journal article·2019-08

Crosslinking of genipin and autoclaving in chitosan-based nanofibrous scaffolds: structural and physiochemical properties

Yi Wah Mak, Wallace Woon-Fong Leung

Journal of Materials Science· Volume 54· Issue 15
Publisher page

Abstract

Chitosan-based electrospun nanofibrous scaffolds have been selected as wound healing/tissue scaffolds because of their extracellular matrix nature and biocompatible properties. However, crosslinking of scaffolds is necessary to avoid lysozyme degradation in an aqueous environment, as a stable scaffold is crucial for the activities of fibroblasts, including adhesion and proliferation during wound healing. Autoclaving (physical) and genipin crosslinking (chemical) methods have been employed to stabilize chitosan-based scaffolds individually. However, the differences in scaffold microstructure induced by the individual or combined crosslinking methods have yet to be investigated systematically. In this study, autoclaving crosslinking improved mainly the structural properties (tensile strength and crystallinity), but it also expanded the chitosan and PEO network by hydrolysis, which enlarged the fiber diameter and caused chitosan chain degradation. Meanwhile, genipin crosslinking improved the physiochemical properties, primarily hydrophilicity. On the other hand, the combined crosslinking significantly improved both the structural and physiochemical properties through the unique reorganization of the polymeric network. The confined geometry of the nanofiber as well as the genipin crosslinks resulted in maximal crystallization of chitosan and amorphization of PEO chains. Unfortunately, the combined crosslinking resulted in the lowest antibacterial activity because of the consumption of amino and protonated amino groups in the crosslinking process. Despite this, the combined crosslinking scaffold achieved the best stability under lysozyme degradation and therefore it is preferred over autoclaving or genipin crosslinking alone. In conclusion, the results show that chemical and physical crosslinking methods induce different changes in crystallinity and hydrophilicity that affect the physicochemical properties. Therefore, crystallinity and hydrophilicity are significant considerations when designing a tissue scaffold.
Source: openaire · Quality: candidate unverified · Source record ↗

Research context

OpenAlex topics: Electrospun Nanofibers in Biomedical Applications · Nanocomposite Films for Food Packaging · Advanced Sensor and Energy Harvesting Materials

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

References (85)

Show source-provided references
1. Jayakumar R, Prabaharan M, Sudheesh Kumar PT, Nair SV, Tamura H (2011) Biomaterials based on chitin and chitosan in wound dressing applications. Biotechnol Adv 29:322–337. https://doi.org/10.1016/j.biotechadv.2011.01.00510.1016/j.biotechadv.2011.01.005 ↗
2. Chung YC, Chen CY (2008) Antibacterial characteristics and activity of acid-soluble chitosan. Bioresour Technol 99:2806–2814. https://doi.org/10.1016/j.biortech.2007.06.04410.1016/j.biortech.2007.06.044 ↗
3. Angelova N, Manolova N, Rashkov I, Maximova V, Bogdanova S, Domard A (1995) Preparation and properties of modified chitosan films for drug release. J Bioact Compat Polym 10:285–298. https://doi.org/10.1177/08839115950100040110.1177/088391159501000401 ↗
