Preparation and properties of calcium sulfate bone cement incorporated with silk fibroin and Sema3A-loaded chitosan microspheres

Abstract

To search for new bioactive materials which can be used as the substitute of bone repairing and drug carriers, Sema3A-loaded chitosan microspheres (SLCM) and silk fibroin (SF) were mixed with calcium sulfate cement (CSC). SEM, particle size analysis and swelling rate determination were performed to study properties of the microspheres. The drug loading, encapsulation efficiency and drug release rate were determined by ELISA. Microspheres with different SLCM weight contents (0.5%, 1% and 5%) were prepared to determine which one has the strongest mechanical properties and the appropriate setting time. It was revealed that CSC/SF/0.5SLCM has satisfactory mechanical properties, and its in vitro biocompatibility was assessed by MTS. Chitosan microspheres (5--18 μm) were globular, the surface was smooth, and the swelling rate is (77.02±5.57)%. With this formula, the setting time was increased with the addition of SLCM in CSC/SF, and the cumulative drug release rate is 44.62% in 28 d. XRD results demonstrate that the main component is calcium sulfate. Also it was found that CSC/SF/0.5SLCM supports the growth of MC3T3 cells. Thus the preparation of CSC/SF/0.5SLCM was reliable, and the products had good structures, physical properties and biocompati-bility, appearing to be a promising bone substitute material.

Keywords

Sema3A / chitosan microsphere / sustained release / silk fibroin (SF) / bone cement

References

[1] Hulme P A, Krebs J, Ferguson S J, . Vertebroplasty and kyphoplasty: a systematic review of 69 clinical studies. Spine, 2006, 31(17): 1983–2001

[2] Taylor R S, Fritzell P, Taylor R J. Balloon kyphoplasty in the management of vertebral compression fractures: an updated systematic review and meta-analysis. European Spine Journal, 2007, 16(8): 1085–1100

[3] Jensen M E, McGraw J K, Cardella J F, . Position statement on percutaneous vertebral augmentation: a consensus statement developed by the American Society of Interventional and Therapeutic Neuroradiology, Society of Interventional Radiology, American Association of Neurological Surgeons/Congress of Neurological Surgeons, and American Society of Spine Radiology. American Journal of Neuroradiology, 2007, 28(8): 1439–1443

[4] Cunin G, Boissonnet H, Petite H, . Experimental vertebroplasty using osteoconductive granular material. Spine, 2000, 25(9): 1070–1076

[5] Lu J X, Huang Z W, Tropiano P, . Human biological reactions at the interface between bone tissue and polymethylmethacrylate cement. Journal of Materials Science: Materials in Medicine, 2002, 13(8): 803–809

[6] Heini P F, Walchli B, Berlemann U. Percutaneous transpedicular vertebroplasty with PMMA: operative technique and early results. European Spine Journal, 2000, 9(5): 445–450

[7] Barralet J E, Gaunt T, Wright A J, . Effect of porosity reduction by compaction on compressive strength and microstructure of calcium phosphate cement. Journal of Biomedical Materials Research, 2002, 63(1): 1–9

[8] Lazáry A, Balla B, Kósa J P, . Effect of gypsum on proliferation and differentiation of MC3T3-E1 mouse osteoblastic cells. Biomaterials, 2007, 28(3): 393–399

[9] Kim S E, Park J H, Cho Y W, . Porous chitosan scaffold containing microspheres loaded with transforming growth factor-beta1: implications for cartilage tissue engineering. Journal of Controlled Release, 2003, 91(3): 365–374

[10] Sadiasa A, Sarkar S K, Franco R A, . Bioactive glass incorporation in calcium phosphate cement-based injectable bone substitute for improved in vitro biocompatibility and in vivo bone regeneration. Journal of Biomaterials Applications, 2014, 28(5): 739–756

[11] Chen C C, Wang C W, Hsueh N S, . Improvement of in vitro physicochemical properties and osteogenic activity of calcium sulfate cement for bone repair by dicalcium silicate. Journal of Alloys and Compounds, 2014, 585: 25–31

