Abstract
Given the recent explosion of mass spectrometric imaging (MSI), it has become easier to assess drug tissue localisation without the use of radiolabeling and other more complex methods (such as PET and MRI). For MSI tissue preparation is of utmost importance, however, the lung in particular does pose some difficulties with imaging since it is made up of a number of air-filled alveoli. These organs are known to collapse when the thoracic cavity is pierced, losing its structural integrity and giving poor histological representation for drug distribution analysis. The use of cryoprotectants as a tissue inflation media will aid in the preservation of the lung’s structural integrity during MSI experiments involving small molecule distribution. Various established cryoprotectants (DMSO, PvP, ethylene glycol, sucrose, DMEM, control serum, OCT) were selected as lung inflation media for MSI analysis of gatifloxacin (GAT). Female Sprague–Dawley rats were treated with GAT (10 mg/kg b.w) via i.p. injection. After 15 min the animals were terminated by halothane overdose, and each set of tissue inflated with a specific agent. Cryosections were made and MSI conducted to determine drug tissue distribution. During the early stages of the experimental procedure some crypreservatives were eliminated due to difficulties with sample preparation. While others displayed excellent preservation of the tissue structure and integrity. Following MSI analysis, some agents showed homogenous drug distribution while some displayed heterogeneous distribution favoring the basal periphery. Taking into account the physiology of the lung and previous MRI investigations of its perfusion, it is expected that a systemically administered drug would localize in the basal areas. DMSO and DMEM proved to display this distribution pattern while keeping structural integrity intact. However, the later was ruled out since it showed complete suppression of GAT in solution. From the cryoprotectants selected for this study, DMSO is the most promising lung inflation media focusing on small molecule distribution via MSI.
Similar content being viewed by others
References
Addie RD, Balluff B, Bovée JV, Morreau H, McDonnell LA (2015) Current State and future challenges of mass spectrometry imaging for clinical research. Anal Chem 87:6426–6433
Aichler M et al (2015) Spatially resolved quantification of gadolinium(III)-based magnetic resonance agents in tissue by MALDI imaging mass spectrometry after in vivo MRI. Angew Chem 127:4353–4357
Baatz JE, Newton DA, Riemer EC, Denlinger CE, Jones EE, Drake RR, Spyropoulos DD (2014) Cryopreservation of viable human lung tissue for versatile post-thaw analyses and culture. In Vivo 28:411–423
Baijnath S et al (2015) Evidence for the presence of clofazimine and its distribution in the healthy mouse brain. J Mol Histol 46:439–442
Berry KAZ et al (2011) MALDI imaging MS of phospholipids in the mouse lung. J Lipid Res 52:1551–1560
Braber S, Verheijden KA, Henricks PA, Kraneveld AD, Folkerts G (2010) A comparison of fixation methods on lung morphology in a murine model of emphysema. Am J Physiol Lung Cell Mol Physiol 299:L843–L851
Brittain HG (2007) Profiles of drug substances, excipients and related methodology: critical compilation of pKa values for pharmaceutical substances, vol 33. Academic Press, Cambridge
Brockbank KGM, Taylor MJ (2007) 8 Tissue preservation. In: Baust JG, Baust JM (eds) Advances in Biopreservation. CRC Press, Boca Raton, pp 157–195
Buckingham KW, Wyder WE (1981) Rapid tracheal infusion method for routine lung fixation using rat and guinea pig. Toxicol Pathol 9:17–20
Costanzo LS (2003) Physiology, philadelphia. Saunders, Elsevier
Deininger S-O et al (2011) Normalization in MALDI-TOF imaging datasets of proteins: practical considerations. Anal Bioanal Chem 401:167–181
Desbenoit N et al (2014) Localized lipidomics in cystic fibrosis: TOF-SIMS imaging of lungs from Pseudomonas aeruginosa-infected mice. Int J Biochem Cell Biol 52:77–82
Dulbecco R, Freeman G (1959) Plaque production by the polyoma virus Virology 8:396–397
Hirsch FR et al (1988) Histopathologic classification of small cell lung cancer changing concepts and terminology. Cancer 62:973–977
Järver P, Langel Ü (2004) The use of cell-penetrating peptides as a tool for gene regulation. Drug Discov Today 9:395–402
