Reinforcing ß-tricalcium phosphate scaffolds for potential applications in bone tissue engineering: impact of functionalized multi-walled carbon nanotubes.

Beta-tricalcium phosphate (ß-TCP) scaffolds manufactured through the foam replication method are widely employed in bone tissue regeneration. The mechanical strength of these scaffolds is a significant challenge, partly due to the rheological properties of the original suspension. Various strategies have been explored to enhance the mechanical properties. In this research, ß-TCP scaffolds containing varying concentrations (0.25-1.00 wt%) of multi-walled carbon nanotubes (MWCNT) were developed. The findings indicate that the addition of MWCNTs led to a concentration-dependent improvement in the viscosity of ß-TCP suspensions. All the prepared slurries exhibited viscoelastic behavior, with the storage modulus surpassing the loss modulus. The three time interval tests revealed that MWCNT-incorporated ß-TCP suspensions exhibited faster structural recovery compared to pure ß-TCP slurries. Introducing MWCNT modified compressive strength, and the optimal improvement was obtained using 0.75 wt% MWCNT. The in vitro degradation of ß-TCP was also reduced by incorporating MWCNT. While the inclusion of carbon nanotubes had a marginal negative impact on the viability and attachment of MC3T3-E1 cells, the number of viable cells remained above 70% of the control group. Additionally, the results demonstrated that the scaffold increased the expression level of osteocalcin, osteoponthin, and alkaline phosphatase genes of adiposed-derived stem cells; however, higher levels of gene expersion were obtained by using MWCNT. The suitability of MWCNT-modified ß-TCP suspensions for the foam replication method can be assessed by evaluating their rheological behavior, aiding in determining the critical additive concentration necessary for a successful coating process.

Wound closure, angiogenesis and antibacterial behaviors of tetracalcium phosphate/hydroxyethyl cellulose/hyaluronic acid/gelatin composite dermal scaffolds.

Polymeric and tetracalcium phosphate (TTCP)-containing polymeric scaffolds were fabricated using a freeze-drying technique, with a homogenous solution of hydroxyethyl cellulose (HEC)/hyaluronic acid (HA)/gelatin (G) or suspension of 15 or 20% TTCP) particles in HEC/HA/G solution. The morphology, phase composition, chemical bands, and swelling behavior of the scaffold were determined. In vitro fibroblast cell viability and migration potential of the scaffolds were determined by MTT, live/dead staining, and scratch assay for wound healing. The in vivo chick embryo angiogenesis test was also carried out. Finally, the initial antibacterial activity of the scaffolds was determined using Staphylococcus aureus. The scaffolds exhibited an enormous porous structure in which the size of pores increased by the presence of TTCP particles. While the polymeric scaffold was amorphous, the formation of low crystalline hydroxyapatite phase and the initial TTCP particles was determined in the composition of TTCP-added scaffolds. TTCP increased swelling behavior of the polymeric scaffold in PBS. The results demonstrated that the amount of TTCP was a crucial factor in cell life. A high concentration of TTCP could restrict cell viability, although all the scaffolds were nontoxic. The scratch assessments determined better cell migration and wound closure in treating with TTCP-containing scaffolds so that after 24 h, a wound closure of 100% was observed. Furthermore, TTCP-incorporated scaffolds significantly improved the angiogenesis, in the chick embryo test. The presence of TTCP had a significant effect on reducing the bacterial activity and 20% TTCP-containing scaffold exhibited better antibacterial activity than the others.

Bioorthogonal hydroxyethyl cellulose-based scaffold crosslinked via click chemistry for cartilage tissue engineering applications.

In this study, azide and alkyne moieties were introduced to the structure of citric acid-modified hydroxyethyl cellulose (HEC) and then through a bioorthogonal click chemistry method: Strain-promoted azide-alkyne cycloaddition, a novel crosslinked HEC scaffold (click sample) was obtained. Chemical modifications and successful crosslinking of the samples were assessed with FTIR and 1H NMR spectroscopy. Lyophilized samples exhibited a porous interconnected microarchitecture with desirable features for commensurate cartilage tissue engineering applications. As the stability of scaffolds improved upon crosslinking, considerable water uptake and swelling degree of ~650% could still be measured for the click sample. Offering Young's modulus of ~10 MPa and tensile strength of ~0.43 MPa, the mechanical characteristics of click sample were comparable with those of normal cartilage tissue. Various in vitro biological assays, including MTT analysis, cellular attachment, histological staining with safranin O, and real-time PCR decisively approved significant biocompatibility, chondrogenic ability, and bioorthogonal features of click sample.

