A new waterborne chitosan-based polyurethane hydrogel as a vehicle to transplant bone marrow mesenchymal cells improved wound healing of ulcers in a diabetic rat model

[EN] Foot ulcers, a common complication of diabetes, can cause physical incapacity and are derived from several factors, including poor wound healing. New therapeutic strategies are needed to minimize this complication for the sake of patients' health. We therefore developed a new chitosan- polyurethane hydrogel membrane (HPUC) and the WA results confirmed that HPUC present low cytotoxicity and improved wound healing when used with mononuclear bone marrow fraction cells in the diabetic rat model. The biodegradable hydrogels were produced in block copolymer networks with a combination of chitosan blocks and biodegradable polyurethane. The membranes were characterized by FTIR, C-13-NMR and thermogravimetry. Swelling and hydrolytic degradation were also evaluated. The non-solubility of the membranes in good solvents and the chemical characterization confirmed that the network structure was formed between the PU and the chitosan through urea/urethane bonds. The findings confirm that the HPUC have interesting properties that make them suitable for wound healing applications.This work was funded by: Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior -doctoral fellowships to Viezzer, C (CAPES/PDSE-BEX: 1408/11-9) and the Spanish Ministry of Economy and Competitiveness (MINECO) through the MAT2016-76039-C4-1-R Project, including FEDER financial support. CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund.Viezzer, C.; Mazzuca, R.; Machado, DC.; Forte, MMDC.; Gómez Ribelles, JL. (2020). A new waterborne chitosan-based polyurethane hydrogel as a vehicle to transplant bone marrow mesenchymal cells improved wound healing of ulcers in a diabetic rat model. Carbohydrate Polymers. 231:1-10. https://doi.org/10.1016/j.carbpol.2019.115734110231Ahmed, R., Tariq, M., Ali, I., Asghar, R., Noorunnisa Khanam, P., Augustine, R., & Hasan, A. (2018). Novel electrospun chitosan/polyvinyl alcohol/zinc oxide nanofibrous mats with antibacterial and antioxidant properties for diabetic wound healing. International Journal of Biological Macromolecules, 120, 385-393. doi:10.1016/j.ijbiomac.2018.08.057Andrade, F., Goycoolea, F., Chiappetta, D. A., das Neves, J., Sosnik, A., & Sarmento, B. (2011). Chitosan-Grafted Copolymers and Chitosan-Ligand Conjugates as Matrices for Pulmonary Drug Delivery. International Journal of Carbohydrate Chemistry, 2011, 1-14. doi:10.1155/2011/865704Baltzis, D., Eleftheriadou, I., & Veves, A. (2014). Pathogenesis and Treatment of Impaired Wound Healing in Diabetes Mellitus: New Insights. Advances in Therapy, 31(8), 817-836. doi:10.1007/s12325-014-0140-xBarikani, M., Honarkar, H., & Barikani, M. (2010). Synthesis and characterization of chitosan-based polyurethane elastomer dispersions. Monatshefte für Chemie - Chemical Monthly, 141(6), 653-659. doi:10.1007/s00706-010-0309-1Boulton, A. J. M. (2013). The Pathway to Foot Ulceration in Diabetes. Medical Clinics of North America, 97(5), 775-790. doi:10.1016/j.mcna.2013.03.007Casettari, L., Vllasaliu, D., Castagnino, E., Stolnik, S., Howdle, S., & Illum, L. (2012). PEGylated chitosan derivatives: Synthesis, characterizations and pharmaceutical applications. Progress in Polymer Science, 37(5), 659-685. doi:10.1016/j.progpolymsci.2011.10.001Chen, L., Tredget, E. E., Wu, P. Y. G., & Wu, Y. (2008). Paracrine Factors of Mesenchymal Stem Cells Recruit Macrophages and Endothelial Lineage Cells and Enhance Wound Healing. PLoS ONE, 3(4), e1886. doi:10.1371/journal.pone.0001886Chen, S.-H., Tsao, C.-T., Chang, C.-H., Wu, Y.