RESUMO
This is the first study that explored the potential use of Zizyphus mauritiana seed extract (ZSE) to synthesize nano-fluorohydroxyapatite/carboxymethyl chitosan nanocomposite scaffolds at different concentrations (CFZ1, CFZ2 and CFZ3) using co-precipitation method. The proposed scaffolds showed presence of intermolecular H bonding interactions between the constituents, according to the FTIR. The mechanical studies revealed shore hardness of 72 ± 4.6 and optimal compressive modulus in case of CFZ3 [1654.48 ± 1.6 MPa], that was comparable with that of human cortical bone. The SEM, TEM and platelet adhesion images corroborated uniformly distributed needle like particles in case of CFZ3 with an average size ranging from 22 to 26 nm, linked rough morphology, and appropriate hemocompatibility. The markedly up regulation in the ALP activity and protein adsorption upon increasing ZSE concentration demonstrated that CFZ nanocomposite scaffolds were compatible with osteoblastic cells relative to CF nanocomposite. The cytotoxicity study indicated that CFZ nanocomposite do not induce toxicity over MG-63 and did not aggravate LDH leakage in contrast to CF. The histopathological investigations on albino rats confirmed significantly improved regeneration of bone in the repair of a critical-size [8 mm] calvarium defect. Therefore, CFZ3 nanocomposite scaffold represents a simple, off-the-shelf solution to the combined challenges associated with bone defects.
Assuntos
Quitosana , Nanocompostos , Ziziphus , Ratos , Animais , Humanos , Alicerces Teciduais , Engenharia Tecidual/métodos , Regeneração Óssea , Durapatita/farmacologiaRESUMO
Nanoparticle-reinforced polymer-based scaffolding matrices as artificial bone-implant materials are potential suitors for bone regenerative medicine as they simulate the native bone. In the present work, a series of bioinspired, osteoconductive tricomposite scaffolds made up of nano-hydroxyapatite (NHA) embedded xanthan gum-chitosan (XAN-CHI) polyelectrolyte complex (PEC) are explored for their bone-regeneration potential. The Fourier transform infrared spectroscopy studies confirmed complex formation between XAN and CHI and showed strong interactions between the NHA and PEC matrix. The X-ray diffraction studies indicated regulation of the nanocomposite (NC) scaffold crystallinity by the physical cues of the PEC matrix. Further results exhibited that the XAN-CHI/NHA5 scaffold, with a 50/50 (polymer/NHA) ratio, has optimized porous structure, appropriate compressive properties, and sufficient swelling ability with slower degradation rates, which are far better than those of CHI/NHA and other XAN-CHI/NHA NC scaffolds. The simulated body fluid studies showed XAN-CHI/NHA5 generated apatite-like surface structures of a Ca/P ratio â¼1.66. Also, the in vitro cell-material interaction studies with MG-63 cells revealed that relative to the CHI/NHA NC scaffold, the cellular viability, attachment, and proliferation were better on XAN-CHI/NHA scaffold surfaces, with XAN-CHI/NHA5 specimens exhibiting an effective increment in cell spreading capacity compared to XAN-CHI/NHA4 and XAN-CHI/NHA6 specimens. The presence of an osteo-friendly environment is also indicated by enhanced alkaline phosphatase expression and protein adsorption ability. The higher expression of extracellular matrix proteins, such as osteocalcin and osteopontin, finally validated the induction of differentiation of MG-63 cells by tricomposite scaffolds. In summary, this study demonstrates that the formation of PEC between XAN and CHI and incorporation of NHA in XAN-CHI PEC developed tricomposite scaffolds with robust potential for use in bone regeneration applications.