[12] Omenetto F G, Kaplan D L. New opportunities for an ancient material. Science, 2010, 329(5991): 528–531

[13] Wang G, Yang H, Li M, . The use of silk fibroin/hydroxyapatite composite co-cultured with rabbit bone-marrow stromal cells in the healing of a segmental bone defect. The Journal of Bone and Joint Surgery British Volume, 2010, 92(2): 320–325

[14] Tian L, Rauvala H, Gahmberg C G. Neuronal regulation of immune responses in the central nervous system. Trends in Immunology, 2009, 30(2): 91–99

[15] Potiron V, Nasarre P, Roche J, . Semaphorin signaling in the immune system. Advances in Experimental Medicine and Biology, 2007, 600: 132–144

[16] Adams R H, Eichmann A. Axon guidance molecules in vascular patterning. Cold Spring Harbor Perspectives in Biology, 2010, 2(5): a001875

[17] Neufeld G, Kessler O. The semaphorins: versatile regulators of tumour progression and tumour angiogenesis. Nature Reviews Cancer, 2008, 8(8): 632–645

[18] Roth L, Koncina E, Satkauskas S, . The many faces of semaphorins: from development to pathology. Cellular and Molecular Life Sciences, 2009, 66(4): 649–666

[19] Hayashi M, Nakashima T, Taniguchi M, . Osteoprotection by semaphorin 3A. Nature, 2012, 485(7396): 69–74

[20] Wang A H, Chen X G, Liu C S, . Preparation and characteristics of chitosan microspheres in different acetylation as drug carrier system. Journal of Microencapsulation, 2009, 26(7): 593–602

[21] Wang Q Z, Chen X G, Li Z X, . Preparation and blood coagulation evaluation of chitosan microspheres. Journal of Materials Science: Materials in Medicine, 2008, 19(3): 1371–1377

[22] Zhang Y, Cheng X, Wang J, . Novel chitosan/collagen scaffold containing transforming growth factor-β1 DNA for periodontal tissue engineering. Biochemical and Biophysical Research Communications, 2006, 344(1): 362–369

[23] Grenha A, Remunan-Lopez C, Carvalho E L S, . Microspheres containing lipid/chitosan nanoparticles complexes for pulmonary delivery of therapeutic proteins. European Journal of Pharmaceutics and Biopharmaceutics, 2008, 69(1): 83–93

[24] Mandal B B, Kundu S C. Cell proliferation and migration in silk fibroin 3D scaffolds. Biomaterials, 2009, 30(15): 2956–2965

[25] Kim U J, Park J, Kim H J, . Three-dimensional aqueous-derived biomaterial scaffolds from silk fibroin. Biomaterials, 2005, 26(15): 2775–2785

[26] Sariibrahimoglu K, Wolke J G, Leeuwenburgh S C, . Injectable biphasic calcium phosphate cements as a potential bone substitute. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2014, 102(3): 415–422

[27] Zhang C, Cheng Y, Qu G, . Preparation and characterization of galactosylated chitosan coated BSA microspheres containing 5-fluorouracil. Carbohydrate Polymers, 2008, 72(3): 390–397

[28] Gupta K, Jabrail F. Effects of degree of deacetylation and cross-linking on physical characteristics, swelling and release behavior of chitosan microspheres. Carbohydrate Polymers, 2006, 66(1): 43–54

[29] Lin Q, Lan X, Li Y, . Anti-washout carboxymethyl chitosan modified tricalcium silicate bone cement: preparation, mechanical properties and in vitro bioactivity. Journal of Materials Science: Materials in Medicine, 2010, 21(12): 3065–3076

[30] Kim S Y, Jeon S H. Setting properties, mechanical strength and in vivo evaluation of calcium phosphate-based bone cements. Journal of Industrial and Engineering Chemistry, 2012, 18(1): 128–136

[31] Habraken W J, Wolke J G, Mikos A G, . Porcine gelatin microsphere/calcium phosphate cement composites: an in vitro degradation study. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2009, 91(2): 555–561