In vitro evaluation of curcumin-loaded chitosan-coated hydroxyapatite nanocarriers as a potential system for effective treatment of cancer.

Nanotechnology has many potential applications in cancer treatment. For example, nano-drug delivery systems (NDDS) with high bioavailability, biodegradability, and biocompatibility have been developed, in order to increase the therapeutic effects of anticancer drugs. Among these NDDS, high-performance hydroxyapatite (HA) nanoparticles are rapidly advancing in the targeted cancer treatment due to their numerous benefits. Curcumin is an herbal metabolite that acts as a chemical inhibitor through the inhibition of tumor cells and the progression of many cancers. However, the poor bioavailability of curcumin is the most important challenge in using this substance. In this study, HA nanoparticles coated by chitosan were used as a pH-sensitive biopolymer to improve the efficiency and bioavailability of curcumin. For this purpose, HA nanoparticles were first synthesized by the sol-gel method. Then, a layer of chitosan was coated on it, and the curcumin drug was encapsulated in the nanocarrier, under controlled conditions. Techniques such as scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) were used to characterize the nanocarriers. In the second part, nano-drugs prepared by various bioassays were examined. For this purpose, the rate of cytotoxicity by the methyl-thiazol-tetrazolium (MTT) assay and the rate of apoptosis induction by the acridine orange and ethidium bromide (AO/EB) staining method on the brain carcinoma U87MG cell line were investigated.

The synergistic effects of SrF2 nanoparticles, YSZ nanoparticles, and poly-ε-l-lysin on physicomechanical, ion release, and antibacterial-cellular behavior of the flowable dental composites.

Resin-based pit-and-fissure sealants (flowable resin composites) were formulated using bisphenol-A-glycerolatedimethacrylate (Bis-GMA)-triethylene glycol dimethacrylate-(TEGDMA)-diurethanedimethacrylate (UDMA) mixed monomers and multiple fillers, including synthetic strontium fluoride (SrF2) nanoparticles as a fluoride-releasing and antibacterial agent, yttria-stabilized zirconia (YSZ) nanoparticles as an auxiliary filler, and poly-ε-l-lysin (ε-PL) as an auxiliary antibacterial agent. Based on the physical, mechanical and initial antibacterial properties, the formulated nano-sealant containing 5 wt% SrF2, 5 wt% YSZ and 0.5 wt% ε-PL was selected as the optimal specimen and examined for ion release and cytotoxicity. The results showed an average release rate of 0.87 µg·cm-2·day-1 in the aqueous medium (pH 6.9) and 1.58 µg·cm-2·day-1 in acidic medium (pH 4.0). The maximum cytotoxicity of 20% toward human bone marrow mesenchymal stem cells (hMSCs) was observed according to the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) cytotoxicity assay and acridine orange staining test. A synergy between SrF2 nanoparticles and ε-PL exhibited a better antibacterial activity in terms of colony reduction compared to the other samples. However, the inclusion of SrF2 and ε-PL caused mechanically weakening of the sealants that was partly compensated by incorporation of YSZ nanoparticles (up to 10 wt%).

Comparative analysis and properties evaluation of gelatin microspheres crosslinked with glutaraldehyde and 3-glycidoxypropyltrimethoxysilane as drug delivery systems for the antibiotic vancomycin.

