To promote bone regeneration in vivo using critical-size calvarial defect model, hybrid hydrogel was prepared by mixing chitosan with hydroxyapatite (HA) and ultraviolet (UV) irradiation in situ. The hydrosoluble, UV-crosslinkable and injectable N-methacryloyl chitosan (N-MAC) was synthesized via single-step N-acylation reaction. The chemical structure was confirmed by 1H-NMR and FTIR spectroscopy. N-MAC hydrogel presented a microporous structure with pore sizes ranging from 10 to 60 μm. Approximately 80% cell viability of N-MAC hydrogel against encapsulated 3T3 cell indicated that N-MAC is an emerging candidate for mimicking native extracellular matrix (ECM). N-MAC hydrogel hybridized with HA was used to accelerate regeneration of calvarial bone using rabbit model. The effects of hybrid hydrogels to promote bone regeneration were evaluated using critical size calvarial bone defect model. The healing effects of injectable hydrogels with/without HA for bone regeneration were investigated by analyzing X-ray image after 4 or 6 weeks. The results showed that the regenerated new bone for N-MAC 100 was significantly greater than N-MAC without HA and untreated controls. The higher HA content in N-MAC/HA hybrid hydrogel benefited the acceleration of bone regeneration. About 50% closure of defect site after 6 weeks postimplantation demonstrated potent osteoinductivity of N-MAC 100 UV-crosslinkable and injectable N-MAC/HA hybrid hydrogel would allow serving as a promising biomaterial for bone regeneration using the critical-size calvarial defect.
Issue Section:
Engineering Cell Microenvironment Using Novel Hydrogels
References
1.
Porter
, J. R.
, Ruckh
, T. T.
, and Popat
, K. C.
, 2009
, “Bone Tissue Engineering: A Review in Bone Biomimetics and Drug Delivery Strategies
,” Biotechnol. Prog.
, 25
(6
), pp. 1539
–1560
.2.
Cho
, Y. R.
, and Gosain
, A. K.
, 2004
, “Biomaterials in Craniofacial Reconstruction
,” Clin. Plast. Surg.
, 31
(3
), pp. 377
–385
.3.
Kretlow
, J. D.
, Young
, S.
, Klouda
, L.
, Wong
, M.
, and Mikos
, A. G.
, 2009
, “Injectable Biomaterials for Regenerating Complex Craniofacial Tissues
,” Adv. Mater.
, 21
(32–33
), pp. 3368
–3393
.4.
Smith
, D. M.
, Cooper
, G. M.
, Mooney
, M. P.
, Marra
, K. G.
, and Losee
, J. E.
, 2008
, “Bone Morphogenetic Protein 2 Therapy for Craniofacial Surgery
,” J. Craniofacial Surg.
, 19
(5
), pp. 1244
–1259
.5.
Schmitz
, J. P.
, and Hollinger
, J. O.
, 1986
, “The Critical Size Defect as an Experimental-Model for Craniomandibulofacial Nonunions
,” Clin. Orthop. Relat. Res.
, 205
, pp. 299
–308
.6.
Zhang
, Y.
, and Zhang
, M. Q.
, 2001
, “Microstructural and Mechanical Characterization of Chitosan Scaffolds Reinforced by Calcium Phosphates
,” J. Non-Cryst. Solids
, 282
(2–3
), pp. 159
–164
.7.
Zhang
, Y.
, and Zhang
, M. Q.
, 2004
, “Cell Growth and Function on Calcium Phosphate Reinforced Chitosan Scaffolds
,” J. Mater. Sci. Mater. Med.
, 15
(3
), pp. 255
–260
.8.
Afifi
, A. M.
, Gordon
, C. R.
, Pryor
, L. S.
, Sweeney
, W.
, Papay
, F. A.
, and Zins
, J. E.
, 2010
, “Calcium Phosphate Cements in Skull Reconstruction: A Meta-Analysis
,” Plast. Reconstr. Surg.
, 126
(4
), pp. 1300
–1309
.9.
Thomson
, R. C.
, Yaszemski
, M. J.
, Powers
, J. M.
, and Mikos
, A. G.
, 1995
, “Fabrication of Biodegradable Polymer Scaffolds to Engineer Trabecular Bone
,” J. Biomater. Sci., Polym. Ed.
, 7
(1
), pp. 23
–38
.10.
Rezwan
, K.
, Chen
, Q. Z.
, Blaker
, J. J.
, and Boccaccini
, A. R.