[32] Ruediger T, Berg A, Guellmar A, . Cytocompatibility of polymer-based periodontal bone substitutes in gingival fibroblast and MC3T3 osteoblast cell cultures. Dental Materials, 2012, 28(10): e239–e249

[33] Cai L, Guinn A S, Wang S. Exposed hydroxyapatite particles on the surface of photo-crosslinked nanocomposites for promoting MC3T3 cell proliferation and differentiation. Acta Biomaterialia, 2011, 7(5): 2185–2199

[34] Zhang M, Wu C, Lin K, . Biological responses of human bone marrow mesenchymal stem cells to Sr–M–Si (M= Zn, Mg) silicate bioceramics. Journal of Biomedical Materials Research Part A, 2012, 100(11): 2979–2990

[35] Puricelli E, Corsetti A, Ponzoni D, . Characterization of bone repair in rat femur after treatment with calcium phosphate cement and autogenous bone graft. Head & Face Medicine, 2010, 6(1): 10

[36] Wang H, Zou Q, Boerman O C, . Combined delivery of BMP-2 and bFGF from nanostructured colloidal gelatin gels and its effect on bone regeneration in vivo. Journal of Controlled Release, 2013, 166(2): 172–181

[37] Zhu C, Chang Q, Zou D, . LvBMP-2 gene-modified BMSCs combined with calcium phosphate cement scaffolds for the repair of calvarial defects in rats. Journal of Materials Science: Materials in Medicine, 2011, 22(8): 1965–1973

[38] Li F, Liu J, Yang G, . Effect of pH and succinic acid on the morphology of α-calcium sulfate hemihydrate synthesized by a salt solution method. Journal of Crystal Growth, 2013, 374: 31–36

[39] Hazra C, Bari S, Kundu D, . Ultrasound-assisted/biosurfactant-templated size-tunable synthesis of nano-calcium sulfate with controllable crystal morphology. Ultrasonics Sonochemistry, 2014, 21(3): 1117–1131

[40] Zhang M, Wu C, Lin K, . Biological responses of human bone marrow mesenchymal stem cells to Sr–M–Si (M= Zn, Mg) silicate bioceramics. Journal of Biomedical Materials Research Part A, 2012, 100(11): 2979–2990

[41] Li Y W, Leong J C, Lu W W, . A novel injectable bioactive bone cement for spinal surgery: a developmental and preclinical study. Journal of Biomedical Materials Research, 2000, 52(1): 164–170

[42] Ginebra M P, Fernández E, De Maeyer E A, . Setting reaction and hardening of an apatitic calcium phosphate cement. Journal of Dental Research, 1997, 76(4): 905–912

[43] Hesaraki S, Moztarzadeh F, Nemati R, . Preparation and characterization of calcium sulfate-biomimetic apatite nanocomposites for controlled release of antibiotics. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2009, 91(2): 651–661

[44] Sugawara A, Asaoka K, Ding S J. Calcium phosphate-based cements: clinical needs and recent progress. Journal of Materials Chemistry B, 2013, 1(8): 1081–1089

[45] Ding S J, Shie M Y, Wang C Y. Novel fast-setting calcium silicate bone cements with high bioactivity and enhanced osteogenesis in vitro. Journal of Materials Chemistry, 2009, 19(8): 1183–1190

[46] Wang B, Xie R J, Huang Y Y. Preparation and characterization of silk fibroin/calcium phosphate composite. Advanced Materials Research, 2011, 332–334: 1655–1658

[47] Wang B, Xie R J, Wan Q, . Effect of silk fibroin on the properties of calcium phosphate cement. Advanced Materials Research, 2011, 175–176: 100–104

[48] Karageorgiou V, Kaplan D. Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials, 2005, 26(27): 5474–5491

Acknowledgements

This work was generously supported by the National Natural Science Foundation of China (Grant Nos. 81071454, 81171689 and 81301559), Department of Orthopedics Clinical Medical Centers in Jiangsu Province (BL2012004), the Natural Science Foundation of Jiangsu Province (BK20130275), and the Key Laboratory of Orthopedic Institute of Soochow University.

RIGHTS & PERMISSIONS

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg