"Physical Characterisation as an Insight into a Gene Delivery System Containing Cyclodextrins with Pluronic® -F127 and Folic acid as Non-Viral Vectors"

Mathew Eng, Amal Elkordy, Paul McCarron, Iman Elkordy, Ahmed Faheem

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Gene delivery into cells offers opportunities to treat various human genetic disease. Effective gene delivery is dependent on its stability and ability to transfect across cells. DNA is susceptible to enzymatic degradation and its negatively charge are barriers towards successful transfection. DNA has to be protected from degradation and neutralised. Non-viral vectors are preferred carrier systems, therefore, the use of cyclodextrins with Pluronic®-F127 and folic acid at different concentrations to stabilise the formulation was investigated. Formulations were characterised in fresh and freeze dried forms. DNA stability in formulations was tested by determining the stability of DNA against enzymatic degradation. Degree of DNA inclusion into cyclodextrins was investigated using fluorescence spectroscopy. Thermal behaviour was studied using Differential Scanning Calorimetry (DSC). Incorporation of Pluronic®-F127 produced most stable formulations regarding enzymatic degradation. These formulations show high percentage inclusion. Shift of peaks in FTIR data, appearance of uniform particulate as detected by SEM and changing in the denaturation temperature as demonstrated by DSC data for Pluronic®-F127 containing formulations confirm clear interaction between Pluronic®-F127 and cyclodextrin/ DNA complex. It was noted that γ-cyclodextrin provide better protection and inclusion compared to β-cyclodextrin. Pluronic®-F127 with cyclodextrins is a promising combination to improve stability.
LanguageEnglish
Pages712-726
JournalCurrent Pharmaceutical Biotechnology
Volume15
Issue number8
Publication statusPublished - 2014

Fingerprint

UCON 50-HB-5100
Poloxamer
Gene Transfer Techniques
Cyclodextrins
Folic Acid
DNA
Differential Scanning Calorimetry
Inborn Genetic Diseases
Fluorescence Spectrometry
Medical Genetics
Fourier Transform Infrared Spectroscopy
Genes
Transfection
Hot Temperature

Keywords

  • Cyclodextrin
  • deoxyribonucleic acid (DNA)
  • DNA degradation
  • gene delivery
  • non-viral vectors
  • Pluronic®-F127

