The development of controlled drug delivery systems for bone regeneration, especially microspheres, has become a research hotspot in recent years. Chitosan and its derivative O-carboxymethyl chitosan have been considered to be an effective way for controlled drug delivery due to their nontoxicity and biodegradability. Currently, most of the studies have researched on synthesizing and characterizing chitosan and O-carboxymethyl chitosan. However, few studies have focused on the differences between chitosan microspheres and O-carboxymethyl chitosan microspheres directly. In this study, chitosan and O-carboxymethyl chitosan microspheres were developed by water-in-oil emulsification cross-linking method using vanillin as the cross-linking agent, and then their physicochemical properties were evaluated by Fourier transform infrared spectroscopy, scanning electron microscopy, and in vitro release testing. The results showed that O-carboxymethyl chitosan was successfully modified by adding carboxymethyl group at the chitosan C6 position.The particle size of chitosan microspheres (50–90 µm) was significantly larger than that of O-carboxymethyl chitosan microspheres (10–50 µm), and the drug release profile of O-carboxymethyl chitosan microspheres showed larger initial burst release within the first day and sustained release at the fourth day, while chitosan microspheres showed sustained release at the seventh day. In addition, Cell Counting Kit-8 assay showed that MC3T3-E1 proliferated well and highly expressed the alkaline phosphatase marker protein on both chitosan and O-carboxymethyl chitosan microspheres. Overall, both chitosan and O-carboxymethyl chitosan microspheres showed good biocompatibility, and chitosan microspheres were superior to O-carboxymethyl chitosan microspheres. Moreover, the different drug release rates suggest that chitosan and O-carboxymethyl chitosan microspheres have the potential to be used for the repair of different bone defects.
JiangTAbdel-FattahWILaurencinCT.In vitro evaluation of chitosan/poly(lactic acid-glycolic acid) sintered microsphere scaffolds for bone tissue engineering. Biomaterials2006; 27: 4894–4903.
2.
BayerEAGottardiRFedorchakMVet al. The scope and sequence of growth factor delivery for vascularized bone tissue regeneration. J Control Release2015; 219: 129–140.
3.
TomicIVidis-MillwardAMueller-ZsigmondyMet al. Setting accelerated dissolution test for PLGA microspheres containing peptide, investigation of critical parameters affecting drug release rate and mechanism. Int J Pharm2016; 505: 42–51.
4.
MuthuMSKuttyRVLuoZet al. Theranostic vitamin E TPGS micelles of transferrin conjugation for targeted co-delivery of docetaxel and ultra bright gold nanoclusters. Biomaterials2015; 39: 234–248.
5.
KumarM.Nano and microparticles as controlled drug delivery devices. J Pharm Pharm Sci2000; 3: 234–258.
6.
AllenTMCullisPR.Liposomal drug delivery systems: from concept to clinical applications. Adv Drug Deliv Rev2013; 65: 36–48.
7.
GuptaLRaoKPChoudharyKet al. Preformulation studies of biodegradable drug implants of meloxicam for orthopedic patient care. J Pharm Sci Technol2011; 3: 494–498.
8.
FreibergSZhuXX.Polymer microspheres for controlled drug release. Int J Pharm2004; 282: 1–18.
9.
WangXJingXZhangXet al. A versatile platform of magnetic microspheres loaded with dual-anticancer drugs for drug release. Mater Chem Phys2016; 177: 213–219.
10.
ShiCFengSLiuPet al. A novel study on the mechanisms of drug release in PLGA-mPEG microspheres with fluorescent drug. J Biomater Sci Polym Ed2016; 27: 854–864.
11.
SawhneyAS.Composite hydrogel drug delivery systems. Patent US9254267 B2, 2016.
12.
LiuQZuoQGuoRet al. Fabrication and characterization of carboxymethyl chitosan/poly(vinyl alcohol) hydrogels containing alginate microspheres for protein delivery. J Bioact Compat Pol2015; 30: 397–411.
13.
VasirJKTambwekarKGargS.Bioadhesive microspheres as a controlled drug delivery system. Int J Pharm2003; 255: 13–32.
14.
WangLZhengLLiCet al. Adrenomedullin delivery in microsphere-scaffold composite for remodeling of the alveolar bone following tooth extraction: an experimental study in the rat. Biomed Eng Online2013; 12: 99.
15.
BaekJJungH-DJangT-Set al. Synthesis and evaluation of bone morphogenetic protein (BMP)-loaded hydroxyapatite microspheres for enhanced bone regeneration. Ceram Int2016; 42: 7748–7756.
16.
LiDDengLYangZet al. Antigen-free bovine cancellous bone loaded with recombinant human bone morphogenetic protein-2 for the repair of tibial bone defects in goat model. J Biomater Appl2016; 30: 1322–1333.
17.
KimSYHwangJ-YShinUS.Preparation of nano/macroporous polycaprolactone microspheres for an injectable cell delivery system using room temperature ionic liquid and camphene. J Colloid Interface Sci2016; 465: 18–25.
18.
LautnerGMeyerhoffMESchwendemanSP.Biodegradable poly (lactic-co-glycolic acid) microspheres loaded with S-nitroso-N-acetyl-D-penicillamine for controlled nitric oxide delivery. J Control Release2016; 225: 133–139.
19.
ShiXFangQDingMet al. Microspheres of carboxymethyl chitosan, sodium alginate and collagen for a novel hemostatic in vitro study. J Biomater Appl2016; 30: 1092–1102.
