More About Fluorescent Products

Fluorescent products have many useful functions in biological studies. The following is a summary of the potential uses.

  1. Permeability studies: fluorescent derivatives can be used for studying biological permeability in cells, tissues, humans (non-clinical), and animals. The permeability of fluorescent derivatives depends on the different product sizes, charges, and fluorescent types.
  2. Sugar metabolic studies: FITC-Trehalose is a good tracer for labeling mycomembranes to study sugar metabolism by bacteria.
  3. pH visualizations: fluorescent dyes can be used as pH-indicators in living cells. They can reflect the changing physiological pH of cells in response to various cellular processes.
  4. Hydrogels: dextran derivatives are useful for hydrogel formation due to their low toxicity and ability to incorporate well with hydrogel structural materials.
  5. Matrix preparation: CM-derivatives can be used as a matrix component for electrochemical immunoassays, regulation of scaffold cell adhesion, enzyme immobilization, construction of polymer membranes, and hydrophilization of substances.

1. Permeability Studies

Biological permeability is the passage of substances through a biological membrane or a barrier which can either be selectively or indiscriminately permeable. The permeability is affected by substance properties such as a polarity, hydrophobicity, charge, size, and shape (1). The properties of the membrane or barrier itself are also important. Fluorescent dextran derivatives or other polysaccharides of various sizes can be used for permeability and transport studies in cells, tissues, and animals. Fluorescent measurements can also provide qualitative data in real time with use of intravital fluorescence microscopy. More specifically, they are ideal for many types of studies such as vascular permeability (2,3), glomerular filtration (4–6), and evaluating the blood-brain barrier (7), internal tissue (14,15), neural stem cells (16), renal tissue (17), keratin permeability (8), and the epithelial (9–11) and mucosal (12,13) layers.  In addition, fluorescent dextran derivatives have been used to study microcirculation, the smallest level of blood circulation in the micro vessels present in all organ tissues (21), for applications such as leukocyte adhesion, macromolecular leakage (22), and intestinal mucosal microcirculation (23). Polysaccharides conjugated with carboxymethyl (CM)-or diethylaminomethyl (DEAE)-groups are useful for studying the effects of charge on permeability (18–20).
 