RESUMO
The present frontiers of bone tissue engineering are being pushed by novel biomaterials that exhibit phenomenal biocompatibility and adequate mechanical strength. In this work, we fabricated a ternary system incorporating nano-hydroxyapatite (n-HA)/gum arabic (GA)/κ-carrageenan (κ-CG) with varying concentrations, i.e., 60/30/10 (CHG1), 60/20/20 (CHG2), and 60/10/30 (CHG3). A binary system with n-HA and GA was also prepared with a ratio of 60/40 (HG) and compared with the ternary system. A rapid mineralization of the apatite layer was observed for the ternary systems after incubation in simulated body fluid (SBF) for 15 days as corroborated by scanning electron microscopy (SEM). CHG2 exhibited the maximum apatite layer deposition. Further, the nanocomposites were physicochemically analyzed by Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and mechanical testing. Their results revealed a substantial interaction among the components, appropriate crystallinity, and significantly enhanced compressive strength and modulus for the ternary nanocomposites. The greatest mechanical strength was achieved by the scaffold containing equal amounts of GA and κ-CG. The cytotoxicity was evaluated by culturing osteoblast-like MG63 cells, which exhibited the highest cell viability for the CHG2 nanocomposite system. It was further supported by confocal microscopy, which revealed the maximum cell proliferation for the CHG2 scaffold. In addition, enhanced antibacterial activity, protein adsorption, biodegradability, and osteogenic differentiation were observed for the ternary nanocomposites. Osteogenic gene markers, such as osteocalcin (OCN), osteonectin (ON), and osteopontin (OPN), were present in higher quantities in the CHG2 and CHG3 nanocomposites as confirmed by western blotting. These results substantiated the pertinence of n-HA-, GA-, and κ-CG-incorporated ternary systems to bone implant materials.
RESUMO
Biomimetic nanocomposites containing hydroxyapatite and natural polymers are promising candidates for bone grafting. In this work, a tricomponent bioactive nanocomposite (nHA-CH-TFSP) assembling nano-hydroxyapatite (nHA), Chitosan (CH) and Trigonella foenum graecum seed polysaccharide (TFSP) was developed using co-precipitation method and investigated against bicomponent nHA-CH nanocomposite for bone tissue engineering. In contrast to nHA-CH nanocomposite, nHA-CH-TFSP nanocomposite exhibited rough and interconnected porous structure as revealed by SEM with the porosity (60.3⯱â¯0.17) lying in the range of cancellous bone. The incorporation of TFSP in nHA-CH substantially enhanced the in vitro water absorption capacity and protein adsorption ability along with appropriate biodegradation rate. Additionally, the nHA-CH-TFSP nanocomposite exhibited superior antibacterial activity. The nHA-CH-TFSP evinced a compressive strength of 6.7⯱â¯0.24â¯MPa and a compressive modulus of 100⯱â¯1.4â¯MPa, which fulfill the strength requisite of cancellous bone and could provide strong support for the growth of osteoblasts cells. Furthermore, the in vitro bioactivity study demonstrated its excellent biomineralization capacity in comparison to nHA-CH. The synthesized nHA-CH-TFSP nanocomposite exhibited better cytocompatibility towards the MG-63 cells along with its haemocompatible nature. Taken together the results of the present study indicate that nHA-CH-TFSP could serve as a prospective analogue for bone tissue engineering.
Assuntos
Materiais Biomiméticos/química , Regeneração Óssea/efeitos dos fármacos , Quitosana/química , Polissacarídeos/química , Materiais Biomiméticos/síntese química , Materiais Biomiméticos/farmacologia , Linhagem Celular , Quitosana/síntese química , Quitosana/farmacologia , Durapatita/síntese química , Durapatita/química , Durapatita/farmacologia , Humanos , Nanocompostos/química , Osteoblastos/efeitos dos fármacos , Polissacarídeos/síntese química , Polissacarídeos/farmacologia , Sementes/química , Engenharia Tecidual , Trigonella/químicaRESUMO
In this work, we have explored the polysaccharide nature of bael fruit gum (BFG) motivated from the current findings about the substantial role of the polysaccharides in bone tissue engineering. The nanocomposite scaffold (CSH-BFG) was prepared by blending BFG, nano-hydroxyapatite (n-HA) and chitosan (CS) by co-precipitation approach and compared with n-HA and CS binary system (CSH). The analysis of different properties was carried out by SEM, TEM, FTIR, XRD and mechanical testing. The CSH-BFG scaffolds revealed a rough morphology and uniform distribution of particles along with strong chemical interactions among different components compared to the CSH scaffold. The incorporation of BFG in the scaffold resulted in significant increase of the compressive strength, compressive modulus, protein adsorption, biodegradation and swelling behaviour. The ternary system exhibited superior antibacterial activity against different bacterial pathogens compared to the binary system. The in vitro biomineralization ability was elucidated from the formation of thick apatite layer complementing the result of ARS study in the CSH-BFG nanocomposite. Our findings also revealed that BFG reinforced CSH nanocomposite exhibited enhanced cell adhesion and proliferation, osteogenic differentiation along with phenomenal cytocompatibility. Overall, our results signified that the fabricated CSH-BFG nanocomposite carries enormous potential to be applied in the bone remodelling procedures.
