title: Bioprintable, cell-laden silk fibroin-gelatin hydrogel supporting multilineage differentiation of stem cells for fabrication of three-dimensional tissue constructs.
abstract: Bioprinting has exciting prospects for printing three-dimensional (3-D) tissue constructs by delivering living cells with appropriate matrix materials. However, progress in this field is currently extremely slow due to limited choices of bioink for cell encapsulation and cytocompatible gelation mechanisms. Here we report the development of clinically relevant sized tissue analogs by 3-D bioprinting, delivering human nasal inferior turbinate tissue-derived mesenchymal progenitor cells encapsulated in silk fibroin-gelatin (SF-G) bioink. Gelation in this bioink was induced via in situ cytocompatible gelation mechanisms, namely enzymatic crosslinking by mushroom tyrosinase and physical crosslinking via sonication. Mechanistically, tyrosinases oxidize the accessible tyrosine residues of silk and/or gelatin into reactive o-quinone moieties that can either condense with each other or undergo nonenzymatic reactions with available amines of both silk and gelatin. Sonication alters the hydrophobic interaction and accelerates self-assembly of silk fibroin macromolecules to form β-sheet crystals, which physically crosslink the hydrogel. However, sonication has no effect on the conformation of gelatin. The effect of optimized rheology, secondary conformations of silk-gelatin bioink, temporally controllable gelation strategies and printing parameters were assessed to achieve maximum cell viability and multilineage differentiation of the encapsulated human nasal inferior turbinate tissue-derived mesenchymal progenitor cells. This strategy offers a unique path forward in the direction of direct printing of spatially customized anatomical architecture in a patient-specific manner.
authors.affiliation: Department of Textile Technology, Indian Institute of Technology, New Delhi 110016, India; Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), 77 Cheong-Am Ro, Nam-gu, Pohang, Kyungbuk 790-784, South Korea; Centre for Biomedical Engineering, Indian Institute of Technology, New Delhi 110016, India.
authors.forename: Sanskrita
authors.initials: S
authors.lastname: Das
authors.affiliation: Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), 77 Cheong-Am Ro, Nam-gu, Pohang, Kyungbuk 790-784, South Korea.
authors.forename: Falguni
authors.initials: F
authors.lastname: Pati
authors.affiliation: Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), 77 Cheong-Am Ro, Nam-gu, Pohang, Kyungbuk 790-784, South Korea.
authors.forename: Yeong-Jin
authors.initials: YJ
authors.lastname: Choi
authors.affiliation: Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), 77 Cheong-Am Ro, Nam-gu, Pohang, Kyungbuk 790-784, South Korea.
authors.forename: Girdhari
authors.initials: G
authors.lastname: Rijal
authors.affiliation: Department of Mechanical Engineering, Korea Polytechnic University, 2121 Jeongwang-dong, Siheung-si, Gyeonggi-do 429-793, South Korea.
authors.forename: Jin-Hyung
authors.initials: JH
authors.lastname: Shim
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul 137-701, South Korea.
authors.forename: Sung Won
authors.initials: SW
authors.lastname: Kim
authors.affiliation: Centre for Biomedical Engineering, Indian Institute of Technology, New Delhi 110016, India.
authors.forename: Alok R
authors.initials: AR
authors.lastname: Ray
authors.affiliation: Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), 77 Cheong-Am Ro, Nam-gu, Pohang, Kyungbuk 790-784, South Korea. Electronic address: dwcho@postech.ac.kr.
authors.forename: Dong-Woo
authors.initials: DW
authors.lastname: Cho
authors.affiliation: Department of Textile Technology, Indian Institute of Technology, New Delhi 110016, India. Electronic address: sghosh08@textile.iitd.ac.in.
authors.forename: Sourabh
authors.initials: S
authors.lastname: Ghosh
mesh_terms: D001672:Biocompatible Materials / Q000138:chemical synthesis; D062028:Bioprinting / Q000295:instrumentation / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005349:Fibroins / Q000737:chemistry*; D005780:Gelatin / Q000737:chemistry; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D008422:Materials Testing; D009924:Organ Culture Techniques / Q000295:instrumentation* / Q000379:methods; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology*; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods; D054457:Tissue Scaffolds
chemical_list: D001672:Biocompatible Materials; D020100:Hydrogels; D005780:Gelatin; D005349:Fibroins
keywords: Bioprinting; Cytocompatible gelation; Multilineage differentiation; Self-standing 3-D construct; Silk–gelatin bioink