Abstract
An in situ injectable chitosan/gelatin hydrogel was formed under slightly acidic conditions (pH 4.0 ~ 4.5) using an acid-tolerant tyrosinase, tyrosinase-CNK. A homogeneous chitosan/tyrosinase-CNK solution was prepared in one part of a dual-barrel syringe, and highly soluble gelatin in distilled water was prepared in the other part of the syringe without any additional crosslinking materials. Chitosan/gelatin hydrogel was formed in situ by simple injection of the solutions at room temperature followed by curing at 37°C. However, conventional mushroom tyrosinase did not catalyze this permanent gel formation. Tyrosinase- CNK-catalyzed glycol chitosan/gelatin hydrogel was similarly formed by this in situ injection approach. The hydrogels exhibited a high swelling ratio of 20-fold their own weight, interconnected micropores with an average diameter of approximately 260 μm and in vitro biodegradability suitable for tissue engineering and drug delivery applications. These results showed that tyrosinase-CNK-mediated chitosan/gelatin hydrogel formation has remarkable potential for the development of novel formulations for in situ injectable gel-forming systems.
Similar content being viewed by others
References
Patterson, J., M. M. Martino, and J. A. Hubbell (2010) Biomimetic materials in tissue engineering. Mater. Today 13: 14–22.
Liu, M., X. Zeng, C. Ma, H. Yi, Z. Ali, X. B. Mou, S. Li, Y. Deng, and N. Y. He (2017) Injectable hydrogels for cartilage and bone tissue engineering. Bone Res. 5: 17014.
Yoon, D. Y. and J. C. Kim (2017) Hydrogel composed of acrylic coumarin and acrylic Pluronic F-127 and its photo- and thermoresponsive release property. Biotechnol. Bioproc. Eng. 22: 481–488.
Karavasili, C. and D. G. Fatouros (2016) Smart materials: in situ gel-forming systems for nasal delivery. Drug Discov. Today 21: 157–166.
Choi, Y. R., E. H. Kim, S. Lim, and Y. S. Choi (2018) Efficient preparation of a permanent chitosan/gelatin hydrogel using an acid-tolerant tyrosinase. Biochem. Eng. J. 129: 50–56.
Periayah, M. H., A. S. Halim, and A. Z. Saad (2016) Chitosan: A promising marine polysaccharide for biomedical research. Pharmacogn. Rev. 10: 39–42.
Kim, H. J., J. N. Jin, E. Kan, K. J. Kim, and S. H. Lee (2017) Bacterial cellulose-chitosan composite hydrogel beads for enzyme immobilization. Biotechnol. Bioproc. Eng. 22: 89–94.
LogithKumar, R., A. KeshavNarayan, S. Dhivya, A. Chawla, S. Saravanan, and N. Selvamurugan (2016) A review of chitosan and its derivatives in bone tissue engineering. Carbohydr. Polym. 151: 172–188.
Song, K. D., L. Y. Li, W. F. Li, Y. X. Zhu, Z. R. Jiao, M. Lim, M. Y. Fang, F. X. Shi, L. Wang, and T. Q. Liu (2015) Threedimensional dynamic fabrication of engineered cartilage based on chitosan/gelatin hybrid hydrogel scaffold in a spinner flask with a special designed steel frame. Mat. Sci. Eng. C-Mater. 55: 384–392.
Su, K. and C. M. Wang (2015) Recent advances in the use of gelatin in biomedical research. Biotechnol. Lett. 37: 2139–2145.
Kim, H., Y. J. Yeon, Y. R. Choi, W. Song, S. P. Pack, and Y. S. Choi (2016) A cold-adapted tyrosinase with an abnormally high monophenolase/diphenolase activity ratio originating from the marine archaeon Candidatus Nitrosopumilus koreensis. Biotechnol. Lett. 38: 1535–1542.
Baker, M. I., S. P. Walsh, Z. Schwartz, B. D. Boyan (2012) A review of polyvinyl alcohol and its uses in cartilage and orthopedic applications, J. Biomed. Mater. Res. Part B 100: 1451–1457.
Kalra, A., A. Lowe, and A. M. Al-Jumaily (2016) Mechanical behavior of skin: a review, J. Mater. Sci. Eng. 5: 1000254.
