Scaffolds based on chitosan/polygalacturonic acid (ChiPgA) complex containing montmorillonite (MMT) clay modified with 5-aminovaleric acid were prepared using freeze-drying technique. The MMT clay was introduced to improve mechanical properties of the scaffold. The microstructure of the scaffolds containing the modified MMT clay was influenced by the incorporation of nanoclays. The MTT assay also indicated that the number of osteoblast cells in ChiPgA scaffolds containing the modified clay was comparable to ChiPgA scaffolds containing hydroxyapatite known for its osteoconductive properties. Overall, the ChiPgA composite scaffolds were found to be biocompatible. This was also indicated by the scanning electron microscopy images of the ChiPgA composite scaffolds seeded with human osteoblast cells. Photoacoustic–Fourier transform infrared (PA-FTIR) experiments on the ChiPgA composite scaffolds indicated formation of a polyelectrolyte complex between chitosan and polygalacturonic acid. PA-FTIR studies also showed that the MMT clay modified with 5-aminovaleric acid was successfully incorporated in the ChiPgA based scaffolds. Swelling studies on ChiPgA composite scaffolds showed the swelling ability of the scaffolds that indicated that the cells and the nutrients would be able to reach the interior parts of the scaffolds. In addition to this, the ChiPgA scaffolds exhibited porosity greater than 90% as appropriate for scaffolds used in tissue engineering studies. High porosity facilitates the nutrient transport throughout the scaffold and also plays a role in the development of adequate vasculature throughout the scaffold. Compressive mechanical tests on the scaffolds showed that the ChiPgA composite scaffolds had compressive elastic moduli in the range of 4–6 MPa and appear to be affected by the high porosity of the scaffolds. Thus, the ChiPgA composite scaffolds containing MMT clay modified with 5-aminovaleric acid are biocompatible. Also, the ChiPgA scaffolds containing the modified MMT clay appears to satisfy some of the basic requirements of scaffolds for tissue engineering applications.
1.
Langer
, R.
, and Vacanti
, J. P.
, 1993, “Tissue Engineering
,” Science
0036-8075, 260
(5110
), pp. 920
–926
.2.
Murugan
, R.
, and Ramakrishna
, S.
, 2005, “Development of Nanocomposites for Bone Grafting
,” Compos. Sci. Technol.
0266-3538, 65
(15–16
), pp. 2385
–2406
.3.
Dorozhkin
, S. V.
, 2007, “Calcium Orthophosphates
,” J. Mater. Sci.
0022-2461, 42
(4
), pp. 1061
–1095
.4.
Salgado
, A. J.
, Coutinho
, O. P.
, and Reis
, R. L.
, 2004, “Bone Tissue Engineering: State of the Art and Future Trends
,” Macromol. Biosci.
1616-5187, 4
(8
), pp. 743
–765
.5.
Goldberg
, M.
, Langer
, R.
, and Jia
, X. Q.
, 2007, “Nanostructured Materials for Applications in Drug Delivery and Tissue Engineering
,” J. Biomater. Sci., Polym. Ed.
0920-5063, 18
(3
), pp. 241
–268
.6.
Barnes
, C. P.
, Sell
, S. A.
, Boland
, E. D.
, Simpson
, D. G.
, and Bowlin
, G. L.
, 2007, “Nanofiber Technology: Designing the Next Generation of Tissue Engineering Scaffolds
,” Adv. Drug Delivery Rev.
0169-409X, 59
(14
), pp. 1413
–1433
.7.
Pirraco
, R. P.
, Marques
, A. P.
, and Reis
, R. L.
, 2010, “Cell Interactions in Bone Tissue Engineering
,” J. Cell. Mol. Med.
, 14
(1–2
), pp. 93
–102
.8.
Langer
, R.
, 1997, “Tissue Engineering: A New Field and Its Challenges
,” Pharmaceutical Research
, 14
(7
), pp. 840
–841
. 0002-78209.
Mano
, J. F.
