Adhesion Of MG-63 Cells Research Articles

3D printed polylactic acid/polyethylene glycol/bredigite nanocomposite scaffold enhances bone tissue regeneration via promoting osteogenesis and angiogenesis

Recently, the fabrication of personalized scaffolds with high accuracy has been developed through 3D printing technology. In the current study, polylactic acid/polyethylene glycol (PLA/PEG) composite scaffolds with varied weight percentages (0, 5, 10, 20 and 30 %) of bredigite nanoparticles (B) were fabricated using the 3D printing and then characterized through scanning electron microscopy and Fourier transform infra-red spectroscopy. The addition of B nanoparticles up to 20 wt% to PLA/PEG scaffold increased the compressive strength (from 7.59 to 13.84 MPa) and elastic modulus (from 142.42 to 268.33 MPa). The apatite formation ability as well as inorganic ion release in simulated body fluid were investigated for 28 days. The MG-63 cells viability and adhesion were enhanced by increasing the amount of B in the PLA/PEG scaffold and the osteogenic differentiation of the rat bone marrow mesenchymal stem cells was confirmed by alkaline phosphatase activity test and alizarin red staining. According to chorioallantoic membrane assay, the highest angiogenesis occurred around the PLA/PEG/B30 scaffold. In vivo experiments on a rat calvarial defect model demonstrated an almost complete recovery in the PLA/PEG/B30 group within 8 weeks. Based on the results, the PLA/PEG/B30 composite scaffold is proposed as an optimal scaffold to repair bone defects.

Multilayer Gelatin-Supported BMP-9 Coating Promotes Osteointegration and Neo-Bone Formation at the n-CDHA/PAA Composite Biomaterial-Bone Interface.

The development of biomaterials capable of accelerating bone wound repair is a critical focus in bone tissue engineering. This study aims to evaluate the osteointegration and bone regeneration potential of a novel multilayer gelatin-supported Bone Morphogenetic Protein 9 (BMP-9) coated nano-calcium-deficient hydroxyapatite/poly-amino acid (n-CDHA/PAA) composite biomaterials, focusing on the material-bone interface, and putting forward a new direction for the research on the interface between the coating material and bone. The BMP-9 recombinant adenovirus (Adenovirus (Ad)-BMP-9/Bone Marrow Mesenchymal Stem Cells (BMSc)) was produced by transfecting BMSc and supported using gelatin (Ad-BMP-9/BMSc/Gelatin (GT). Multilayer Ad-BMP-9/BMSc/GT coated nano-calcium deficient hydroxyapatite/polyamino acid (n-CDHA/PAA) composite biomaterials were then prepared and co-cultured with MG63 cells for 10 days, with biocompatibility assessed through microscopy, Cell Counting Kit-8 (CCK-8), and alkaline phosphatase (ALP) assays. Subsequently, multilayer Ad-BMP-9/BMSc/GT coated n-CDHA/PAA composite biomaterial screws were fabricated, and the adhesion of the coating to the substrate was observed using scanning electron microscopy (SEM). In vivo studies were conducted using a New Zealand White rabbit intercondylar femoral fracture model. The experimental group was fixed with screws featuring multilayer Ad-BMP-9/BMSc/GT coatings, while the control groups used medical metal screws and n-CDHA/PAA composite biomaterial screws. Fracture healing was monitored at 1, 4, 12, and 24 weeks, respectively, using X-ray observation, Micro-CT imaging, and SEM. Integration at the material-bone interface and the condition of neo-tissue were assessed through these imaging techniques. The Ad-BMP-9/GT coating significantly enhanced MG63 cell adhesion, proliferation, and differentiation, while increasing BMP-9 expression in vitro. In vivo studies using a rabbit femoral fracture model confirmed the biocompatibility and osteointegration potential of the multilayer Ad-BMP-9/BMSc/GT coated n-CDHA/PAA composite biomaterial screws. Compared to control groups (medical metal screws and n-CDHA/PAA composite biomaterial screws), this material demonstrated faster fracture healing, stronger osteointegration, and facilitated new bone tissue formation with increased calcium deposition at the material-bone interface. The multilayer GT-supported BMP-9 coated n-CDHA/PAA composite biomaterials have demonstrated favorable osteogenic cell interface performance, both in vitro and in vivo. This study provides a foundation for developing innovative bone repair materials, holding promise for significant advancements in clinical applications.