Cite this

@article{bfba847085d6494fbd685c9ca776fc3a,
title = "{"}Physical Characterisation as an Insight into a Gene Delivery System Containing Cyclodextrins with Pluronic{\circledR} -F127 and Folic acid as Non-Viral Vectors{"}",
abstract = "Gene delivery into cells offers opportunities to treat various human genetic disease. Effective gene delivery is dependent on its stability and ability to transfect across cells. DNA is susceptible to enzymatic degradation and its negatively charge are barriers towards successful transfection. DNA has to be protected from degradation and neutralised. Non-viral vectors are preferred carrier systems, therefore, the use of cyclodextrins with Pluronic{\circledR}-F127 and folic acid at different concentrations to stabilise the formulation was investigated. Formulations were characterised in fresh and freeze dried forms. DNA stability in formulations was tested by determining the stability of DNA against enzymatic degradation. Degree of DNA inclusion into cyclodextrins was investigated using fluorescence spectroscopy. Thermal behaviour was studied using Differential Scanning Calorimetry (DSC). Incorporation of Pluronic{\circledR}-F127 produced most stable formulations regarding enzymatic degradation. These formulations show high percentage inclusion. Shift of peaks in FTIR data, appearance of uniform particulate as detected by SEM and changing in the denaturation temperature as demonstrated by DSC data for Pluronic{\circledR}-F127 containing formulations confirm clear interaction between Pluronic{\circledR}-F127 and cyclodextrin/ DNA complex. It was noted that γ-cyclodextrin provide better protection and inclusion compared to β-cyclodextrin. Pluronic{\circledR}-F127 with cyclodextrins is a promising combination to improve stability.",
keywords = "Cyclodextrin, deoxyribonucleic acid (DNA), DNA degradation, gene delivery, non-viral vectors, Pluronic{\circledR}-F127",
author = "Mathew Eng and Amal Elkordy and Paul McCarron and Iman Elkordy and Ahmed Faheem",
note = "Reference text: [1] Anchordoquy, J.T., Allison, S.D., Molina, M.d.C., Girouard ,L.G., Carson, T.K. Physical stabilization of DNA-based therapeutics. Drug Discovery Today. 2001; 6: 463-470. [2] Niidome, T., Huang, L. Gene Therapy Progress and Prospects: Nonviral Vectors. Gene Therapy (Nature) 2002; 9: 1647-1652. [3] Wong, S.Y., Pelet, J.M., Putnam, D. Polymer systems for gene delivery- past, present and future. Progress in Polymer Science. 2007; 32: 799-837. [4] Park, T.G., Jeong, J.H., Kim, S.W. Current status of polymeric gene delivery systems. Advanced Drug Delivery Reviews. 2006; 58: 467- 486. [5] Abdelhady, H.G., Allen, S., Davies, M.C., Roberts, C.J., Tendler, S.J., Williams, P.M. Direct real-time molecular scale visualisation of the degradation of condensed DNA complexes exposed to DNase I. Nucleic Acids Research. 2003; 31: 4001-4005. [6] Lechardeur, D., Sohn, K-J., Haardt, M., Joshi, P.B., Monck, M., Graham, R.W., Beatty, B., Squire, J. H., O’Brodovich, Lukacs, G.L. Metabolic instability of plasmid DNA in the cytosol: a potential barrier to gene transfer. Gene Therapy. 1999; 6: 482-497. [7] Brown, M.D., Schatzlein, A.G., Uchegbu, I.F. Gene delivery with synthetic (non viral) carriers. Int. J. Pharm. 2001; 229: 1-21. [8] Schaffer, D.V., Lauffenburger, D.A. Optimization of Cell Surface Binding Enhances Efficiency and Specificity of Molecular Conjugate Gene Delivery. J. Biol. Chem. 1998; 273: 28004-28009. [9] Herweijer, H., Wolff, J.A. Progress and prospects: naked DNA gene transfer and therapy. Gene Therapy (Nature). 20003; 10: 453- 458. [10] Williamson, J.F. Advances and Perspectives in intracellular biomolecule delivery. Pharmaceutical Technology Europe 2008. [11] Well, D.J. Review Gene Therapy Progress and Prospects: Electroporation and other physical methods. Gene Therapy (Nature). 2004; 11: 1363-1369. [12] Huang, L., Li, S. Non-viral gene therapy: promises and challenges. Gene Therapy (Nature). 2000; 7: 31-34. [13] David, M.E. Non-viral gene delivery systems. Curr. Opin. Biotech. 2002; 13: 28-131. [14] Lv, H., Zhang, S., Wang, B., Cui, S., Yan, J. Toxicity of cationic lipids and cationic polymers in gene delivery. J. Cont. Release. 2006; 114: 100-109. [15] Del Valle, E.M.M. Cyclodextrins and their uses: a review. Process Biochem. 