-M., Liu, Z.-W., Lin, C.-P., … Hsieh, K.-H. (2012). Synthesis and characterization of thermal-responsive chitin-based polyurethane copolymer as a smart material. Carbohydrate Polymers, 88(4), 1483-1487. doi:10.1016/j.carbpol.2012.01.055Ching Ting Tsao, Chih Hao Chang, Yu Dar Li, Ming Fung Wu, Chun Pin Lin, Jin Lin Han, … Kuo Huang Hsieh. (2011). Development of chitosan/ dicarboxylic acid hydrogels as wound dressing materials. Journal of Bioactive and Compatible Polymers, 26(5), 519-536. doi:10.1177/0883911511422627De Britto, D., & Campana-Filho, S. P. (2007). Kinetics of the thermal degradation of chitosan. Thermochimica Acta, 465(1-2), 73-82. doi:10.1016/j.tca.2007.09.008De Moura, M. R., Aouada, F. A., & Mattoso, L. H. C. (2008). Preparation of chitosan nanoparticles using methacrylic acid. Journal of Colloid and Interface Science, 321(2), 477-483. doi:10.1016/j.jcis.2008.02.006Dinh, T., Tecilazich, F., Kafanas, A., Doupis, J., Gnardellis, C., Leal, E., … Veves, A. (2012). Mechanisms Involved in the Development and Healing of Diabetic Foot Ulceration. Diabetes, 61(11), 2937-2947. doi:10.2337/db12-0227Eming, S. A., Martin, P., & Tomic-Canic, M. (2014). Wound repair and regeneration: Mechanisms, signaling, and translation. Science Translational Medicine, 6(265). doi:10.1126/scitranslmed.3009337Escobar Ivirico, J. L., Salmerón-Sánchez, M., Gómez Ribelles, J. L., & Monleón Pradas, M. (2009). Poly(l-lactide) networks with tailored water sorption. Colloid and Polymer Science, 287(6), 671-681. doi:10.1007/s00396-009-2026-zFlory, P. J., & Rehner, J. (1943). Statistical Mechanics of Cross‐Linked Polymer Networks II. Swelling. The Journal of Chemical Physics, 11(11), 521-526. doi:10.1063/1.1723792Gámiz-González, M. A., Vidaurre, A., & Gómez Ribelles, J. L. (2017). Biodegradable chitosan-poly(Ɛ-caprolactone) dialdehyde copolymer networks for soft tissue engineering. Polymer Degradation and Stability, 138, 47-54. doi:10.1016/j.polymdegradstab.2017.02.005García-Pacios, V., Costa, V., Colera, M., & Martín-Martínez, J. M. (2011). Waterborne polyurethane dispersions obtained with polycarbonate of hexanediol intended for use as coatings. Progress in Organic Coatings, 71(2), 136-146. doi:10.1016/j.porgcoat.2011.01.006García Cruz, D. M., Gomez Ribelles, J. L., & Salmerón Sánchez, M. (2008). Blending polysaccharides with biodegradable polymers. I. Properties of chitosan/polycaprolactone blends. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 85B(2), 303-313. doi:10.1002/jbm.b.30947Heux, L., Brugnerotto, J., Desbrières, J., Versali, M.-F., & Rinaudo, M. (2000). Solid State NMR for Determination of Degree of Acetylation of Chitin and Chitosan. Biomacromolecules, 1(4), 746-751. doi:10.1021/bm000070yHilfiker, A., Kasper, C., Hass, R., & Haverich, A. (2011). Mesenchymal stem cells and progenitor cells in connective tissue engineering and regenerative medicine: is there a future for transplantation? Langenbeck’s Archives of Surgery, 396(4), 489-497. doi:10.1007/s00423-011-0762-2Mohd Hilmi, A. B., Halim, A. S., Hassan, A., Lim, C. K., Noorsal, K., & Zainol, I. (2013). In vitro characterization of a chitosan skin regenerating template as a scaffold for cells cultivation. SpringerPlus, 2(1). doi:10.1186/2193-1801-2-79Hu, Y., Liu, Y., Qi, X., Liu, P., Fan, Z., & Li, S. (2011). Novel bioresorbable hydrogels prepared from chitosan-graft-polylactide copolymers. Polymer International, 61(1), 74-81. doi:10.1002/pi.3150Jiang, T., Nukavarapu, S. P., Deng, M., Jabbarzadeh, E., Kofron, M. D., Doty, S. B., … Laurencin, C. T. (2010). Chitosan–poly(lactide-co-glycolide) microsphere-based scaffolds for bone tissue engineering: In vitro degradation and in vivo bone