Atomic layer deposition of biocompatible multifunctional ZnO-TiO2 nanocoatings on the surface of additively manufactured nitinol

In order to enhance the corrosion resistance and bioactivity of the additively manufactured nitinol alloy, ZnO-TiO2 (ZTO) nanocoatings have been synthesised by atomic layer deposition. The supercycle approach was employed to deposit 40 nm coatings with varying ZnO/TiO2 ratios: 5/1, 1/1, 1/5 and 1/20. The results demonstrated that this approach permitted the coating composition to be varied over a wide range. The study of anticorrosion properties in physiological Ringer’s solution revealed that the coatings, depending on their composition, are capable of reducing the corrosion rate of nitinol by a value ranging from 4 to 58 times. In vitro studies have demonstrated high viability, good adhesion and spreading of human osteoblast-like MG-63 and mesenchymal stem FetMSC cells on the surface of all samples except those with high zinc content (ZnO and ZTO-5/1). All coatings induced differentiation of both cell lines in the osteogenic direction and demonstrated antibacterial activity against multi-drug resistant A. baumannii (>96 %) and P. aeruginosa (>90 %) strains. The results indicate the considerable potential of the developed methodology for the ALD of ZTO that combine the biocompatibility of titanium oxide, the antibacterial properties of zinc oxide and their overall stability and effectiveness for the protection of nitinol against biocorrosion.

Mechanical, physical, and biological properties of polycaprolactone/ Mg-doped SrFe12O19 nanocomposite scaffolds for bone tissue engineering applications

Nowadays, the utilization of 3D printing has become common in the construction of composite scaffolds. In this research, the co-precipitation method was applied to synthesize pure strontium hexaferrite oxide (SrFe12O19) and magnesium-doped strontium hexaferrite oxide nanoparticles (Mg–SrFe12O19). The synthesized SrFe12O19 nanoparticles had a spherical morphology with a mean size of 60 nm and magnetization (Ms) value of 54.38 emu/g. The Mg–SrFe12O19 also had a spherical morphology with a mean particle size of 46 nm and a magnetization value of 29.89 emu/g. The produced polycaprolactone composite scaffolds containing different amounts (0, 25, 50, and 75 wt%) of reinforcement were fabricated by the 3D robocasting printing method. Based on the results of the compression test, the 25 wt% addition of the pure SrFe12O19 nanoparticles to the polycaprolactone scaffold increased the compressive strength from 2.87 to 11.68 MPa compared to the 100 wt% pure polycaprolactone scaffold. Furthermore, the polycaprolactone nanocomposite scaffold containing 25 wt% Mg–SrFe12O19 also increased the compressive strength compared to the pure polycaprolactone scaffold to 12.94 MPa. Therefore, the 25 wt% reinforced scaffold was chosen as the optimal sample. The contact angles of the pure polycaprolactone SrFe12O19 and Mg– SrFe12O19 reinforced polycaprolactone scaffolds were 77.17, 68.53, and 35.58°, respectively. The values indicate the significant effect of doping magnesium on the wettability. The degradability of the 3D-printed scaffolds containing 25 wt% of the two different reinforcements was evaluated for 28 days immersing in phosphate-buffered saline (PBS) solution. The results revealed that the degradability was higher for the Mg–SrFe12O19 after 28 days compared to the composite scaffold reinforced by pure SrFe12O19 reinforcement. Moreover, it was shown that the bioactivity and cell adhesion of MG63 cells for the scaffolds reinforced by Mg–SrFe12O19 have increased in laboratory conditions. Based on the MTT test, it was observed that both of the scaffolds were non-toxic. The results obtained in this study, suggest that polycaprolactone nanocomposite scaffolds containing 25 wt% of Mg–SrFe12O19, made by the 3D robocasting printing method, are suitable for bone tissue engineering.