4. Kumar MNVR, Muzzarelli RAA, Muzzarelli C, Sashiwa H, Domb AJ (2005) Chitosan chemistry and pharmaceutical perspectives. Chem Rev 104:6017–6084. https://doi.org/10.1021/cr030441b10.1021/cr030441b ↗
5. Paul W, Sharma CP (2004) Chitosan and alginate wound dressings: a short review. Trends Biomater Artif Organs 18:18–23
6. Guillaume C, Erik S (2014) Physical influences of the extracellular environment on cell migration. Nat Rev Mol Cell Biol 15:813–824. https://doi.org/10.1038/nrm389710.1038/nrm3897 ↗
7. Liu M, Duan X-P, Li Y-M, Yang D-P, Long Y-Z (2017) Electrospun nanofibers for wound healing. Mater Sci Eng C 76:1413–1423. https://doi.org/10.1016/j.msec.2017.03.03410.1016/j.msec.2017.03.034 ↗
8. Nae Gyune R, Choongsoo SS, Heungsoo S (2013) Current approaches to electrospun nanofibers for tissue engineering. Biomed Mater 8:014102. https://doi.org/10.1088/1748-6041/8/1/01410210.1088/1748-6041/8/1/014102 ↗
9. Jia Y-T, Gong J, Gu X-H, Kim H-Y, Dong J, Shen X-Y (2007) Fabrication and characterization of poly(vinyl alcohol)/chitosan blend nanofibers produced by electrospinning method. Carbohydr Polym 67:403–409. https://doi.org/10.1016/j.carbpol.2006.06.01010.1016/j.carbpol.2006.06.010 ↗
10. Ignatova M, Manolova N, Rashkov I (2007) Novel antibacterial fibers of quaternized chitosan and poly(vinyl pyrrolidone) prepared by electrospinning. Eur Polym J 43:1112–1122. https://doi.org/10.1016/j.eurpolymj.2007.01.01210.1016/j.eurpolymj.2007.01.012 ↗
11. Bhattarai N, Edmondson D, Veiseh O, Matsen FA, Zhang M (2005) Electrospun chitosan-based nanofibers and their cellular compatibility. Biomaterials 26:6176–6184. https://doi.org/10.1016/j.biomaterials.2005.03.02710.1016/j.biomaterials.2005.03.027 ↗
12. Muzzarelli RAA (1997) Human enzymatic activities related to the therapeutic administration of chitin derivatives CMLS. Cell Mol Life Sci 53:131–140. https://doi.org/10.1007/PL0000058410.1007/PL00000584 ↗
13. Lim LY, Khor E, Ling CE (1999) Effects of dry heat and saturated steam on the physical properties of chitosan. J Biomed Mater Res 48:111–116. https://doi.org/10.1002/(SICI)1097-4636(1999)48:2%3c111:AID-JBM3%3e3.0.CO;2-W10.1002/(SICI)1097-4636(1999)48:2<111::AID-JBM3>3.0.CO;2-W ↗
14. Ritthidej CC, Phaechamud T, Koizumi T (2002) Moist heat treatment on physicochemical change of chitosan salt films. Int J Pharm 232:11–2210.1016/S0378-5173(01)00894-8 ↗
15. Mayol L, De Stefano D, Campani V, De Falco F, Ferrari E, Cencetti C, Matricardi P, Maiuri L, Carnuccio R, Gallo A, Maiuri M, De Rosa G (2014) Design and characterization of a chitosan physical gel promoting wound healing in mice. Eur Soc Biomater 25:1483–1493. https://doi.org/10.1007/s10856-014-5175-710.1007/s10856-014-5175-7 ↗
16. Schiffman JD, Schauer CL (2007) Cross-linking chitosan nanofibers. Biomacromolecules 8:594–601. https://doi.org/10.1021/bm060804s10.1021/bm060804s ↗
17. Jawad AH, Nawi MA (2012) Oxidation of crosslinked chitosan-epichlorohydrine film and its application with TiO2 for phenol removal. Carbohydr Polym 90:87–94. https://doi.org/10.1016/j.carbpol.2012.04.06610.1016/j.carbpol.2012.04.066 ↗
18. Yue W (2014) Prevention of browning of depolymerized chitosan obtained by gamma irradiation. Carbohydr Polym 101:857–863. https://doi.org/10.1016/j.carbpol.2013.10.01110.1016/j.carbpol.2013.10.011 ↗
