and Hsp 27 inducible components
During the iGEM project, we designed, built and tested OptiCare LensCase, a multifunctional smart contact lens case that monitors the physicochemical properties of the care solution in real time, features a user-friendly touchscreen interactive interface and a UV disinfection module in case of emergencies, and is designed to improve user safety and compatibility with specially formulated care solutions. Together, our team validated the problem, designed the treatment, and validated the solution to make the therapy desirable, feasible, safe and accountable to the world. The proof-of-principle was developed and tested in vitro in the lab, and our goal was to demonstrate that DMBT1 could inhibit the twitching motility of Pseudomonas aeruginosa, with the proper introduction and collaboration of the various plasmid components: i.e., whether engineered bacterial colony sensing would allow for the proper performance of suicidality, whether the toxin-antitoxin system would function, and whether the Hsp27 system would be efficiently induced. In addition, we built a simulation model of colony growth and evolution of acting proteins and implemented colony identification counting using openCV to support experimental optimization. By simulating the behavior of the system and predictions can be adjusted in the lab to optimize the parameters of the system and experimental conditions to support future lab work.
To achieve the function of engineered bacteria to produce DMBT 1 protein to inhibit Pseudomonas aeruginosa, our goal was to construct four-part systems separately for the regulation of engineered bacteria. Plasmids capable of producing DMBT 1 protein, systems with a toxin antitoxin system, control of population density by quorum sensing, and Hsp 27 inducible components.
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[3] M. Green, S. Sara, I. Hughes, A. Apel, F. Stapleton.Trends in contact lens microbial keratitis 1999 to 2015: a retrospective clinical review.Clin Exp Ophthalmol, 47 (6) (2019), pp. 726-732, 10.1111/ceo.13484
[4] L. Bennett, H. YH, S. Tai, B. Ernst, E.J. Schmidt, R. Parihar, et al.Contact lens versus non-contact lens-related corneal ulcers at an academic center.Eye Contact Lens, 45 (5) (2019), pp. 301-305
[5]Lim, Chris H. L. B.Sc. (Med) (Hons), B.Med., M.D.; Stapleton, Fiona B.Sc. (Hons), M.C.Optom., M.Sc., Ph.D.; Mehta, Jodhbir S. B.Sc. (Hons), M.B.B.S., F.R.C.Ophth., F.R.C.S.(Ed), F.A.M.S.. Review of Contact Lens-Related Complications. Eye & Contact Lens: Science & Clinical Practice 44():p S1-S10, November 2018. | DOI: 10.1097/ICL.0000000000000481
[6] F. Stapleton, T. Naduvilath, L. Keay, C. Radford, J. Dart, K. Edwards, et al.Risk factors and causative organisms in microbial keratitis in daily disposable contact lens wear.PLoS One, 12 (8) (2017),Article e0181343, 10.1371/journal.pone.0181343
[7] Fleiszig, S. M. J., Kroken, A. R., Nieto, V., Grosser, M. R., Wan, S. J., Metruccio, M. M. E., & Evans, D. J. (2020). Contact lens-related corneal infection: Intrinsic resistance and its compromise. Progress in retinal and eye research, 76, 100804.
[8] Bradley CS, Sicks LA, Pucker AD. Common Ophthalmic Preservatives in Soft Contact Lens Care Products: Benefits, Complications, and a Comparison to Non-Preserved Solutions. Clin Optom (Auckl). 2021 Sep 7;13:271-285. doi: 10.2147/OPTO.S235679. PMID: 34522149; PMCID: PMC8434857.
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[11]Li J, Wan SJ, Metruccio MME, Ma S, Nazmi K, Bikker FJ, Evans DJ, Fleiszig SMJ. DMBT1 inhibition of Pseudomonas aeruginosa twitching motility involves its N-glycosylation and cannot be conferred by the Scavenger Receptor Cysteine-Rich bacteria-binding peptide domain. Sci Rep. 2019 Sep 11;9(1):13146. doi: 10.1038/s41598-019-49543-w. PMID: 31511582; PMCID: PMC6739395.
[12] Somayajulu M, Ekanayaka S, McClellan SA, Bessert D, Pitchaikannu A, Zhang K, Hazlett LD. Airborne Particulates Affect Corneal Homeostasis and Immunity. Invest Ophthalmol Vis Sci. 2020 Apr 9;61(4):23. doi: 10.1167/iovs.61.4.23. PMID: 32301974; PMCID: PMC7401652.
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