A rapid freezing/lyophilizing/reinforcing process is suggested to fabricate reinforced keratin/hydroxyapatite (HA) scaffold with improved mechanical property and biocompatibility for tissue engineering. The keratin, extracted from human hair, and HA mixture were rapidly frozen with liquid nitrogen and then lyophilized to prepare keratin/HA laminar scaffold. The scaffold was then immersed in PBS for reinforcement treatment, and followed by a second lyophilization to prepare the reinforced keratin/HA scaffold. The morphology, mechanical, chemical, crystal and thermal property of the keratin/HA scaffold were investigated by SEM, FTIR, XRD, DSC, respectively. The results showed that the keratin/HA scaffold had a high porosity of 76.17 ± 3%. The maximum compressive strength and compressive modulus of the reinforced scaffold is 0.778 and 3.3 MPa respectively. Subcutaneous implantation studies in mice showed that in vivo the scaffold was biocompatible since the foreign body reaction seen around the implanted scaffold samples was moderate and became minimal upon increasing implantation time. These results demonstrate that the keratin/HA reinforced scaffold prepared here is promising for biomedical utilization.
Keratin Hydroxyapatite Scaffold Biomaterial Hair
This is a preview of subscription content, log in to check access
The present work is supported by National Natural Science Foundation of China under Grant (No. 51573133), A Foundation for the Author of National Excellent Doctoral Dissertation of PR China (No. 201255), Program for New Century Excellent Talents in University (NCET-12-1063), Tianjin Natural Science Foundation (14JCYBJC17600), and National Training Program of Innovation and Entrepreneurship for Undergraduates (201510058056).
Compliance with ethical standards
Conflict of interest
We wish to confirm that there are no known conflicts of interest associated with this publication.
This study was approved by the laboratory animal center of North china university of science and technology (XYXK (冀) 2015-0038).
Thompson ZS, Rijal NP, Jarvis D, Edwards A, Bhattarai N. Synthesis of keratin-based nanofiber for biomedical engineering. J Vis Exp. 2016;108:e53381.Google Scholar
Tachibana A, Furuta Y, Takeshima H, Tanabe T, Yamauchi K. Fabrication of wool keratin sponge scaffolds for long-term cell cultivation. J Biotechnol. 2002;93:165–70.CrossRefPubMedGoogle Scholar
Katoh K, Tanabe T, Yamauchi K. Novel approach to fabricate keratin sponge scaffolds with controlled pore size and porosity. Biomaterials. 2004;25:4255–62.CrossRefPubMedGoogle Scholar
Yamauchi K, Khoda A. Novel proteinous microcapsules from wool keratins. Colloids Surf B Biointerfaces. 1997;9:117–9.CrossRefGoogle Scholar
Reichl S. Films based on human hair keratin as substrates for cell culture and tissue engineering. Biomaterials. 2009;30:6854–66.CrossRefPubMedGoogle Scholar
Fujii T, Tanaka T, Ohkawa K. Biomineralization of calcium phosphate on human hair protein film and formation of a novel hydroxyapatite-protein composite material. J Biomed Mater Res B Appl Biomater. 2009;91:528–36.CrossRefPubMedGoogle Scholar
Lee H, Noh K, Lee SC, Kwon IK, Han DW, Lee IS, et al. Human hair keratin and its-based biomaterials for biomedical applications. Tissue Eng Regen Med. 2014;11:255–65.CrossRefGoogle Scholar
Tachibana A, Kaneko S, Tanabe T, Yamauchi K. Rapid fabrication of keratin–hydroxyapatite hybrid sponges toward osteoblast cultivation and differentiation. Biomaterials. 2005;26:297–302.CrossRefPubMedGoogle Scholar
Verma V, Verma P, Ray P, Ray AR. Preparation of scaffolds from human hair proteins for tissue-engineering applications. Biomed Mater. 2008;3:025007.CrossRefPubMedGoogle Scholar
Elliott JC. Structure and chemistry of the apatites and other calcium orthophosphates. Amsterdam: Elsevier; 1994. p. 380–97.Google Scholar
Lu SZ, Liu JJ, Yan SQ, Liu JB, Li MZ. Preparation and characterization of silk fibroin/hydroxyapatite porous composite materials. J Clin Rehabil Tissue Eng Res. 2009;13:6789–92.Google Scholar
Cox SC, Thornby JA, Gibbons GJ, Williams MA, Mallick KK. 3D printing of porous hydroxyapatite scaffolds intended for use in bone tissue engineering applications. Mater Sci Eng C Mater Biol Appl. 2015;47:237–47.CrossRefPubMedGoogle Scholar
Dias GJ, Mahoney P, Swain M, Kelly RJ, Smith RA, Ali MA. Keratin-hydroxyapatite composites: biocompatibility, osseointegration, and physical properties in an ovine model. J Biomed Mater Res A. 2010;95:1084–95.CrossRefPubMedGoogle Scholar
Yamauchi K, Yamauchi A, Kusunoki T, Kohda A, Konishi Y. Preparation of stable aqueous solution of keratins, and physiochemical and biodegradational properties of films. J Biomed Mater Res. 1996;31:439–44.CrossRefPubMedGoogle Scholar
Aluigi A, Corbellini A, Rombaldoni F, Zoccola M, Canetti M. Morphological and structural investigation of wool-derived keratin nanofibres crosslinked by thermal treatment. Int J Biol Macromol. 2013;57:30–7.CrossRefPubMedGoogle Scholar
Kang HW, Tabata Y, Ikada Y. Fabrication of porous gelatin scaffolds for tissue engineering. Biomaterials. 1999;20:1339–44.CrossRefPubMedGoogle Scholar
Xu X, He H, Zhang Y, Zhang D, Yang Z. Influence of position on the microstructure of carbon black/polyvinyl alcohol composite obtained by the directional freeze-drying process. J Macromol Sci Phys. 2014;53:568–74.CrossRefGoogle Scholar
Li J, Li Y, Li L, Mak AFT, Ko F, Qin L. Fabrication and degradation of poly (l-lactic acid) scaffolds with wool keratin. Compos B Eng. 2009;40:664–7.CrossRefGoogle Scholar
Prasong S, Wasan T. Preparation and characterization of hair keratin/gelatin blend films. Pak J Biol Sci. 2011;14:351–6.CrossRefPubMedGoogle Scholar
Liu Y, Yu X, Li J, Fan J, Wang M, Lei TD, et al. Fabrication and properties of high-content keratin/poly (ethylene oxide) blend nanofibers using two-step cross-linking process. J Nanomater. 2015;2015:803937.Google Scholar
Chen YM, Xi TF, Zheng YD, Zheng YF, Wan YZ. In vitro degradation performance of nano-hydroxyapatite/bacterial cellulose for bone tissue engineering. Acta Sci Nat Univ Pekin. 2012;48:524–32.Google Scholar
Li J, Liu X, Zhang J, Zhang Y, Han Y, Hu J, et al. Synthesis and characterization of wool keratin/hydroxyapatite nanocomposite. J Biomed Mater Res B Appl Biomater. 2012;100:896–902.CrossRefPubMedGoogle Scholar
Mohandes F, Salavati-Niasari M, Fathi M, Fereshteh Z. Hydroxyapatite nanocrystals: simple preparation, characterization and formation mechanism. Mater Sci Eng C Mater Biol Appl. 2014;45:29–36.CrossRefPubMedGoogle Scholar
Cao J. Melting study of the alpha-form crystallites in human hair keratin by DSC. Thermochim Acta. 1999;335:5–9.CrossRefGoogle Scholar