19. Cheng YX, Ramos D, Lee P, Liang DN, Yu XJ, Kumbar SG (2014) Collagen functionalized bioactive nanofiber matrices for osteogenic differentiation of mesenchymal stem cells: bone tissue engineering. J Biomed Nanotechnol 10:287–298. https://doi.org/10.1166/jbn.2014.175310.1166/jbn.2014.1753 ↗
20. Pauliukaite R, Ghica ME, Fatibello-Filho O, Brett CMA (2009) Comparative study of different cross-linking agents for the immobilization of functionalized carbon nanotubes within a chitosan film supported on a graphite–epoxy composite electrode. Anal Chem 81:5364–5372. https://doi.org/10.1021/ac900464z10.1021/ac900464z ↗
21. Huang LLH, Sung HW, Tsai CC, Huang DM (1998) Biocompatibility study of a biological tissue fixed with a naturally occurring crosslinking reagent. J Biomed Mater Res 42:568–576. https://doi.org/10.1002/(sici)1097-4636(19981215)42:4%3c568:aid-jbm13%3e3.0.co;2-710.1002/(SICI)1097-4636(19981215)42:4<568::AID-JBM13>3.0.CO;2-7 ↗
22. Norowski PA, Fujiwara T, Clem WC, Adatrow PC, Eckstein EC, Haggard WO, Bumgardner JD (2012) Novel naturally crosslinked electrospun nanofibrous chitosan mats for guided bone regeneration membranes: material characterization and cytocompatibility. J Tissue Eng Regen Med 9:577–583. https://doi.org/10.1002/term.164810.1002/term.1648 ↗
23. Mi F-L, Tan Y-C, Liang H-C, Huang R-N, Sung H-W (2001) In vitro evaluation of a chitosan membrane cross-linked with genipin. J Biomater Sci Polym Ed 12:835–850. https://doi.org/10.1163/15685620175311305110.1163/156856201753113051 ↗
24. Mekhail M, Jahan K, Tabrizian M (2014) Genipin-crosslinked chitosan/poly-l-lysine gels promote fibroblast adhesion and proliferation. Carbohydr Polym 108:91–98. https://doi.org/10.1016/j.carbpol.2014.03.02110.1016/j.carbpol.2014.03.021 ↗
25. Sung H-W, Huang R-N, Huang LLH, Tsai C-C (1999) In vitro evaluation of cytotoxicity of a naturally occurring cross-linking reagent for biological tissue fixation. J Biomater Sci Polym Ed 10:63–78. https://doi.org/10.1163/156856299X0028910.1163/156856299X00289 ↗
26. Hc L, Chang W, Hf L, Lee M, Hw S (2004) Crosslinking structures of gelatin hydrogels crosslinked with genipin or a water-soluble carbodiimide. J Appl Polym Sci 91:4017–4026. https://doi.org/10.1002/app.1356310.1002/app.13563 ↗
27. Muzzarelli RAA (2009) Genipin-crosslinked chitosan hydrogels as biomedical and pharmaceutical aids. Carbohydr Polym 77:1–9. https://doi.org/10.1016/j.carbpol.2009.01.01610.1016/j.carbpol.2009.01.016 ↗
28. Ma B, Wang X, Wu C, Chang J (2014) Crosslinking strategies for preparation of extracellular matrix-derived cardiovascular scaffolds. Regen Biomater 1:81–89. https://doi.org/10.1093/rb/rbu00910.1093/rb/rbu009 ↗
29. Silva SS, Motta A, Rodrigues MT, Pinheiro AFM, Gomes ME, Mano JF, Reis RL, Migliaresi C (2008) Novel genipin-cross-linked chitosan/silk fibroin sponges for cartilage engineering strategies. Biomacromolecules 9:2764–2774. https://doi.org/10.1021/bm800874q10.1021/bm800874q ↗
30. Gomes SR, Rodrigues G, Martins GG, Henriques CMR, Silva JC (2012) In vitro evaluation of crosslinked electrospun fish gelatin scaffolds. Mater Sci Eng C 33:1219–1227. https://doi.org/10.1016/j.msec.2012.12.01410.1016/j.msec.2012.12.014 ↗
31. Norowski PA, Mishra S, Adatrow PC, Haggard WO, Bumgardner JD (2012) Suture pullout strength and in vitro fibroblast and RAW 264.7 monocyte biocompatibility of genipin crosslinked nanofibrous chitosan mats for guided tissue regeneration. J Biomed Mater Res Part A 100:2890–2896. https://doi.org/10.1002/jbm.a.3422410.1002/jbm.a.34224 ↗
