Improving the crosslinking of collagen casing and glutaraldehyde by facilitating the formation of conjugate structure via pH

Abstract

Glutaraldehyde (GTA) crosslinking is commonly used to improve the thermal stability and mechanical strength of collagen casings. The aim of this research was to determine the optimal pH of the crosslinking between GTA and collagen as well as the crosslinking mechanisms. The weakly alkaline environment could facilitate the generation of GTA polymerization through the rapid generation of -C = C-C = O and -N = C-C = C- conjugated structures, and enhance the crosslinking reaction of GTA polymers with collagen amino groups. In the pH range of 8–10, the fibril diameter and d-space value declined significantly in the self-assembled collagen fibril-GTA system. Meanwhile, collagen casing films crosslinked with GTA in weakly alkaline conditions exhibited higher mechanical strength and thermal stability. These results suggest that the crosslinking of collagen casings and GTA can be improved by adjusting the pH. Possible crosslinking mechanisms related to the formation of conjugated long chains have also been proposed. This study could provide guidance on the appropriate use of GTA in the production process of collagen casings.

Graphical Abstract

References

[1] Luo QY, Hossen MA, Zeng YB, Dai JW, Li SQ, Qin W, et al. . Gelatin-based composite films and their application in food packaging: a review. J Food Eng, 2022, 313: 110762. PubMed PMID: WOS:000693999800009

[2] Irastorza A, Zarandona I, Andonegi M, Guerrero P, de la Caba K. The versatility of collagen and chitosan: from food to biomedical applications. Food Hydrocoll, 2021, 116: 106633. PubMed PMID: WOS:000626462900001

[3] Chen H, Xue L, Gong G, Pan J, Wang X, Zhang Y, et al. Collagen-based materials in reproductive medicine and engineered reproductive tissues. J Leather Sci Eng. 2022;4:3. https://doi.org/10.1186/s42825-021-00075-y.

[4] Rico-Llanos GA, Borrego-Gonzalez S, Moncayo-Donoso M, Becerra J, Visser R. Collagen type I biomaterials as scaffolds for bone tissue engineering. Polymers, 2021, 13(4): 20. PubMed PMID: WOS:000624253800001

[5] Han Y, Hu J, Sun G. Recent advances in skin collagen: functionality and non-medical applications. J Leather Sci Eng. 2021;3:4. https://doi.org/10.1186/s42825-020-00046-9.

[6] Adamiak K, Sionkowska A. Current methods of collagen cross-linking: review. Int J Biol Macromol, 2020, 161: 550-560. PubMed PMID: WOS:000572333900009

[7] Damink LHHO, Dijkstra PJ, Luyn MJAV. Glutaraldehyde as a crosslinking agent for collagen-based biomaterials. J Mater Sci: Mater Med, 1995, 6(8): 460-72.

[8] Hopwood D, Callen CR, McCabe M. The reactions between glutaraldehyde and various proteins. An investigation of their kinetics. Histochem J, 1970, 2(2): 137-50.

[9] Tian Z, Shen L, Liu W, Li G. Construction of collagen gel with high viscoelasticity and thermal stability via combining cross-linking and dehydration. J Biomed Mater Res A, 2020, 108(9): 1934-43.

[10] Peng YY, Glattauer V, Ramshaw JAM. Stabilisation of collagen sponges by glutaraldehyde vapour crosslinking. Int J Biomater, 2017, 2017(1): 8947823.

[11] Chen XF, Meng J, Xu HZ, Shinoda M, Kishimoto M, Sakurai S, et al. . Fabrication and properties of electrospun collagen tubular scaffold crosslinked by physical and chemical treatments. Polymers, 2021, 13(5): 16. PubMed PMID: WOS:000628408000001

[12] FDA. Food additive status list. 2022. 2021. https://www.fda.gov/food/food-additives-petitions/food-additive-status-list.

[13] Uragami T, Matsuda T, Okuno H, Miyata T. Structure of chemically modified chitosan membranes and their characteristics of permeation and separation of aqueous ethanol solutions. J Membr Sci, 1994, 88(2–3): 243-51.

[14] Migneault I, Dartiguenave C, Bertrand MJ, Waldron KC. Glutaraldehyde: behavior in aqueous solution, reaction with proteins, and application to enzyme crosslinking. Biotechniques, 2004, 37(5): 790-802.

[15] Lubig R, Kusch P, Röper K, Zahn H. On the mechanism of protein crosslinking with glutaraldehyde. Monatsh Chem, 1981, 112(11): 1313-23.