Assuntos
Adesivos/química , Aegle/química , Materiais Biocompatíveis , Quitosana/química , Durapatita/química , Frutas/química , Engenharia Tecidual , Biomineralização , Sobrevivência Celular , Fenômenos Químicos , Humanos , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
It is the first report where different amounts of resol resin (RS) were incorporated with chitosan-hydroxyapatite (CHA) to develop a triconstituent nanoensemble CHA-RS(0.5,1,2), via simple co-precipitation method. The results of SEM, TEM, TGA and mechanical analysis revealed irregular interconnected rough morphology with homogenous distribution of needle shaped particles having average size ranging between 12 and 19nm, possessing higher thermal stability and mechanical strength, respectively relative to CHA (binary) nanocomposite. The CHA-1RS nanocomposite showed enhanced protein adsorption and ALP activity with excellent apatite formation ability compared to CHA-RS(0.5,2) and CHA nanocomposites. Thus, CHA-1RS nanocomposite was selectively tested as bare implant in the repair of critical-size calvarium defect (8mm) in albino rat. The histopathological and radiological investigations indicated that CHA-1RS prompted the bone regeneration ability as early as 2 weeks postimplantation demonstrating remarkably faster healing of calvarial defect relative to Cerabone. These findings have placed CHA-1RS on the pedestal to be employed as a potential alternative biomaterial for bone tissue engineering.
RESUMO
Herein, we report the synthesis of a novel tri-component nanocomposite system incorporating ß-cyclodextrin (ß-CD) with nano-hydroxyapatite (n-HA) and chitosan (CS), (n-HA/ß-CD/CS) at three different temperatures via co-precipitation method. The chemical interactions and surface morphology have been evaluated by TEM, SEM and AFM techniques revealing the agglomerated nanoparticles in CS/n-HA-HA binary system whereas the ternary systems produced needle shaped nanoparticles dispersed homogeneously at low temperature with more porous and rougher surface. The addition of ß-CD in CS/n-HA at low temperature decreased the particle size and raised the thermal stability as compared to CS/n-HA. The comparative hemolytic, protein adsorption and platelet adhesion studies confirmed the better hemocompatibility of n-HA/ß-CD/CS-(RT,HT,LT) nanocomposites relative to CS/n-HA. The cell viability has been evaluated in vitro using MG-63 cell line which revealed superior non toxicity of n-HA/ß-CD/CS-LT nanocomposite in comparison to n-HA/ß-CD/CS-(RT,HT) and CS/n-HA nanocomposites. Thus it may be concluded that the orchestrated organic/inorganic n-HA/ß-CD/CS-(RT,HT,LT) nanocomposites exhibited relatively higher cell viability of human osteoblast cells, stimulated greater osteogenesis, controlled biodegradation, enhanced antibacterial activity with excellent in-vitro biomineralization and remarkable mechanical parameters as compared to CS/n-HA nanocomposite and thus may provide opportunities for potential use as an alternative biomaterial for Bone tissue engineering applications.