, Silva
, G. A.
, Azevedo
, H. S.
, Malafaya
, P. B.
, Sousa
, R. A.
, Silva
, S. S.
, Boesel
, L. F.
, Oliveira
, J. M.
, Santos
, T. C.
, Marques
, A. P.
, Neves
, N. M.
, and Reis
, R. L.
, 2007, “Natural Origin Biodegradable Systems in Tissue Engineering and Regenerative Medicine: Present Status and Some Moving Trends
,” J. R. Soc., Interface
1742-5689, 4
(17
), pp. 999
–1030
.10.
Zhang
, Y.
, Venugopal
, J. R.
, El-Turki
, A.
, Ramakrishna
, S.
, Su
, B.
, and Lim
, C. T.
, 2008, “Electrospun Biomimetic Nanocomposite Nanofibers of Hydroxyapatite/Chitosan for Bone Tissue Engineering
,” Biomaterials
0142-9612, 29
(32
), pp. 4314
–4322
.11.
Kong
, L.
, Gao
, Y.
, Lu
, G.
, Gong
, Y.
, Zhao
, N.
, and Zhang
, X.
, 2006, “A Study on the Bioactivity of Chitosan/Nano-Hydroxyapatite Composite Scaffolds for Bone Tissue Engineering
,” Eur. Polym. J.
0014-3057, 42
(12
), pp. 3171
–3179
.12.
Kikuchi
, M.
, Matsumoto
, H. N.
, Yamada
, T.
, Koyama
, Y.
, Takakuda
, K.
, and Tanaka
, J.
, 2004, “Glutaraldehyde Cross-Linked Hydroxyapatite/Collagen Self-Organized Nanocomposites
,” Biomaterials
0142-9612, 25
(1
), pp. 63
–69
.13.
Kim
, H. W.
, Gu
, H. J.
, and Lee
, H. H.
, 2007, “Microspheres of Collagen-Apatite Nanocomposites With Osteogenic Potential for Tissue Engineering
,” Tissue Eng.
1076-3279, 13
(5
), pp. 965
–973
.14.
Svensson
, A.
, Nicklasson
, E.
, Harrah
, T.
, Panilaitis
, B.
, Kaplan
, D. L.
, Brittberg
, M.
, and Gatenholm
, P.
, 2005, “Bacterial Cellulose as a Potential Scaffold for Tissue Engineering of Cartilage
,” Biomaterials
0142-9612, 26
(4
), pp. 419
–431
.15.
Müller
, F. A.
, Müller
, L.
, Hofmann
, I.
, Greil
, P.
, Wenzel
, M. M.
, and Staudenmaier
, R.
, 2006, “Cellulose-Based Scaffold Materials for Cartilage Tissue Engineering
,” Biomaterials
0142-9612, 27
(21
), pp. 3955
–3963
.16.
Santos
, M. I.
, Fuchs
, S.
, Gomes
, M. E.
, Unger
, R. E.
, Reis
, R. L.
, and Kirkpatrick
, C. J.
, 2007, “Response of Micro- and Macrovascular Endothelial Cells to Starch-Based Fiber Meshes for Bone Tissue Engineering
,” Biomaterials
0142-9612, 28
(2
), pp. 240
–248
.17.
Sundaram
, J.
, Durance
, T. D.
, and Wang
, R.
, 2008, “Porous Scaffold of Gelatin-Starch With Nanohydroxyapatite Composite Processed via Novel Microwave Vacuum Drying
,” Acta Biomater.
1742-7061, 4
(4
), pp. 932
–942
.18.
Hofmann
, S.
, Hagenmüller
, H.
, Koch
, A. M.
, Müller
, R.
, Vunjak-Novakovic
, G.
, Kaplan
, D. L.
, Merkle
, H. P.
, and Meinel
, L.
, 2007, “Control of in Vitro Tissue-Engineered Bone-Like Structures Using Human Mesenchymal Stem Cells and Porous Silk Scaffolds
,” Biomaterials
0142-9612, 28
(6
), pp. 1152
–1162
.19.