Injectable macroporous naturally-derived apatite bone cement as a potential trabecular bone substitute.

In this study, we have formulated a novel apatite bone cements derived from natural sources (i.e. eggshell and fishbone) with improved qualities that is, porosity, resorbability, biological activity, and so forth. The naturally-derived apatite bone cement (i.e. FBDEAp) was prepared by mixing hydroxyapatite (synthesized from fishbone) and tricalcium phosphate (synthesized from eggshell) as a solid phase with a liquid phase (a dilute acidic blend of cement binding accelerator and biopolymers like gelatin and chitosan) with polysorbate (as liquid porogen) to get a desired bone cement paste. The prepared cement paste sets within the clinically acceptable setting time (≤20 min), easily injectable (>85%) through hands and exhibits physiological pH stability (7.3-7.4). The pure apatite phased bone cement was confirmed by x-ray diffraction and Fourier transform infrared spectroscopy analyses. The FBDEAp bone cement possesses acceptable compressive strength (i.e. 5-7 MPa) within trabecular bone range and is resorbable up to 28% in simulated body fluid solution within 12 weeks of incubation at physiological conditions. The FBDEAp is macroporous in nature (average pore size ~50-400 μm) with interconnected pores verified by SEM and micro-CT analyses. The FBDEAp showed significantly increased MG63 cell viability (>125% after 72 h), cell adhesion, proliferation, and key osteogenic genes expression levels (up to 5-13 folds) compared to the synthetically derived, synthetic and eggshell derived as well as synthetic and fishbone derived bone cements. Thus, we strongly believe that our prepared FBDEAp bone cement can be used as potential trabecular bone substitute in orthopedics.

Characterization and In Vitro Evaluation of Porous Polymer-Blended Scaffolds Functionalized with Tricalcium Phosphate.

Bone tissue is one of the most transplanted tissues. The ageing population and bone diseases are the main causes of the growing need for novel treatments offered by bone tissue engineering. Three-dimensional (3D) scaffolds, as artificial structures that fulfil certain characteristics, can be used as a temporary matrix for bone regeneration. In this study, we aimed to fabricate 3D porous polymer scaffolds functionalized with tricalcium phosphate (TCP) particles for applications in bone tissue regeneration. Different combinations of poly(lactic acid) (PLA), poly(ethylene glycol) (PEG with molecular weight of 600 or 2000 Da) and poly(ε-caprolactone) (PCL) with TCP were blended by a gel-casting method combined with rapid heating. Porous composite scaffolds with pore sizes from 100 to 1500 µm were obtained. ATR-FTIR, DSC, and wettability tests were performed to study scaffold composition, thermal properties, and hydrophilicity, respectively. The samples were observed with the use of optical and scanning electron microscopes. The addition of PCL to PLA increased the hydrophobicity of the composite scaffolds and reduced their susceptibility to degradation, whereas the addition of PEG increased the hydrophilicity and degradation rates but concomitantly resulted in enhanced creation of rounded mineral deposits. The scaffolds were not cytotoxic according to an indirect test in L929 fibroblasts, and they supported adhesion and growth of MG-63 cells when cultured in direct contact.

Degradation protection and enhanced biocompatibility of Mg alloys pretreated with plasma proteins.

After implantation of the Mg alloy in the human body, the adsorption of plasma protein on surface will cause a series of cell reactions and affect the degradation of Mg alloys. Herein, in vitro biological reactions of the ZK60 and AZ31 Mg alloys are analyzed in plasma protein environment. Combined with mass spectrometry analysis of the type of adsorbed proteins, it is shown that proteins such as fibrinogen, vitronectin, fibronectin, and prothrombin are prone to get adsorbed on the surface of the alloys than other proteins, leading to the promotion of MG63 cell adhesion and proliferation. The effect of selected proteins (fibrinogen, fibronectin, and prothrombin) on degradation of ZK60 and AZ31 Mg alloys is investigated using immersion tests. The degradation of AZ31 Mg alloy is significantly restrained with the presence of proteins. This is due to the protein adsorption effect on the sample surface. The molecular dynamics simulation results indicate that both fibrinogen and fibronectin tend to adsorb onto the AZ31 rather than ZK60, forming a stable protein layer on the AZ31 Mg alloy retarding the degradation of the samples. As to ZK60 alloy, the addition of protein inhibits the degradation in the short term, however, the degradation increases after a long time of immersion. This phenomenon is particularly pronounced in fibronectin solution.

