MG63 Osteoblast-like Cells Research Articles

One-Pot Hydrothermal Preparation of Hydroxyapatite/Zinc Oxide Nanorod Nanocomposites and Their Cytotoxicity Evaluation against MG-63 Osteoblast-like Cells.

In the present study, HAp-ZnO nanorod nanocomposites were successfully prepared using a customized hydrothermal reactor and studied for their compatibility against MG-63 osteoblast-like cells. The crystallinity, morphology, presence of chemical elements, and surface area properties were studied by XRD (X-ray diffraction), FE-SEM (field emission scanning electron microscopy), TEM (transmission electron microscopy), EDS (energy dispersive spectrum) and N2 adsorption/desorption isotherm techniques, respectively. Further, the mechanical strength and thermal analysis were carried out using the nanoindentation method and thermogravimetric/differential scanning calorimeter (TG/DSC) methods, respectively. Moreover, in vitro biocompatibility studies for the prepared samples were carried out against human osteosarcoma cell lines (MG-63). The crystalline nature of the samples without any impurity phases was notified from XRD results. The formation of composites with the morphology of nanorods and the presence of desired elements in the intended ratio were verified using FE-SEM and EDS spectra, respectively. The TG/DSC results revealed the improved thermal stability of the HAp matrix, promoted by the reinforcement of the ZnO nanorods. The nanoindentation study ensured a significant enhancement in the mechanical stability of the prepared composite material. Finally, it demonstrated that the HAp matrix's mechanical strength and thermal stability were improved by the reinforcement of ZnO, and the cytotoxicity evaluation affirmed the biocompatible nature of the biomimetic hydroxyapatite in the composite.

Biphasic monolithic osteochondral scaffolds obtained by diffusion-limited enzymatic mineralization of gellan gum hydrogel

Biphasic monolithic materials for the treatment of osteochondral defects were produced from polysaccharide hydrogel, gellan gum (GG). GG was enzymatically mineralized by alkaline phosphatase (ALP) in the presence of calcium glycerophosphate (CaGP). The desired distribution of the calcium phosphate (CaP) mineral phase was achieved by limiting the availability of CaGP to specific parts of the GG sample. Therefore, mineralization of GG was facilitated by the diffusion of CaGP, causing the formation of the CaP gradient. The distribution of CaP was analyzed along the cross section of the GG. The formation of a CaP gradient was mainly affected by the mineralization time and the ALP concentration. The formation of CaP was confirmed by Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy and mapping, as well as energy-dispersive X-ray spectroscopy (EDX) mapping of the interphase. The microstructure of mineralized and non-mineralized parts of the material was characterized by scanning electron microscopy (SEM) and atomic force microscopy (AFM) showing sub-micrometer CaP crystal formation, resulting in increased surface roughness. Compression tests and rheometric analyzes showed a 10-fold increase in stiffness of the GG mineralized part. Concomitantly, micromechanical tests performed by AFM showed an increase of Young's modulus from 17.8 to more than 200 kPa. In vitro evaluation of biphasic scaffolds was performed in contact with osteoblast-like MG-63 cells. The mineralized parts of GG were preferentially colonized by the cells over the non-mineralized parts. The results showed that osteochondral scaffolds of the desired structure and properties can be made from GG using a diffusion-limited enzymatic mineralization method.

3D printing of PLA:CaP:GO scaffolds for bone tissue applications.