In the present comparative study, gelatin microspheres (GMs) were prepared by emulsification-solvent-extraction method using well-known crosslinker: glutaraldehyde (GA) and biocompatible silane-coupling agent: glycidoxypropyltrimethoxysilane (GPTMS). Crosslinking with GA was done by a definite and common procedure, while GPTMS crosslinking potency was investigated after 5, 10, 24, and 48 h synthesis periods and the fabrication method was adjusted in order for preparation of GMs with optimized morphological and compositional characteristics. The prepared GMs were then evaluated and compared as drug delivery systems for the antibiotic vancomycin (Vm). Morphological observations, FTIR, ninhydrin assay, swelling behavior evaluation and Hydrolytic degradation analysis proved successful modification of GMs and revealed that increasing synthesis time from 5 h to 24 h and 48 h, when using GPTMS as crosslinker, led to formation of morphologically-optimized GMs with highest crosslinking degree (∼50%) and the slowest hydrolytic degradation rate. Such GMs also exhibited most sustained release period of Vm. The antibacterial test results against gram-positive bacterium Staphylococcus aureus, were in accordance with the release profiles of Vm, as well. Together, GPTMS-crosslinked GMs with their preferable characteristics and known as biocompatible gelatin-siloxane hybrids, could act as proper drug delivery systems for the sustained release of the antibiotic vancomycin.

Synthesis, physicochemical, rheological and in-vitro characterization of double-crosslinked hyaluronic acid hydrogels containing dexamethasone and PLGA/dexamethasone nanoparticles as hybrid systems for specific medical applications.

Injectable hydrogels and biodegradable nanoparticles are using in tissue engineering applications and drug delivery systems. To improve physiochemical properties of biomaterials and to develop their applications, hybrid systems consist of hydrogels, and biodegradable nanoparticles were synthesized. In this study, hybrid systems based on double crosslinked hyaluronic acid and PLGA/Dexamethasone sodium phosphate (PLGADEX) nanoparticles are designed and synthesized in several steps. At the first step, poly(l-lactide-co-glycolide) (PLGA) in a ratio of LLA:GA = 85:15 mol% was synthesized via ring-opening polymerization. Then, PLGADEX nanoparticles were synthesized in different ratios using the partially modified emulsification-diffusion method and fully characterized, and desirable nanoparticle was selected (PLGADEX20). At the second step, a double cross-linked hyaluronic acid (XHA) was prepared by mixing various ratios of amino-hyaluronic acid and aldehyde-hyaluronic acid in the presence of genipin. Finally, by mixing of various ratios of PLGADEX20 and Dexamethasone sodium phosphate (DEX) with different ratios of XHA, hybrid systems were prepared. Based on the characterization of hybrid samples and the release studies, hydrogels containing nanoparticles showed a controlled drug release, while the best sample with 3% of optimized nanoparticle was chosen. According to physiochemical and biological properties, these hybrid systems can be good candidates for anti-adhesion barriers, wound dressings, and novel drug delivery systems.

Characterization and biological properties of a novel synthesized silicon-substituted hydroxyapatite derived from eggshell.

In the present study, the effect of adding different concentrations of silicon on physical, mechanical and biological properties of a synthesized aqueous precipitated eggshell-derived hydroxyapatite (e-HA) was evaluated. No secondary phases were detected by X-ray diffraction for the specimens e-HA and e-HA containing silicon (Si-e-HAs) before and after heating at 1200°C. A reduction in the crystallite size and a-axis as well as an increase in c-axis was occurred when silicon replacement was happened in the structure of e-HA. The presence of Si-O vibrations and carbonate modes for Si-e-HAs was confirmed by Fourier transform infrared spectroscopy analysis. The range of porosity and density was varied from 25% and 2.4 g cm-3 to 7% and 2.8 g cm-3 for e-HA and Si-e-HAs. The values of Young's modulus ( E) and compressive strength were varied for e-HA and Si-e-HAs. The porous structure of the samples was reduced when they were heated as e-HA kept the porous microstructure containing some dense areas and Si-e-HAs possessed a rough surface including slight levels of microporosity. The acellular in vitro bioactivity represented different apatite morphologies for e-HA and Si-e-HAs. The G-292 osteoblastic cells were stretched well on the surface with polygon-shaped morphology for 0.8Si-e-HA after 7 days of culture. According to MTT assay and alkaline phosphatase test, the maximum cell activity was related to 0.8Si-e-HA. The minimum inhibitory concentration for 0.8Si-e-HA and e-HA was estimated to be about 3.2 and 4.4 mg/mL, respectively. In overall, the sample 0.8Si-e-HA exhibited a higher bacteriostatic effect than e-HA against gram-negative bacterial strain Escherichia coli.

The Effective Role of Hydroxyapatite Based Composites in Anticancer Drug Delivery Systems.

