Biocompatibility and osteoinductive activity of nano-hydroxyapatite/chitosan/ poly(lactide-co-glycolide) scaffolds in vitro

doi:10.3969/j.issn.2095-4344.2014.08.009

Abstract

Abstract:

BACKGROUND: Studies have found that combination of two of chitosan (CS), nano-hydroxyapatite (nHA) and poly(lactide-co-glycolide) (PLGA) can improve the mechanical properties and biocompatibility of the composite stent in certain extent as well as improve osteogenic differentiation of the cells, but there is a certain distance from the ideal bone tissue engineering scaffolds.
OBJECTIVE: To study biocompatibility and osteoinductive activity of nHA/CS/PLGA scaffolds with different proportions in vitro.
METHODS: nHA/CS/PLGA scaffolds were prepared at mass ratio of 10:10:80, 10:20:70, 20:10:70 respectively by particle leaching method. And human bone marrow stem cells (hBMSCs) were co-cultured with these scaffolds in vitro. Adhesion, proliferation, and osteoinductive activity of these scaffolds were examined qualitatively and quantitatively by growth curve of hBMSCs on scaffolds. Gene expression of alkaline phosphatase activity and osteocalcin was detected by RT-PCR.
RESULTS AND CONCLUSION: hBMSCs could be attached, proliferated, and osteoinduced better on the nHA/CS/PLGA scaffold with the mass ratio of 20:10:70, compared to the other two groups of scaffolds. The differences were significant statistically (P < 0.05). Alkaline phosphatase and osteocalcin expressions were respectively higher in the scaffold with the mass ratio of 20:10:70 after 9-27 days of co-culture and 15-27 days of co-culture, in comparison with the other two groups of scaffolds. These findings indicate that the nHA/CS/PLGA scaffolds with the mass ratio of 20:10:70 demonstrated preferable biocompatibility and osteogenic inductivity, which is expected to be a promising scaffold material for bone tissue engineering.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: biocompatible materials, nanoparticles, chitosan, hydroxyapatites

CLC Number:

R318

Wang Fei, Zhou Hong, Guo Yu-cheng, Su Xiao-xia, Rao Guo-zhou, Zhao Xiao-peng. Biocompatibility and osteoinductive activity of nano-hydroxyapatite/chitosan/ poly(lactide-co-glycolide) scaffolds in vitro[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(8): 1198-1204.

Figures/Tables 4

References

[1] Baskin JZ,Eppell SJ.A selected review of the recent advances in craniomaxillofacial bone tissue engineering.Curr Opin Otolaryngol Head Neck Surg.2013;21(4):389-395.

[2] Jin GZ,Kim TH,Kim JH,et al.Bone tissue engineering of induced pluripotent stem cells cultured with macrochanneled polymer scaffold.J Biomed Mater Res A. 2013;101(5): 1283- 1291.

[3] Thibault RA,Mikos AG,Kasper FK.Scaffold/Extracellular matrix hybrid constructs for bone-tissue engineering.Adv Healthc Mater.2013;2(1):13-24.

[4] Bao TQ,Franco RA,Lee BT.Preparation and characterization of a novel 3D scaffold from poly(?-caprolactone)/biphasic calcium phosphate hybrid composite microspheres adhesion. Biochem Eng J.2012;64:76-83.

[5] Sadiasa A,Kim MS,Lee BT.Poly(lactide-co-glycolide acid)/biphasic calcium phosphate composite coating on a porous scaffold to deliver simvastatin for bone tissue engineering. J Drug Target.2013;21(8):719-729.

[6] Li B,Yang J,Ma L,et al.Fabrication of poly(lactide-co-glycolide) scaffold filled with fibrin gel, mesenchymal stem cells, and poly(ethylene oxide)-b-poly(L-lysine)/TGF-beta1 plasmid DNA complexes for cartilage restoration in vivo.J Biomed Mater Res A.2013.

[7] Kavya KC,Jayakumar R,Nair S,et al.Fabrication and characterization of chitosan/gelatin/nSiO2 composite scaffold for bone tissue engineering. Int J Biol Macromol.2013;59: 255-263.