Graphene oxide (GO) has attracted increasing interest for biomedical applications owing to its outstanding properties such as high specific surface area, ability to bind functional molecules for therapeutic purposes and solubility, together with mechanical resistance and good thermal conductivity. The combination of GO with other biomaterials, such as calcium phosphate (CaP) and biodegradable polymers, presents a promising strategy for bone tissue engineering. Presently, the development of these advanced biomaterials benefits from the use of additive manufacturing techniques, such as 3D printing. In this study, we develop a 3D printed PLA:CaP:GO scaffold for bone tissue engineering. First, GO was characterised alone by XPS to determine its main bond contributions and C : O ratio. Secondly, we determined the GO dose which ensures the absence of toxicity, directly exposed in vitro (human osteoblast-like cells MG-63) and in vivo (zebrafish model). In addition, GO was microinjected in the zebrafish to evaluate its effect on immune cells, quantifying the genetic expression of the main markers. Results indicated that the GO tested (C : O of 2.14, 49.50% oxidised, main bonds: C-OH, C-O-C) in a dose ≤0.25 mg mL-1 promoted MG63 cells viability percentages above 70%, and in a dose ≤0.10 mg mL-1 resulted in the absence of toxicity in zebrafish embryos. The immune response evaluation reinforced this result. Finally, the optimised GO dose (0.10 mg mL-1) was combined with polylactic acid (PLA) and CaP to obtain a 3D printed PLA:CaP:GO scaffold. Physicochemical characterisation (SEM/EDS, XRD, FT-Raman, nano-indentation) was performed and in vivo tests confirmed its biocompatibility, enabling a novel approach for bone tissue-related applications.

25-Hydroxycholesterol-induced Osteoblast Oxiapoptophagy Is Involved in the Pathophysiological Process of Osteoporosis.

25-hydroxycholesterol (25-HC) plays important roles in lipid metabolism, inflammatory responses, and apoptosis, but its pathophysiological association with osteoporosis (OP) has not been verified in osteoblasts. Hence, we studied the pathophysiological linkage and underlying cellular mechanisms of 25-HC in human osteoblast-like MG-63 cells and an ovariectomy-induced osteoporotic mouse model. To investigate the pathophysiological linkage between 25-HC-induced osteoblast oxiapoptophagy and OP, 25-HC ELISA assay, MTT assay, cell live/dead staining, hematoxylin and eosin staining, DAPI staining, flow cytometry analysis, western blot, caspase-3 staining, reactive oxygen species (ROS) assay, autophagy staining, immunocytochemistry, Micro-CT image analysis and immunocytochemistry were performed in MG-63 cells and ovariectomy-induced OP animals. The expression of cholesterol-25-hydroxylase (CH25H), an enzyme catalyzing the conversion of cholesterol to 25-HC, and the production of 25-HC were increased by lipopolysaccharide in MG-63 cells. Cytotoxicity was increased by 25-HC in MG-63 cells. Apoptosis with condensed chromatin and altered morphology was induced by 25-HC through cleavage of caspases-8, -9, and -3 in MG-63 cells. 25-HC induced oxidative stress in MG-63 cells via elevation of ROS production, cyclooxygenase-2, and inducible nitric oxide synthase. Furthermore, the expression of autophagy biomarkers, including beclin-1 and microtubule-associated protein 1A/1B-light chain 3, was elevated by 25-HC in MG-63 cells. In addition, p53 expression was increased, whereas Akt phosphorylation was suppressed in 25-HC-incubated MG-63 cells. The expression of CH25H, cleaved caspase-3, and beclin-1 were up-regulated in the femoral bone of ovariectomy-induced mouse osteoporotic animals. 25-HC plays a role in OP via the induction of oxiapoptophagic osteoblast death.

Evaluation of indocyanine-mediated photodynamic therapy cytotoxicity in human osteoblast-like cells: an in vitro study.

Antimicrobial photodynamic therapy (aPDT) is an adjunctive non-invasive procedure for the management of periodontal tissue infection and deep periodontal pocket decontamination. However, the effects of this procedure on periodontal cells like osteoblasts that play a role in periodontal tissue repair and regeneration is not yet clear.

Doxycycline-doped collagen membranes accelerate in vitro osteoblast proliferation and differentiation.

