Zn ion-incorporated injected hydrogels with reactive oxygen species and glucose scavenging capacity for diabetic wound healing

Abstract

Background: Patients with diabetic wounds often experience challenges in the repair process, owing to increased concentration of glucose and reactive oxygen species (ROS). In addition, high glucose levels usually result in bacterial infections, which in turn worsen wound healing. This study aims to develop a multifunctional hydrogel with integrated antibacterial activity, ROS scavenging, and glucose-responsive properties to accelerate healing of infected diabetic wounds. Methods: A Zn ion-incorporated injected hydrogel was prepared using 4-carboxyphenylboronic acid-modified gelatine, tannic acid, and zinc ions. The spectra were detected using a Fourier transform infrared spectrometer and surface morphologies of hydrogels were obtained using a scanning electron microscopy. The release behavior of Zn ions was investigated using an inductively coupled plasma mass spectrometry instrument. To evaluate the antimicrobial properties of the GPT and GPT@Zn hydrogels, strains of Escherichia coli and Staphylococcus aureus were utilized. Cytocompatibility was evaluated using mouse fibroblasts (L929 cells) and human umbilical vein endothelial cells (HUVECs). Finally, diabetic wound models were constructed in rats to evaluate the effects of hydrogels on wound healing. Results: The results show that the hydrogels are injectable and have self-healing properties. Moreover, borate ester bonds are formed in the hydrogels, which are responsive to H₂O₂ and glucose and can eliminate them. At the same time, zinc ions were released, giving the hydrogels good antibacterial efficacy, with antibacterial rates of 99.7% and 99.9% against S. aureus and E. coli, respectively. Furthermore, the hydrogels demonstrated good cell compatibility with L929 cells and HUVECs and increased the gene expression of VEGF, COL I, and COL III because of the addition of zinc ions. Based on the ROS, glucose scavenging capacity, and biological functions of zinc ions, the hydrogels advanced the recovery of S. aureus-contaminated whole skin wounds in diabetic rats. Conclusions: This study provides a novel treatment strategy for diabetic wound healing by constructing Zn ion-incorporated injected hydrogels with reactive oxygen species and glucose-scavenging capacity.

Keywords

Hydrogel / Borate ester bond / Reactive oxygen species / Antibacterial / Zn ions

References

[1] Schreml S, Szeimies RM, Karrer S, Heinlin J, Landthaler M, Babilas P. The impact of the pH value on skin integrity and cutaneous wound healing. J Eur Acad Dermatol Venereol. 2010; 24: 373-8. https://doi.org/10.1111/j.1468-3083.2009.03413.x.

[2] Werdin F, Tenenhaus M, Rennekampff HO. Chronic wound care. Lancet. 2008; 372: 1860-2. https://doi.org/10.1016/S0140-6736(08)61793-6.

[3] Morton LM, Phillips TJ. Wound healing and treating wounds differential diagnosis and evaluation of chronic wounds. J Am Acad Dermatol. 2016; 74: 589-605. https://doi.org/10.1016/j.jaad.2015.08.068.

[4] Ma Z, Song W, He Y, Li H. Multilayer injectable hydrogel system sequentially delivers bioactive substances for each wound healing stage. ACS Appl Mater Interfaces. 2020; 12: 29787-806. https://doi.org/10.1021/acsami.0c06360.

[5] Wang H, Xu Z, Zhao M, Liu G, Wu J. Advances of hydrogel dressings in diabetic wounds. Biomater Sci. 2021; 9: 1530-46. https://doi.org/10.1039/D0BM01747G.

[6] Kang SM, Cho H, Jeon D, Park SH, Shin DS, Heo CY. A matrix metalloproteinase sensing biosensor for the evaluation of chronic wounds. Biochip J. 2019; 13: 323-32. https://doi.org/10.1007/s13206-019-3403-4.

[7] Kim YE, Kim J. ROS-scavenging therapeutic hydrogels for modulation of the inflammatory response. ACS Appl Mater Interfaces. 2022; 14: 23002-21. https://doi.org/10.1021/acsami.1c18261.

[8] Eming SA, Martin P, Tomic-Canic M. Wound repair and regeneration: mechanisms, signaling, and translation. Sci Transl Med. 2014; 6: 3009337. https://doi.org/10.1126/scitranslmed.3009337.

[9] Liu K, Kang Y, Dong X, Li Q, Wang Y, Wu X, et al. A simple yet effective hydrogel dressing for advanced microenvironmental management of diabetic wounds with intrinsic regulation. Chem Eng J. 2023; 470: 143987. https://doi.org/10.1016/j.cej.2023.143987.

[10] Zhang Y, Zhu Y, Ma P, Wu H, Xiao D, Zhang Y, et al. Functional carbohydrate-based hydrogels for diabetic wound therapy. Carbohydr Polym. 2023; 312: 120823. https://doi.org/10.1016/j.carbpol.2023.120823.