Graphene oxide-encapsulated baghdadite nanocomposite improved physical, mechanical, and biological properties of a vancomycin-loaded PMMA bone cement

Polymethyl methacrylate (PMMA) bone cement is commonly used in orthopedic surgeries to fill the bone defects or fix the prostheses. These cements are usually containing amounts of a nonbioactive radiopacifying agent such as barium sulfate and zirconium dioxide, which does not have a good interface compatibility with PMMA, and the clumps formed from these materials can scratch metal counterfaces. In this work, graphene oxide encapsulated baghdadite (GOBgh) nanoparticles were applied as radiopacifying and bioactive agent in a PMMA bone cement containing 2 wt.% of vancomycin (VAN). The addition of 20 wt.% of GOBgh (GOBgh20) nanoparticles to PMMA powder caused a 33.6% increase in compressive strength and a 70.9% increase in elastic modulus compared to the Simplex® P bone cement, and also enhanced the setting properties, radiopacity, antibacterial activity, and the apatite formation in simulated body fluid. In vitro cell assessments confirmed the increase in adhesion and proliferation of MG-63 cells as well as the osteogenic differentiation of human adipose-derived mesenchymal stem cells on the surface of PMMA-GOBgh20 cement. The chorioallantoic membrane assay revealed the excellent angiogenesis activity of nanocomposite cement samples. In vivo experiments on a rat model also demonstrated the mineralization and bone integration of PMMA-GOBgh20 cement within four weeks. Based on the promising results obtained, PMMA-GOBgh20 bone cement is suggested as an optimal sample for use in orthopedic surgeries.

Preparation of a Zn/hydroxyapatite/chitosan composite coating with an antibacterial ability and cytocompatibility on a NiTi alloy

Zn has been extensively used to prevent microbial infections in orthopedic implants; however, high amounts of Zn inhibit cell proliferation. This study investigated the ability to decrease the cytotoxicity of Zn-doped coatings on NiTi shape memory alloys (NiTi SMAs) while maintaining their antibacterial activity. The Zn3(PO4)2 distributed in the chitosan coating, annotated as ZHAP, was obtained by electrophoretic deposition in a chitosan solution with Zn acetate acid and hydroxyapatite particles. The ZHAP inhibited bacterial growth and exhibited a good antibacterial activity (86 %) against Staphylococcus aureus. In addition, the MG63-cell adhesion and cytocompatibility with ZHAP were better than those with the Zn-doped coating due to the precipitation of Zn3(PO4)2 which decreased the release of Zn2+ ions from the chitosan matrix. The coatings did not affect neither the superelasticity (< 0.5 % residual strain) nor the shape memory ability (100 % recovery rate) of the NiTi SMAs.

Advantages of agarose on alginate for the preparation of polysaccharide/hydroxyapatite porous bone scaffolds compatible with a proline-rich antimicrobial peptide

The optimized proline-rich antimicrobial peptide B7-005 was loaded on bone scaffolds based on polysaccharides and hydroxyapatite. Alginate was firstly chosen in order to exploit its negative charges, which allowed an efficient B7-005 loading but hindered its release, due to the strong interactions with the positive charged peptide. Hence, alginate was substituted with agarose which allowed to prepare scaffolds with similar structure, porosity, and mechanical performance than the ones prepared with alginate and hydroxyapatite. Moreover, agarose scaffolds could release B7-005 within the first 24 h of immersion in aqueous environment. The peptide did not impaired MG-63 cell adhesion and proliferation in the scaffold, and a positive cell proliferation trend was observed up to two weeks. The released B7-005 was effective against the pathogens E. coli, K. pneumoniae, and A. baumannii, but not against S. aureus and P. aeruginosa, thus requiring further tuning of the system to improve its antimicrobial activity.