Tumors consist of a heterogeneous population of cancer cells carrying multiple genetic mutations. During the past few decades, efforts have focused on curing cancer using various methods. However, traditional cancer therapies still carry some drawbacks, such as limited application for only a few cancer types, killing of normal cells, poor specificity, and associated toxicity. To overcome these disadvantages, drug-delivery methods that emphasize biomaterials have been developed and applied to optimize cancer treatments. Hydroxyapatite (HAP) is a biocompatible inorganic material that can be applied in biomedical drug-delivery applications. This review discusses the features and properties of HAP that make it an effective biomaterial and provides a comprehensive summary of recent studies in which HAP and composites containing HAP were applied as anticancer drug carriers. We believe that HAP-based composites show great promise for cancer treatment using controlled release of therapeutic agents, leading to enhanced efficiency, selective release of drugs, and prohibition of cancer cell proliferation.

Detection and qualification of optimum antibacterial and cytotoxic activities of silver-doped bioactive glasses.

This study aims to detect the optimum antibacterial activity of silver-doped bioactive glasses (Ag-BGs) for prevention of post-transplant infections in tissue engineering. The results have shown that the Ag-BG samples had broad-spectrum antibacterial efficacy in an Ag concentration-dependent manner. The 2% Ag-BG had the highest effect during the first 10 min to 72 h. The minimum inhibitory concentration of 2% Ag-BG was estimated to be 2 mg/ml for Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aeruginosa) and 2.66 mg/ml for Staphylococcus aureus (S. aureus). A concentration of 0.5% Ag-BG repressed growth of E. coli after 1 h, but did not have any detectable antibacterial effect for longer periods. Evaluation of the effects of prepared Ag-BG on human osteoblast cells viability showed that 1 and 2% samples changed the cell proliferation rate in masses of more than 3.33 and 2 mg/ml, respectively. Moreover, in a typical manner, the release of Ag ions from the glass structure started immediately, continued steadily and affected bacterial growth when it reached its critical concentration in the medium. This systematic study can illustrate the optimum antibacterial property of the Ag-BG samples in masses of 3.33 and 2 mg/ml for 1 and 2% Ag, respectively, for prevention of post-transplant infections.

Effect of adding nano-titanium dioxide on the microstructure, mechanical properties and in vitro bioactivity of a freeze cast merwinite scaffold.

In the present research, merwinite (M) scaffolds with and without nano-titanium dioxide (titania) were synthesized by water-based freeze casting method. Two different amounts (7.5 and 10 wt%) of n-TiO2 were added to M scaffolds. They were sintered at temperature of 1573.15°K and at cooling rate of 4°K/min. The changes in physical and mechanical properties were investigated. The results showed that although M and M containing 7.5 wt% n-TiO2 (MT7.5) scaffolds had approximately the same microstructures in terms of pore size and wall thickness, these factors were different for sample MT10. In overall, the porosity, volume and linear shrinkage were decreased by adding different weight ratios of n-TiO2 into the M structure. According to the obtained mechanical results, the optimum mechanical performance was related to the sample MT7.5 (E = 51 MPa and σ = 2 MPa) with respect to the other samples, i.e.: M (E = 47 MPa and σ = 1.8 MPa) and MT10 (E = 32 MPa and σ = 1.4 MPa). The acellular in vitro bioactivity experiment confirmed apatite formation on the surfaces of all samples for various periods of soaking time. Based on cell study, the sample which possessed favorable mechanical behavior (MT7.5) supported attachment and proliferation of osteoblastic cells. These results revealed that the MT7.5 scaffold with improved mechanical and biological properties could have a potential to be used in bone substitute.

Efficient removal of lead (II) ions and methylene blue from aqueous solution using chitosan/Fe-hydroxyapatite nanocomposite beads.

Chitosan is a well-known sorbent and effective in the uptake of anionic or reactive dyes, but it has deficiency in adsorption of basic dyes. In this work, chitosan/Fe-substituted hydroxyapatite composite beads were prepared in a different ratio via embedding of hydroxyapatite into chitosan solution for removal of basic dye and heavy metal from aqueous solution. The composite beads were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy in order to reveal their composition and surface morphology. In this particular study, methylene blue (MB) and lead (Pb (II)) ions were selected as representatives of dye and a heavy metal, respectively. The various experimental conditions affecting dye adsorption were explored to achieve maximum adsorption capacity. Moreover, the kinetic, thermodynamic and adsorption isotherm models were employed for the description of the heavy metal and dye adsorption processes. The results indicated that the prepared hydrogel is an efficient adsorbent for the aforementioned dye and metal concomitant with the ability of regeneration without losing the original activity and stability for water treatment applications.