[11] Boateng J, Catanzano O. Advanced therapeutic dressings for effective wound HealingA review. J Pharm Sci. 2015; 104: 3653-80. https://doi.org/10.1002/jps.24610.

[12] Brumberg V, Astrelina T, Malivanova T, Samoilov A. Modern wound dressings: hydrogel dressings. Biomedicines. 2021; 9: 9091235. https://doi.org/10.3390/biomedicines9091235.

[13] Tavakoli S, Klar AS. Advanced hydrogels as wound dressings. Biomol Ther. 2020; 10: 10081169. https://doi.org/10.3390/biom10081169.

[14] Yao H, Wu M, Lin L, Wu Z, Bae M, Park S, et al. Design strategies for adhesive hydrogels with natural antibacterial agents as wound dressings: status and trends. Mater Today Bio. 2022; 16: 100429. https://doi.org/10.1016/j.mtbio.2022.100429.

[15] Wang Y, Wu Y, Long L, Yang L, Fu D, Hu C, et al. Inflammation-responsive drug-loaded hydrogels with sequential Hemostasis, antibacterial, and anti-inflammatory behavior for chronically infected diabetic wound treatment. ACS Appl Mater Interfaces. 2021; 13: 33584-99. https://doi.org/10.1021/acsami.1c09889.

[16] Chen J, He J, Yang Y, Qiao L, Hu J, Zhang J, et al. Antibacterial adhesive self-healing hydrogels to promote diabetic wound healing. Acta Biomater. 2022; 146: 119-30. https://doi.org/10.1016/j.actbio.2022.04.041.

[17] Xu G, Geng Y, Hu L, Wang J, Pan P, Chen J. One-pot preparation of polysaccharide-based antibacterial hydrogel for skin wound repair. Macromol Mater Eng. 2022; 307: 202100739. https://doi.org/10.1002/mame.202100739.

[18] Liu J, Qu M, Wang C, Xue Y, Huang H, Chen Q, et al. A dual-cross-linked hydrogel patch for promoting diabetic wound healing. Small. 2022; 18: 202106172. https://doi.org/10.1002/smll.202106172.

[19] Zhao H, Huang J, Li Y, Lv X, Zhou H, Wang H, et al. ROS-scavenging hydrogel to promote healing of bacteria infected diabetic wounds. Biomaterials. 2020; 258: 120286. https://doi.org/10.1016/j.biomaterials.2020.120286.

[20] Zhao L, Niu L, Liang H, Tan H, Liu C, Zhu F. pH and glucose dual-responsive injectable hydrogels with insulin and fibroblasts as bioactive dressings for diabetic wound healing. ACS Appl Mater Interfaces. 2017; 9: 37563-74. https://doi.org/10.1021/acsami.7b09395.

[21] Zhang Y, Chen H, Li J. Recent advances on gelatin methacrylate hydrogels with controlled microstructures for tissue engineering. Int J Biol Macromol. 2022; 221: 91-107. https://doi.org/10.1016/j.ijbiomac.2022.08.171.

[22] Huang Y, Bai L, Yang Y, Yin Z, Guo B. Biodegradable gelatin/silver nanoparticle composite cryogel with excellent antibacterial and antibiofilm activity and hemostasis for Pseudomonas aeruginosa-infected burn wound healing. J Colloid Interface Sci. 2022; 608: 2278-89. https://doi.org/10.1016/j.jcis.2021.10.131.

[23] Jafari H, Ghaffari-Bohlouli P, Niknezhad SV, Abedi A, Izadifar Z, Mohammadinejad R, et al. Tannic acid: a versatile polyphenol for design of biomedical hydrogels. J Mater Chem B. 2022; 10: 5873-912. https://doi.org/10.1039/D2TB01056A.

[24] Jing W, Xiaolan C, Yu C, Feng Q, Haifeng Y. Pharmacological effects and mechanisms of tannic acid. Biomed Pharmacother. 2022; 154: 113561. https://doi.org/10.1016/j.biopha.2022.113561.

[25] Wei Q, Zhao Y, Wei Y, Wang Y, Jin Z, Ma G, et al. Facile preparation of polyphenol-crosslinked chitosan-based hydrogels for cutaneous wound repair. Int J Biol Macromol. 2023; 228: 99-110. https://doi.org/10.1016/j.ijbiomac.2022.12.215.

[26] Shi W, Kong Y, Su Y, Kuss MA, Jiang X, Li X, et al. Tannic acid-inspired, self-healing, and dual stimuli responsive dynamic hydrogel with potent antibacterial and anti-oxidative properties. J Mater Chem B. 2021; 9: 7182-95. https://doi.org/10.1039/D1TB00156F.

[27] Deng CC, Brooks WLA, Abboud KA, Sumerlin BS. Boronic acid-based hydrogels undergo self-healing at neutral and acidic pH. ACS Macro Lett. 2015; 4: 220-4. https://doi.org/10.1021/acsmacrolett.5b00018.