Liu X et al (2013) Molecular imaging of drug transit through the blood-brain barrier with MALDI mass spectrometry imaging. Sci Rep. doi:10.1038/srep02859
Mäe M, Langel Ü (2006) Cell-penetrating peptides as vectors for peptide, protein and oligonucleotide delivery. Curr Opin Pharmacol 6:509–514
Mancia A, Spyropoulos DD, McFee WE, Newton DA, Baatz JE (2012) Cryopreservation and in vitro culture of primary cell types from lung tissue of a stranded pygmy sperm whale (Kogia breviceps). Comp Biochem Physiol Part C Toxicol Pharmacol 155:136–142
Munyeza CF et al (2015) Rapid and widespread distribution of doxycycline in rat brain: a mass spectrometric imaging study. Xenobiotica. doi:10.3109/00498254.2015.10851307
Nikolaou K et al (2004) Quantification of pulmonary blood flow and volume in healthy volunteers by dynamic contrast-enhanced magnetic resonance imaging using a parallel imaging technique. Invest Radiol 39:537–545
Nunn JF (2013) Applied respiratory physiology. Butterworth-Heinemann, Oxford
Oakes JM, Scadeng M, Breen EC, Prisk GK, Darquenne C (2013) Regional distribution of aerosol deposition in rat lungs using magnetic resonance imaging. Ann Biomed Eng 41:967–978
Oakes JM, Breen EC, Scadeng M, Tchantchou GS, Darquenne C (2014a) MRI-based measurements of aerosol deposition in the lung of healthy and elastase-treated rats. J Appl Physiol 116:1561–1568
Oakes JM, Marsden AL, Grandmont C, Shadden SC, Darquenne C, Vignon-Clementel IE (2014b) Airflow and particle deposition simulations in health and emphysema: from in vivo to in silico animal experiments. Ann Biomed Eng 42:899–914
Ohno Y et al (2004) Quantitative assessment of regional pulmonary perfusion in the entire lung using three-dimensional ultrafast dynamic contrast-enhanced magnetic resonance imaging: preliminary experience in 40 subjects. J Magn Reson Imaging 20:353–365
Porta T, Lesur A, Varesio E, Hopfgartner G (2015) Quantification in MALDI-MS imaging: What can we learn from MALDI-selected reaction monitoring and what can we expect for imaging? Anal Bioanal Chem 407:2177–2187
Prideaux B et al (2011) High-sensitivity MALDI-MRM-MS imaging of moxifloxacin distribution in tuberculosis-infected rabbit lungs and granulomatous lesions. Anal Chem 83:2112–2118
Prideaux B et al (2015) The association between sterilizing activity and drug distribution into tuberculosis lesions. Nat Med 21:1223–1227
Shariatgorji M, Källback P, Gustavsson L, Schintu N, Svenningsson P, Goodwin RJ, Andren PE (2012) Controlled-pH tissue cleanup protocol for signal enhancement of small molecule drugs analyzed by MALDI-MS imaging. Anal Chem 84:4603–4607
Shobo A, Bratowska D, Naiker S, Somboro AM, Bester LA, Singh S, Naicker T, Kruger HG, Govender T (2015a) MALDI MSI and LC‐MS/MS: towards preclinical determination of the neurotoxic potential of fluoroquinolones. Drug Test Anal. doi:10.1002/dta.1862
Shobo A et al (2015b) Tissue distribution of pretomanid in rat brain via mass spectrometry imaging. Xenobiotica. doi:10.3109/00498254.2015.1067935
Stock KW, Chen Q, Levin D, Hatabu H, Edelman RR (1999) Demonstration of gravity-dependent lung perfusion with contrast-enhanced magnetic resonance imaging. J Magn Reson Imaging 9:557–561
Tomlinson L, Fuchser J, Fütterer A, Baumert M, Hassall DG, West A, Marshall PS (2014) Using a single, high mass resolution mass spectrometry platform to investigate ion suppression effects observed during tissue imaging. Rapid Commun Mass Spectrom 28:995–1003
Wiseman JM, Ifa DR, Zhu Y, Kissinger CB, Manicke NE, Kissinger PT, Cooks RG (2008) Desorption electrospray ionization mass spectrometry: imaging drugs and metabolites in tissues. Proc Natl Acad Sci 105:18120–18125
Acknowledgments
The authors would like to thank the National Research Foundation, SA; Aspenpharmacare, SA; and the University of KwaZulu-Natal, Durban, SA for having funded this project.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Baijnath, S., Shobo, A., Bester, L.A. et al. Small molecule distribution in rat lung: a comparison of various cryoprotectants as inflation media and their applicability to MSI. J Mol Hist 47, 213–219 (2016). https://doi.org/10.1007/s10735-016-9658-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10735-016-9658-3