regeneration studies. Acta Biomaterialia, 6(9), 3457-3470. doi:10.1016/j.actbio.2010.03.023Jiang, X., Li, J., Ding, M., Tan, H., Ling, Q., Zhong, Y., & Fu, Q. (2007). Synthesis and degradation of nontoxic biodegradable waterborne polyurethanes elastomer with poly(ε-caprolactone) and poly(ethylene glycol) as soft segment. European Polymer Journal, 43(5), 1838-1846. doi:10.1016/j.eurpolymj.2007.02.029Jovanovic, D., Engels, G. E., Plantinga, J. A., Bruinsma, M., van Oeveren, W., Schouten, A. J., … Harmsen, M. C. (2010). Novel polyurethanes with interconnected porous structure induce in vivo tissue remodeling and accompanied vascularization. Journal of Biomedical Materials Research Part A, 95A(1), 198-208. doi:10.1002/jbm.a.32817Koehler, J., Brandl, F. P., & Goepferich, A. M. (2018). Hydrogel wound dressings for bioactive treatment of acute and chronic wounds. European Polymer Journal, 100, 1-11. doi:10.1016/j.eurpolymj.2017.12.046Kuo, Y.-R., Wang, C.-T., Cheng, J.-T., Wang, F.-S., Chiang, Y.-C., & Wang, C.-J. (2011). Bone Marrow–Derived Mesenchymal Stem Cells Enhanced Diabetic Wound Healing through Recruitment of Tissue Regeneration in a Rat Model of Streptozotocin-Induced Diabetes. Plastic and Reconstructive Surgery, 128(4), 872-880. doi:10.1097/prs.0b013e3182174329Leventis, N., Sotiriou-Leventis, C., Chandrasekaran, N., Mulik, S., Larimore, Z. J., Lu, H., … Mang, J. T. (2010). Multifunctional Polyurea Aerogels from Isocyanates and Water. A Structure−Property Case Study. Chemistry of Materials, 22(24), 6692-6710. doi:10.1021/cm102891dMarchant, R. E., Zhao, Q., Anderson, J. M., & Hiltner, A. (1987). Degradation of a poly(ether urethane urea) elastomer: infra-red and XPS studies. Polymer, 28(12), 2032-2039. doi:10.1016/0032-3861(87)90037-1Marcos-Fernández, A., Abraham, G. A., Valentín, J. L., & Román, J. S. (2006). Synthesis and characterization of biodegradable non-toxic poly(ester-urethane-urea)s based on poly(ε-caprolactone) and amino acid derivatives. Polymer, 47(3), 785-798. doi:10.1016/j.polymer.2005.12.007Masson-Meyers, D. S., Bumah, V. V., & Enwemeka, C. S. (2016). A comparison of four methods for determining viability in human dermal fibroblasts irradiated with blue light. Journal of Pharmacological and Toxicological Methods, 79, 15-22. doi:10.1016/j.vascn.2016.01.001Merchant, Z., Taylor, K. M. G., Stapleton, P., Razak, S. A., Kunda, N., Alfagih, I., … Somavarapu, S. (2014). Engineering hydrophobically modified chitosan for enhancing the dispersion of respirable microparticles of levofloxacin. European Journal of Pharmaceutics and Biopharmaceutics, 88(3), 816-829. doi:10.1016/j.ejpb.2014.09.005Moise, M., Şunel, V., Holban, M., Popa, M., Desbrieres, J., Peptu, C., & Lionte, C. (2012). Double crosslinked chitosan and gelatin submicronic capsules entrapping aminoacid derivatives with potential antitumoral activity. Journal of Materials Science, 47(23), 8223-8233. doi:10.1007/s10853-012-6719-1Neto, C. G. T., Giacometti, J. A., Job, A. E., Ferreira, F. C., Fonseca, J. L. C., & Pereira, M. R. (2005). Thermal Analysis of Chitosan Based Networks. Carbohydrate Polymers, 62(2), 97-103. doi:10.1016/j.carbpol.2005.02.022Ouyang, Q.-Q., Hu, Z., Lin, Z.-P., Quan, W.-Y., Deng, Y.-F., Li, S.-D., … Chen, Y. (2018). Chitosan hydrogel in combination with marine peptides from tilapia for burns healing. International Journal of Biological Macromolecules, 112, 1191-1198. doi:10.1016/j.ijbiomac.2018.01.217Page, J. M., Prieto, E. M., Dumas, J. E., Zienkiewicz, K. J., Wenke, J. C., Brown-Baer, P., & Guelcher, S. A. (2012). Biocompatibility and chemical reaction kinetics of injectable, settable polyurethane/allograft bone biocomposites. Acta Biomaterialia, 8(12), 4405-4416. doi:10.1016/j.actbio.2012.07.037Park, W. S., Ahn, S. Y., Sung, S. I., Ahn, J.