Assuntos
Osso e Ossos/citologia , Quitosana/química , Durapatita/química , Nanocompostos/química , Engenharia Tecidual , beta-Ciclodextrinas/química , Fosfatase Alcalina/metabolismo , Antibacterianos/química , Antibacterianos/metabolismo , Antibacterianos/farmacologia , Antibacterianos/toxicidade , Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Materiais Biocompatíveis/farmacologia , Materiais Biocompatíveis/toxicidade , Linhagem Celular , Força Compressiva , Dureza , Hemólise/efeitos dos fármacos , Humanos , Nanocompostos/toxicidade , Alicerces Teciduais/químicaRESUMO
A novel ternary nanocomposite system incorporating hydroxyapatite, chitosan and starch (n-HA/CS-ST) has been synthesized by co-precipitation method at room temperature, addressing the issues of biocompatibility, mechanical strength and cytotoxicity required for bone tissue engineering. The interactions, crystallite size, surface morphology and thermal stability against n-HA/CS nanocomposite have been obtained by comparing the results of FTIR, SEM, TEM, DLS, XRD and TGA/DTA. A comparative study of bioactivity and thermal stability of n-HA/CS and n-HA/CS-ST nanocomposites revealed that the incorporation of starch as templating agent enhanced these properties in n-HA/CS-ST nanocomposite. A lower swelling rate of n-HA/CS-ST relative to n-HA/CS indicates a higher mechanical strength supportive of bone tissue ingrowths. The MTT assay on murine fibroblast L929 and human osteoblasts-like MG-63 cells and in vitro bioactivity of n-HA/CS-ST matrix referred superior non-toxic nature of n-HA/CS-ST nanocomposite and greater possibility of osteointegration in vivo respectively. Furthermore n-HA/CS-ST exhibited improved antibacterial property against both Gram-positive and Gram-negative bacteria relative to n-HA/CS.
Assuntos
Materiais Biocompatíveis/síntese química , Materiais Biocompatíveis/farmacologia , Osso e Ossos/efeitos dos fármacos , Quitosana/química , Durapatita/química , Nanocompostos/química , Amido/química , Animais , Antibacterianos/síntese química , Antibacterianos/química , Antibacterianos/farmacologia , Antibacterianos/toxicidade , Materiais Biocompatíveis/química , Materiais Biocompatíveis/toxicidade , Biomimética , Osso e Ossos/citologia , Linhagem Celular , Técnicas de Química Sintética , Escherichia coli/efeitos dos fármacos , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Humanos , Fenômenos Mecânicos , Camundongos , Testes de Sensibilidade Microbiana , Staphylococcus aureus/efeitos dos fármacos , Alicerces Teciduais/químicaRESUMO
A polycarbazole-titanium dioxide (PCz/TiO2-8) nanocomposite was synthesized for the first time by in-situ oxidative polymerization with APS as an oxidizing agent in the presence of TiO2 nanoparticles which was characterized by SEM, TEM, FT-IR, DTA and TGA techniques. The SEM and TEM results showed that PCz has polymerized on the surface of the TiO2. The results confirmed the incorporation of TiO2 in PCz indicating the formation of nanocomposite due to strong interaction between TiO2 and PCz matrix affecting the properties as compared to pristine PCz. The PCz/TiO2-8 nanocomposite was tested for antimicrobial activity and found to exhibit activity against gram negative and gram positive strains at micromolar concentrations. The four types of 3D molecular field descriptors or field points as extrema of electrostatic, steric, and hydrophobic fields were explained. These field points were used to define the properties necessary for a molecule to bind in a characteristic way into a specified active site. A molecular docking simulation was used to predict the modes of interactions of the drugs (PCz and PCz/TiO2-8) with DNA. The molecular docking conclusion indicated that the modes of interactions between two (PCz and PCz/TiO2-8) and DNA helix can be regarded as minor groove binder. The K value (3.55 × 10(4) M(-1) and 2.13 × 10(5) M(-1)) obtained from fluorescence data are indicative of binding of PCz and PCz/TiO2-8 with DNA helix.