Controlling the Release of Proteins from Therapeutic Nanofibers: The Effect of Fabrication Modalities on Biocompatibility and Antimicrobial Activity of Lysozyme

Salem Seif; Viktoria Planz; Maike Windbergs

doi:10.1055/s-0042-109715

Planta Medica, Inhaltsverzeichnis

Planta Med 2017; 83(05): 445-452
DOI: 10.1055/s-0042-109715

Formulation and Delivery Systems of Natural Products

Original Papers

Georg Thieme Verlag KG Stuttgart · New York

Controlling the Release of Proteins from Therapeutic Nanofibers: The Effect of Fabrication Modalities on Biocompatibility and Antimicrobial Activity of Lysozyme

Autor*innen

Salem Seif

¹Saarland University, Department of Biopharmaceutics and Pharmaceutical Technology, Saarbrücken, Germany

²PharmBioTec GmbH, Saarbrücken, Germany
Viktoria Planz

¹Saarland University, Department of Biopharmaceutics and Pharmaceutical Technology, Saarbrücken, Germany

³Helmholtz Centre for Infection Research (HZI) and Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Department of Drug Delivery (DDEL), Saarbrücken, Germany
Maike Windbergs

¹Saarland University, Department of Biopharmaceutics and Pharmaceutical Technology, Saarbrücken, Germany

²PharmBioTec GmbH, Saarbrücken, Germany

³Helmholtz Centre for Infection Research (HZI) and Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Department of Drug Delivery (DDEL), Saarbrücken, Germany

Abstract

Therapeutic application of pharmacologically active proteins requires advanced drug delivery systems for stabilizing their activity and preventing denaturation during storage and patient treatment. Depending on their clinical target, controlled drug release is often required to achieve the intended therapeutic effect. In this context, electrospun nanofibers gained considerable attention. However, even though immediate drug release from such fibers can easily be realized, fiber mat fabrication providing long-term controlled protein release still bares challenges.

In this study, lysozyme was encapsulated in poly(vinyl alcohol) fibers followed by post-modification with MeOH, glutaraldehyde vapor, or UV light. Subsequently, a systematic investigation of the effect of these post-modification treatments on the physicochemical properties of the fibers and the stability and release kinetics of lysozyme was performed. MeOH treatment did not affect lysozyme release kinetics compared to untreated fibers, whereas glutaraldehyde vapor and UV light treatment prolonged the drug release. Infrared spectroscopy revealed cross-linking of the polymer by glutaraldehyde vapor, which reduced the lysozyme release from the fibers. Further, protein activity was significantly reduced for fibers treated with glutaraldehyde vapor and UV light. In addition, reduced viability was identified for cells in contact with glutaraldehyde vapor-treated fibers and, to a lesser extent, for UV light-treated fibers, whereas MeOH-treated fibers did not affect cell viability. These results elucidated the effects of fiber post-modification on the release kinetics, activity, and biocompatibility of protein drugs and can serve as guidance for rational development of nanomedicines for safe and effective therapeutic delivery of natural proteins.

Key words

lysozyme - electrospinning - nanomedicines - controlled release - protein activity - antimicrobials

Volltext

Referenzen

References
1 Watkins R, Wu L, Zhang C, Davis RM, Xu B. Natural product-based nanomedicine: recent advances and issues. Int J Nanomedicine 2015; 10: 6055-6074
2 Malik NN. Drug discovery: past, present and future. Drug Discov Today 2008; 13: 909-912
3 Leader B, Baca QJ, Golan DE. Protein therapeutics: a summary and pharmacological classification. Nat Rev Drug Discov 2008; 7: 21-39
4 Fleming A. Lysozyme: Presidentʼs Address. Proc R Soc Med 1932; 26: 71-84
5 Gorin G, Wang SF, Papapavlou L. Assay of lysozyme by its lytic action on M. lysodeikticus cells. Anal Biochem 1971; 39: 113-127
6 Meyer K, Hahnel E, Steinberg A. Lysozyme of plant origin. J Biol Chem 1946; 163: 733-740
7 Guenther F, Melzig MF. Investigation of lysozyme from the latices of Euphorbia coerulescens and Euphorbia fortissima . Planta Med 2015; 81: PW149
8 Wang SY, Shao B, Chang JL, Rao PF. Isolation and identification of a plant lysozyme from Momordica charantia L. Eur Food Res Technol 2011; 232: 613-619
9 Osman H, Darwish S, El-Difrawy E, Debever J. The inhibitory effect of lysozyme on the growth of some pathogenic and spoilage bacteria. Alexandria J Agricult Res 1995; 40: 169-176
10 Jollès P. Lysozymes: Model Enzymes in Biochemistry and Biology. Basel: Birkhäuser; 1996
11 Kuijpers AJ, van Wachem PB, van Luyn MJA, Engbers GHM, Krijgsveld J, Zaat SAJ, Dankert J, Feijen J. In vivo and in vitro release of lysozyme from cross-linked gelatin hydrogels: a model system for the delivery of antibacterial proteins from prosthetic heart valves. J Control Release 2000; 67: 323-336
12 Jiang G, Woo BH, Kang FR, Singh J, DeLuca PP. Assessment of protein release kinetics, stability and protein polymer interaction of lysozyme encapsulated poly (D,L-lactide-co-glycolide) microspheres. J Control Release 2002; 79: 137-145
13 Vermonden T, Censi R, Hennink WE. Hydrogels for protein delivery. Chem Rev 2012; 112: 2853-2888
14 Bysell H, Mansson R, Hansson P, Malmsten M. Microgels and microcapsules in peptide and protein drug delivery. Adv Drug Deliv Rev 2011; 63: 1172-1185
15 Caon T, Jin L, Simões CMO, Norton RS, Nicolazzo JA. Enhancing the buccal mucosal delivery of peptide and protein therapeutics. Pharm Res 2015; 32: 1-21
16 Tessmar JK, Gopferich AM. Matrices and scaffolds for protein delivery in tissue engineering. Adv Drug Deliv Rev 2007; 59: 274-291
17 Sill TJ, von Recum HA. Electrospinning: applications in drug delivery and tissue engineering. Biomaterials 2008; 29: 1989-2006
18 Meinel AJ, Germershaus O, Luhmann T, Merkle HP, Meinel L. Electrospun matrices for localized drug delivery: current technologies and selected biomedical applications. Eur J Pharm Biopharm 2012; 81: 1-13
19 Greiner A, Wendorff JH. Electrospinning: a fascinating method for the preparation of ultrathin fibres. Angew Chem Int Ed Engl 2007; 46: 5670-5703
20 Agarwal S, Wendorff JH, Greiner A. Use of electrospinning technique for biomedical applications. Polymer (Guildf) 2008; 49: 5603-5621
21 Ignatious F, Sun LH, Lee CP, Baldoni J. Electrospun nanofibers in oral drug delivery. Pharm Res 2010; 27: 576-588
22 Vrbata P, Berka P, Stránská D, Doležal P, Musilová M, Čižinská L. Electrospun drug loaded membranes for sublingual administration of sumatriptan and naproxen. Int J Pharm 2013; 457: 168-176
23 Zahedi P, Rezaeian I, Ranaei-Siadat SO, Jafari SH, Supaphol P. A review on wound dressings with an emphasis on electrospun nanofibrous polymeric bandages. Polym Adv Technol 2010; 21: 77-95
24 Said SS, El-Halfawy OM, El-Gowelli HM, Aloufy AK, Boraei NA, El-Khordagui LK. Bioburden-responsive antimicrobial PLGA ultrafine fibers for wound healing. Eur J Pharm Biopharm 2012; 80: 85-94
25 Chen P, Wu QS, Ding YP, Chu M, Huang ZM, Hu W. A controlled release system of titanocene dichloride by electrospun fiber and its antitumor activity in vitro . Eur J Pharm Biopharm 2010; 76: 413-420
26 Ji W, Yang F, van den Beucken JJ, Bian Z, Fan M, Chen Z, Jansen JA. Fibrous scaffolds loaded with protein prepared by blend or coaxial electrospinning. Acta Biomater 2010; 6: 4199-4207
27 Maretschek S, Greiner A, Kissel T. Electrospun biodegradable nanofiber nonwovens for controlled release of proteins. J Control Release 2008; 127: 180-187
28 Sahoo S, Ang LT, Goh JCH, Toh SL. Growth factor delivery through electrospun nanofibers in scaffolds for tissue engineering applications. J Biomed Mater Res A 2010; 93: 1539-1550
29 Choi JS, Leong KW, Yoo HS. In vivo wound healing of diabetic ulcers using electrospun nanofibers immobilized with human epidermal growth factor (EGF). Biomaterials 2008; 29: 587-596
30 Yang Y, Li X, Qi M, Zhou S, Weng J. Release pattern and structural integrity of lysozyme encapsulated in core-sheath structured poly(DL-lactide) ultrafine fibers prepared by emulsion electrospinning. Eur J Pharm Biopharm 2008; 69: 106-116
31 Nie H, Soh BW, Fu YC, Wang CH. Three-dimensional fibrous PLGA/HAp composite scaffold for BMP-2 delivery. Biotechnol Bioeng 2008; 99: 223-234
32 Jiang HL, Hu YQ, Li Y, Zhao PC, Zhu KJ, Chen WL. A facile technique to prepare biodegradable coaxial electrospun nanofibers for controlled release of bioactive agents. J Control Release 2005; 108: 237-243
33 Zhang YZ, Wang X, Feng Y, Li J, Lim CT, Ramakrishna S. Coaxial electrospinning of (fluorescein isothiocyanate-conjugated bovine serum albumin)-encapsulated poly(epsilon-caprolactone) nanofibers for sustained release. Biomacromolecules 2006; 7: 1049-1057
34 Zeng J, Aigner A, Czubayko F, Kissel T, Wendorff JH, Greiner A. Poly(vinyl alcohol) nanofibers by electrospinning as a protein delivery system and the retardation of enzyme release by additional polymer coatings. Biomacromolecules 2005; 6: 1484-1488
35 Taepaiboon P, Rungsardthong U, Supaphol P. Effect of cross-linking on properties and release characteristics of sodium salicylate-loaded electrospun poly(vinyl alcohol) fibre mats. Nanotechnology 2007; 18: 175102
36 Kenawy ER, Abdel-Hay FI, El-Newehy MH, Wnek GE. Controlled release of ketoprofen from electrospun poly(vinyl alcohol) nanofibers. Mat Sci Eng A-Struct 2007; 459: 390-396
37 Yao L, Haas TW, Guiseppi-Elie A, Bowlin GL, Simpson DG, Wnek GE. Electrospinning and stabilization of fully hydrolyzed poly(vinyl alcohol) fibers. Chem Mater 2003; 15: 1860-1864
38 Alves MH, Jensen BE, Smith AA, Zelikin AN. Poly(vinyl alcohol) physical hydrogels: new vista on a long serving biomaterial. Macromol Biosci 2011; 11: 1293-1313
39 Baker MI, Walsh SP, Schwartz Z, Boyan BD. A review of polyvinyl alcohol and its uses in cartilage and orthopedic applications. J Biomed Mater Res B Appl Biomater 2012; 100?B: 1451-1457
40 Gao Q, Takizawa J, Kimura M. Hydrophilic non-wovens made of cross-linked fully-hydrolyzed poly(vinyl alcohol) electrospun nanofibers. Polymer (Guildf) 2013; 54: 120-126
41 Zhang YZ, Venugopal J, Huang ZM, Lim CT, Ramakrishna S. Crosslinking of the electrospun gelatin nanofibers. Polymer (Guildf) 2006; 47: 2911-2917
42 Tang C, Saquing CD, Harding JR, Khan SA. In situ cross-linking of electrospun poly(vinyl alcohol) nanofibers. Macromolecules 2010; 43: 630-637
43 Choi SS, Hong JP, Seo YS, Chung SM, Nah C. Fabrication and characterization of electrospun polybutadiene fibers crosslinked by UV light. J Appl Polym Sci 2006; 101: 2333-2337
44 Yoo HS, Kim TG, Park TG. Surface-functionalized electrospun nanofibers for tissue engineering and drug delivery. Adv Drug Deliv Rev 2009; 61: 1033-1042
45 Griebenow K, Klibanov AM. On protein denaturation in aqueous-organic mixtures but not in pure organic solvents. J Am Chem Soc 1996; 118: 11695-11700
46 Mansur HS, Sadahira CM, Souza AN, Mansur AA. FTIR spectroscopy characterization of poly (vinyl alcohol) hydrogel with different hydrolysis degree and chemically crosslinked with glutaraldehyde. Mater Sci Eng C 2008; 28: 539-548
47 Liao YH, Brown MB, Martin GP. Turbidimetric and HPLC assays for the determination of formulated lysozyme activity. J Pharm Pharmacol 2001; 53: 549-554
48 Shugar D. The measurement of lysozyme activity and the ultra-violet inactivation of lysozyme. Biochim Biophys Acta 1952; 8: 302-309
49 Bellomo G. Cell-damage by oxygen free-radicals. Cytotechnology 1991; 5: 71-73
50 Walker JM. The bicinchoninic acid (BCA) assay for protein quantitation. In: Walker JM. editor Basic Protein and Peptide Protocols. Totowa, NJ: Humana Press; 1994: 5-8
51 Miranda TMR, Gonçalves AR, Amorim MTP. Ultraviolet-induced crosslinking of poly(vinyl alcohol) evaluated by principal component analysis of FTIR spectra. Polym Int 2001; 50: 1068-1072