The aim of the study was to evaluate the effect of doxycycline- and dexamethasone-doped collagen membranes on the proliferation and differentiation of osteoblasts. Collagen barrier membranes are frequently used to promote bone regeneration and to boost this biological activity their functionalization with antibacterial and immunomodulatory substances has been suggested. The design included commercially available collagen membranes doped with doxycycline (Dox-Col-M) or dexamethasone (Dex-Col-M), as well as undoped membranes (Col-M) as controls, which were placed in contact with cultured MG63 osteoblast-like cells (ATCC). Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay and differentiation by measuring the alkaline phosphatase (ALP) activity using spectrophotometry. Real-time quantitative polymerase chain reaction was used to study the expression of the genes: Runx-2, OSX, ALP, OSC, OPG, RANKL, Col-I, BMP-2, BMP-7, TGF-β1, VEGF, TGF-βR1, TGF-βR2, and TGF-βR3. Scanning electron microscopy was used to study osteoblast morphology. Data were assessed using one-way analysis of variance or Kruskal-Wallis tests, once their distribution normality was assessed by Kolmogorov-Smirnov tests (p > .05). Bonferroni for multiple comparisons were carried out (p < .05). Osteoblast proliferation was significantly enhanced in the functionalized membranes as follows: (Col-M < Dex-Col-M < Dox-Col-M). ALP activity was significantly higher on cultured osteoblasts on Dox-Col-M. Runx-2, OSX, ALP, OSC, BMP-2, BMP-7, TGF-β1, VEGF, TGF-βR1, TGF-βR2, and TGF-βR3 were overexpressed, and RANKL was down-regulated in osteoblasts cultured on Dox-Col-M. The osteoblasts cultured in contact with the functionalized membranes demonstrated an elongated spindle-shaped morphology. The functionalization of collagen membranes with Dox promoted an increase in the proliferation and differentiation of osteoblasts.

Xianling Gubao attenuates high glucose-induced bone metabolism disorder in MG63 osteoblast-like cells.

Diabetes mellitus (DM) patients are prone to osteoporosis, and high glucose (HG) can affect bone metabolism. In the present study, we investigated the protective effects of traditional Chinese herbal formulation Xianling Gubao (XLGB) on HG-treated MG63 osteoblast-like cells. MG63 cells were incubated with control (mannitol), HG (20 mM glucose) or HG + XLGB (20 mM glucose+200 mg/L XLGB) mediums. Cell proliferation, apoptosis, migration and invasion were examined using CCK8, colony-formation, flow cytometry, Hoechst/PI staining, wound-healing and transwell assays, respectively. ELISA, RT-PCR and western blot analysis were used to detect the levels of osteogenesis differentiation-associated markers such as ALP, OCN, OPN, RUNX2, OPG, and OPGL in MG63 cells. The levels of the PI3K/Akt signaling pathway related proteins, cell cycle-related proteins, and mitochondrial apoptosis-related proteins were detected using western blot analysis. In HG-treated MG63 cells, XLGB significantly attenuated the suppression on the proliferation, migration and invasion of MG63 cells caused by HG. HG downregulated the activation of the PI3K/Akt signaling pathway and the expressions of cell cycle-related proteins, while XLGB reversed the inhibition of HG on MG63 cells. Moreover, XLGB significantly reduced the promotion on the apoptosis of MG63 cells induced by HG, the expressions of mitochondrial apoptosis-related proteins were suppressed by XLGB treatment. In addition, the expressions of osteogenesis differentiation-associated proteins were also rescued by XLGB in HG-treated MG63 cells. Our data suggest that XLGB rescues the MG63 osteoblasts against the effect of HG. The potential therapeutic mechanism of XLGB partially attributes to inhibiting the osteoblast apoptosis and promoting the bone formation of osteoblasts.

Surface modification of polyether ether ketone implant with a novel nanocomposite coating containing poly (vinylidene fluoride) toward improving piezoelectric and bioactivity performance

Polyether ether ketone (PEEK) is an appropriate biomaterial for orthopedic implant applications due to its superior mechanical properties, chemical resistance, nontoxicity, and Magnetic resonance imaging (MRI) compatibility. Unfortunately, the inherent bio-inertness of PEEK restricted its application and required some modification to provide better bioactivity. Besides it, the generated electrical signals in the bone due to its piezoelectricity features have a vital role in regulating bone repair and regeneration. We aimed to modify the surface of PEEK with a dual-functionality nanocomposite that provides surface bioactivity and simulates the piezoelectricity of bone. So, we introduced a novel piezoelectric-bioactive nanocomposite of dispersed poly (vinylidene fluoride) (PVDF) in a sulfonated PEEK (SPEEK) matrix containing Nanohydroxyapatite (nHA) and Carbon nanofiber (CNF) fillers for coating on PEEK substrate to improve its biological activity and simulate the electrical microenvironment for bone tissue. Furthermore, sulfonation of the PEEK surface was conducted as an intermediate layer to prepare better adhesion between the coating nanocomposite and the PEEK sublayer. Surface and cross-section morphology, apatite formation, and cell attachment were investigated on the different treated PEEK surfaces using field-emission scanning electron microscopy (FESEM) and energy dispersive X-ray analysis (EDX). Also, piezoelectric performance, electrical conductivity, contact angle, and mechanical properties were examined on the prepared samples. Moreover, cell viability and cell morphology were investigated for biological evaluation with human osteoblast-like MG-63 cells. Collectively, the hydrophilicity of modified PEEK (mPEEK) coated with nanocomposite was improved due to the synergistic effects of SPEEK functional groups and nHA. Also, comprehensive investigation on the mPEEK treated with nanocomposite indicated a noticeably better bone-like apatite formation, cell proliferation, and cell attachments in the presence of nHA. The transfer of induced piezoelectric charges from dispersed PVDF in the matrix to the surface of nanocomposite containing 2 wt% of CNF increased output voltage to 1893 mV. On the other hand, the presence of CNF in nanocomposites enhanced tensile strength and Young's modulus by 92% and 117%, respectively.

TiO2/HA and Titanate/HA Double-Layer Coatings on Ti6Al4V Surface and Their Influence on In Vitro Cell Growth and Osteogenic Potential.

Hydroxyapatite (HA) layers are appropriate biomaterials for use in the modification of the surface of implants produced inter alia from a Ti6Al4V alloy. The issue that must be solved is to provide implants with appropriate biointegration properties, enabling the permanent link between them and bone tissues, which is not so easy with the HA layer. Our proposition is the use of the intermediate layer ((IL) = TiO2, and titanate layers) to successfully link the HA coating to a metal substrate (Ti6Al4V). The morphology, structure, and chemical composition of Ti6Al4V/IL/HA systems were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectrometry (EDS). We evaluated the apatite-forming ability on the surface of the layer in simulated body fluid. We investigated the effects of the obtained systems on the viability and growth of human MG-63 osteoblast-like cells, mouse L929 fibroblasts, and adipose-derived human mesenchymal stem cells (ADSCs) in vitro, as well as on their osteogenic properties. Based on the obtained results, we can conclude that both investigated systems reflect the physiological environment of bone tissue and create a biocompatible surface supporting cell growth. However, the nanoporous TiO2 intermediate layer with osteogenesis-supportive activity seems most promising for the practical application of Ti6Al4V/TiO2/HA as a system of bone tissue regeneration.

Nanocomposites containing ZnO-TiO2-Chitosan and berbamine promote osteoblast differentiation, proliferation, and calcium mineralization in MG63 osteoblasts

Recently, biomedical research has focused on bio-nanocomposites that are capable of regeneration of tissues, as well as treating osteomyelitis, osteosarcoma, and other diseases with therapeutic medications. A novel osteoconductive nanocomposite will be synthesized and used to enhance the mineralization of injured bone tissues, as well as their antimicrobial properties. The unique structural characteristics of ZnO-TiO2-Chitosan-berbamine nanocomposites have been described by UV-Vis, PL, FTIR, XRD, FESEM, and HR-TEM experiments, while antimicrobial activity against pathogenic microorganisms has been evaluated by the agar well diffusion method. MTT assays and levels of ALP enzymes were used to test nanocomposites on osteosarcoma cell lines (MG-63) and to assess the osteosarcoma cell line's physical characteristics, bioactivity, and calcium deposition by staining with Alizarin Red S. In addition to showing significant Zones of Inhibition (ZOI) against bacterial and fungal strains, structural characterization revealed the antimicrobial properties of ZnO-TiO2-Chitosan-berbamine composites. According to research conducted on osteoblast-like MG-63 cells grown on ZnO-TiO2-Chitosan-berbamine composites, the composites do not cause toxicity and significantly increase osteoblast proliferation, ALP activity, calcium deposition, and bone growth, making ZnO-TiO2-Chitosan-berbamine a potential therapy for bone-related diseases. Understanding the druggable mechanism of ZnO-TiO2-Chitosan-berbamine will require further studies in the future, including in vivo testing and clinical trials.

Novel Application of 3D Scaffolds of Poly(E-Caprolactone)/Graphene as Osteoinductive Properties in Bone Defect.

Scaffolds provided a surface on which cells could attach, proliferate, and differentiate. Nowadays, bone tissue engineering offers hope for treating bone cancer. Poly(e-caprolactone) (PCL)/graphene have capability as an osteogenic and regenerative therapy. It could be used to produce bone tissue engineering scaffolds. The purpose of this study was to investigate the ability of PCL/graphene to enhance the osteoinductive mechanism. The PCL/graphene scaffold was developed utilizing a particulate-leaching process and cultured with osteoblast-like cells MG63 at 0.5, 1.5, and 2.5 wt% of graphene. We evaluated the porosity, pore size, migratory cells, and cell attachment of the scaffold. Data was expressed as the mean ± standard error of the mean and statistical analyses were performed using one-way analysis of variance and Tukey's post hoc at a level of p-value < 0.05. Porosity of scaffold with various percentage of graphene was nonsignificant (p > 0.05). There were differences in the acceleration of cell migration following wound closure between groups at 24 hours (p < 0.01) and 48 hours (p < 0.00). Adding the graphene on the scaffolds enhanced migration of osteoblast cells culture and possibility to attach. Graphene on 2.5 wt% exhibited good characteristics over other concentrations. This finding suggests that PCL/graphene composites may have potential applications in bone tissue engineering.

Anabolic Peptide-Enriched Stealth Nanoliposomes for Effective Anti-Osteoporotic Therapy.

The objective of the present work was to develop PTH (1-34)-loaded stealth nanoliposomes (PTH-LPs) by employing the use of the Quality by Design (QbD) approach. Risk identification was carried out using the Ishikawa fishbone diagram. PTH-LPs were optimized using Box Behnken Design, a type of response surface methodology to examine the effect of independent variables on dependent variables such as particle size and percentage entrapment efficiency (%EE). Design space was generated for PTH-LPs to reduce interbatch variability during the formulation development process. Furthermore, a cytotoxicity assay, cell proliferation assay, calcium calorimetric assay, mineralized nodule formation, and cellular uptake assay were carried out on MG-63 osteoblast-like cells. The results obtained from these procedures demonstrated that lipid concentration had a significant positive impact on particle size and %EE, whereas cholesterol concentration showed a reduction in %EE. The particle size and %EE of optimized formulation were found to be 147.76 ± 2.14 nm and 69.18 ± 3.62%, respectively. Optimized PTH-LPs showed the sustained release profile of the drug. In vitro cell evaluation studies showed PTH-LPs have good biocompatibility with MG-63 cells. The cell proliferation study revealed that PTH-LPs induced osteoblast differentiation which improved the formation of mineralized nodules in MG-63 cells. The outcome of the present study conclusively demonstrated the potential of the QbD concept to build quality in PTH-LPs with improved osteoanabolic therapy in osteoporosis.

Dual pulsed laser deposition of Ag nanoparticles on calcium phosphate coatings for biomedical applications

Pulsed laser deposition (PLD) represents a promising bottom-up methodology for the synthesis and transference of nanoparticles to the surface of a biomedical device. Silver (Ag) nanoparticles directly incorporated on the metallic implant emerge as an alternative strategy for local action against prosthetic joint-associated infections. In the present research, a dual sequential PLD process is proposed to obtain a bilayer coating with (1) a bio-derived calcium phosphate (CaP) layer, to provide osteointegrative properties and (2) the controlled growth of the Ag nanoparticles over it, ranging the number of laser pulses from 100 to 500. The characterization by SEM, EDS, TEM, XPS and AFM revealed the uniform deposit of Ag rounded nanoparticles, with a narrow mean size distribution, in the original non-oxidized metallic state. Moreover, given the evidences from XPS and AFM techniques, the occurrence of a coalescence phenomenon from 400 pulses onwards was proposed together with the expected positive linear relation between the number of pulses and Ag contribution with a deposition rate of 0.05 at. % of Ag per pulse. Conversely, the decrease in roughness as the Ag content increased was also verified. Finally, the expected bacteriostatic activity for these PLD deposited metallic state Ag nanoparticles against the bacterial strain Staphylococcus aureus was confirmed. Moreover, the evaluation of the osteoblast-like MG-63 cells viability on the Ag(100–500)-CaP coatings revealed a significant increased proliferation (p > 0.05) on the Ag100-CaP coating compared to the control (Ag0-CaP). When same coating was evaluated against S. aureus the effect was not significant. The possibility of modulating the amount of nanoparticles in the bilayer coating to obtain a greater or lesser effect in combination with CaP was revealed.

Mechanical and morphological responses of osteoblast-like cells to two-photon polymerized microgrooved surfaces.

Microgrooved surfaces are recognized as an important strategy of tissue engineering to promote the alignment of bone cells. In this work, we have investigated the mechanical and morphological aspects of osteoblasts cells after interaction with different micro-structured polymeric surfaces. Femtosecond laser writing technique was used for the construction of circular and parallel microgrooved patterns in biocompatible polymeric surfaces based on pentaerythritol triacrylate. Additionally, we have studied the influence of the biocompatible TiO2 nanocrystals (NCs) related to the cell behavior, when incorporated to the photoresin. The atomic force microscopy technique was used to investigate the biomechanical reaction of the human osteoblast-like MG-63 cells for the different microgroove. It was demonstrated that osteoblasts grown on circular microgrooved surfaces exhibited significantly larger Young's modulus compared to cells sown on flat films. Furthermore, we could observe that TiO2 NCs improved the circular microgrooves effects, resulting in more populated sites, 34% more elongated cells, and increasing the cell stiffness by almost 160%. These results can guide the design and construction of effective scaffold surfaces with circular microgrooves for tissue engineering and bone regeneration.

Comparison of Different Chemical and Mechanical Modalities for Implant Surface Decontamination: Activity against Biofilm and Influence on Cellular Regrowth-An In Vitro Study.

ObjectivesThe aim of this in vitro study was to compare the efficacy of chemical and mechanical methods for decontamination of titanium dental implant surfaces previously infected with polymicrobial biofilms in a model simulating a peri-implant defect. Furthermore, the effect of each decontamination protocol on MG-63 osteoblast-like cells morphology and adhesion to the treated implants was assessed.BackgroundPeri-implantitis is a growing issue in dentistry, and evidence about implant surface decontamination procedures is lacking and inconclusive.MethodsA total of 40 previously biofilm-contaminated implants were placed into a custom-made model simulating a peri-implant defect and randomly assigned to five treatment groups: (C) control (no treatment); (AW) air abrasion without any powder; (ESC) air abrasion with powder of erythritol, amorphous silica, and 0.3% chlorhexidine; (HBX) decontamination with a sulfonic/sulfuric acid solution in gel; and (HBX + ESC) a combination of HBX and ESC. Microbiological analysis was performed on five implants per treatment group, and the residual viable bacterial load measured in log 10 CFU/mL was counted for each bacterial strain and for the total number of colonies. The remaining three implants per group and three noncontaminated (NC) implants were used to assess surface biocompatibility using a scanning electron microscope and a backscattered electron microscope after seeding with MG-63 cells.ResultsA significant decontaminant effect was achieved using HBX or HBX + ESC, while no differences were observed among other groups. The percentage of implant surface covered by adherent MG-63 cells was influenced by the treatment method. Progressive increases in covered surfaces were observed in groups C, AW, ESC, HBX, HBX + ESC, and NC.ConclusionsA combination of mechanical and chemical decontamination may provide more predictable results than mechanical cleaning alone.