[16] Wine Y, Cohen-Hadar N, Freeman A, Frolow F. Elucidation of the mechanism and end products of glutaraldehyde crosslinking reaction by X-ray structure analysis. Biotechnol Bioeng, 2007, 98(3): 711-8.

[17] Chandran PL, Paik DC, Holmes JW. Structural mechanism for alteration of collagen gel mechanics by glutaraldehyde crosslinking. Connect Tissue Res, 2012, 53(4): 285-297. PubMed PMID: WOS:000306190400002

[18] Blauer G, Zvilichovsky B, Yanai P, Meir E, Swenson MK. The interaction of glutaraldehyde with poly(, L-lysine), n-butylamine, and collagen. II. Hydrodynamic, electron microscopic, and optical investigations on the reaction products. Biopolymers, 1975, 14(12): 2599-612.

[19] Singh RS, Singh T. Glutaraldehyde functionalization of halloysite nanoclay enhances immobilization efficacy of endoinulinase for fructooligosaccharides production from inulin. Food Chem, 2022, 381: 132253.

[20] Liu YS, Jiang JZ. Preparation of β-ionone microcapsules by gelatin/pectin complex coacervation. Carbohydr Polym, 2023, 312: 120839. PubMed PMID: WOS:000968381600001

[21] Khan S, Anwar N. Gelatin/carboxymethyl cellulose based stimuli-responsive hydrogels for controlled delivery of 5-fluorouracil, development, in vitro characterization, in vivo safety and bioavailability evaluation. Carbohydr Polym, 2021, 257: 117617. PubMed PMID: WOS:000617786800002

[22] Jones GJ. Letter: Polymerization of glutaraldehyde at fixative pH. J Histochem Cytochem, 1974, 22(9): 911-913. PubMed PMID: MEDLINE:4213220

[23] Xu J, Liu F, Wang T, Goff HD, Zhong F. Fabrication of films with tailored properties by regulating the swelling of collagen fiber through pH adjustment. Food Hydrocoll, 2020, 108: 106016.

[24] Farris S, Song JH, Huang QR. Alternative reaction mechanism for the cross-linking of gelatin with glutaraldehyde. J Agric Food Chem, 2010, 58(2): 998-1003. PubMed PMID: WOS:000273671900045

[25] Wang P, Zou Y, Li Y, Qin Z, Liu X, Zhang H. pH-sensitive self-assembled nanofibers based on electrostatic interaction and Schiff base bonding for controlled release of curcumin. Food Hydrocoll, 2022, 131: 107805.

[26] Lin J, Pan D, Sun Y, Ou C, Wang Y, Cao J. The modification of gelatin films: based on various cross-linking mechanism of glutaraldehyde at acidic and alkaline conditions. Food Sci Nutr, 2019, 7(12): 4140-4146. PubMed PMID: WOS:000501181100001

[27] Akbar I, Jaswir I, Jamal P. Optimisation of the production of fish gelatine nanoparticles as a carrier for sunflower-derived biopeptide. Int Food Res J, 2020, 27(1): 171-81 PubMed PMID: WOS:000538056000018

[28] Slusarewicz P, Zhu K, Hedman T. Kinetic characterization and comparison of various protein crosslinking reagents for matrix modification. J Mater Sci Mater Med, 2010, 21(4): 1175-81.

[29] Mauri AN, Añón MC. Effect of solution pH on solubility and some structural properties of soybean protein isolate films. J Sci Food Agric, 2006, 86(7): 1064-72.

[30] Kawahara J-I, Ishikawa K, Uchimaru T, Takaya H. Chemical cross-linking by glutaraldehyde between amino groups: its mechanism and effects. Polym Modif. 1997:119–31. https://doi.org/10.1007/978-1-4899-1477-4_11.

[31] Kawahara J, Ohmori T, Ohkubo T, Hattori S, Kawamura M. The structure of glutaraldehyde in aqueous solution determined by ultraviolet absorption and light scattering. Anal Biochem, 1992, 201(1): 94-8.

[32] Zuo P, Zhang J, Zhou Y, Xie N, Xiao D. Spontaneous formation of fluorescent carbon nanoparticles in glutaraldehyde solution and their fluorescence mechanism. J Fluoresc, 2021, 31(2): 509-16.

