Human Eye Lens Fluid Analysis Using High-Performance Liquid Chromatography-LED-Induced Fluorescence to Explore Cataract: A Pilot Study

Sulatha V. Bhandary; Sphurti S. Adigal; Jijo Lukose; Reena V. John; Alisha Rizvi; Santhosh Chidangil

doi:10.18287/JBPE24.10.030302

Human Eye Lens Fluid Analysis Using High-Performance Liquid Chromatography-LED-Induced Fluorescence to Explore Cataract: A Pilot Study

Sulatha V. Bhandary
Kasturba Medical College, Manipal Academy of Higher Education, India

Sphurti S. Adigal
Centre of Excellence for Biophotonics, Manipal Academy of Higher Education, India

Jijo Lukose
Centre of Excellence for Biophotonics, Manipal Academy of Higher Education, India

Reena V. John
Centre of Excellence for Biophotonics, Manipal Academy of Higher Education, India

Alisha Rizvi
Kasturba Medical College, Manipal Academy of Higher Education, India

Santhosh Chidangil (Login required)
Centre of Excellence for Biophotonics, Manipal Academy of Higher Education, India

Paper #9009 received 4 Aug 2023; revised manuscript received 20 Mar 2024; accepted for publication 20 May 2024; published online 17 Aug 2024.

DOI: 10.18287/JBPE24.10.030302

Abstract

Cataract is the common cause of reversible blindness and visual impairment. It is common in low socioeconomic groups in developing countries. This paper describes the application of an assembled High-Performance Liquid Chromatography-LED-Induced Fluorescence (HPLC-LED-IF) system to study the protein profiles of lens fluid samples from cataract patients with and without diabetes. The HPLC-LED-IF system assembled in our laboratory was used to record protein profiles of lens fluid samples obtained from volunteers having cataract with diabetes and without diabetes who have undergone cataract surgery. Statistical analyses such as Descriptive Statistics and Principal Component Analysis (PCA) were conducted to observe any kind of changes in the protein pattern of the two classes of samples. High-Performance Liquid Chromatography combined with LED-Induced Fluorescence (HPLC-LIF) generated lens fluid protein profile data of the two categories (cataract with and without diabetes) of samples have shown considerable changes in their pattern. These changes are confirmed by Descriptive Statistics and PCA. Protein profiles of lens fluid using an assembled HPLC-LED-IF system have shown significant differences in relative intensities of crystalline proteins of cataract with diabetes and without diabetes. UV absorption and fluorescence spectroscopy of the lens fluid have shown valuable information on the absorption and fluorescence intensity variations. The origin of the changes has to be further studied to get more information on this observation.

Keywords

cataract lens fluid; UV-absorption; fluorescence; HPLC-LED-IF; protein profile

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References

1. J. Thompson, N. Lakhani, “Cataracts,” Primary Care: Clinics in Office Practice 42(3), 409−423 (2015).

2. K. Pietrowska, D. A. Dmuchowska, P. Krasnicki, A. Bujalska, P. Samczuk, E. Parfieniuk, T. Kowalczyk, M. Wojnar, Z. Mariak, A. Kretowski, and M. Ciborowski, “An exploratory LC-MS-based metabolomics study reveals differences in aqueous humor composition between diabetic and non-diabetic patients with cataract,” Electrophoresis 39(9−10), 1233-1240 (2018).

3. Z. Zhuang, M. Zhu, Y. Huang, J. Liu, Z. Guo, K. Xiong, N. Li, S. Chen, and X. Qiu, “Study of molecule variation in various grades of human nuclear cataracts by confocal micro-Raman spectroscopy,” Applied Physics Letters 101(17), 173701 (2012).

4. N. Pescosolido, A. Barbato, R. Giannotti, C. Komaiha, and F. Lenarduzzi, “Age-related changes in the kinetics of human lenses: prevention of the cataract,” International Journal of Ophthalmology 9(10), 1506−1517 (2016).

5. A. Gakamsky, R.R. Duncan, N.M. Howarth, B. Dhillon, K. K. Buttenschön, D. J. Daly, and D. Gakamsky, “Tryptophan and Non-Tryptophan Fluorescence of the Eye Lens Proteins Provides Diagnostics of Cataract at the Molecular Level,” Scientific Reports 7(1), 40375 (2017).

6. W.-C. Chang, C.-H. Lee, S.-H. Chiou, C.-C. Liao, and C.-W. Cheng, “Proteomic Analysis of Aqueous Humor Proteins in Association with Cataract Risks: Diabetes and Smoking,” Journal of Clinical Medicine 10(24), 5731 (2021).

7. R. Augustine, N. Kalarikkal, and S. Thomas, “Role of wound dressings in the management of chronic and acute diabetic wounds,” in Diabetes mellitus and human health care: a holistic approach to diagnosis and treatment, A. George, R. Augustine, and M. Sebastian, Apple Academic Press, Canada, 273−314 (2014). ISBN: 9780429159343.