[8] Zhou Y,Xu L,Zhang X,et al.Radiation synthesis of gelatin/CM-chitosan/β-tricalcium phosphate composite scaffold for bone tissue engineering. Materials Science and Engineering: C.2012;32(4):994-1000.

[9] Fricain JC,Schlaubitz S,Le Visage C,et al.A nano-hydroxyapatite--pullulan/dextran polysaccharide composite macroporous material for bone tissue engineering.Biomaterials.2013;34(12):2947-2959.

[10] Cho JS,Rhee SH.Formation mechanism of nano-sized hydroxyapatite powders through spray pyrolysis of a calcium phosphate solution containing polyethylene glycol.J Eur Ceram Soc.2013;33(2):233-241.

[11] Liu G,Li Y,Sun J,et al.In vitro and in vivo evaluation of osteogenesis of human umbilical cord blood-derived mesenchymal stem cells on partially demineralized bone matrix.Tissue Eng Part A.2010;16(3):971-982.

[12] Lee RW,Volz RG,Sheridan DC.The role of fixation and bone quality on the mechanical stability of tibial knee components.Clin Orthop Relat Res.1991;(273):177-183.

[13] Croisier F,Jérôme C.Chitosan-based biomaterials for tissue engineering. Eur Polymer J.2013;49(4):780-792.

[14] Kim IY,Seo SJ,Moon HS,et al.Chitosan and its derivatives for tissue engineering applications.Biotechnol Adv.2008;26(1): 1-21.

[15] Xu M,Li Y,Suo H,et al.Fabricating a pearl/PLGA composite scaffold by the low-temperature deposition manufacturing technique for bone tissue engineering. Biofabrication. 2010; 2(2):025002.

[16] Lin ZY,Duan ZX,Guo XD,et al.Bone induction by biomimetic PLGA-(PEG-ASP)n copolymer loaded with a novel synthetic BMP-2-related peptide in vitro and in vivo.J Controlled Release.2010;144(2):190-195.

[17] Minamiguchi S,Takechi M,Yuasa T,et al.Basic research on aw-AC/PLGA composite scaffolds for bone tissue engineering.J Mater Sci Mater Med.2008;19(3):1165-1172.

[18] Kim SS,Sun Park M,Jeon O,et al. Poly(lactide-co-glycolide)/hydroxyapatite composite scaffolds for bone tissue engineering.Biomaterials.2006;27(8): 1399- 1409.

[19] Li M,Liu W,Sun J,et al.Culturing primary human osteoblasts on electrospun poly(lactic-co-glycolic acid) and poly(lactic-co-glycolic acid)/nanohydroxyapatite scaffolds for bone tissue engineering.ACS Appl Mater Interfaces. 2013; 5(13):5921-5926.

[20] Kavlock KD,Whang K,Guelcher SA,et al.Degradable segmented polyurethane elastomers for bone tissue engineering: effect of polycaprolactone content.J Biomater Sci Polym Ed.2013;24(1):77-93.

[21] Zhang P,Hong Z,Yu T,et al.In vivo mineralization and osteogenesis of nanocomposite scaffold of poly(lactide-co-glycolide) and hydroxyapatite surface-grafted with poly(L-lactide).Biomaterials.2009;30(1):58-70.

[22] Ngiam M,Liao S,Patil AJ,et al.The fabrication of nano-hydroxyapatite on PLGA and PLGA/collagen nanofibrous composite scaffolds and their effects in osteoblastic behavior for bone tissue engineering. Bone.2009; 45(1):4-16.

[23] Jose MV,Thomas V,Johnson KT,et al.Aligned PLGA/HA nanofibrous nanocomposite scaffolds for bone tissue engineering. Acta Biomater.2009;5(1):305-315.

[24] Huang YX,Ren J,Chen C,et al.Preparation and properties of poly(lactide-co-glycolide) (PLGA)/ nano-hydroxyapatite (NHA) scaffolds by thermally induced phase separation and rabbit MSCs culture on scaffolds.J Biomater Appl.2008;22(5): 409-432.