32. Li Q, Wang X, Lou X, Yuan H, Tu H, Li B, Zhang Y (2015) Genipin-crosslinked electrospun chitosan nanofibers: determination of crosslinking conditions and evaluation of cytocompatibility. Carbohydr Polym 130:166–174. https://doi.org/10.1016/j.carbpol.2015.05.03910.1016/j.carbpol.2015.05.039 ↗
33. Fessel G, Cadby J, Wunderli S, van Weeren R, Snedeker JG (2014) Dose- and time-dependent effects of genipin crosslinking on cell viability and tissue mechanics—toward clinical application for tendon repair. Acta Biomater 10:1897–1906. https://doi.org/10.1016/j.actbio.2013.12.04810.1016/j.actbio.2013.12.048 ↗
34. Luna SM, Silva SS, Gomes ME, Mano JF, Reis RL (2011) Cell adhesion and proliferation onto chitosan-based membranes treated by plasma surface modification. J Biomater Appl 26:101–116. https://doi.org/10.1177/088532821036292410.1177/0885328210362924 ↗
35. Jhala D, Rather H, Vasita R (2016) Polycaprolactone–chitosan nanofibers influence cell morphology to induce early osteogenic differentiation. Biomater Sci 4:1584–1595. https://doi.org/10.1039/c6bm00492j10.1039/C6BM00492J ↗
36. Galante R, Rediguieri CF, Kikuchi IS, Vasquez PAS, Colaço R, Serro AP, Pinto TJA (2016) About the sterilization of chitosan hydrogel nanoparticles. PLoS ONE 11:e0168862. https://doi.org/10.1371/journal.pone.016886210.1371/journal.pone.0168862 ↗
37. Rao SB, Sharma CP (1995) Sterilization of chitosan: implications. J Biomater Appl 10:136–143. https://doi.org/10.1177/08853282950100020410.1177/088532829501000204 ↗
38. Rivero S, García MA, Pinotti A (2013) Physical and chemical treatments on chitosan matrix to modify film properties and kinetics of biodegradation. J Mater Phys Chem 1:51–57. https://doi.org/10.12691/jmpc-1-3-510.12691/jmpc-1-3-5 ↗
39. Chiono V, Pulieri E, Vozzi G, Ciardelli G, Ahluwalia A, Giusti P (2008) Genipin-crosslinked chitosan/gelatin blends for biomedical applications. J Mater Sci Mater Med 19:889–898. https://doi.org/10.1007/s10856-007-3212-510.1007/s10856-007-3212-5 ↗
40. Kim CH, Park HS, Gin YJ, Son Y, Lim SH, Choi YJ, Park KS, Park CW (2004) Improvement of the biocompatibility of chitosan dermal scaffold by rigorous dry heat treatment. Macromol Res 12:367–37310.1007/BF03218413 ↗
41. Krishnakumar GS, Sampath S, Muthusamy S, John MA (2019) Importance of crosslinking strategies in designing smart biomaterials for bone tissue engineering: a systematic review. Mater Sci Eng C Mater Biol Appl 96:941–954. https://doi.org/10.1016/j.msec.2018.11.08110.1016/j.msec.2018.11.081 ↗
42. Donius AE, Kiechel MA, Schauer CL, Wegst UG (2013) New crosslinkers for electrospun chitosan fibre mats. Part II: mechanical properties. J R Soc Interface 10:20120946. https://doi.org/10.1098/rsif.2012.094610.1098/rsif.2012.0946 ↗
43. Vetten MA, Yah CS, Singh T, Gulumian M (2014) Challenges facing sterilization and depyrogenation of nanoparticles: effects on structural stability and biomedical applications. Nanomed Nanotechnol Biol Med 10:1391–1399. https://doi.org/10.1016/j.nano.2014.03.01710.1016/j.nano.2014.03.017 ↗
44. Focher B, Beltrame PL, Naggi A, Torri G (1990) Alkaline N-deacetylation of chitin enhanced by flash treatments. Reaction kinetics and structure modifications. Carbohydr Polym 12:405–418. https://doi.org/10.1016/0144-8617(90)90090-F10.1016/0144-8617(90)90090-F ↗
45. Lu C, Chiang SW, Du H, Li J, Gan L, Zhang X, Chu X, Yao Y, Li B, Kang F (2017) Thermal conductivity of electrospinning chain-aligned polyethylene oxide (PEO). Polymer 115:52–59. https://doi.org/10.1016/j.polymer.2017.02.02410.1016/j.polymer.2017.02.024 ↗
46. Osorio-Madrazo A, David L, Trombotto S, Lucas J-M, Peniche-Covas C, Domard A (2010) Kinetics study of the solid-state acid hydrolysis of chitosan: evolution of the crystallinity and macromolecular structure. Biomacromolecules 11:1376–1386. https://doi.org/10.1021/bm100168510.1021/bm1001685 ↗
47. Wojdyr M (2010) Fityk : a general-purpose peak fitting program. J Appl Crystallogr 43:1126–1128. https://doi.org/10.1107/S002188981003049910.1107/S0021889810030499 ↗
48. Langford JI, Wilson AJC (1978) Scherrer after sixty years: a survey and some new results in the determination of crystallite size. J Appl Crystallogr 11:102–113. https://doi.org/10.1107/S002188987801284410.1107/S0021889878012844 ↗
49. Chen ZG, Wang PW, Wei B, Mo XM, Cui FZ (2010) Electrospun collagen-chitosan nanofiber: a biomimetic extracellular matrix for endothelial cell and smooth muscle cell. Acta Biomater 6:372–382. https://doi.org/10.1016/j.actbio.2009.07.02410.1016/j.actbio.2009.07.024 ↗
50. Jett BD, Hatter KL, Huycke MM, Gilmore MS (1997) Simplified agar plate method for quantifying viable bacteria. Biotechniques 23:648–650. https://doi.org/10.2144/97234bm2210.2144/97234bm22 ↗
51. Duarte ML, Ferreira MC, Marvão MR, Rocha J (2002) An optimised method to determine the degree of acetylation of chitin and chitosan by FTIR spectroscopy. Int J Biol Macromol 31:1–8. https://doi.org/10.1016/S0141-8130(02)00039-910.1016/S0141-8130(02)00039-9 ↗
52. Rakkapao N, Vao-Soongnern V, Masubuchi Y, Watanabe H (2011) Miscibility of chitosan/poly(ethylene oxide) blends and effect of doping alkali and alkali earth metal ions on chitosan/PEO interaction. Polymer 52:2618–2627. https://doi.org/10.1016/j.polymer.2011.03.04410.1016/j.polymer.2011.03.044 ↗
53. Lagaron JM, Fernandez-Saiz P, Ocio MJ (2007) Using ATR-FTIR spectroscopy to design active antimicrobial food packaging structures based on high molecular weight chitosan polysaccharide. J Agric Food Chem 55:2554–2562. https://doi.org/10.1021/jf063110j10.1021/jf063110j ↗
54. Butler MF, Ng YF, Pudney PDA (2003) Mechanism and kinetics of the crosslinking reaction between biopolymers containing primary amine groups and genipin. J Polym Sci Part A Polym Chem 41:3941–3953. https://doi.org/10.1002/pola.1096010.1002/pola.10960 ↗
55. Osman Z, Arof AK (2003) FTIR studies of chitosan acetate based polymer electrolytes. Electrochim Acta 48:993–999. https://doi.org/10.1016/S0013-4686(02)00812-510.1016/S0013-4686(02)00812-5 ↗
56. Toffey A, Samaranayake G, Frazier CE, Glasser WG (1996) Chitin derivatives. I. Kinetics of the heat-induced conversion of chitosan to chitin. J Appl Polym Sci 60:75–85. https://doi.org/10.1002/(SICI)1097-4628(19960404)60:1%3c75:AID-APP9%3e3.0.CO;2-S10.1002/(SICI)1097-4628(19960404)60:1<75::AID-APP9>3.0.CO;2-S ↗
57. Arteche Pujana M, Pérez-Álvarez L, Cesteros Iturbe LC, Katime I (2013) Biodegradable chitosan nanogels crosslinked with genipin. Carbohydr Polym 94:836–842. https://doi.org/10.1016/j.carbpol.2013.01.08210.1016/j.carbpol.2013.01.082 ↗
58. Martinová L, Lubasová D (2008) Electrospun chitosan based nanofibers. Res J Text Appar 12:72–79. https://doi.org/10.1108/RJTA-12-02-2008-B00910.1108/RJTA-12-02-2008-B009 ↗
59. Pakravan M, Heuzey M-C, Ajji A (2011) A fundamental study of chitosan/PEO electrospinning. Polymer 52:4813–4824. https://doi.org/10.1016/j.polymer.2011.08.03410.1016/j.polymer.2011.08.034 ↗
60. Liu H, Du Y, Wang X, Sun L (2004) Chitosan kills bacteria through cell membrane damage. Int J Food Microbiol 95:147–155. https://doi.org/10.1016/j.ijfoodmicro.2004.01.02210.1016/j.ijfoodmicro.2004.01.022 ↗
61. Mauricio-Sánchez RA, Salazar R, Luna-Bárcenas JG, Mendoza-Galván A (2018) FTIR spectroscopy studies on the spontaneous neutralization of chitosan acetate films by moisture conditioning. Vib Spectrosc 94:1–6. https://doi.org/10.1016/j.vibspec.2017.10.00510.1016/j.vibspec.2017.10.005 ↗
62. Demarger-Andre S, Domard A (1994) Chitosan carboxylic acid salts in solution and in the solid state. Carbohydr Polym 23:211–219. https://doi.org/10.1016/0144-8617(94)90104-X10.1016/0144-8617(94)90104-X ↗
63. Fl M, Ss S, Ck P (2005) Characterization of ring-opening polymerization of genipin and pH-dependent cross-linking reactions between chitosan and genipin. J Polym Sci Part A Polym Chem 43:1985–2000. https://doi.org/10.1002/pola.2066910.1002/pola.20669 ↗
64. Zivanovic S, Li J, Davidson PM, Kit K (2007) Physical, mechanical, and antibacterial properties of chitosan/PEO blend films. Biomacromolecules 8:1505–1510. https://doi.org/10.1021/bm061140p10.1021/bm061140p ↗
65. Naito P-K, Ogawa Y, Kimura S, Iwata T, Wada M (2015) Crystal transition from hydrated chitosan and chitosan/monocarboxylic acid complex to anhydrous chitosan investigated by X-ray diffraction. J Polym Sci Part B Polym Phys 53:1065–1069. https://doi.org/10.1002/polb.2374810.1002/polb.23748 ↗
66. Szymanska E, Winnicka K (2015) Stability of chitosan-A challenge for pharmaceutical and biomedical applications. Mar Drugs 13:1819–1846. https://doi.org/10.3390/md1304181910.3390/md13041819 ↗
67. Kawada J, Abe Y, Yui T, Okuyama K, Ogawa K (1999) Crystalline transformation of chitosan from hydrated to anhydrous polymorph via chitosan monocarboxylic acid salts. J Carbohydr Chem 18:559–571. https://doi.org/10.1080/0732830990854401910.1080/07328309908544019 ↗
68. Mejía A, García N, Guzmán J, Tiemblo P (2013) Confinement and nucleation effects in poly(ethylene oxide) melt-compounded with neat and coated sepiolite nanofibers: modulation of the structure and semicrystalline morphology. Eur Polym J 49:118–129. https://doi.org/10.1016/j.eurpolymj.2012.09.01410.1016/j.eurpolymj.2012.09.014 ↗
69. Desai K, Kit K, Li J, Zivanovic S (2008) Morphological and surface properties of electrospun chitosan nanofibers. Biomacromolecules 9:1000–1006. https://doi.org/10.1021/bm701017z10.1021/bm701017z ↗
70. Peniche-Covas C, Argüelles-Monal W, San Román J (1993) A kinetic study of the thermal degradation of chitosan and a mercaptan derivative of chitosan. Polym Degrad Stab 39:21–28. https://doi.org/10.1016/0141-3910(93)90120-810.1016/0141-3910(93)90120-8 ↗
71. Neto CGT, Giacometti JA, Job AE, Ferreira FC, Fonseca JLC, Pereira MR (2005) Thermal analysis of chitosan based networks. Carbohydr Polym 62:97–103. https://doi.org/10.1016/j.carbpol.2005.02.02210.1016/j.carbpol.2005.02.022 ↗
72. Klein MP, Hackenhaar CR, Lorenzoni ASG, Rodrigues RC, Costa TMH, Ninow JL, Hertz PF (2016) Chitosan crosslinked with genipin as support matrix for application in food process: support characterization and β-d-galactosidase immobilization. Carbohydr Polym 137:184–190. https://doi.org/10.1016/j.carbpol.2015.10.06910.1016/j.carbpol.2015.10.069 ↗
73. Juan AS, Montembault A, Gillet D, Say JP, Rouif S, Bouet T, Royaud I, David L (2012) Degradation of chitosan-based materials after different sterilization treatments. IOP Conf Ser Mater Sci Eng 31:012007. https://doi.org/10.1088/1757-899X/31/1/01200710.1088/1757-899X/31/1/012007 ↗
74. Aguirre-Chagala YE, Pavón-Pérez LB, Altuzar V, Domínguez-Chávez JG, Muñoz-Aguirre S, Mendoza-Barrera C (2017) Comparative study of one-step cross-linked electrospun chitosan-based membranes. J Nanomater 2017:14. https://doi.org/10.1155/2017/198071410.1155/2017/1980714 ↗
75. Andersson RL, Ström V, Gedde UW, Mallon PE, Hedenqvist MS, Olsson RT (2014) Micromechanics of ultra-toughened electrospun PMMA/PEO fibres as revealed by in situ tensile testing in an electron microscope. Sci Rep 4:6335. https://doi.org/10.1038/srep0633510.1038/srep06335 ↗
76. Kohsari I, Shariatinia Z, Pourmortazavi SM (2016) Antibacterial electrospun chitosan-polyethylene oxide nanocomposite mats containing ZIF-8 nanoparticles. Int J Biol Macromol 91:778–788. https://doi.org/10.1016/j.ijbiomac.2016.06.03910.1016/j.ijbiomac.2016.06.039 ↗
77. Moghadas B, Dashtimoghadam E, Mirzadeh H, Seidi F, Hasani-Sadrabadi MM (2016) Novel chitosan-based nanobiohybrid membranes for wound dressing applications. RSC Adv 6:7701–7711. https://doi.org/10.1039/c5ra23875g10.1039/C5RA23875G ↗
78. Bots JGF, van der Does L, Bantjes A (1986) Small diameter blood vessel prostheses from blends of polyethylene oxide and polypropylene oxide. Biomaterials 7:393–399. https://doi.org/10.1016/0142-9612(86)90011-610.1016/0142-9612(86)90011-6 ↗
79. Vårum KM, Myhr MM, Hjerde RJN, Smidsrød O (1997) In vitro degradation rates of partially N-acetylated chitosans in human serum. Carbohydr Res 299:99–101. https://doi.org/10.1016/S0008-6215(96)00332-110.1016/S0008-6215(96)00332-1 ↗
80. Hasmann A, Wehrschuetz-Sigl E, Kanzler G, Gewessler U, Hulla E, Schneider KP, Binder B, Schintler M, Guebitz GM (2011) Novel peptidoglycan-based diagnostic devices for detection of wound infection. Diagn Microbiol Infect Dis 71:12–23. https://doi.org/10.1016/j.diagmicrobio.2010.09.00910.1016/j.diagmicrobio.2010.09.009 ↗
81. Frohm M, Gunne H, Bergman AC, Agerberth B, Bergman T, Boman A, Lidén S, Jörnvall H, Boman HG (1996) Biochemical and antibacterial analysis of human wound and blister fluid. Eur J Biochem 237:86–92. https://doi.org/10.1111/j.1432-1033.1996.0086n.x10.1111/j.1432-1033.1996.0086n.x ↗
82. Friedman M, Sigel CW (1966) A kinetic study of the ninhydrin reaction. Biochemistry 5:478–485. https://doi.org/10.1021/bi00866a01210.1021/bi00866a012 ↗
83. Wang QZ, Chen XG, Liu N, Wang SX, Liu CS, Meng XH, Liu CG (2006) Protonation constants of chitosan with different molecular weight and degree of deacetylation. Carbohydr Polym 65:194–201. https://doi.org/10.1016/j.carbpol.2006.01.00110.1016/j.carbpol.2006.01.001 ↗
84. Roberts GAF (1992) Chemical behaviour of chitin and chitosan. In: Roberts GAF (ed) Chitin chemistry. Macmillan Education, London, pp 203–273. https://doi.org/10.1007/978-1-349-11545-7_510.1007/978-1-349-11545-7_5 ↗
85. Li K, Xing R, Liu S, Qin Y, Yu H, Li P (2014) Size and pH effects of chitooligomers on antibacterial activity against Staphylococcus aureus. Int J Biol Macromol 64:302–305. https://doi.org/10.1016/j.ijbiomac.2013.11.03710.1016/j.ijbiomac.2013.11.037 ↗