2004; 39: 1033-1046. [16] Loftsson, T. Cyclodextrins I: Physicochemical Properties and in vitro Evaluation. AAPS Webinar 2011. [17] Rasheed, A., Kumar, A., Sravanthi, V. V. N. S. S. Cyclodextrins as Drug Carrier Molecule: A Review. Sci Pharm. 2008; 76: 567-598. [18] Szejtli, J. Medicinal applications of cyclodextrins. Medicinal Research Reviews. 1994; 14: 353-386. [19] Stella, V.J., Rajeswski, R.A. Cyclodextrins: Their future in drug formulations and delivery. Pharm. Res. 1997; 14: 556-567. [20] Burckbuchler, V., Wintgens, V., Leborgne, C., Lecomte, S., Leygue, N., Scherman, D., Kichler, A., Amiel, C. Development and Characterization of New Cyclodextrin Polymer-Based DNA Delivery Systems. Bioconjugate Chemistry. 2008; 19(12): 2311-2320. [21] Strappe, P.M., Hampton, D.W., Cachon-Gonzalez, B., Fawcett, J.W., Lever, A. Delivery of a lentiviral vector in a Pluronic F127 gel to cells of the central nervous system. Eur. J. Pharm. Biopharm. 2005; 61: 126-133. [22] Guo, W., Lee, R.L. Receptor-targeted gene delivery via folate conjugated polyethylenimine. AAPS PharmSci. 1999; 1(4). [23] Lyscov, V., N., Moshkovsky, Y.,S. DNA cryolysis. Biochim. Biophy. Acta. 1969; 190: 101-110. [24] Maitani, Y., Asob, Y., Yamadaa, A., Yoshioka, S. Effect of sugars on storage stability of lyophilized liposome/DNA complexes with high transfection efficiency. Int. J. Pharm. 2008; 356: 69-75. [25] Anchordoquy, T.J., Giourourard, L.G., Carpenter, J.F., Kroll, D.J. Stability of lipid/DNA complexes during agitation and freezethawing. J. Pharm. Sci. 1988; 87: 1046-1051. [26] Shikama, K. Effect of freezing and thawing on the stability of double helix of DNA. Nature 1965; 207: 529-530. [27] Talsma, H., Cherng, J.Y., Lehrmann, H., Kursa, M., Ogris, M., Hennink, W.E., Cotton, M., Wagner, E. Stabilization of gene delivery systems by freeze-drying. Int. J. Pharm. 1997; 157: 233-238. [28] Cryan,S.A., Holohan, A., Donhue, R., Darcy, R., O’Driscoll. Cell transfection with polycationic cyclodextrin vectors. Eur. J. Pharm. Sci. 2004; 21: 625-633. [29] Cooper, A., Nutley, M.A., Wadood, A. Differential scanning microcalorimetry. In S. E. Harding and B. Z. Chowdhry (Eds.), Protein- Ligand Interactions: hydrodynamics and calorimetry. Oxford University Press, Oxford New York, 2000; 287-318. [30] Rengarajan, K., Cristol, S.M., Mehta, M., Nickerson, J.M. Quantifying DNA concentrations using fluorometry: A comparison of fluorophores. Molecular Vision. 2002; 8: 416-421. [31] Li, B., Li, S., Tan, Y., Stolz, D.B., Watkins, S.C., Block, L.H. Lyophilization of cationic lipid-protamine- DNA (LPD) complexes. J. Pharm. Sci. 2000; 89: 355-364. [32] Pozo-Rodr{\'i}guez, A.D., Solin{\'i}s, M.A., Gasc{\'o}n, A.R., PedrazShort, J.L. Short and long-term stability study of lyophilized solid lipid nanoparticles for gene therapy. Eur. J. Pharm. Biopharm. 2009; 71: 181-189. [33] Beba, H.M., Elkordy, A.A. Carrier systems in gene delivery - progressive research in mental health pharmacy. Clinical Pharmacist. 2011; 3 (3): S2-S3. [34] Zhang, Y., Lam, Y.M. Study of Mixed Micelles and Interaction Parameters for Polymeric Nonionic and Normal Surfactants, American Scientific Publishers. 2006; 6: 1-5. [35] Linse, P. Micellization of poly(ethylene oxide)-poly(propylene oxide) triblock copolymers in aqueous solution. Macromolecules. 1993; 26: 4437-4449 [36] Bohorqueza, M., Kocha, M., Trygstadb, T., abd Pandit, N. Study of the Temperature-Dependent Micellization of Pluronic F127. J. Colloid and Interface Science. 1999; 216: 34-40. [37] Mao, Y., Daniel, L.N., Whittaker, N., Saffiottil, U. DNA Binding to Crystalline Silica Characterized by Fourier-Transform Infrared Spectroscopy. Environmental Health Perspectives 1993. [38] Ruiz-Chica, A.J., Khomutov, A.R., Medina, M.A., Sanchez- Jimenez, F., Ramirez, F.J. Interaction of DNA with an aminooxy analogue of spermidine {\DH} an FT-IR and FT-Raman approach. J. Molecular Structure. 2001; 565; 253-258. [39] Ouameur, A.A., Tajmir-Riah, H.A. Structural Analysis of DNA Interactions with Biogenic Polyamines and Cobalt(III) hexamine Studied by Fourier Transform Infrared and Capillary Electrophoresis. J. Biol. Chem. 2004; 279(40): 42041-42054.",
year = "2014",
language = "English",
volume = "15",
pages = "712--726",
journal = "Current Pharmaceutical Biotechnology",
issn = "1389-2010",
number = "8",

