Skip to main content
SpringerLink
Log in

An untargeted metabolomics approach to study changes of the medium during human cornea culture

  • Original Article
  • Published:
Metabolomics Aims and scope Submit manuscript

Abstract

Introduction

Two main approaches (organ culture and hypothermia) for the preservation and storage of human donor corneas are globally adopted for corneal preservation before the transplant. Hypothermia is a hypothermic storage which slows down cellular metabolism while organ culture, a corneal culture performed at 28–37 °C, maintains an active corneal metabolism. Researchers, till now, have just studied the impact of organ culture on human cornea after manipulating and disrupting tissues.

Objectives

The aim of the current work was to optimize an analytical procedure which can be useful for discovering biomarkers capable of predicting tissue health status. For the first time, this research proposed a preliminary metabolomics study on medium for organ culture without manipulating and disrupting the valuable human tissues which could be still used for transplantation.

Methods

In particular, the present research proposed a method for investigating changes in the medium, over a storage period of 20 days, in presence and absence of a human donor cornea. An untargeted metabolomics approach using UHPLC-QTOF was developed to deeply investigate the differences on metabolites and metabolic pathways and the influence of the presence of the cornea inside the medium.

Results

Differences in the expression of some compounds emerged from this preliminary metabolomics approach, in particular in medium maintained for 10 and 20 days in presence but also in the absence of cornea. A total of 173 metabolites have been annotated and 36 pathways were enriched by pathway analysis.

Conclusion

The results revealed a valuable untargeted metabolomics approach which can be applied in organ culture metabolomics.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

No datasets were generated or analysed during the current study.

References

  • Akram, M. (2014). Citric acid cycle and role of its intermediates in metabolism. Cell biochemistry and biophysics, 68(3), 475–478.

    Article  CAS  PubMed  Google Scholar 

  • Armitage, W. J. (2011). Preservation of human cornea. Transfusion Medicine and Hemotherapy, 38(2), 143–147.

    Article  PubMed  PubMed Central  Google Scholar 

  • Benavente, M. C. R., & Argüeso, P. (2018). Glycosylation pathways at the ocular surface. Biochemical Society Transactions, 46(2), 343.

    Article  Google Scholar 

  • Borderie, V. M., Baudrimont, M., Lopez, M., Carvajal, S., & Laroche, L. (1997). Evaluation of the deswelling period in dextran-containing medium after corneal organ culture. Cornea, 16(2), 215–223.

    Article  CAS  PubMed  Google Scholar 

  • Bucolo, C., Maugeri, G., Giunta, S., D’Agata, V., Drago, F., & Romano, G. L. (2023). Corneal wound healing and nerve regeneration by novel ophthalmic formulations based on cross-linked sodium hyaluronate, taurine, vitamin B6, and vitamin B12. Frontiers in Pharmacology, 14, 1109291.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bull, A. T. (2010). The renaissance of continuous culture in the post-genomics age. Journal of Industrial Microbiology and Biotechnology, 37(10), 993–1021.

    Article  CAS  PubMed  Google Scholar 

  • Calabrese, E. J. (2001). Apoptosis: biphasic dose responses. Critical Reviews in Toxicology, 31(4–5), 607–613.

    Article  CAS  PubMed  Google Scholar 

  • Chen, X., Zhang, C., Tian, L., Wu, L., Jie, Y., Wang, N., et al. (2022). In situ metabolic profile and spatial distribution of ocular tissues: New insights into dry eye disease. The Ocular Surface, 24, 51–63.

    Article  PubMed  Google Scholar 

  • Elisabeth, P., Hilde, B., & Ilse, C. (2008). Eye bank issues: II. Preservation techniques: warm versus cold storage. International ophthalmology, 28, 155–163.

    Article  Google Scholar 

  • Gain, P., Jullienne, R., He, Z., Aldossary, M., Acquart, S., Cognasse, F., & Thuret, G. (2016). Global survey of corneal transplantation and eye banking. JAMA ophthalmology, 134(2), 167–173.

    Article  PubMed  Google Scholar 

  • Hao, T., Ma, H. W., Zhao, X. M., & Goryanin, I. (2012). The reconstruction and analysis of tissue specific human metabolic networks. Molecular BioSystems, 8(2), 663–670.

    Article  CAS  PubMed  Google Scholar 

  • Jeng, B. H. (2006). Preserving the cornea: corneal storage media. Current opinion in ophthalmology, 17(4), 332–337.

    Article  PubMed  Google Scholar 

  • Kryczka, T. (2014). Cellular Stress after Transferring Human Cornea from in vivo to in vitro Milieu-A Metabolomic Approach. J Cytol Tissue Biol, 1(005).

  • Kryczka, T., Ehlers, N., Nielsen, K., & Midelfart, A. (2012). Impact of organ culturing on metabolic profile of human corneas: preliminary results. Acta ophthalmologica, 90(8), 761–767.

    Article  CAS  PubMed  Google Scholar 

  • Kryczka, T., Szaflik, J. P., Szaflik, J., & Midelfart, A. (2015). Metabolic changes in donor corneas during cold storage–challenging stereotypes. Integrative Mol Med, 2, 238–242.

    Google Scholar 

  • Kumar, A., & Bachhawat, A. K. (2012). Pyroglutamic acid: throwing light on a lightly studied metabolite. Current Science, 288–297.

  • Land, W. G. (2012a). Emerging role of innate immunity in organ transplantation part III: the quest for transplant tolerance via prevention of oxidative allograft injury and its consequences. Transplantation Reviews, 26(2), 88–102.

    Article  PubMed  Google Scholar 

  • Land, W. G. (2012b). Emerging role of innate immunity in organ transplantation: part I: evolution of innate immunity and oxidative allograft injury. Transplantation Reviews, 26(2), 60–72.

    Article  PubMed  Google Scholar 

  • Li, S., Park, Y., Duraisingham, S., Strobel, F. H., Khan, N., Soltow, Q. A. (2013). Predicting network activity from high throughput metabolomics. PLoS computational biology, 9(7), e1003123.

  • Ma, J., Shen, Z., Peng, R., Li, C., Hu, B., & Hong, J. (2021). Tear lipid metabolites as potential diagnostic biomarkers for ocular chronic graft-versus-host disease. Transplantation and Cellular Therapy, 27(3), 232–e1.

    Article  CAS  Google Scholar 

  • Pang, Z., Chong, J., Zhou, G., De Lima Morais, D. A., Chang, L., Barrette, M., et al. (2021). MetaboAnalyst 5.0: narrowing the gap between raw spectra and functional insights. Nucleic Acids Research, 49(W1), W388–W396. https://doi.org/10.1093/NAR/GKAB382.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pedley, A. M., & Benkovic, S. J. (2017). A new view into the regulation of purine metabolism: the purinosome. Trends in biochemical sciences, 42(2), 141–154.

    Article  CAS  PubMed  Google Scholar 

  • Pellegrino, R. M., Blasi, F., Angelini, P., Ianni, F., Alabed, H. B. R., Emiliani, C., et al. (2022). LC/MS Q-TOF metabolomic investigation of amino acids and dipeptides in Pleurotus ostreatus grown on different substrates. Journal of Agricultural and Food Chemistry, 70(33), 10371–10382.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Priyadarsini, S., McKay, T. B., Sarker-Nag, A., Allegood, J., Chalfant, C., Ma, J. X., & Karamichos, D. (2016). Complete metabolome and lipidome analysis reveals novel biomarkers in the human diabetic corneal stroma. Experimental eye research, 153, 90–100.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Robaczewska, J., Kedziora-Kornatowska, K., Kozakiewicz, M., Zary-Sikorska, E., Pawluk, H., Pawliszak, W., & Kedziora, J. (2016). Role of glutathione metabolism and glutathione-related antioxidant defense systems in hypertension. Journal Of Physiology And Pharmacology, 67(3), 331–337.

    CAS  PubMed  Google Scholar 

  • Salek, R. M., Steinbeck, C., Viant, M. R., Goodacre, R., & Dunn, W. B. (2013). The role of reporting standards for metabolite annotation and identification in metabolomic studies. GigaScience, 2(1), 2047–217X.

    Article  Google Scholar 

  • Schumann, S., Dietrich, E., Kruse, C., Grisanti, S., & Ranjbar, M. (2021). Establishment of a robust and simple corneal organ culture model to monitor wound healing. Journal of Clinical Medicine, 10(16), 3486.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Snytnikova, O. A., Yanshole, L. V., Iskakov, I. A., Yanshole, V. V., Chernykh, V. V., Stepakov, D. A., et al. (2017). Quantitative metabolomic analysis of the human cornea and aqueous humor. Metabolomics, 13, 1–9.

    Google Scholar 

  • Stanley, P., Taniguchi, N., & Aebi, M. (2017). N-glycans. Essentials of Glycobiology [Internet]. 3rd edition.

  • Teo, A. W. J., Zhang, J., Zhou, L., & Liu, Y. C. (2023). Metabolomics in Corneal Diseases: A Narrative Review from Clinical Aspects. Metabolites, 13(3), 380.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tsugawa, H., Cajka, T., Kind, T., Ma, Y., Higgins, B., Ikeda, K., et al. (2015). MS-DIAL: data-independent MS/MS deconvolution for comprehensive metabolome analysis. Nature Methods, 12(6), 523–526. https://doi.org/10.1038/nmeth.3393.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vaz, F. M., & Ferdinandusse, S. (2017). Bile acid analysis in human disorders of bile acid biosynthesis. Molecular aspects of medicine, 56, 10–24.