Mandal
, B. B.
, and Kundu
, S. C.
, 2009, “Non-Mulberry Silk Gland Fibroin Protein 3-D Scaffold for Enhanced Differentiation of Human Mesenchymal Stem Cells Into Osteocytes
,” Acta Biomater.
1742-7061, 5
(7
), pp. 2579
–2590
.20.
Liu
, H.
, Slamovich
, E. B.
, and Webster
, T. J.
, 2006, “Less Harmful Acidic Degradation of Poly(Lactic-Co-Glycolic Acid) Bone Tissue Engineering Scaffolds Through Titania Nanoparticle Addition
,” Int. J. Nanomedicine
, 1
(4
), pp. 541
–545
.21.
Nejati
, E.
, Mirzadeh
, H.
, and Zandi
, M.
, 2008, “Synthesis and Characterization of Nano-Hydroxyapatite Rods/Poly(L-Lactide Acid) Composite Scaffolds for bone Tissue Engineering
,” Composites, Part A
1359-835X, 39
(10
), pp. 1589
–1596
.22.
Serrano
, M. C.
, Pagani
, R.
, Vallet-Regí
, M.
, Peña
, J.
, Rámila
, A.
, Izquierdo
, I.
, and Portolés
, M. T.
, 2004, “In Vitro Biocompatibility Assessment of Poly(ε-Caprolactone) Films Using L929 Mouse Fibroblasts
,” Biomaterials
0142-9612, 25
(25
), pp. 5603
–5611
.23.
Nukavarapu
, S. P.
, Kumbar
, S. G.
, Brown
, J. L.
, Krogman
, N. R.
, Weikel
, A. L.
, Hindenlang
, M. D.
, Nair
, L. S.
, Allcock
, H. R.
, and Laurencin
, C. T.
, 2008, “Polyphosphazene/Nano-Hydroxyapatite Composite Microsphere Scaffolds for Bone Tissue Engineering
,” Biomacromolecules
1525-7797, 9
(7
), pp. 1818
–1825
.24.
Mistry
, A. S.
, Cheng
, S. H.
, Yeh
, T.
, Christenson
, E.
, Jansen
, J. A.
, and Mikos
, A. G.
, 2009, “Fabrication and In Vitro Degradation of Porous Fumarate-Based Polymer/Alumoxane Nanocomposite Scaffolds for Bone Tissue Engineering
,” J. Biomed. Mater. Res. Part A
1549-3296, 89A
(1
), pp. 68
–79
.25.
Christenson
, E. M.
, Anseth
, K. S.
, Van den Beucken
, L.
, Chan
, C. K.
, Ercan
, B.
, Jansen
, J. A.
, Laurencin
, C. T.
, Li
, W. J.
, Murugan
, R.
, Nair
, L. S.
, Ramakrishna
, S.
, Tuan
, R. S.
, Webster
, T. J.
, and Mikos
, A. G.
, 2007, “Nanobiomaterial Applications in Orthopedics
,” J. Orthop. Res.
0736-0266, 25
(1
), pp. 11
–22
.26.
Stevens
, M. M.
, and George
, J. H.
, 2005, “Exploring and Engineering the Cell Surface Interface
,” Science
0036-8075, 310
(5751
), pp. 1135
–1138
.27.
Okada
, A.
, Kawasumi
, M.
, Usuki
, A.
, Kojima
, Y.
, Kurauchi
, T.
, and Kamigaito
, O.
, 1990, “Synthesis and Properties of Nylon-6/Clay Hybrids
,” Polymer Based Molecular Composites, MRS Symposium Proceedings
, D. W.
Schaefer
and J. E.
Mark
, eds., Vol. 171
, pp. 45
–50
.28.
Giannelis
, E. P.
, 1996, “Polymer Layered Silicate Nanocomposites
,” Adv. Mater.
0935-9648, 8
(1
), pp. 29
–35
.29.