}

"Physical Characterisation as an Insight into a Gene Delivery System Containing Cyclodextrins with Pluronic® -F127 and Folic acid as Non-Viral Vectors". / Eng, Mathew; Elkordy, Amal; McCarron, Paul; Elkordy, Iman; Faheem, Ahmed.

In: Current Pharmaceutical Biotechnology, Vol. 15, No. 8, 2014, p. 712-726.

Research output: Contribution to journalArticle

TY - JOUR

T1 - "Physical Characterisation as an Insight into a Gene Delivery System Containing Cyclodextrins with Pluronic® -F127 and Folic acid as Non-Viral Vectors"

AU - Eng, Mathew

AU - Elkordy, Amal

AU - McCarron, Paul

AU - Elkordy, Iman

AU - Faheem, Ahmed

N1 - Reference text: [1] Anchordoquy, J.T., Allison, S.D., Molina, M.d.C., Girouard ,L.G., Carson, T.K. Physical stabilization of DNA-based therapeutics. Drug Discovery Today. 2001; 6: 463-470. [2] Niidome, T., Huang, L. Gene Therapy Progress and Prospects: Nonviral Vectors. Gene Therapy (Nature) 2002; 9: 1647-1652. [3] Wong, S.Y., Pelet, J.M., Putnam, D. Polymer systems for gene delivery- past, present and future. Progress in Polymer Science. 2007; 32: 799-837. [4] Park, T.G., Jeong, J.H., Kim, S.W. Current status of polymeric gene delivery systems. Advanced Drug Delivery Reviews. 2006; 58: 467- 486. [5] Abdelhady, H.G., Allen, S., Davies, M.C., Roberts, C.J., Tendler, S.J., Williams, P.M. Direct real-time molecular scale visualisation of the degradation of condensed DNA complexes exposed to DNase I. Nucleic Acids Research. 2003; 31: 4001-4005. [6] Lechardeur, D., Sohn, K-J., Haardt, M., Joshi, P.B., Monck, M., Graham, R.W., Beatty, B., Squire, J. H., O’Brodovich, Lukacs, G.L. Metabolic instability of plasmid DNA in the cytosol: a potential barrier to gene transfer. Gene Therapy. 1999; 6: 482-497. [7] Brown, M.D., Schatzlein, A.G., Uchegbu, I.F. Gene delivery with synthetic (non viral) carriers. Int. J. Pharm. 2001; 229: 1-21. [8] Schaffer, D.V., Lauffenburger, D.A. Optimization of Cell Surface Binding Enhances Efficiency and Specificity of Molecular Conjugate Gene Delivery. J. Biol. Chem. 1998; 273: 28004-28009. [9] Herweijer, H., Wolff, J.A. Progress and prospects: naked DNA gene transfer and therapy. Gene Therapy (Nature). 20003; 10: 453- 458. [10] Williamson, J.F. Advances and Perspectives in intracellular biomolecule delivery. Pharmaceutical Technology Europe 2008. [11] Well, D.J. Review Gene Therapy Progress and Prospects: Electroporation and other physical methods. Gene Therapy (Nature). 2004; 11: 1363-1369. [12] Huang, L., Li, S. Non-viral gene therapy: promises and challenges. Gene Therapy (Nature). 2000; 7: 31-34. [13] David, M.E. Non-viral gene delivery systems. Curr. Opin. Biotech. 2002; 13: 28-131. [14] Lv, H., Zhang, S., Wang, B., Cui, S., Yan, J. Toxicity of cationic lipids and cationic polymers in gene delivery. J. Cont. Release. 2006; 114: 100-109. [15] Del Valle, E.M.M. Cyclodextrins and their uses: a review. Process Biochem. 2004; 39: 1033-1046. [16] Loftsson, T. Cyclodextrins I: Physicochemical Properties and in vitro Evaluation. AAPS Webinar 2011. [17] Rasheed, A., Kumar, A., Sravanthi, V. V. N. S. S. Cyclodextrins as Drug Carrier Molecule: A Review. Sci Pharm. 2008; 76: 567-598. [18] Szejtli, J. Medicinal applications of cyclodextrins. Medicinal Research Reviews. 1994; 14: 353-386. [19] Stella, V.J., Rajeswski, R.A. Cyclodextrins: Their future in drug formulations and delivery. Pharm. Res. 1997; 14: 556-567. [20] Burckbuchler, V., Wintgens, V., Leborgne, C., Lecomte, S., Leygue, N., Scherman, D., Kichler, A., Amiel, C. Development and Characterization of New Cyclodextrin Polymer-Based DNA Delivery Systems. Bioconjugate Chemistry. 