, 2006
, “Biodegradable and Bioactive Porous Polymer/Inorganic Composite Scaffolds for Bone Tissue Engineering
,” Biomaterials
, 27
(18
), pp. 3413
–3431
.11.
Swetha
, M.
, Sahithi
, K.
, Moorthi
, A.
, Srinivasan
, N.
, Ramasamy
, K.
, and Selvamurugan
, N.
, 2010
, “Biocomposites Containing Natural Polymers and Hydroxyapatite for Bone Tissue Engineering
,” Int. J. Biol. Macromol.
, 47
(1
), pp. 1
–4
.12.
Lee
, K. Y.
, Jeong
, L.
, Kang
, Y. O.
, Lee
, S. J.
, and Park
, W. H.
, 2009
, “Electrospinning of Polysaccharides for Regenerative Medicine
,” Adv. Drug Delivery Rev.
, 61
(12
), pp. 1020
–1032
.13.
Bhattarai
, N.
, Gunn
, J.
, and Zhang
, M. Q.
, 2010
, “Chitosan-Based Hydrogels for Controlled, Localized Drug Delivery
,” Adv. Drug Delivery Rev.
, 62
(1
), pp. 83
–99
.14.
Vermonden
, T.
, Censi
, R.
, and Hennink
, W. E.
, 2012
, “Hydrogels for Protein Delivery
,” Chem. Rev.
, 112
(5
), pp. 2853
–2888
.15.
Liu
, X. H.
, and Ma
, P. X.
, 2004
, “Polymeric Scaffolds for Bone Tissue Engineering
,” Ann. Biomed. Eng.
, 32
(3
), pp. 477
–486
.16.
Di Martino
, A.
, Sittinger
, M.
, and Risbud
, M. V.
, 2005
, “Chitosan: A Versatile Biopolymer for Orthopaedic Tissue-Engineering
,” Biomaterials
, 26
(30
), pp. 5983
–5990
.17.
Muzzarelli
, R. A. A.
, 2009
, “Chitins and Chitosans for the Repair of Wounded Skin, Nerve, Cartilage and Bone
,” Carbohydr. Polym.
, 76
(2
), pp. 167
–182
.18.
Delange
, G. L.
, and Donath
, K.
, 1989
, “Interface Between Bone Tissue and Implants of Solid Hydroxyapatite or Hydroxyapatite-Coated Titanium Implants
,” Biomaterials
, 10
(2
), pp. 121
–125
.19.
Suchanek
, W.
, and Yoshimura
, M.
, 1998
, “Processing and Properties of Hydroxyapatite-Based Biomaterials for Use as Hard Tissue Replacement Implants
,” J. Mater. Res.
, 13
(1
), pp. 94
–117
.20.
Oshima
, A.
, Tadokoro
, M.
, Hirose
, M.
, Takakura
, Y.
, and Ohgushi
, H.
, 2006
, “Stability of Cultured Osteoblasts on Calcium Phosphate Ceramics
,” Key Eng. Mater.
, 309–311
, pp. 93
–96
.21.
Li
, B. Q.
, Jia
, D. C.
, Zhou
, Y.
, Hu
, Q. L.
, and Cai
, W.
, 2006
, “In Situ Hybridization to Chitosan/Magnetite Nanocomposite Induced by the Magnetic Field
,” J. Magn. Magn. Mater.
, 306
(2
), pp. 223
–227
.22.
Hu
, Q. L.
, Li
, B. Q.
, Wang
, M.
, and Shen
, J. C.
, 2004
, “Preparation and Characterization of Biodegradable Chitosan/Hydroxyapatite Nanocomposite Rods Via In Situ Hybridization: A Potential Material as Internal Fixation of Bone Fracture
,” Biomaterials
, 25
(5
), pp. 779
–785
.23.
Chenite
, A.
, Chaput
, C.
, Wang
, D.
, Combes
, C.
, Buschmann
, M. D.
, Hoemann
, C. D.
, Leroux
, J. C.
, Atkinson
, B. L.
, Binette
, F.
, and Selmani
, A.
, 2000
, “Novel Injectable Neutral Solutions of Chitosan Form Biodegradable Gels In Situ
,” Biomaterials
, 21
(21
), pp. 2155
–2161
.24.
Ruel-Gariepy
, E.
, Shive
, M.
, Bichara
, A.
, Berrada
, M.
, Le Garrec
, D.
, Chenite
, A.
, and Leroux
, J. C.
, 2004
, “A Thermosensitive Chitosan-Based Hydrogel for the Local Delivery of Paclitaxel
,” Eur. J. Pharm. Biopharm.