[28] Xie W, Zhang Y, Zhang J, Chen X, Pan J, Zhu X, et al. Dynamically crosslinked protien hydrogel composite as multifunctional wound dressing for cutaneous infection. Colloid Interface Sci Commun. 2022; 50: 100654. https://doi.org/10.1016/j.colcom.2022.100654.

[29] Yuan Z, Wan Z, Tian Z, Han Y, Huang X, Feng Y, et al. In situ fused granular hydrogels with ultrastretchability, strong adhesion, and mutli-bioactivities for efficient chronic wound care. Chem Eng J. 2022; 450: 138076. https://doi.org/10.1016/j.cej.2022.138076.

[30] Yang F, Xue Y, Wang F, Guo D, He Y, Zhao X, et al. Sustained release of magnesium and zinc ions synergistically accelerates wound healing. Bioact Mater. 2023; 26: 88-101. https://doi.org/10.1016/j.bioactmat.2023.02.019.

[31] Tao B, Chen M, Lin C, Lu L, Yuan Z, Liu J, et al. Zn-incorporation with graphene oxide on Ti substrates surface to improve osteogenic activity and inhibit bacterial adhesion. J Biomed Mater Res Part A. 2019; 107: 2310-26. https://doi.org/10.1002/jbm.a.36740.

[32] Nguyen HD, Sun X, Yokota H, Lin CC. Probing osteocyte functions in Gelatin hydrogels with Tunable viscoelasticity. Biomacromolecules. 2021; 22: 1115-26. https://doi.org/10.1021/acs.biomac.0c01476.

[33] Liu L, Zhang X, Zhang Y, Pu Y, Yin L, Liu H, et al. Conjugation of antibody on carboxyl-SPIO nanoparticles based on EDC/sulfo-NHS. Acta Univ Med Nanjing. 2013; 33: 480-4.

[34] Li C, Wu G, Ma R, Liu Y, Liu Y, Lv J, et al. Nitrilotriacetic acid (NTA) and Phenylboronic acid (PBA) functionalized Nanogels for efficient encapsulation and controlled release of insulin. ACS Biomater Sci Eng. 2018; 4: 2007-17. https://doi.org/10.1021/acsbiomaterials.7b00546.

[35] Jayakumar A, Jose VK, Lee JM.Hydrogels for medical and environmental applications. Small Methods. 2020; 4: 201900735. https://doi.org/10.1002/smtd.201900735.

[36] Roberts MC, Hanson MC, Massey AP, Karren EA, Kiser PF. Dynamically restructuring hydrogel networks formed with reversible covalent crosslinks. Adv Mater. 2007; 19: 2503-7. https://doi.org/10.1002/adma.200602649.

[37] Chen MH, Wang LL, Chung JJ, Kim YH, Atluri P, Burdick JA. Methods to assess shear-thinning hydrogels for application As injectable biomaterials. ACS Biomater Sci Eng. 2017; 3: 3146-60. https://doi.org/10.1021/acsbiomaterials.7b00734.

[38] Henkelman S, Rakhorst G, Blanton J, van Oeveren W. Standardization of incubation conditions for hemolysis testing of biomaterials. Mater Sci Eng C. 2009; 29: 1650-4. https://doi.org/10.1016/j.msec.2009.01.002.

[39] Zhu Y, Matsumura Y, Velayutham M, Foley LM, Hitchens TK, Wagner WR. Reactive oxygen species scavenging with a biodegradable, thermally responsive hydrogel compatible with soft tissue injection. Biomaterials. 2018; 177: 98-112. https://doi.org/10.1016/j.biomaterials.2018.05.044.

[40] Dhall S, Do DC, Garcia M, Kim J, Mirebrahim SH, Lyubovitsky J, et al. Generating and reversing chronic wounds in diabetic mice by manipulating wound redox parameters. J Diabetes Res. 2014; 2014: 1-18. https://doi.org/10.1155/2014/562625.

[41] Korntner S, Lehner C, Gehwolf R, Wagner A, Grütz M, Kunkel N, et al. Limiting angiogenesis to modulate scar formation. Adv Drug Deliv Rev. 2019; 146: 170-89. https://doi.org/10.1016/j.addr.2018.02.010.

[42] Zhang S, Ge G, Qin Y, Li W, Dong J, Mei J, et al. Recent advances in responsive hydrogels for diabetic wound healing. Mater Today Bio2023. 2022; 18: 100508. https://doi.org/10.1016/j.mtbio.2022.100508.

[43] Chen Y, Wang X, Tao S, Wang Q, Ma PQ, Li ZB, et al. Research advances in smart responsive-hydrogel dressings with potential clinical diabetic wound healing properties. Mil Med Res. 2023; 10:s40779-023-00473-9. https://doi.org/10.1186/s40779-023-00473-9.

[44] Falanga V, Isseroff RR, Soulika AM, Romanelli M, Margolis D, Kapp S, et al. Chronic wounds. Nat Rev Dis Primers. 2022; 8:s41572-022-00377-3. https://doi.org/10.1038/s41572-022-00377-3.