References  

  1.  J. Reece, N. Campbell. Campbell Biology (Benjamin Cummings/Pearson., 2008).
  2. C. Bulant, P. Blanco, L. Müller, J. Scharfstein, E. Svensjö, Computer-aided quantification of microvascular networks: Application to alterations due to pathological angiogenesis in the hamster. Microvasc Res 112, 53-64 (2017).
  3. C. Nascimento, D. Andrade, C. Carvalho-Pinto, R. Serra, L. Vellasco, et al., Mast Cell Coupling to the Kallikrein-Kinin System Fuels Intracardiac Parasitism and Worsens Heart Pathology in Experimental Chagas Disease. Front Immunol 8, 840 (2017).
  4. D. Asgeirsson, D. Venturoli, E. Fries, B. Rippe, C. Rippe, Glomerular sieving of three neutral polysaccharides, polyethylene oxide and bikunin in rat. Effects of molecular size and conformation. Acta Physiol (Oxf) 191, 237-46 (2007).
  5. J. Dolinina, K. Sverrisson, A. Rippe, C. Öberg, B. Rippe, Nitric oxide synthase inhibition causes acute increases in glomerular permeability in vivo, dependent upon reactive oxygen species. Am J Physiol Renal Physiol 311, F984-F990 (2016).
  6. C. Rippe, D. Asgeirsson, D. Venturoli, A. Rippe, B. Rippe, Effects of glomerular filtration rate on Ficoll sieving coefficients (theta) in rats. Kidney Int 69, 1326-32 (2006).
  7. S. Gustafsson, T. Gustavsson, S. Roshanbin, G. Hultqvist, M. Hammarlund-Udenaes, et al., Blood-brain barrier integrity in a mouse model of Alzheimer's disease with or without acute 3D6 immunotherapy. Neuropharmacology 143, 1-9 (2018).
  8. J. Navarro, J. Swayambunathan, M. Lerman, M. Santoro, J. Fisher, Development of keratin-based membranes for potential use in skin repair. Acta Biomater 83, 177-188 (2019).
  9. R. Bücker, S. Krug, V. Moos, C. Bojarski, M. Schweiger, et al., Campylobacter jejuni impairs sodium transport and epithelial barrier function via cytokine release in human colon. Mucosal Immunol 11, 474-485 (2018).
  10. D. Propheter, A. Chara, T. Harris, K. Ruhn, L. Hooper, Resistin-like molecule ? is a bactericidal protein that promotes spatial segregation of the microbiota and the colonic epithelium. Proc Natl Acad Sci U S A 114, 11027-11033 (2017).
  11. R. Bowie, S. Donatello, C. Lyes, M. Owens, I. Babina, et al., Lipid rafts are disrupted in mildly inflamed intestinal microenvironments without overt disruption of the epithelial barrier. Am J Physiol Gastrointest Liver Physiol 302, G781-93 (2012).
  12. S. Lee, H. Kim, K. Kim, H. Lee, S. Lee, D. Lee, et al., Arhgap17, a RhoGTPase activating protein, regulates mucosal and epithelial barrier function in the mouse colon. Sci Rep 6, 26923 (2016).
  13. T. Dolowschiak, A. Mueller, L. Pisan, R. Feigelman, B. Felmy, et al., IFN-? Hinders Recovery from Mucosal Inflammation during Antibiotic Therapy for Salmonella Gut Infection. Cell Host Microbe 20, 238-49 (2016).
  14. A. Torge, G. Pavone, M. Jurisic, K. Lima-Engelmann, & M. Schneider, A comparison of spherical and cylindrical microparticles composed of nanoparticles for pulmonary application. Aerosol Sci. Technol. 53, 53–62 (2019).
  15. H. Epple, et al., Architectural and functional alterations of the small intestinal mucosa in classical Whipple’s disease. Mucosal Immunol. 10, 1542–1552 (2017).
  16. C. Zhu, S. Mahesula, S. Temple, E. Kokovay, Heterogeneous Expression of SDF1 Retains Actively Proliferating Neural Progenitors in the Capillary Compartment of the Niche. Stem Cell Reports 12, 6-13 (2019).
  17. O. Palygin, V. Levchenko, D. Ilatovskaya, T. Pavlov, O. Pochynyuk, et al., Essential role of Kir5.1 channels in renal salt handling and blood pressure control. JCI Insight 2, e92331 (2017).
  18. N. Srikantha, F. Mourad, K. Suhling, N. Elsaid, J. Levitt, et al., Influence of molecular shape, conformability, net surface charge, and tissue interaction on transscleral macromolecular diffusion. Exp Eye Res 102, 85-92 (2012).
  19. D. Asgeirsson, D. Venturoli, B. Rippe, C. Rippe, Increased glomerular permeability to negatively charged Ficoll relative to neutral Ficoll in rats. Am J Physiol Renal Physiol 291, F1083-9 (2006).
  20. S. Stewart, S. Kondos, A. Matthews, M. D'Angelo, M. Dunstone, et al., The perforin pore facilitates the delivery of cationic cargos. J Biol Chem 289, 9172-81 (2014).
  21. W. Jackson, “Chapter 89 – Microcirculation” in Muscle, J. Hill & E. Olson, Eds.(Academic Press, 2012), pp. 1197–1206
  22. C. Simões, E. Svensjö, E. Bouskela, Effects of cromakalim and glibenclamide on arteriolar and venular diameters and macromolecular leakage in the microcirculation during ischemia/reperfusion. J Cardiovasc Pharmacol 39, 340-6 (2002).
  23. C. Schmidt, C. Lautenschläger, B. Petzold, Y. Sakr, G. Marx, A. Stallmach, et al., Confocal laser endomicroscopy reliably detects sepsis-related and treatment-associated changes in intestinal mucosal microcirculation. Br J Anaesth 111, 996-1003 (2013).

2. Sugar Metabolic Studies

In nature, trehalose can be found in animals, plants, and microorganisms. Fluorescein isothiocyanate trehalose (FITC-Trehalose), which is a fluorescent derivative of trehalose, proves to be a useful functional and imaging probe in studies of trehalose uptake by culture cells (1) or bacteria (2), and sugar metabolization in bacteria (3). 
 

References

  1. S. A. Mercado, N. K. H. Slater, Increased cryosurvival of osteosarcoma cells using an amphipathic pH-responsive polymer for trehalose uptake. Cryobiology. 73, 175–180 (2016).
  2. K. M. Backus, H. I. Boshoff, C. S. Barry, O. Boutureira, M. K. Patel, F. D’Hooge, S. S. Lee, L. E. Via, K. Tahlan, C. E. Barry 3rd, B. G. Davis, Uptake of unnatural trehalose analogs as a reporter for Mycobacterium tuberculosis. Nat. Chem. Biol. 7, 228–235 (2011).
  3. B. J. Ignacio, T. Bakkum, K. M. Bonger, N. I. Martin, S. I. van Kasteren, Metabolic labeling probes for interrogation of the host–pathogen interaction. Org. Biomol. Chem. 19, 2856–2870 (2021).