, 57
(1
), pp. 53
–63
.25.
Mukhopadhyay
, P.
, Sarkar
, K.
, Bhattacharya
, S.
, Bhattacharyya
, A.
, Mishra
, R.
, and Kundu
, P. P.
, 2014
, “pH Sensitive N-Succinyl Chitosan Grafted Polyacrylamide Hydrogel for Oral Insulin Delivery
,” Carbohydr. Polym.
, 112
, pp. 627
–637
.26.
Ifkovits
, J. L.
, and Burdick
, J. A.
, 2007
, “Review: Photopolymerizable and Degradable Biomaterials for Tissue Engineering Applications
,” Tissue Eng.
, 13
(10
), pp. 2369
–2385
.27.
Zhang
, Y.
, Zhan
, F.
, and Shi
, W. F.
, 2011
, “Photopolymerization Behavior and Properties of Highly Branched Poly(Thioether-Urethane) Acrylates Used for UV-Curing Coatings
,” Prog. Org. Coat.
, 71
(4
), pp. 399
–405
.28.
Li
, B. Q.
, Wang
, L.
, Xu
, F.
, Gang
, X. M.
, Demirci
, U.
, Wei
, D. Q.
, Li
, Y.
, Feng
, Y. J.
, Jia
, D. C.
, and Zhou
, Y.
, 2015
, “Hydrosoluble, UV-Crosslinkable and Injectable Chitosan for Patterned Cell-Laden Microgel and Rapid Transdermal Curing Hydrogel In Vivo
,” Acta Biomater.
, 22
, pp. 59
–69
.29.
Naik
, S.
, Bhattacharjya
, G.
, Talukdar
, B.
, and Patel
, B. K.
, 2004
, “Chemoselective Acylation of Amines in Aqueous Media
,” Eur. J. Org. Chem.
, 2004
(6
), pp. 1254
–1260
.30.
Ma
, G. P.
, Zhang
, X. D.
, Han
, J.
, Song
, G. Q.
, and Nie
, J.
, 2009
, “Photo-Polymeriable Chitosan Derivative Prepared by Michael Reaction of Chitosan and Polyethylene Glycol Diacrylate (PEGDA)
,” Int. J. Biol. Macromol.
, 45
(5
), pp. 499
–503
.31.
Valmikinathan
, C. M.
, Mukhatyar
, V. J.
, Jain
, A.
, Karumbaiah
, L.
, Dasari
, M.
, and Bellamkonda
, R. V.
, 2012
, “Photocrosslinkable Chitosan Based Hydrogels for Neural Tissue Engineering
,” Soft Matter
, 8
(6
), pp. 1964
–1976
.32.
Lee
, J. I.
, Kim
, H. S.
, and Yoo
, H. S.
, 2009
, “DNA Nanogels Composed of Chitosan and Pluronic With Thermo-Sensitive and Photo-Crosslinking Properties
,” Int. J. Pharm.
, 373
(1–2
), pp. 93
–99
.33.
Lin
, R. Z.
, Chen
, Y. C.
, Moreno-Luna
, R.
, Khademhosseini
, A.
, and Melero-Martin
, J. M.
, 2013
, “Transdermal Regulation of Vascular Network Bioengineering Using a Photopolymerizable Methacrylated Gelatin Hydrogel
,” Biomaterials
, 34
(28
), pp. 6785
–6796
.34.
Gaharwar
, A. K.
, Dammu
, S. A.
, Canter
, J. M.
, Wu
, C. J.
, and Schmidt
, G.
, 2011
, “Highly Extensible, Tough, and Elastomeric Nanocomposite Hydrogels From Poly(Ethylene Glycol) and Hydroxyapatite Nanoparticles
,” Biomacromolecules
, 12
(5
), pp. 1641
–1650
.35.
Guvendiren
, M.
, and Burdick
, J. A.
, 2012
, “Stiffening Hydrogels to Probe Short- and Long-Term Cellular Responses to Dynamic Mechanics
,” Nat. Commun.
, 3
, p. 792
.36.
Rihova
, B.
, 2000
, “Immunocompatibility and Biocompatibility of Cell Delivery Systems
,” Adv. Drug Delivery Rev.
, 42
(1–2
), pp. 65
–80
.37.
Anderson
, J. M.
, 2001
, “Biological Responses to Materials
,” Annu. Rev. Mater. Res.
, 31
(1
), pp. 81
–110
.Copyright © 2016 by ASME
You do not currently have access to this content.