In vitro biocompatibility of chitosan/hyaluronic acid-containing calcium phosphate bone cements.

The need for bone repair has increased as the population ages. In this research, calcium phosphate cements, with and without chitosan (CS) and hyaluronic acid (HA), were synthesized. The composition and morphological properties of cements were evaluated by X-ray diffraction and scanning electron microscopy. The acellular in vitro bioactivity revealed that different apatite morphologies were formed on the surfaces of cements after soaking in simulated body fluid. The in vitro osteoblastic cell biocompatibility of in situ forming cements was evaluated and compared with those of conventional calcium phosphate cements (CPCs). The viability and growth rate of the cells were similar for all CPCs, but better alkaline phosphatase activity was observed for CPC with CS and HA. Calcium phosphate cements supported attachment of osteoblastic cells on their surfaces. Spindle-shaped osteoblasts with developed cytoplasmic membrane were found on the surfaces of cement samples after 7 days of culture. These results reveal the potential of the CPC-CS/HA composites to be used in bone tissue engineering.

Preparation of laminated poly(ε-caprolactone)-gelatin-hydroxyapatite nanocomposite scaffold bioengineered via compound techniques for bone substitution.

In this research, new bioactive nanocomposite scaffolds were successfully developed using poly(ε-caprolactone) (PCL), cross-linked gelatin and nanoparticles of hydroxyapatite (HAp) after testing different solvents and methods. First, HAp powder was synthesized via a chemical precipitation technique and characterized. Then, the nanocomposites were prepared through layer solvent casting combined with freeze-drying and lamination techniques. According to the results, the increasing of the PCL weight in the scaffolds led to the improvement of the mechanical properties. The amount of ultimate stress, stiffness and also elastic modulus increased from 8 MPa for 0% wt PCL to 23.5 MPa for 50% wt PCL. The biomineralization study revealed the formation of an apatite layer on the scaffolds after immersion in simulated body fluid (SBF). The Ca-P ratios were in accordance to nonstoichiometric biological apatite, which was approximately 1.67. The in vitro biocompatibility and cytocompatibility of the scaffolds were tested using mesenchymal stem cells (MSCs), and the results indicated no sign of toxicity, and cells were found to be attached to the scaffold walls. The in vivo biocompatibility and osteogenesis of these scaffolds in the animal experiments is also under investigation, and the result will be published at the end of the study.

Preparation and characterization of calcium sulfate-biomimetic apatite nanocomposites for controlled release of antibiotics.

In the present study, release properties of antibiotic-loaded cement-type nanocomposites of biomimetic apatite and calcium sulfate were studied. Nanocrystalline component of the nanocomposite was synthesized by soaking a mixture of calcium phosphate reactants in tris-buffered simulated body fluid (SBF). The release patterns of cephalexin and gentamicin from both pure calcium sulfate and nanocomposite cements into SBF were collected up to 144 h and fitted by Higuchi and Weibull equations. The effect of loaded antibiotics on physical properties of the cements was also evaluated. Fast release behavior of both antibiotics was obtained from calcium sulfate matrix, in which 80-85% of the loaded antibiotics were liberated during the first 10 h of elution. In contrast, an administered elution was acquired from nanonocomposite materials so that the release was controlled, in all cases, by a combined mechanism; major mechanism was drug diffusion through the matrix and the minor was matrix dissolution. The results showed that the initial setting time and injectability of cements were increased from 7 min and 71% for pure calcium sulfate cement (powder-to-liquid ratio = 2.5 g/mL) to 33 min and 95% for the nanocomposite cement containing 60 wt % apatite, respectively. The compressive strength of nanocomposite was about 0.9 MPa, nearly four times lower than that of pure calcium sulfate. In addition, the use of cephalexin monohydrate did not influence the setting time and compressive strength of the cements, whereas (adding) gentamicin sulfate significantly improved these properties.