2008; 19(12): 2311-2320. [21] Strappe, P.M., Hampton, D.W., Cachon-Gonzalez, B., Fawcett, J.W., Lever, A. Delivery of a lentiviral vector in a Pluronic F127 gel to cells of the central nervous system. Eur. J. Pharm. Biopharm. 2005; 61: 126-133. [22] Guo, W., Lee, R.L. Receptor-targeted gene delivery via folate conjugated polyethylenimine. AAPS PharmSci. 1999; 1(4). [23] Lyscov, V., N., Moshkovsky, Y.,S. DNA cryolysis. Biochim. Biophy. Acta. 1969; 190: 101-110. [24] Maitani, Y., Asob, Y., Yamadaa, A., Yoshioka, S. Effect of sugars on storage stability of lyophilized liposome/DNA complexes with high transfection efficiency. Int. J. Pharm. 2008; 356: 69-75. [25] Anchordoquy, T.J., Giourourard, L.G., Carpenter, J.F., Kroll, D.J. Stability of lipid/DNA complexes during agitation and freezethawing. J. Pharm. Sci. 1988; 87: 1046-1051. [26] Shikama, K. Effect of freezing and thawing on the stability of double helix of DNA. Nature 1965; 207: 529-530. [27] Talsma, H., Cherng, J.Y., Lehrmann, H., Kursa, M., Ogris, M., Hennink, W.E., Cotton, M., Wagner, E. Stabilization of gene delivery systems by freeze-drying. Int. J. Pharm. 1997; 157: 233-238. [28] Cryan,S.A., Holohan, A., Donhue, R., Darcy, R., O’Driscoll. Cell transfection with polycationic cyclodextrin vectors. Eur. J. Pharm. Sci. 2004; 21: 625-633. [29] Cooper, A., Nutley, M.A., Wadood, A. Differential scanning microcalorimetry. In S. E. Harding and B. Z. Chowdhry (Eds.), Protein- Ligand Interactions: hydrodynamics and calorimetry. Oxford University Press, Oxford New York, 2000; 287-318. [30] Rengarajan, K., Cristol, S.M., Mehta, M., Nickerson, J.M. Quantifying DNA concentrations using fluorometry: A comparison of fluorophores. Molecular Vision. 2002; 8: 416-421. [31] Li, B., Li, S., Tan, Y., Stolz, D.B., Watkins, S.C., Block, L.H. Lyophilization of cationic lipid-protamine- DNA (LPD) complexes. J. Pharm. Sci. 2000; 89: 355-364. [32] Pozo-Rodríguez, A.D., Solinís, M.A., Gascón, A.R., PedrazShort, J.L. Short and long-term stability study of lyophilized solid lipid nanoparticles for gene therapy. Eur. J. Pharm. Biopharm. 2009; 71: 181-189. [33] Beba, H.M., Elkordy, A.A. Carrier systems in gene delivery - progressive research in mental health pharmacy. Clinical Pharmacist. 2011; 3 (3): S2-S3. [34] Zhang, Y., Lam, Y.M. Study of Mixed Micelles and Interaction Parameters for Polymeric Nonionic and Normal Surfactants, American Scientific Publishers. 2006; 6: 1-5. [35] Linse, P. Micellization of poly(ethylene oxide)-poly(propylene oxide) triblock copolymers in aqueous solution. Macromolecules. 1993; 26: 4437-4449 [36] Bohorqueza, M., Kocha, M., Trygstadb, T., abd Pandit, N. Study of the Temperature-Dependent Micellization of Pluronic F127. J. Colloid and Interface Science. 1999; 216: 34-40. [37] Mao, Y., Daniel, L.N., Whittaker, N., Saffiottil, U. DNA Binding to Crystalline Silica Characterized by Fourier-Transform Infrared Spectroscopy. Environmental Health Perspectives 1993. [38] Ruiz-Chica, A.J., Khomutov, A.R., Medina, M.A., Sanchez- Jimenez, F., Ramirez, F.J. Interaction of DNA with an aminooxy analogue of spermidine Ð an FT-IR and FT-Raman approach. J. Molecular Structure. 2001; 565; 253-258. [39] Ouameur, A.A., Tajmir-Riah, H.A. Structural Analysis of DNA Interactions with Biogenic Polyamines and Cobalt(III) hexamine Studied by Fourier Transform Infrared and Capillary Electrophoresis. J. Biol. Chem. 2004; 279(40): 42041-42054.