Morphological and fractal features of cancer cells anchored on composite layers based on magnesium-doped hydroxyapatite loaded 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, containing human osteosarcoma MG63 cells anchored. Studies regarding the biocompatibility of the composite layers were performed with the aid of a MTT (3–4,5-Dimethylthiazol 2,5-diphenyltetrazolium bromide) assay. The data determined that the composite layers did not inhibit the growth and adhesion of MG63 cells to their surfaces exhibiting good biocompatibility properties. Furthermore, the attachment and development of MG63 cells on the surface of MgHApCh composite layers were investigated using atomic force microscopy (AFM). AFM topographical maps emphasized that the HApCh and 8MgHApCh composite layers surface promoted the attachment and proliferation of MG63 cells on their surface. Meanwhile, in the case of 30MgHApCh layers incubated for 48 h, a slight modification of the morphological features of the MG63 cells. In addition, the effects of the composite layers against Candida albicans ATCC 10231 were also evaluated. The data results from the in vitro antifungal assay depicted that the composite layers successfully inhibited the growth of the fungal cells onto their surface. Morphological and fractal analyses unveil cancer cell surfaces on Mg-containing composite layers with intricate 3D patterns, driven by high-frequency components. Their remarkable complexity and roughness arises from a strong multifractal nature, supporting more effective vertical growth compared to Si and HApCh surfaces. The cell viability reduced of uncoated Si surface is highlighted by its less intense 3D pattern growth. Our results show that the uncoated Si surface promotes lower viability of MG63 cancer cells, with less rough and complex 3D spatial patterns.

Prunetin Triggers ROS-mediated Apoptosis through the Suppression of MAPKs/STAT-3/NF-κB-p65 Signaling Pathway in Human Osteosarcoma Cells

Background Osteosarcoma (OS) is a highly metastatic primary bone malignancy, and the treatment options remain inadequate. Hence, exploring innovative natural medications is required. Prunetin (PRU) is an isoflavone that has been a proven anticancer agent in numerous cancer cell lines. However, the activity of PRU against OS remains uncertain. Materials and Methods Here, we studied the anticancer activity of PRU (20 and 25 µM) on human OS cells MG-63 and investigated its latent mechanism. The PRU activity of MG-63 cells cytotoxicity, intracellular ROS, metastasis, apoptosis, anti-apoptotic proteins, MAPK/STAT-3, and AKT signaling pathways was assessed by MTT assay, DCFH-DA, DAPI, PI, AO/EB, cell adhesion, and RT-PCR analysis. Findings unveiled that PRU could constrain MG-63 cell viability and adhesion through elevated intracellular ROS and elicited apoptosis. Results Likewise, PRU (20 and 25 µM) avert the MG-63 cell proliferation, which stimulates apoptosis by the enhancement of Bax and caspases, while it diminishes Bcl-2 in a dose-dependent way. Furthermore, PRU could reduce Pin-1, and anti-apoptotic elements, as well as trigger apoptotic signaling pathways. Our data established that PRU alleviates MG-63 cell proliferation and metastasis via ROS-mediated apoptosis, which triggers MAPKs/STAT3 and AKT pathways, suggesting that PRU is a promising natural remedy for OS. In order to comprehend the therapeutic target for cancer, we assessed the effect of PRU on the expression of Pin1, which is thought to be over-expressed in many human malignancies. According to our findings, PRU specifically suppressed Pin1 expression to reduce the expression of Akt, STAT3, P38, JNK, P65, and IL-6. We evaluated the impact of PRU on the expression of Pin1, which is allegedly over-expressed in many human malignancies, to better understand the therapeutic target for cancer. Researchers state that PRU inhibited the expression of Akt, STAT3, P38, JNK, P65, and IL-6 in particular, by suppressing Pin1 expression. Conclusion Together, these results suggest that PRU may be an effective treatment for bone cancer in people by preventing Pin1 expression.

Extrusion-based 3D printing of osteoinductive scaffolds with a spongiosa-inspired structure.