3. pH Visualizations

Fluorescent dyes have the ability of changing color in response to pH-changes, utilized for measuring pH in living cells. Changes in cellular pH can reflect a range of physiological processes, including muscle contraction, endocytosis, cell proliferation, apoptosis, and ion transport (1). Compared to microelectrode techniques, fluorescent pH-indicators also have greater spatial sampling capability (1). Fluorescent pH-indicators can be coupled with macromolecules, such as dextran. The advantage of using fluorescent dextran derivatives is that the molecules can be accumulated into specific intracellular compartments and don’t bind to cellular proteins (2). Dextran derivatives involving dye-entities like FITC, TRITC or Texas Red™ (3) can be combined in a single dextran in order to achieve more accurate results for a more information-dence readout.
 

References

  1. J. Han, K. Burgess, Fluorescent indicators for intracellular pH. Chem Rev 110, 2709-28 (2010).
  2. S. Takahashi, Y. Kagami, K. Hanaoka, T. Terai, T. Komatsu, et al., Development of a Series of Practical Fluorescent Chemical Tools To Measure pH Values in Living Samples. J Am Chem Soc 140, 5925-5933 (2018).
  3. R. Weigert, Ed. Advances in Intravital Microscopy From Basic to Clinical Research (Springer, Heidelberg, 2014).

4. Hydrogels

Dextran derivatives can be incorporated in hydrogels which can be considered matrices for the controlled release of drug molecules. Ongoing research aimed at improving the fabrication efficiency as well as the drug delivery capability of various anti-cancer drugs can utilize dextran derivatives as they exhibit high hydrogel formation capacity, low toxicity combined with high biocompatibility, and biodegradability (1). Fluorescent dextran derivatives, blue dextran, and other polysaccharides have been used for studying drug delivery with hydrogel scaffolds (2), drug release with microneedles arrays (3,4), drug loading features of nano-erythrocytes (5), and biphasic pulsatile drug release (6).
 

References

  1. S. Thompson, K. Cass, E. Stellwagen, Blue dextran-sepharose: an affinity column for the dinucleotide fold in proteins. Proc Natl Acad Sci U S A 72, 669-72 (1975).
  2. J. Grenier, et al. Mechanisms of pore formation in hydrogel scaffolds textured by freezedrying. Acta Biomaterialia 94, 195-203 (2019).
  3. E. Larrañeta, S. Stewart, S. Fallows, L. Birkhäuer, M. McCrudden, et al., A facile system to evaluate in vitro drug release from dissolving microneedle arrays. Int J Pharm 497, 62-9 (2016).
  4. R. Thakur, I. Tekko, F. Al-Shammari, A. Ali, H. McCarthy, R. Donnelly, et al., Rapidly dissolving polymeric microneedles for minimally invasive intraocular drug delivery. Drug Deliv Transl Res 6, 800-815 (2016).
  5. X. Dong, Y. Niu, Y. Ding, Y. Wang, J. Zhao, et al., Formulation and Drug Loading Features of Nano-Erythrocytes. Nanoscale Res Lett 12, 202 (2017).
  6. M. Beugeling, N. Grasmeijer, P. Born, d. van, d. van, et al., The mechanism behind the biphasic pulsatile drug release from physically mixed poly(dl-lactic(-co-glycolic) acid)-based compacts. Int J Pharm 551, 195-202 (2018).

5. Matrix Preparation

CM-dextran and CM-polysucrose can be used as a matrix component in order to prepare surfaces for cell adherence and culture in studies of magnetic nanoparticles targeted to cancer cells (1) and P-selectin (2), electrochemical immunoassays (3), regulation of scaffold cell adhesion (4) and enzyme immobilization (5), and more. CM-dextran can also be used for the construction of polymer membranes. Phenyl-dextran can use for coating medical devices to impart a more hydrophilic character.
 

References

  1. B. Ficko, C. NDong, P. Giacometti, K. Griswold, S. Diamond, A Feasibility Study of Nonlinear Spectroscopic Measurement of Magnetic Nanoparticles Targeted to Cancer Cells. IEEE Trans Biomed Eng 64, 972-979 (2017).
  2. M. Juenet, R. Aid-Launais, B. Li, A. Berger, J. Aerts, et al., Thrombolytic therapy based on fucoidan-functionalized polymer nanoparticles targeting P-selectin. Biomaterials 156, 204-216 (2018).
  3. H. Hwang, et al. MESIA: Magnetic force-assisted electrochemical sandwich immunoassays for quantification of prostate-specific antigen in human serum. Analytica Chimica Acta 1061, 92–100 (2019).
  4. M. Burke, J. Armstrong, A. Goodwin, R. Deller, B. Carter, et al., Regulation of Scaffold Cell Adhesion Using Artificial Membrane Binding Proteins. Macromol Biosci 17, (2017).
  5. T. Terentyeva, et al., Bioactive flakeshell capsules: soft silica nanoparticles for efficient enzyme immobilization. J. Mater. Chem. B 1, 3248–3256 (2013).

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