Chen
, G. X.
, Hao
, G. J.
, Guo
, T. Y.
, Song
, M. D.
, and Zhang
, B. H.
, 2002, “Structure and Mechanical Properties of Poly(3-Hydroxybutyrateco-3-Hydroxyvalerate) (PHBV)/Clay Nanocomposites
,” J. Mater. Sci. Lett.
0261-8028, 21
, pp. 1587
–1589
.30.
Lim
, S. T.
, Lee
, C. H.
, Choi
, H. J.
, and Jhon
, M. S.
, 2003, “Solidlike Transition of Melt-Intercalated Biodegradable Polymer/Clay Nanocomposites
,” J. Polym. Sci., Part B: Polym. Phys.
0887-6266, 41
(17
), pp. 2052
–2061
.31.
Ma
, C. -C. M.
, Kuo
, C. -T.
, Kuan
, H. -C.
, and Chiang
, C. -L.
, 2003, “Effects of Swelling Agents on the Crystallization Behavior and Mechanical Properties of Polyamide 6/Clay Nanocomposites
,” J. Appl. Polym. Sci.
0021-8995, 88
(7
), pp. 1686
–1693
.32.
Park
, H. -M.
, Lee
, W. -K.
, Park
, C. -Y.
, Cho
, W. -J.
, and Ha
, C. -S.
, 2003, “Environmentally Friendly Polymer Hybrids Part I Mechanical, Thermal, and Barrier Properties of Thermoplastic Starch/Clay Nanocomposites
,” J. Mater. Sci.
0022-2461, 38
, pp. 909
–915
.33.
Yano
, K.
, Usuki
, A.
, Okada
, A.
, Kurauchi
, T.
, and Kamigaito
, O.
, 1993, “Synthesis and Properties of Polyimide-Clay Hybrid
,” J. Polym. Sci., Part A: Polym. Chem.
0887-624X, 31
(10
), pp. 2493
–2498
.34.
Messersmith
, P. B.
, and Giannelis
, E. P.
, 1995, “Synthesis and Barrier Properties of Poly(ε-Caprolactone)-Layered Silicate Nanocomposites
,” J. Polym. Sci., Part A: Polym. Chem.
0887-624X, 33
(7
), pp. 1047
–1057
.35.
Gilman
, J. W.
, Jackson
, C. L.
, Morgan
, A. B.
, and Harris
, R.
, Jr., 2000, “Flammability Properties of Polymer–Layered-Silicate Nanocomposites. Polypropylene and Polystyrene Nanocomposites
,” Chem. Mater.
0897-4756, 12
, pp. 1866
–1873
.36.
Gilman
, J. W.
, 1999, “Flammability and Thermal Stability Studies of Polymer Layered-Silicate (Clay) Nanocomposites
,” Appl. Clay Sci.
0169-1317, 15
(1–2
), pp. 31
–49
.37.
Sinha Ray
, S.
, and Okamoto
, M.
, 2003, “Polymer/Layered Silicate Nanocomposites: A Review From Preparation to Processing
,” Prog. Polym. Sci.
0079-6700, 28
(11
), pp. 1539
–1641
.38.
Sikdar
, D.
, Pradhan
, S. M.
, Katti
, D. R.
, Katti
, K. S.
, and Mohanty
, B.
, 2008, “Altered Phase Model for Polymer Clay Nanocomposites
,” Langmuir
0743-7463, 24
(10
), pp. 5599
–5607
.39.
Sikdar
, D.
, Katti
, D. R.
, and Katti
, K. S.
, 2008, “The Role of Interfacial Interactions on the Crystallinity and Nanomechanical Properties of Clay-Polymer Nanocomposites: A Molecular Dynamics Study
,” J. Appl. Polym. Sci.
0021-8995, 107
(5
), pp. 3137
–3148
.40.
Sikdar
, D.
, Katti
, D.
, Katti
, K.
, and Mohanty
, B.