Critical-sized bone defects resulting from trauma, inflammation, and tumor resections are individual in their size and shape. Implants for the treatment of such defects have to consider biomechanical and biomedical factors, as well as the individual conditions within the implantation site. In this context, 3D printing technologies offer new possibilities to design and produce patient-specific implants reflecting the outer shape and internal structure of the replaced bone tissue. The selection or modification of materials used in 3D printing enables the adaption of the implant, by enhancing the osteoinductive or biomechanical properties. In this study, scaffolds with bone spongiosa-inspired structure for extrusion-based 3D printing were generated. The computer aided design process resulted in an up scaled and simplified version of the bone spongiosa. To enhance the osteoinductive properties of the 3D printed construct, polycaprolactone (PCL) was combined with 20% (wt) calcium phosphate nano powder (CaP). The implants were designed in form of a ring structure and revealed an irregular and interconnected porous structure with a calculated porosity of 35.2% and a compression strength within the range of the natural cancellous bone. The implants were assessed in terms of biocompatibility and osteoinductivity using the osteosarcoma cell line MG63 and patient-derived mesenchymal stem cells in selected experiments. Cell growth and differentiation over 14days were monitored using confocal laser scanning microscopy, scanning electron microscopy, deoxyribonucleic acid (DNA) quantification, gene expression analysis, and quantitative assessment of calcification. MG63 cells and human mesenchymal stem cells (hMSC) adhered to the printed implants and revealed a typical elongated morphology as indicated by microscopy. Using DNA quantification, no differences for PCL or PCL-CaP in the initial adhesion of MG63 cells were observed, while the PCL-based scaffolds favored cell proliferation in the early phases of culture up to 7days. In contrast, on PCL-CaP, cell proliferation for MG63 cells was not evident, while data from PCR and the levels of calcification, or alkaline phosphatase activity, indicated osteogenic differentiation within the PCL-CaP constructs over time. For hMSC, the highest levels in the total calcium content were observed for the PCL-CaP constructs, thus underlining the osteoinductive properties.

Use of Various UVC Photofunctionalization Times to Modify Surface Characteristics and Enhance the Biologic Activity of SLA Titanium.

To evaluate the effect of ultraviolet-C light (UVC) photofunctionalization treatment time on the biologic activity of airborne particle-abraded and acid-etched (SLA) titanium surfaces and to analyze its physical and chemical mechanisms. SLA titanium was treated with UVC light for different lengths of time (10 minutes, 20 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, and 24 hours), and then changes to its surface characteristics were evaluated via electron microscope scanning, X-ray photoelectron spectroscopy (XPS), water contact angle measurement, and zeta potential measurement. The effect of UVC photofunctionalization on the biologic processes of SLA titanium surfaces was assessed by analyzing the bovine serum albumin adsorption, adhesion, proliferation, morphology, and alkaline phosphatase activity of MG-63 cells. UVC irradiation did not change the topography of SLA titanium surfaces. As treatment times increased, the water contact angle decreased from 120 degrees to 0 degrees, and the hydrocarbon content decreased. Zeta potential testing showed increased surface potential of photofunctionalized titanium. In vitro testing showed that cell adhesion, proliferation, morphology, and alkaline phosphate (ALP) activity on titanium surfaces were significantly improved by UVC photofunctionalization. UVC photofunctionalization can improve the biologic activity of SLA titanium surfaces by removing hydrocarbons and increasing the surface potential of titanium.

Incorporation of graphene oxide as a coupling agent in a 3D printed polylactic acid/hardystonite nanocomposite scaffold for bone tissue regeneration applications

3D printing fabrication has become a dominant approach for the creation of tissue engineering constructs as it is accurate, fast, reproducible and can produce patient-specific templates. In this study, 3D printing is applied to create nanocomposite scaffold of polylactic acid (PLA)/hardystonite (HT)-graphene oxide (GO). GO is utilized as a coupling agent of alkaline treated HT nanoparticles within PLA matrix. The addition of HT-GO nanoparticles of up to 30 wt% to PLA matrix was found to increase the degradability from 7.33 ± 0.66 to 16.03 ± 1.47 % during 28 days. Also, the addition of 20 wt% of HT-GO nanoparticles to PLA scaffold (PLA/20HTGO sample) significantly increased the compressive strength (from 7.65 ± 0.86 to 14.66 ± 1.01 MPa) and elastic modulus (from 94.46 ± 18.03 to 189.15 ± 10.87 MPa). The apatite formation on the surface of nanocomposite scaffolds in simulated body fluid within 28 days confirmed the excellent bioactivity of nanocomposite scaffolds. The MG63 cell adhesion and proliferation and, also, the rat bone marrow mesenchymal stem cells osteogenic differentiation were highly stimulated on the PLA/20HTGO scaffold. According to the sum of results obtained in the current study, the optimized PLA/20HTGO nanocomposite scaffold is highly promising for hard tissue engineering applications.

Preparation, characterization, and bioactivity of reinforced monetite with chitosan-gelatin electrospun composite scaffold for bone tissue engineering

In this study, chitosan-gelatin-monetite (CGM)-based electrospun scaffolds have been developed that closely mimicked the microstructure and chemical composition of the extracellular matrix of natural bone. CGM-based nanofibrous composite scaffolds were prepared with the help of the electrospinning technique, post-cross-linked using ethyl(dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide solution to improve their stability in an aqueous environment. The prepared chitosan/gelatin (CG) scaffold showed an average fiber diameter of 308 ± 17 nm, whereas 5 and 7 wt% monetite containing CGM5 and CGM7 scaffolds, exhibited an average fiber diameter of 287 ± 13 and 265 ± 9 nm, respectively, revealing the fine distribution of monetite particles on the fibrous surface. The distribution of monetite nanoparticles onto the CG nanofibrous surface was confirmed using x-ray diffraction, Fourier transform infrared, and EDAX. Moreover, the addition of 7 wt% monetite into the CG electrospun matrix increased their ultimate tensile strength from 7.62 ± 0.13 MPa in the CG scaffold to 14.34 ± 0.39 MPa in the CGM7 scaffold. Simulated body fluid study and staining with alizarin red S (ARS) confirmed the higher mineralization ability of monetite-containing scaffolds compared to that revealed by the CG scaffold. The monetite incorporation into the CG matrix improved its osteogenic properties, including pre-osteoblast MG-63 cell adhesion, proliferation, and differentiation, when seeded with the cells. A higher degree of cellular adhesion, spreading, and migration was observed on the monetite-incorporated CG scaffold than that on the CG scaffold. From 3-(4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide) MTT assay, alkaline phosphatase activity, ARS staining, and immunocytochemistry study, the cultured cells discovered a more conducive microenvironment to proliferate and subsequently differentiate into osteoblast lineage in contact with CGM7 nanofibers rather than that in CGM0 and CGM5. In-vitro results indicated that electrospun CGM-based composite scaffolds could be used as a potential candidate to repair and regenerate new bone tissues.

Chitosan/Graphene Oxide Nanocomposite Coatings on Magnesium Alloy: Corrosion and Biocompatibility Properties

Abstract In the present study, optimal conditions for the fabrication of chitosan (CS)/graphene oxide (GO) nanocomposite coatings were evaluated by the pulse electrodeposition (PED) process on Mg–2wt%Zn scaffolds. The size distribution of CS, GO suspension, and CS/GO composite was evaluated using dynamic light scattering. The coatings’ microstructure and morphologies were investigated by Fourier-transform infrared spectroscopy, ultraviolet-visible spectroscopy, Raman spectroscopy, thermogravimetric analysis, derivative thermogravimetric analysis, differential thermal analysis, and scanning electron microscopy. The Taguchi statistical method was used to optimize PED parameters, including peak current density (j), duty cycle, and GO nanosheets content (1, 2, and 3 percent by weight [wt%]). Results showed that optimal coatings were produced under the conditions of 2 wt% GO, j = 20 mA/cm2, duty cycle = 0.5, and pH = 5. The process’s time, temperature, and frequency were 20 min, 37°C, and 1,000 Hz. The biocompatibility of coatings was assessed by in vitro test. The results of cell viability and adhesion of MG63 cells on optimal coating are promising for application in bone tissue engineering.

Composite microgranular scaffolds with surface modifications for improved initial osteoblastic cell proliferation

Polyester-based granular scaffolds are a potent material for tissue engineering due to their porosity, controllable pore size, and potential to be molded into various shapes. Additionally, they can be produced as composite materials, e.g., mixed with osteoconductive β-tricalcium phosphate or hydroxyapatite. Such polymer-based composite materials often happen to be hydrophobic, which disrupts cell attachment and decreases cell growth on the scaffold, undermining its primary function. In this work, we propose the experimental comparison of three modification techniques for granular scaffolds to increase their hydrophilicity and cell attachment. Those techniques include atmospheric plasma treatment, polydopamine coating, and polynorepinephrine coating. Composite polymer/β-tricalcium phosphate granules have been produced in a solution-induced phase separation (SIPS) process using commercially available biomedical polymers: poly(lactic acid), poly(lactic-co-glycolic acid), and polycaprolactone. We used thermal assembly to prepare cylindrical scaffolds from composite microgranules. Atmospheric plasma treatment, polydopamine coating, and polynorepinephrine coating showed similar effects on polymer composites' hydrophilic and bioactive properties. All modifications significantly increased human osteosarcoma MG-63 cell adhesion and proliferation in vitro compared to cells cultured on unmodified materials. In the case of polycaprolactone/β-tricalcium phosphate scaffolds, modifications were the most necessary, as unmodified polycaprolactone-based material disrupted the cell attachment. Modified polylactide/β-tricalcium phosphate scaffold supported excellent cell growth and showed ultimate compressive strength exceeding this of human trabecular bone. This suggests that all investigated modification techniques can be used interchangeably for increasing wettability and cell attachment properties of various scaffolds for medical applications, especially those with high surface and volumetric porosity, like granular scaffolds.

Fabrication of polycaprolactone/heparinized nano fluorohydroxyapatite scaffold for bone tissue engineering uses

In this study, some polycaprolactone (PCL)/heparinized nano fluorohydroxyapatite (H-nFHA) scaffolds were fabricated with different amounts of H-nFHA using solid-liquid phase separation and freeze-drying methods. The scaffolds were characterized using Fourier transform infrared spectroscopy, x-ray diffraction, scanning electron microscopy techniques, and compressive test. Their biological behavior also was studied using Alkaline phosphatase activity and MTT assay. The results showed that adding H-nFHA to the PCL matrix improves MG-63 cells adhesion and proliferation and they also showed that heparinizing the nFHA particles improves the mechanical properties and dispersion of the nFHA particles in the PCL matrix for bone tissue engineering uses.

Gelatin/monetite electrospun scaffolds to regenerate bone tissue: Fabrication, characterization, and in-vitro evaluation

This work is dedicated to combining nanotechnology with bone tissue engineering to prepare and characterize electrospun gelatin/monetite nanofibrous scaffold with improved physicochemical, mechanical, and biological properties. Nanofibrous scaffolds possessing fiber diameter in the range of 242–290 nm were prepared after incorporating varying content of monetite nanoparticles up to 7 wt % into the gelatin matrix using the electrospinning technique. Cross-linking of gelatin chains in the scaffold was performed using 0.25 wt% glutaraldehyde as indicated by imine (-CN-) bond formation in the FTIR analysis. With an increase in monetite addition up to 7 wt%, a decrease in swelling ratio and bio-degradability of cross-linked gelatin scaffolds was observed. Gelatin scaffold with 7 wt% monetite content registered the highest values of tensile strength and tensile modulus of 18.8 MPa and 170 MPa, as compared to 0% and 5 wt% monetite containing scaffolds respectively. Cell viability and differentiation were studied after culturing MG-63 cells onto the scaffolds from confocal microscopy of live and dead cells images, MTT assay, and alkaline phosphatase assay for a cell culture period of up to 21 days. It was observed that 7 wt % monetite containing gelatin scaffold exhibited better MG-63 cell adhesion, proliferation, higher biomineralization, and ALP activity compared to 0% and 5 wt% monetite containing electrospun scaffolds studied here.