The preparation and cytocompatibility of injectable thermosensitive chitosan/poly(vinyl alcohol) hydrogel

Abstract

In order to investigate the strength, structure and cell cytocompatibility of injectable thermosensitive chitosan (CS)/poly(vinyl alcohol) (PVA) composite hydrogel, chitosan hydrochloride solution was transferred to a neutral pH and mixed with different proportions of PVA, then the gelation time and strength of these different hydrogels were tested and spatial structures were observed under a scanning electron microscopy (SEM) after freeze-drying. The cytocompatibility of the hydrogels was evaluated through cytotoxicity test and three-dimensional culture with bone marrow mesenchymal stem cells. The results showed that the CS/PVA solution kept in liquid state at low temperature (0–4°C) and turned into transparent elastomer about 15–20 min at 37°C. Gelation time was prolonged, the strength increased and porous structure became dense with the PVA content increased in the mixed hydrogel. The cytotoxicity grades of these gels were from 0 to 1. Rabbit bone marrow mesenchymal stem cells could survive and proliferate in the gel within 3 weeks, and the gel had good cytocompatibility. It was concluded that thermosensitive CS/PVA composite hydrogel not only has interpenetrating network structure and better mechanical strength, but also has good cytocompatibility, and may be used as an injectable scaffold for tissue engineering.

Keywords

hydrogel / chitosan / polyvinyl alcohol / mesenchymal stem cells

References

[1] KimI.Y., SeoS.J., MoonH.S., et al.. Chitosan and its derivatives for tissue engineering applications. Biotechnol Adv, 2008, 26(1): 1-21

[2] KumarM.N., MuzzarelliR.A., MuzzarelliC., et al.. Chitosan chemistry and pharmaceutical perspectives. Chem Rev, 2004, 104(12): 6017-6084

[3] KhorE., LimL.Y.. Implantable applications of chitin and chitosan. Biomaterials, 2003, 24(13): 2339-2349

[4] SezerA.D., CevherE., HatipoǧluF., et al.. Preparation of fucoidan-chitosan hydrogel and its application as burn healing accelerator on rabbits. Biol Pharm Bull, 2008, 31(12): 2326-2333

[5] GutowskaA., JeongB., JasionowskiM.. Injectable gels for tissue engineering. Anat Rec, 2001, 263(4): 342-349

[6] ShuX.Z., LiuY.C., PalumboF.S., et al.. In situ crosslinkable hyaluronan hydrogels for tissue engineering. Biomaterials, 2004, 25(7–8): 1339-1348

[7] JeongB., KimS.W., BaebY.H.. Thermosensitive sol-gel reversible hydrogels. Adv Drug Deliv Rev, 2002, 54(1): 37-51

[8] ParkJ., KimD.. Release behavior of amoxicillin from glycol chitosan superporous hydrogels. J Biomater Sci Polym Ed, 2009, 20(5–6): 853-862

[9] de Souza Costa-JúniorE., PereiraM.M., MansurH.S.. Properties and biocompatibility of chitosan films modified by blending with PVA and chemically crosslinked. J Mater Sci Mater Med, 2009, 20(2): 553-561

[10] MoriY., TokuraH., YoshikawaM.. Properties of hydrogels synthesized by freezing and thawing aqueous polyvinyl alcohol solutions and their applications. J Materials Sci, 1997, 32(2): 491-496

[11] KurkuriM.D., AminabhaviT.M.. Poly(vinyl alcohol) and poly(acrylic acid) sequential interpenetrating network pH-sensitive microspheres for the delivery of diclofenac sodium to the intestine. J Control Release, 2004, 96(1): 9-20

[12] ParkH., KimD.. Swelling and mechanical properties of glycolchitosan/poly(vinyl alcohol) IPN-type superporous hydrogels. J Biomed Mater Res A, 2006, 78(4): 662-667

[13] TangY., DuY., LiY., et al.. A thermosensitive chitosan/ poly(vinyl alcohol) hydrogel containing hydroxyapatite for protein delivery. J Biomed Mater Res A, 2009, 91(4): 953-963

[14] GongZ., DingS.S., YinY.J., et al.. Optimized design of hy drogels for tissue engineering. Hua Xue Gongye Yu Gong Cheng Jin Zhan (Chinese)., 2008, 27(11): 1743-1748

[15] Bureau of Technical Supervision.The People’s Republic of China National Standards: Biological evaluation of medical silicone materials test methods (GB/T 16175-1996)[S], 1996, Beijing, China Standard Press: 17-18

[16] KretlowJ.D., KloudaL., MikosA.G.. Injectable matrices and scaffolds for drug delivery in tissue engineering. Adv Drug Deliv Rev, 2007, 59(4–5): 236-273

[17] BalakrishnanB., JayakrishnanA.. Self-cross-linking biopolymers as injectable in situ forming biodegradable scaffolds. Biomaterials, 2005, 26(18): 3941-3951

[18] CheniteA., ChaputC., WangD., et al.. Novel injectable neutral solutions of chitosan form biodegradable gels in situ. Biomaterials, 2000, 21(21): 2155-2161

[19] CuiY.L., QiA.D., LiuW.G., et al.. Biomimetic surface modification of poly(L-lactic acid) with chitosan and its effects on articular chondrocytes in vitro. Biomaterials, 2003, 24(21): 3859-3868

[20] DonatiI., StredanskaS., SilvestriniG., et al.. The aggregation of pig articular chondrocyte and synthesis of extracellular matrix by a lactose-modified chitosan. Biomaterials, 2005, 26(9): 987-998

[21] HongY., SongH., GongY., et al.. Covalentlycrosslinked chitosan hydrogel: properties of in vitro degradation and chondrocyte encapsulation. Acta Biomaterialia, 2007, 3(1): 23-31

[22] BhattaraiN., RamayH.R., GunnJ., et al.. PEG-grafted chitosan as an injectable thermosensitive hydrogel for sustained protein release. J Control Release, 2005, 103(3): 609-624

[23] DruryJ.L., MooneyD.J.. Hydrogels for tissue engineering: scaffold design variables and applications. Biomaterials, 2003, 24(24): 4337-4351

[24] TanH., GongY., LaoL., et al.. Gelatin/chitosan/hyaluronan ternary complex scaffold containing basic fibroblast growth factor for cartilage tissue engineering. J Mater Sci Mater Med, 2007, 18(10): 1961-1968

[25] CromptonK.E., GoudJ.D., BellamkondaR.V., et al.. Polylysine-functionalised thermoresponsive chitosan hydrogel for neural tissue engineering. Biomaterials, 2007, 28(3): 441-449