Schneider, C. A., W. S. Rasband, and K. W. Eliceiri (2012) NIH image to ImageJ: 25 years of image analysis. Nat. Methods 9: 671–675.
Chen, T. H., H. D. Embree, E. M. Brown, M. M. Taylor, and G. F. Payne (2003) Enzyme-catalyzed gel formation of gelatin and chitosan: potential for in situ applications. Biomaterials 24: 2831–2841.
Do, H., E. Kang, B. Yang, H. J. Cha, and Y. S. Choi (2017) A tyrosinase, mTyr-CNK, that is functionally available as a monophenol monooxygenase. Sci. Rep. 7: 17267.
Choi, Y. R., H. Do, D. Jeong, J. Park, Y. S. Choi (2016) Reaction stability of the recombinant tyorisinase-CNK originating from the psychrophilic marine microorganism Candidatus Nitrosopumilus Koreensis. Clean Tech. 22: 175–180.
Hubbell, J. A. (1995) Biomaterials in tissue engineering. Biotechnology (NY) 13: 565–576.
Guo, L., R. H. Colby, C. P. Lusignan, and A. M. Howe (2003) Physical gelation of gelatin studied with rheo-optics. Macromolecules 36: 10009–10020.
Alizadeh, M., F. Abbasi, A. B. Khoshfetrat, and H. Ghaleh (2013) Microstructure and characteristic properties of gelatin/ chitosan scaffold prepared by a combined freeze-drying/leaching method. Mat. Sci. Eng. C-Mater. 33: 3958–3967.
Vlierberghe, S. V., V. Cnudde, P. Dubruel, B. Masschaele, A. Cosijns, I. D. Paepe, P. J. S. Jacobs, L. V. Hoorebeke, J. P. Remon, and E. Schacht (2007) Porous gelatin hydrogels: 1. Cryogenic formation and structure analysis. Biomacromolecules. 8: 331–337.
Qiao, C., X. Cao, and F. Wang (2012) Swelling behavior study of physically crosslinked gelatin hydrogels. Polym. Polym. Compos. 20: 53–58.
Li, J. and F. Yao (2012) Environment-stimuli response of chitosan-based hydrogels. In: K. Yao, J. Li, F. Yao, Y. Yin (eds.). Chitosan-based hydrogels. CRC Press,6000 Broken Sound Parkway, NW, USA.
Shen, Z. S., X. Cui, R. X. Hou, Q. Li, H. X. Deng, and J. Fu (2015) Tough biodegradable chitosan-gelatin hydrogels via in situ precipitation for potential cartilage tissue engineering. Rsc. Adv. 5: 55640–55647.
Nieto-Suarez, M., M. A. Lopez-Quintela, and M. Lazzari (2016) Preparation and characterization of cross-linked chitosan/gelatin scaffolds by ice segregation induced self-assembly. Carbohyd. Polym. 141: 175–183.
Gariboldi, M. I. and S. M. Best (2015) Effect of ceramic scaffold architectural parameters on biological response. Front Bioeng. Biotechnol. 3: 151.
Saraiva, S. M., S. P. Miguel, M. P. Ribeiro, P. Coutinho, and I. J. Correia (2015) Synthesis and characterization of a photocrosslinkable chitosan-gelatin hydrogel aimed for tissue regeneration. Rsc. Adv. 5: 63478–63488.
Yang, C., L. Xu, Y. Zhou, X. M. Zhang, X. Huang, M. Wang, Y. Han, M. L. Zhai, S. C. Wei, and J. Q. Li (2010) A green fabrication approach of gelatin/CM-chitosan hybrid hydrogel for wound healing. Carbohyd. Polym. 82: 1297–1305.
Correia, C. R., L. S. Moreira-Teixeira, L. Moroni, R. L. Reis, C. A. van Blitterswijk, M. Karperien, and J. F. Mano (2011) Chitosan scaffolds containing hyaluronic acid for cartilage tissue engineering. Tissue Eng. Part. C-Me 17: 717–730.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kim, E.H., Lim, S., Kim, T.E. et al. Preparation of in situ Injectable Chitosan/Gelatin Hydrogel Using an Acid-tolerant Tyrosinase. Biotechnol Bioproc E 23, 500–506 (2018). https://doi.org/10.1007/s12257-018-0315-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12257-018-0315-4