PY - 2014

Y1 - 2014

N2 - Gene delivery into cells offers opportunities to treat various human genetic disease. Effective gene delivery is dependent on its stability and ability to transfect across cells. DNA is susceptible to enzymatic degradation and its negatively charge are barriers towards successful transfection. DNA has to be protected from degradation and neutralised. Non-viral vectors are preferred carrier systems, therefore, the use of cyclodextrins with Pluronic®-F127 and folic acid at different concentrations to stabilise the formulation was investigated. Formulations were characterised in fresh and freeze dried forms. DNA stability in formulations was tested by determining the stability of DNA against enzymatic degradation. Degree of DNA inclusion into cyclodextrins was investigated using fluorescence spectroscopy. Thermal behaviour was studied using Differential Scanning Calorimetry (DSC). Incorporation of Pluronic®-F127 produced most stable formulations regarding enzymatic degradation. These formulations show high percentage inclusion. Shift of peaks in FTIR data, appearance of uniform particulate as detected by SEM and changing in the denaturation temperature as demonstrated by DSC data for Pluronic®-F127 containing formulations confirm clear interaction between Pluronic®-F127 and cyclodextrin/ DNA complex. It was noted that γ-cyclodextrin provide better protection and inclusion compared to β-cyclodextrin. Pluronic®-F127 with cyclodextrins is a promising combination to improve stability.

AB - Gene delivery into cells offers opportunities to treat various human genetic disease. Effective gene delivery is dependent on its stability and ability to transfect across cells. DNA is susceptible to enzymatic degradation and its negatively charge are barriers towards successful transfection. DNA has to be protected from degradation and neutralised. Non-viral vectors are preferred carrier systems, therefore, the use of cyclodextrins with Pluronic®-F127 and folic acid at different concentrations to stabilise the formulation was investigated. Formulations were characterised in fresh and freeze dried forms. DNA stability in formulations was tested by determining the stability of DNA against enzymatic degradation. Degree of DNA inclusion into cyclodextrins was investigated using fluorescence spectroscopy. Thermal behaviour was studied using Differential Scanning Calorimetry (DSC). Incorporation of Pluronic®-F127 produced most stable formulations regarding enzymatic degradation. These formulations show high percentage inclusion. Shift of peaks in FTIR data, appearance of uniform particulate as detected by SEM and changing in the denaturation temperature as demonstrated by DSC data for Pluronic®-F127 containing formulations confirm clear interaction between Pluronic®-F127 and cyclodextrin/ DNA complex. It was noted that γ-cyclodextrin provide better protection and inclusion compared to β-cyclodextrin. Pluronic®-F127 with cyclodextrins is a promising combination to improve stability.

KW - Cyclodextrin

KW - deoxyribonucleic acid (DNA)

KW - DNA degradation

KW - gene delivery

KW - non-viral vectors

KW - Pluronic®-F127

M3 - Article

VL - 15

SP - 712

EP - 726

JO - Current Pharmaceutical Biotechnology

T2 - Current Pharmaceutical Biotechnology

JF - Current Pharmaceutical Biotechnology

SN - 1389-2010

IS - 8

ER -