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Raman microspectroscopy analysis in the treatment of acanthamoeba keratitis
Giulia Rusciano1, Paola Capriglione, Giuseppe Pesce
1Dipartimento di Fisica, Università 'Federico II', Complesso Universitario Monte S. Angelo, Napoli, Italy.
Abstract:
Acanthamoeba keratitis is a rare but serious corneal disease, often observed in contact lens wearers. Clinical treatment of infected patients frequently involves the use of polyhexamethylene biguanide (PHMB), a polymer used as a disinfectant and antiseptic, which is toxic also for the epithelial cells of the cornea. Prompt and effective diagnostic tools are hence highly desiderable for both starting early therapy and timely suspension of the treatment. In this work we use Raman microspectroscopy to analyse in vitro a single Acanthamoeba cell in cystic phase. In particular, we investigate the effect of PHMB at the single-cell level, providing useful information on both the underlying biochemical mechanism and the time frame for Acanthamoeba eradication in ocular infections. Furthermore, we demonstrate that Raman spectroscopy, in conjunction with standard multivariate analysis methods, allows discriminating between live and dead Acanthamoebas, which is fundamental to optimizing patients' treatment.
Insights
Raman spectroscopy distinguishes live from dead Acanthamoeba cells, aiding contact lens-related keratitis treatment. This method monitors polyhexamethylene biguanide (PHMB) effectiveness, crucial for corneal health.
Area of Science:
- Ophthalmology
- Microbiology
- Biophysics
Background:
- Acanthamoeba keratitis is a severe eye infection linked to contact lens use.
- Current treatments like polyhexamethylene biguanide (PHMB) can harm corneal cells.
- Effective diagnostics are needed to guide treatment duration and minimize toxicity.
Purpose of the Study:
- To investigate the effects of PHMB on Acanthamoeba at a single-cell level using Raman microspectroscopy.
- To establish Raman spectroscopy as a tool for monitoring Acanthamoeba eradication in ocular infections.
- To differentiate between live and dead Acanthamoeba cells for optimized patient treatment.
Main Methods:
- In vitro analysis of single Acanthamoeba cells in the cystic phase.
- Application of Raman microspectroscopy to study cellular responses to PHMB.
- Utilizing multivariate analysis techniques for data interpretation.
Main Results:
- Raman microspectroscopy revealed biochemical changes in Acanthamoeba upon PHMB exposure.
- The study provides insights into the mechanism and timeframe of PHMB's action against Acanthamoeba.
- Raman spectroscopy successfully discriminated between live and dead Acanthamoeba cells.
Conclusions:
- Raman spectroscopy is a valuable tool for assessing the efficacy of PHMB in treating Acanthamoeba keratitis.
- This technique enables real-time monitoring of parasite viability, crucial for adjusting treatment.
- Optimizing treatment by distinguishing live from dead parasites can improve patient outcomes and reduce corneal damage.
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