8. R. Anjana, M. Ali, R. Pradeepa, M. Deepa, M. Datta, R. Unnikrishnan, M. Rema, and V. Mohan, “The need for obtaining accurate nationwide estimates of diabetes prevalence in India-rationale for a national study on diabetes,” Indian Journal of Medical Research 133(4), 369−380 (2011).

9. Z. L. Teo, Y.-C. Tham, M. Yu, M. L. Chee, T. H. Rim, N. Cheung, M. M. Bikbov, Y. X. Wang, Y. Tang, Y. Lu, I. Y. Wong, D. S. W. Ting, G. S. W. Tan, J. B. Jonas, C. Sabanayagam, T. Y. Wong, and C.-Y. Cheng, “Global Prevalence of Diabetic Retinopathy and Projection of Burden through 2045: Systematic Review and Meta-analysis,” Ophthalmology 128(11), 1580–1591 (2021).

10. S. Chandra, S. Sivaprasad, P. G. Ursell, K. Naderi, D. O’Brart, A. Alwitry, Zahra Ashena, and M. A. Nanavaty, “Recurring themes during cataract assessment and surgery,” Eye 35(9), 2482–2498 (2021).

11. S.-L. Nyeo, R. R. Ansari, “Early cataract detection by dynamic light scattering with sparse bayesian learning,” Journal of Innovative Optical Health Sciences 02(03), 303–313 (2009).

12. Z. Kyselova, “Mass spectrometry-based proteomics approaches applied in cataract research,” Mass Spectrometry Reviews 30(6), 1173−1184 (2011).

13. R. J. W. Truscott, M. G. Friedrich, “Old proteins and the A chilles heel of mass spectrometry. The role of proteomics in the etiology of human cataract,” PROTEOMICS–Clinical Applications 8(3–4), 195–203 (2014)..

14. B. N. Zhang, X. Wu, Y. Dai, B. Qi, C. Fan, and Y. Huang, “Proteomic analysis of aqueous humor from cataract patients with retinitis pigmentosa,” Journal Cellular Physiology 236(4), 2659–2668 (2021).

15. K. L. Schey, Z. Wang, M. G. Friedrich, D. L. Garland, and R. J. W. Truscott, “Spatiotemporal changes in the human lens proteome: Critical insights into long-lived proteins,” Progress in Retinal and Eye Research 76, 100802 (2020).

16. K. Venkatakrishna, K. M. Pai, C. M. Krishna, O. Ravikiran, G. Ullas, and V. B. Kartha, “HPLC-LIF for early detection of oral cancer,” Current Science 84(4), 551−557 (2003).

17. S. Bhat, A. Patil, L. Rai, V. B. Kartha, and S. Chidangil, “Application of HPLC Combined with Laser Induced Fluorescence for Protein Profile Analysis of Tissue Homogenates in Cervical Cancer,” The Scientific World Journal 2012, 1–7 (2012).

18. A. Patil, S. Bhat, K. M. Pai, L. Rai, V. B. Kartha, and S. Chidangil, “Ultra-sensitive high performance liquid chromatography–laser-induced fluorescence based proteomics for clinical applications,” Journal of Proteomics 127, 202–210 (2015).

19. R. V. John, T. Devasia, S. S. Adigal, J. Lukose, V. B. Kartha, and S. Chidangil, “Serum protein profile study of myocardial infarction using a LED induced fluorescence based HPLC system,” Journal of Chromatography B 1217, 123616 (2023).

20. S. S. Adigal, N. I. R. Kuzhuppilly, N. Hegde, N. V. R., A. Rizvi, R. V. John, S. D. George, V. B. Kartha, S. V. Bhandary, and S. Chidangil, “HPLC-LED-Induced Fluorescence Analysis of Tear Fluids: An Objective Method for Primary Open Angle Glaucoma Diagnosis,” Current Eye Research 49(3), 260−269 (2023).

21. S. S. Adigal, S. V. Bhandary, N. Hegde, V. R. Nidheesh, R. V. John, A. Rizvi, S. D. George, V. B. Kartha, and S. Chidangil, “Protein profile analysis of tear fluid with hyphenated HPLC-UV LED-induced fluorescence detection for the diagnosis of dry eye syndrome,” RSC Advances 13(32), 22559–22568 (2023).

22. S. Menon, S. Raja, K. K. Kumar, J. Kurien, B. R. Krishnanand, K. K. Mahato, S. D. George, V. B. Kartha, and S. Chidangil, “Protein profile study of breast cancer tissues using HPLC-LIF: a pilot study,” Proceedings of SPIE 6430, 64300W (2007).

23. S. Rao, R. John, A. Patil, U. V. K., S. D. George, K. M. Pai, R. Ongole, T. Devasia, V. B. Kartha, and S. Chidangil, “Optical Biopsy and Optical Pathology: Affordable Health Care Under Low-Resource Settings,” Journal of Biomedical Photonics & Engineering 6(2), 020309 (2020).