    Article  CAS  PubMed  Google Scholar 

  • Vlaanderen, J. J., Janssen, N. A., Hoek, G., Keski-Rahkonen, P., Barupal, D. K., Cassee, F. R., et al. (2017). The impact of ambient air pollution on the human blood metabolome. Environmental research, 156, 341–348.

    Article  CAS  PubMed  Google Scholar 

  • Wahlig, S., Peh, G. S. L., Adnan, K., Ang, H. P., Lwin, C. N., Morales-Wong, F., et al. (2020). Optimisation of storage and transportation conditions of cultured corneal endothelial cells for cell replacement therapy. Scientific Reports, 10(1), 1681.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Walker, V. (2009). Ammonia toxicity and its prevention in inherited defects of the urea cycle. Diabetes Obesity and Metabolism, 11(9), 823–835.

    Article  CAS  Google Scholar 

  • Wojcik, G., Ferrari, S., Romano, V., Ponzin, D., Ahmad, S., & Parekh, M. (2021). Corneal storage methods: considerations and impact on surgical outcomes. Expert Review of Ophthalmology, 16(1), 1–9.

    Article  CAS  Google Scholar 

  • Zhang, W., Li, H., Ogando, D. G., Li, S., Feng, M., Price, F. W., et al. (2017). Glutaminolysis is essential for energy production and ion transport in human corneal endothelium. EBioMedicine, 16, 292–301.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang, X., Li, Q., & Dou, J. (2020). Mass spectrometry-based metabolomics in health and medical science: A systematic review. RSC advances, 10(6), 3092–3104.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhao, C., Li, W., Duan, H., Li, Z., Jia, Y., Zhang, S., et al. (2020). NAD + precursors protect corneal endothelial cells from UVB-induced apoptosis. American Journal of Physiology-Cell Physiology, 318(4), C796–C805.

    Article  CAS  PubMed  Google Scholar 

  • Zheng, X., Baker, H., Hancock, W. S., Fawaz, F., McCaman, M., & Pungor, E. Jr. (2006). Proteomic analysis for the assessment of different lots of fetal bovine serum as a raw material for cell culture. Part IV. Application of proteomics to the manufacture of biological drugs. Biotechnology progress, 22(5), 1294–1300.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Chemistry Interdisciplinary Project (ChIP), School of Pharmacy, University of Camerino, I-62032, Camerino, Italy

    Massimo Ricciutelli, Simone Angeloni, Giovanni Caprioli & Gianni Sagratini

  2. The Marche Region Eye Bank, AST Ancona - E. Profili Hospital, 60044, Fabriano, Italy

    Silvia Conforti & Massimiliano Corneli

  3. Department of Chemistry, Biology and Biotechnology, University of Perugia, 06100, Perugia, Italy

    Husam B.R. Alabed & Roberto Maria Pellegrino

  4. Research and Development, AL.CHI.MI.A. S.R.L,, Ponte San Nicolò, Italy

    Jana D’Amato Tóthová

Authors
  1. Massimo Ricciutelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Simone Angeloni
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Silvia Conforti
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Massimiliano Corneli
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Giovanni Caprioli
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gianni Sagratini
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Husam B.R. Alabed
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jana D’Amato Tóthová
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Roberto Maria Pellegrino
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

MR and MC, designed the experiments; SA, MR, MC and SC performed the experiments, analyzed the data, and wrote the manuscript; SA and MR developed the untargeted metabolomics approach; MC and SC provided the samples and conducted the human cornea preservation and storage; SA and MR analyzed, elaborated and processed the data; SA, MR, MC and SC conceived the study and participated in its design and coordination. GC, GS, RMP, HBRA and JDT supervised and reviewed the manuscript. All authors have read and agreed to the final version of the manuscript.

Corresponding author

Correspondence to Simone Angeloni.

Ethics declarations

Conflict of interest

All authors declare no conflict of interest.

Research involving human and animal rights

This article does not contain any studies with animal and human performed by any of the authors. The cornea used in the present work was not suitable for transplantation and for tissues appointed as not suitable for transplantation the characterization of medium is allowed for validating the medium capacity in maintaining active and operative the tissue metabolism, as it has been stated in the note of 23/12/2021 of Centro Nazionale Trapianti (CNT). Furthermore, even with suitable tissue, the developed analytical procedure does not require ethical committee approval. The methodology involves only the collection of small quantities of medium (the cornea is not manipulated and disrupted), which do not interfere with the organ culture.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ricciutelli, M., Angeloni, S., Conforti, S. et al. An untargeted metabolomics approach to study changes of the medium during human cornea culture. Metabolomics 20, 44 (2024). https://doi.org/10.1007/s11306-024-02102-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11306-024-02102-5

Keywords

Search

Navigation