, 2009, “Influence of Backbone Chain Length and Functional Groups of Organic Modifiers on Crystallinity and Nanomechanical Properties of Intercalated Clay-Polycaprolactam Nanocomposites
,” Int. J. Nanotechnol.
1475-7435, 6
(5/6
), pp. 468
–492
.41.
Katti
, D. R.
, Ghosh
, P.
, Schmidt
, S.
, and Katti
, K. S.
, 2005, “Mechanical Properties of the Sodium Montmorillonite Interlayer Intercalated With Amino Acids
,” Biomacromolecules
1525-7797, 6
(6
), pp. 3276
–3282
.42.
Forni
, F.
, Iannuccelli
, V.
, Coppi
, G.
, and Bernabei
, M. T.
, 1989, “Effect of Montmorillonite on Drug Release From Polymeric Matrices
,” Arch. Pharm. (Weinheim)
0365-6233, 322
(11
), pp. 789
–793
.43.
Lee
, W. -F.
, and Fu
, Y. -T.
, 2003, “Effect of Montmorillonite on the Swelling Behavior and Drug-Release Behavior of Nanocomposite Hydrogels
,” J. Appl. Polym. Sci.
0021-8995, 89
(13
), pp. 3652
–3660
.44.
Dong
, Y.
, and Feng
, S. -S.
, 2005, “Poly(D,L-Lactide-Co-Glycolide)/Montmorillonite Nanoparticles for Oral Delivery of Anticancer Drugs
,” Biomaterials
0142-9612, 26
(30
), pp. 6068
–6076
.45.
Lin
, F. -H.
, Chen
, C. -H.
, Cheng
, W. T. K.
, and Kuo
, T. -F.
, 2006, “Modified Montmorillonite as Vector for Gene Delivery
,” Biomaterials
0142-9612, 27
(17
), pp. 3333
–3338
.46.
Viseras
, C.
, Aguzzi
, C.
, Cerezo
, P.
, and Lopez-Galindo
, A.
, 2007, “Uses of Clay Minerals in Semisolid Health Care and Therapeutic Products
,” Appl. Clay Sci.
0169-1317, 36
(1–3
), pp. 37
–50
.47.
Takahashi
, T.
, Yamada
, Y.
, Kataoka
, K.
, and Nagasaki
, Y.
, 2005, “Preparation of a Novel PEG-Clay Hybrid as a DDS Material: Dispersion Stability and Sustained Release Profiles
,” J. Controlled Release
0168-3659, 107
(3
), pp. 408
–416
.48.
des Rieux
, A.
, Fievez
, V.
, Garinot
, M.
, Schneider
, Y. -J.
, and Préat
, V.
, 2006, “Nanoparticles as Potential Oral Delivery Systems of Proteins and Vaccines: A Mechanistic Approach
,” J. Controlled Release
0168-3659, 116
(1
), pp. 1
–27
.49.
Sun
, B.
, Ranganathan
, B.
, and Feng
, S. -S.
, 2008, “Multifunctional Poly(D,L-Lactide-Co-Glycolide)/Montmorillonite (PLGA/MMT) Nanoparticles Decorated by Trastuzumab for Targeted Chemotherapy of Breast Cancer
,” Biomaterials
0142-9612, 29
(4
), pp. 475
–486
.50.
Wang
, X.
, Du
, Y.
, and Luo
, J.
, 2008, “Biopolymer/Montmorillonite Nanocomposite: Preparation, Drug-Controlled Release Property and Cytotoxicity
,” Nanotechnology
0957-4484, 19
(6
), p. 065707
.51.
Depan
, D.
, Kumar
, A. P.
, and Singh
, R. P.
, 2009, “Cell Proliferation and Controlled Drug Release Studies of Nanohybrids Based on Chitosan-G-Lactic Acid and Montmorillonite
,” Acta Biomater.
1742-7061, 5
(1
), pp. 93
–100
.52.
Carretero
, M. I.
, 2002, “Clay Minerals and Their Beneficial Effects Upon Human Health. A Review
,” Appl. Clay Sci.