-Y., & Chang, Y. S. (2017). Strategies to enhance paracrine potency of transplanted mesenchymal stem cells in intractable neonatal disorders. Pediatric Research, 83(1-2), 214-222. doi:10.1038/pr.2017.249Pérez-Limiñana, M. A., Arán-Aís, F., Torró-Palau, A. M., César Orgilés-Barceló, A., & Miguel Martín-Martínez, J. (2005). Characterization of waterborne polyurethane adhesives containing different amounts of ionic groups. International Journal of Adhesion and Adhesives, 25(6), 507-517. doi:10.1016/j.ijadhadh.2005.02.002Pretsch, T., Jakob, I., & Müller, W. (2009). Hydrolytic degradation and functional stability of a segmented shape memory poly(ester urethane). Polymer Degradation and Stability, 94(1), 61-73. doi:10.1016/j.polymdegradstab.2008.10.012Rowlands, A. S., Lim, S. A., Martin, D., & Cooper-White, J. J. (2007). Polyurethane/poly(lactic-co-glycolic) acid composite scaffolds fabricated by thermally induced phase separation. Biomaterials, 28(12), 2109-2121. doi:10.1016/j.biomaterials.2006.12.032Abdel-Rahman, R. M., Abdel-Mohsen, A. M., Hrdina, R., Burgert, L., Fohlerova, Z., Pavliňák, D., … Jancar, J. (2016). Wound dressing based on chitosan/hyaluronan/nonwoven fabrics: Preparation, characterization and medical applications. International Journal of Biological Macromolecules, 89, 725-736. doi:10.1016/j.ijbiomac.2016.04.087Subramani, S., Park, Y.-J., Lee, Y.-S., & Kim, J.-H. (2003). New development of polyurethane dispersion derived from blocked aromatic diisocyanate. Progress in Organic Coatings, 48(1), 71-79. doi:10.1016/s0300-9440(03)00118-8Tao, Z., Shi, A., & Zhao, J. (2015). Epidemiological Perspectives of Diabetes. Cell Biochemistry and Biophysics, 73(1), 181-185. doi:10.1007/s12013-015-0598-4Ünlü, C., Pollet, E., & Avérous, L. (2018). Original Macromolecular Architectures Based on poly(ε-caprolactone) and poly(ε-thiocaprolactone) Grafted onto Chitosan Backbone. International Journal of Molecular Sciences, 19(12), 3799. doi:10.3390/ijms19123799Velazquez-Morales, P., Le Nest, J.-F., & Gandini, A. (1998). Polymer electrolytes derived from chitosan/polyether networks. Electrochimica Acta, 43(10-11), 1275-1279. doi:10.1016/s0013-4686(97)10030-5Wang, W., Ping, P., Chen, X., & Jing, X. (2006). Polylactide-based polyurethane and its shape-memory behavior. European Polymer Journal, 42(6), 1240-1249. doi:10.1016/j.eurpolymj.2005.11.029Yamane, T., Nakagami, G., Yoshino, S., Shimura, M., Kitamura, A., Kobayashi-Hattori, K., … Sanada, H. (2015). Hydrocellular foam dressings promote wound healing associated with decrease in inflammation in rat periwound skin and granulation tissue, compared with hydrocolloid dressings. Bioscience, Biotechnology, and Biochemistry, 79(2), 185-189. doi:10.1080/09168451.2014.968088Zawadzki, J., & Kaczmarek, H. (2010). Thermal treatment of chitosan in various conditions. Carbohydrate Polymers, 80(2), 394-400. doi:10.1016/j.carbpol.2009.11.037Zhang, S., Cheng, L., & Hu, J. (2003). NMR studies of water-borne polyurethanes. Journal of Applied Polymer Science, 90(1), 257-260. doi:10.1002/app.12696Zhang, S., Ren, Z., He, S., Zhu, Y., & Zhu, C. (2007). FTIR spectroscopic characterization of polyurethane-urea model hard segments (PUUMHS) based on three diamine chain extenders. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 66(1), 188-193. doi:10.1016/j.saa.2006.02.041Zuber, M., Zia, K. M., Mahboob, S., Hassan, M., & Bhatti, I. A. (2010). Synthesis of chitin–bentonite clay based polyurethane bio-nanocomposites. International Journal of Biological Macromolecules, 47(2), 196-200. doi:10.1016/j.ijbiomac.2010.04.02