[25] Wu YC,Shaw SY,Lin HR,et al.Bone tissue engineering evaluation based on rat calvaria stromal cells cultured on modified PLGA scaffolds. Biomaterials.2006;27(6):896-904.

[26] Hu Q,Li B,Wang M,et al.Preparation and characterization of biodegradable chitosan/hydroxyapatite nanocomposite rods via in situ hybridization: a potential material as internal fixation of bone fracture.Biomaterials.2004;25(5):779-785.

[27] Thein-Han WW,Misra RDK.Biomimetic chitosan–nanohydroxyapatite composite scaffolds for bone tissue engineering.Acta Biomater.2009;5(4):1182-1197.

[28] Teng X,Ren J,Gu S.Preparation and characterization of porous PDLLA/HA composite foams by supercritical carbon dioxide technology.J Biomed Mater Res B Appl Biomater. 2007; 81(1):185-193.

[29] Mooney DJ,Mazzoni CL,Breuer C,et al.Stabilized polyglycolic acid fibre-based tubes for tissue engineering.Biomaterials. 1996;17(2):115-124.

[30] Whang K,Thomas CH,Healy KE,et al.A novel method to fabricate bioabsorbable scaffolds.Polymer. 1995;36(4): 837-842.

[31] Harris LD,Kim BS,Mooney DJ.Open pore biodegradable matrices formed with gas foaming.J Biomed Mater Res. 1998;42(3):396-402.

[32] Porter JR,Ruckh TT,Popat KC.Bone tissue engineering: a review in bone biomimetics and drug delivery strategies. Biotechnol Prog.2009;25(6):1539-1560.

[33] Caverzasio J,Manen D.Essential role of Wnt3a-mediated activation of mitogen-activated protein kinase p38 for the stimulation of alkaline phosphatase activity and matrix mineralization in C3H10T1/2 mesenchymal cells. Endocrinology.2007;148(11):5323-5330.

[34] Rey A,Manen D,Rizzoli R,et al.Evidences for a role of p38 MAP kinase in the stimulation of alkaline phosphatase and matrix mineralization induced by parathyroid hormone in osteoblastic cells.Bone.2007;41(1):59-67.

[35] Nakamura A,Dohi Y,Akahane M,et al.Osteocalcin secretion as an early marker of in vitro osteogenic differentiation of rat mesenchymal stem cells.Tissue Eng Part C Methods.2009; 15(2):169-180.

[36] Akahane M,Ueha T,Dohi Y,et al.Secretory osteocalcin as a nondestructive osteogenic marker of tissue-engineered bone.J Orthop Sci. 2011;16 (5): 622-628.

[37] Pandit V,Zuidema JM,Venuto KN,et al.Evaluation of Multifunctional Polysaccharide Hydrogels with Varying Stiffness for Bone Tissue Engineering. Tissue Eng Part A.2013;19(21-22);2452-2463.

[38] Vozzi G,Corallo C,Carta S,et al. Collagen-gelatin-genipin-

hydroxyapatite composite scaffolds colonized by human primary osteoblasts are suitable for bone tissue engineering applications: In vitro evidences.J Biomed Mater Res A.2013. doi;10.1002/jbm.a.34823.

[39] Kim J,Jeong SY,Ju YM,et al.In vitro osteogenic differentiation of human amniotic fluid-derived stem cells on a poly(lactide-co-glycolide) (PLGA)-bladder submucosa matrix (BSM) composite scaffold for bone tissue engineering.Biomed Mater.2013;8(1):014107.

[40] Pavlin D,Zadro R,Gluhak-Heinrich J.Temporal pattern of stimulation of osteoblast-associated genes during mechanically-induced osteogenesis in vivo: early responses of osteocalcin and type I collagen.Connect Tissue Res.2001; 42(2):135-148.

[41] Ito M.In vitro properties of a chitosan-bonded hydroxyapatite bone-filling paste.Biomaterials.1991;12(1):41-45.