0169-1317, 21
(3–4
), pp. 155
–163
.53.
Katti
, K. S.
, Ambre
, A. H.
, Peterka
, N.
, and Katti
, D. R.
, 2010, “Use of Unnatural Amino Acids for Design of Novel Organomodified Clays as Components of Nanocomposite Biomaterials
,” Philos. Trans. R. Soc. London, Ser. A
0962-8428, 368
, pp. 1963
–1980
.54.
Lin
, K. -F.
, Hsu
, C. -Y.
, Huang
, T. -S.
, Chiu
, W. -Y.
, Lee
, Y. -H.
, and Young
, T. -H.
, 2005, “A Novel Method to Prepare Chitosan/Montmorillonite Nanocomposites
,” J. Appl. Polym. Sci.
0021-8995, 98
(5
), pp. 2042
–2047
.55.
Marras
, S. I.
, Kladi
, K. P.
, Tsivintzelis
, I.
, Zuburtikudis
, I.
, and Panayiotou
, C.
, 2008, “Biodegradable Polymer Nanocomposites: The Role of Nanoclays on the Thermomechanical Characteristics and the Electrospun Fibrous Structure
,” Acta Biomater.
1742-7061, 4
(3
), pp. 756
–765
.56.
Zheng
, J. P.
, Wang
, C. Z.
, Wang
, X. X.
, Wang
, H. Y.
, Zhuang
, H.
, and Yao
, K. D.
, 2007, “Preparation of Biomimetic Three-Dimensional Gelatin/Montmorillonite-Chitosan Scaffold for Tissue Engineering
,” React. Funct. Polym.
1381-5148, 67
(9
), pp. 780
–788
.57.
Katti
, K. S.
, Katti
, D. R.
, and Dash
, R.
, 2008, “Synthesis and Characterization of a Novel Chitosan/Montmorillonite/Hydroxyapatite Nanocomposite for Bone Tissue Engineering
,” Biomed. Mater.
, 3
(3
), p. 034122
.58.
Krayukhina
, M. A.
, Samoilova
, N. A.
, and Yamskov
, I. A.
, 2008, “Polyelectrolyte Complexes of Chitosan: Formation, Properties and Applications
,” Russ. Chem. Rev.
0036-021X, 77
(9
), p. 799
.59.
Verma
, D.
, Katti
, K. S.
, and Katti
, D. R.
, 2008, “Effect of Biopolymers on Structure of Hydroxyapatite and Interfacial Interactions in Biomimetically Synthesized Hydroxyapatite/Biopolymer Nanocomposites
,” Ann. Biomed. Eng.
0090-6964, 36
(6
), pp. 1024
–1032
.60.
Verma
, D.
, Katti
, K. S.
, and Katti
, D. R.
, 2009, “Polyelectrolyte-Complex Nanostructured Fibrous Scaffolds for Tissue Engineering
,” Mater. Sci. Eng., C
0928-4931, 29
(7
), pp. 2079
–2084
.61.
Verma
, D.
, Katti
, K. S.
, and Katti
, D. R.
, 2010, “Osteoblast Adhesion, Proliferation and Growth on Polyelectrolyte Complex-Hydroxyapatite Nanocomposites
,” Philos. Trans. R. Soc. London, Ser. A
0962-8428, 368
(1917
), p. 2083
–2097
.62.
Verma
, D.
, Katti
, K. S.
, Katti
, D. R.
, and Mohanty
, B.
, 2008, “Mechanical Response and Multilevel Structure of Biomimetic Hydroxyapatite/Polygalacturonic/Chitosan Nanocomposites
,” Mater. Sci. Eng., C
0928-4931, 28
, pp. 399
–405
.63.
Gibson
, L. J.
, and Ashby
, M. F.
, 1999, Cellular Solids: Structure and Properties
, Cambridge University Press
, New York
.Copyright © 2010
by American Society of Mechanical Engineers
You do not currently have access to this content.