Influence of the Components and Orientation of Hydroxyapatite Fibrous Substrates on Osteoblast Behavior.

Synthetic hydroxyapatite has good biocompatibility, bioactivity and osteoconductive ability because its chemical properties and biological properties are similar to those of bioapatite in bone tissue. Strontium-substituted hydroxyapatite has better degradability than hydroxyapatite and can both promote osteogenesis and inhibit adipogenesis in mesenchymal stem cells. Hence, hydroxyapatite and strontium-substituted hydroxyapatite are widely used as bone graft materials, cell carriers and drug/gene delivery carriers. In addition, osteoblasts cultured on aligned nanofibrous substrates had higher expression of osteogenesis-related genes than did those cultured on random nanofibrous substrates. However, to date, no study has explored the effects of the components and orientation of hydroxyapatite nanofibrous substrates on osteoblastic behavior. In this study, a random hydroxyapatite nanofibrous substrate (R-HANF), a random strontium-substituted hydroxyapatite nanofibrous substrate (R-SrHANF), an aligned hydroxyapatite nanofibrous substrate (A-HANF) and an aligned strontium-substituted hydroxyapatite nanofibrous substrate (A-SrHANF) were successfully fabricated by using the electrospinning technique. The effect of fiber composition on osteoblast-like MG63 cells was assessed by evaluating cell morphology, cell proliferation and osteogenesis-related gene expression. The results showed that MG63 cells cultured on A-SrHANF had higher osteogenesis-related gene expression than those cultured on A-HANF. Additionally, MG63 cells were cultured on R-SrHANF and A-SrHANF to evaluate the effects of fiber orientation on cell behavior. On A-SrHANF, the cells aligned along the direction of the nanofibers, with typical bipolar morphologies, and exhibited higher osteogenesis-related gene expression than cells on R-SrHANF. Hence, the components and orientation of hydroxyapatite nanofibrous substrates are critical parameters affecting the osteogenesis process.

Highly effective electrospun polycaprolactone/ layered double hydroxide nanofibrous scaffold for bone tissue engineering

Bone tissue engineering is considered a promising approach for repairing, regenerating and orthopedic implants. In this regard, polymeric supportive scaffolds reinforced with bioactive nanomaterials provide unique functions help in substitution of impaired bone tissues. In this constant, various layered double hydroxide (LDH) concentrations (ranging from 0.1 wt% to 10 wt%.) within the poly (ε-caprolactone) (PCL) scaffolds were fabricated successfully through electrospinning. The scaffolds were investigated by XRD, SEM, FTIR, and EDX analyses to study the phase identification, morphology, chemical properties, and elemental composition, respectively. The findings demonstrated that the inclusion of LDH to PCL scaffold increased mechanical tensile strength from 1.63 ± 0.4 to 1.920 ± 0.3 Mpa for the pure PCL and PCL + LDH (1 wt%) scaffolds, respectively. A decrease in the average diameter of PCL nanofibers was observed with the addition of LDH. The enrichment of PCL scaffold with LDH promotes in vitro biomineralization when subjected to simulated body fluid (SBF), indicating the bioactive features of the nanocomposite scaffolds. In addition, increased of LDH leads to improvement in protein adsorption capacity compared to the pure one. Furthermore, the cytocompatibility and cytotoxicity of nanocomposite fibers was assessed using human MG-63 osteoblast-like cells. The LDH/PCL scaffold induced mineral deposition and osteogenic differentiation of MG-63 as evident by an increase in ALP activity in comparison with the pure PCL. Overall, our results revealed that electrospun PCL-LDH nanofiber might be an ideal choice for bone tissue engineering.