KucharskaMWalenkoKLewandowska-SzumiełMet al. Chitosan and composite microsphere-based scaffold for bone tissue engineering: evaluation of tricalcium phosphate content influence on physical and biological properties. J Mater Sci Mater Med2015; 26: 1–12.
22.
ChenPXiaCMeiSet al. Intra-articular delivery of sinomenium encapsulated by chitosan microspheres and photo-crosslinked GelMA hydrogel ameliorates osteoarthritis by effectively regulating autophagy. Biomaterials2016; 81: 1–13.
23.
SinhaVRSinglaAKWadhawanSet al. Chitosan microspheres as a potential carrier for drugs. Int J Pharm2004; 274: 1–33.
KenawyER.Biologically active polymers: modification and anti-microbial activity of chitosan derivatives. J Bioact Compat Pol2005; 20: 95–111.
26.
JainRA.The manufacturing techniques of various drug loaded biodegradable poly (lactide-co-glycolide)(PLGA) devices. Biomaterials2000; 21: 2475–2490.
27.
MuzzarelliCStanicVGobbiLet al. Spray-drying of solutions containing chitosan together with polyuronans and characterisation of the microspheres. Carbohyd Polym2004; 57: 73–82.
28.
LiuZCheungRWuXYet al. A study of doxorubicin loading onto and release from sulfopropyl dextran ion-exchange microspheres. J Control Release2001; 77: 213–224.
29.
AndrianovAKChenJPayneLG.Preparation of hydrogel microspheres by coacervation of aqueous polyphosphazene solutions. Biomaterials1998; 19: 109–115.
30.
ShuXZhuK.Chitosan/gelatin microspheres prepared by modified emulsification and ionotropic gelation. J Microencapsul2001; 18: 237–245.
31.
ZhouGYuXTaiJet al. Research on a novel chitosan microsphere/scaffold system by double crosslinkers. Dent Mater J2016.
32.
WalkeSSrivastavaGNikaljeMet al. Fabrication of chitosan microspheres using vanillin/TPP dual crosslinkers for protein antigens encapsulation. Carbohyd Polym2015; 128: 188–198.
33.
LiuXFGuanYLYangDZet al. Antibacterial action of chitosan and carboxymethylated chitosan. J Appl Polym Sci2001; 79: 1324–1335.
34.
ChenX-GParkH-J.Chemical characteristics of O-carboxymethyl chitosans related to the preparation conditions. Carbohyd Polym2003; 53: 355–359.
35.
UpadhyayaLSinghJAgarwalVet al. The implications of recent advances in carboxymethyl chitosan based targeted drug delivery and tissue engineering applications. J Control Release2014; 186: 54–87.
JeonYJKamilJYShahidiF.Chitosan as an edible invisible film for quality preservation of herring and Atlantic cod. J Agric Food Chem2002; 50: 5167–5177.
38.
AnithaADivya RaniVVKrishnaRet al. Synthesis, characterization, cytotoxicity and antibacterial studies of chitosan, O-carboxymethyl and N,O-carboxymethyl chitosan nanoparticles. Carbohyd Polym2009; 78: 672–677.
39.
LiGYZhongMZhouZDet al. Formulation optimization of chelerythrine loaded O-carboxymethylchitosan microspheres using response surface methodology. Int J Biol Macromol2011; 49: 970–978.
40.
SáezMIBarrosAMVizcaínoAJet al. Effect of alginate and chitosan encapsulation on the fate of BSA protein delivered orally to gilthead sea bream (Sparus aurata). Anim Feed Sci Tech2015; 210: 114–124.
41.
WangHBoermanOCSariibrahimogluKet al. Comparison of micro- vs. nanostructured colloidal gelatin gels for sustained delivery of osteogenic proteins: bone morphogenetic protein-2 and alkaline phosphatase. Biomaterials2012; 33: 8695–8703.
42.
KempenDHLuLHeijinkAet al. Effect of local sequential VEGF and BMP-2 delivery on ectopic and orthotopic bone regeneration. Biomaterials2009; 30: 2816–2825.
43.
TerukinaTNaitoYTagamiTet al. The effect of the release behavior of simvastatin from different PLGA particles on bone regeneration in vitro and in vivo: comparison of simvastatin-loaded PLGA microspheres and nanospheres. J Drug Deliv Sci Tec2016; 33: 136–142.
44.
LeeEJShinDSKimHEet al. Membrane of hybrid chitosan-silica xerogel for guided bone regeneration. Biomaterials2009; 30: 743–750.
45.
JiangZHanBLiHet al. Carboxymethyl chitosan represses tumor angiogenesis in vitro and in vivo. Carbohyd Polym2015; 129: 1–8.
46.
EmbuscadoMEHuberKC (eds). Edible films and coatings for food applications. Berlin: Springer Science+Business Media, 2009, pp. 367–390.
47.
LinS-JPascallMA.Incorporation of vitamin E into chitosan and its effect on the film forming solution (viscosity and drying rate) and the solubility and thermal properties of the dried film. Food Hydrocoll2014; 35: 78–84.
48.
NiuXFengQWangMet al. In vitro degradation and release behavior of porous poly(lactic acid) scaffolds containing chitosan microspheres as a carrier for BMP-2-derived synthetic peptide. Polym Degrad Stabil2009; 94: 176–182.
49.
BerklandCKipperMJNarasimhanBet al. Microsphere size, precipitation kinetics and drug distribution control drug release from biodegradable polyanhydride microspheres. J Control Release2004; 94: 129–141.
50.
RejmanJOberleVZuhornISet al. Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis. Biochem J2004; 377: 159–169.