[33] Zhang T, Yu Z, Ma Y, Chiou BS, Liu F, Zhong F. Modulating physicochemical properties of collagen films by cross-linking with glutaraldehyde at varied pH values. Food Hydrocoll, 2022, 124: 107270. PubMed PMID: WOS:000708639600005

[34] Chen C, Liu F, Yu Z, Ma Y, Goff HD, Zhong F. Improvement in physicochemical properties of collagen casings by glutaraldehyde cross-linking and drying temperature regulating. Food Chem, 2020, 318: 126404. PubMed PMID: WOS:000520858000034

[35] He L, Lan W, Cen L, Chen S, Liu S, Liu Y, et al. . Improving catalase stability by its immobilization on grass carp (Ctenopharyngodon idella) scale collagen self-assembly films. Mater Sci Eng C, 2019, 105: 110024.

[36] Sun L, Li B, Song W, Si L, Hou H. Characterization of Pacific cod (Gadus macrocephalus) skin collagen and fabrication of collagen sponge as a good biocompatible biomedical material(Article). Process Biochem, 2017, 63: 229-235.

[37] Yan M, Li B, Zhao X, Qin S. Effect of concentration, pH and ionic strength on the kinetic self-assembly of acid-soluble collagen from walleye pollock (Theragra chalcogramma) skin. Food Hydrocoll, 2012, 29(1): 199-204.

[38] Zhang Y, Ingham B, Leveneur J, Cheong S, Yao Y, Clarke DJ, et al. . Can sodium silicates affect collagen structure during tanning? Insights from small angle X-ray scattering (SAXS) studies. RSC Adv, 2017, 7(19): 11665-71.

[39] Liao J, Yang L, Grashow J, Sacks MS. Molecular orientation of collagen in intact planar connective tissues under biaxial stretch. Acta Biomater. 2005;1(1):45–54. https://doi.org/10.1016/j.actbio.2004.09.007.

[40] Miao KY, Li XY, Yang D, Xu YB, Mu CD, Li DF, et al. . Hydrothermal shrinkage behavior of pigskin. Thermochim Acta, 2021, 699: 178896. PubMed PMID: WOS:000659820000007

[41] Aktaş N. The effects of pH, NaCl and CaCl₂ on thermal denaturation characteristics of intramuscular connective tissue. Thermochim Acta. 2003;407(1–2):105. https://doi.org/10.1016/s0040-6031(03)00306-x.

[42] Shi R, He J, Jirimutu. Ultrasonic-assisted extraction and structure characterization of collagen from camel skin. J Chin Inst Food Sci Technol. 2022;22(2):213–23. https://doi.org/10.16429/j.1009-7848.2022.02.023. (in Chinese).

[43] Wang Y-N, Hu L. Essential role of isoelectric point of skin/leather in leather processing. J Leather Sci Eng. 2022;4:25. https://doi.org/10.1186/s42825-022-00099-y.

[44] Gray RE, Seng N, Mackay IR, Rowley MJ. Measurement of antibodies to collagen II by inhibition of collagen fibril formation in vitro. J Immunol Methods. 2004;285(1):55–61. https://doi.org/10.1016/j.jim.2003.11.010.

[45] Jingmin W, Fei L, Zhe Y, Yun M, Douglas GH, Jianguo M, et al. . Facile preparation of collagen fiber-glycerol-carboxymethyl cellulose composite film by immersing method. Carbohydr Polym, 2020, 229: 115429.

[46] Nilsuwan K, Fusang K, Pripatnanont P, Benjakul S. Properties and characteristics of acid-soluble collagen from salmon skin defatted with the aid of ultrasonication. Fishes (MDPI AG), 2022, 7(1): 51.

[47] Zhang Y, Snow T, Smith AJ, Holmes G, Prabakar S. A guide to high-efficiency chromium (III)-collagen cross-linking: synchrotron SAXS and DSC study. Int J Biol Macromol, 2019, 126: 123-129.

[48] Sizeland KH, Edmonds RL, Basil-Jones MM, Kirby N, Hawley A, Mudie S, et al. . Changes to collagen structure during leather processing. J Agric Food Chem, 2015, 63(9): 2499-505.

[49] Gillett R, Gull K. Glutaraldehyde–its purity and stability. Histochemie, 1972, 30(2): 162-7.

[50] Cheung DT, Nimni ME. Mechanism of crosslinking of proteins by glutaraldehyde I: reaction with model compounds. Connect Tissue Res, 2009, 10(2): 187-99.

[51] Ruijgrok JM, Boon ME, Wijn JRD. The effect of heating by microwave irradiation and by conventional heating on the aldehyde concentration in aqueous glutaraldehyde solutions. Histochem J, 1990, 22(6–7): 389-93.

Funding

Collaborative Innovationcenter of Food Safety and Quality Control in Jiangsu Province,

National Key R&D Program of China,(2023YFF1104302)