Biological and Physico-Chemical Properties of Composite Layers Based on Magnesium-Doped Hydroxyapatite in Chitosan Matrix.

In the present study, we report the development and characterization of composite layers (by spin coating) based on magnesium-doped hydroxyapatite in a chitosan matrix, (Ca10−xMgx(PO4)6(OH)2; xMg = 0, 0.08 and 0.3; HApCh, 8MgHApCh and 30MgHApCh). The MgHApCh composite layers were investigated using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray photoelectron spectroscopy (XPS) techniques. The in vitro biological evaluation included the assessment of their cytotoxicity on MG63 osteoblast-like cells and antifungal activity against Candida albicans ATCC 10231 fungal cell lines. The results of the physico-chemical characterization highlighted the obtaining of uniform and homogeneous composite layers. In addition, the biological assays demonstrated that the increase in the magnesium concentration in the samples enhanced the antifungal effect but also decreased their cytocompatibility. However, for certain optimal magnesium ion concentrations, the composite layers presented both excellent biocompatibility and antifungal properties, suggesting their promising potential for biomedical applications in both implantology and dentistry.

Graphene oxide reinforced SrHAP composite as a drug carrier in bone regeneration

Strontium substituted HAP (SrHAP), with a 10 mol% substitution, was mineralized on increasing weight percentages of graphene oxide (2, 4 and 6). The GS composites were comprehensively characterized for drug delivery in bone reconstruction. The formation of SrHAP was verified by XRD and FT-IR results. The apatite crystallization was influenced by graphene oxide content and strontium. The EDS results confirmed the presence of strontium and HR-SEM depicted rod shape apatite, of length between 58 and 135 nm, uniformly embedded on graphene oxide. The reinforcement of graphene oxide increased the surface area, porosity, microhardness (upto 0.59 GPa), protein adsorption (upto 18.16 μg/mg), water uptake and degradation properties. Also, the increase in graphene oxide fraction significantly enhanced the curcumin encapsulation efficiency (upto 80.16%) and the drug release was considerably retarded over SrHAP. The in vitro studies using human osteoblast-like MG-63 cells demonstrated that curcumin-loaded composite was biocompatible and promoted proliferation, differentiation and matrix mineralization. The results highlight the combinational therapy of osteogenic ion (strontium) and osteogenic drug (curcumin) as a promising platform in bone tissue engineering.

Evaluation of quaternization effect on chitosan-HAP composite for bone tissue engineering application

This study attempts to improve the aqueous solubility of chitosan and utilizes it in the fabrication of composites with hydroxyapatite (HAP). The composites were evaluated as a curcumin delivery vehicle for bone regeneration. The chitosan was modified by quaternization, with a quaternization degree of 5 % for low quaternized chitosan (LQC) and 11 % for high quaternized chitosan (HQC). The modified chitosan, at alkaline pH 11, facilitated in situ HAP growth and formed LQC-HAP and HQC-HAP composites. The quaternization weakens intermolecular hydrogen bonds, facilitates interaction with the apatite precursor ions and promotes the growth of HAP. The modification significantly improved drug encapsulation (2.6 fold) but at the cost of a slight decrease in mechanical strength and increase in drug release. The in vitro studies with human osteoblast-like MG-63 cells established that the curcumin-loaded composites, LQC-HAP-C and HQC-HAP-C are biocompatible, encourage proliferation and promote a 2-fold increase in calcium mineralization over drug-free composites. The study exemplifies the reciprocity between quaternization degree and drug load/release properties and also illustrates that the magnitude of the latter reflects bioactivity. Thus, the quaternized chitosan-based HAP composite with tailorable bio-physicochemical properties becomes an interesting drug delivery system in bone regeneration.