Functionalization of poly (lactic-co-glycolic acid) nano­calcium sulphate and fucoidan 3D scaffold using human bone marrow mesenchymal stromal cells for bone tissue engineering application.

This study aimed to functionalize a novel porous PLGA (Poly lactic-co-glycolic acid) composite scaffold in combination with nano­calcium sulphate (nCS) and/or fucoidan (FU) to induce osteogenic differentiation of human bone marrow stromal cells. The composite scaffolds (PLGA-nCS-FU, PLGA-nCS or PLGA-FU) were fabricated and subjected to characterization using Fourier-transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), Scanning electron microscopy (SEM) and Energy Dispersive X-Ray (EDX). The biocompatibility and osteogenic induction potential of scaffolds on seeded human bone marrow derived mesenchymal stromal cells (hBMSCs) were studied using cell attachment and alamar blue cell viability and alkaline phosphatase (ALP), osteocalcin and osteogenic gene expression, respectively. The composition of different groups was reflected in FTIR, XRD and EDX. The SEM micrographs revealed a difference in the surface of the scaffold before and after FU addition. The confocal imaging and SEM micrographs confirmed the attachment of cells onto all three composite scaffolds. However, the AB assay indicated a significant increase (p < 0.05) in cell viability/proliferation seeded on PLGA-nCS-FU on day 21 and 28 as compared with other combinations. A 2-fold significant increase (p < 0.05) in ALP and OC secretion of seeded hBMSCs onto PLGA-nCS-FU was observed when compared with other combinations. A significant increase in RUNX2, OPN, COL-I and ALP genes were observed in the cells seeded on PLGA-nCS-FU on day 14 and 28 as compared with day 0. In conclusion, the incorporation of both Fucoidan and Nano­calcium sulphate with PLGA showed a promising improvement in the osteogenic potential of hBMSCs. Therefore, PLGA-nCS-FU could be the ideal candidate for subsequent pre-clinical studies to develop a successful bone substitute to repair critical bone defects.

Optimization of Metal Ion/Fuel Ratio for an Effective Combustion of Monticellite and Investigation of Its Microbial and Hemolytic Activity for Biomedical Applications.

Bioactive silicates have gained popularity as bone graft substitutes in recent years due to their exceptional ability to bind to host tissues. The current study investigates the effect of changing the metal ion-to-fuel ratio on the properties and biological activity of monticellite prepared via the sol-gel connived combustion technique. Single-phasic monticellite was obtained at 900 °C, without any secondary-phase contaminants for the fuel-lean, stoichiometric, and fuel-rich conditions. SEM and TEM micrographs revealed the porous, spongy morphology of the materials. Because of the reduced crystallite size and higher surface area, the biomineralization of monticellite prepared under fuel-lean conditions resulted in more apatite deposition than those of the other two samples. The results show that the material has a good compressive strength comparable to natural bone, while its brittleness is equivalent to the lower moduli of bone. In terms of antibacterial and antifungal activities, the monticellite bioceramics outperformed the clinical pathogens. It can be used for bone tissue engineering and other biological applications due to its excellent anti-inflammatory and hemolysis inhibitory properties.

Agronomic Investigation of Spray Dispersion of Metal-Based Nanoparticles on Sunflowers in Real-World Environments.

In environmental and agronomic settings, even minor imbalances can trigger a range of unpredicted responses. Despite the widespread use of metal-based nanoparticles (NPs) and new bio-nanofertilizers, their impact on crop production is absent in the literature. Therefore, our research is focused on the agronomic effect of spray application of gold nanoparticles anchored to SiO2 mesoporous silica (AuSi-NPs), zinc oxide nanoparticles (ZnO-NPs), and iron oxide nanoparticles (Fe3O4-NPs) on sunflowers under real-world environments. Our findings revealed that the biosynthetically prepared AuSi-NPs and ZnO-NPs were highly effective in enhancing sunflower seasonal physiology, e.g., the value of the NDVI index increased from 0.012 to 0.025 after AuSi-NPs application. The distribution of leaf trichomes improved and the grain yield increased from 2.47 t ha-1 to 3.29 t ha-1 after ZnO-NPs application. AuSi-NPs treatment resulted in a higher content of essential linoleic acid (54.37%) when compared to the NPs-free control (51.57%), which had a higher determined oleic acid. No NPs or residual translocated metals were detected in the fully ripe sunflower seeds, except for slightly higher silica content after the AuSi-NPs treatment. Additionally, AuSi-NPs and NPs-free control showed wide insect biodiversity while ZnO-NPs treatment had the lowest value of phosphorus as anti-nutrient. Contradictory but insignificant effect on physiology, yield, and insect biodiversity was observed in Fe3O4-NPs treatment. Therefore, further studies are needed to fully understand the long-term environmental and agricultural sustainability of NPs applications.

Synthesis and characterization of calcium and magnesium based oxides and titanates for photocatalytic degradation of rhodamine B: a comparative study.

The current investigation deals with the simple and ecological synthesis of CaO, MgO, CaTiO3, and MgTiO3 for the photocatalytic dilapidation of rhodamine B dye. CaO was procured from chicken eggshell waste by calcination process, while MgO was produced by solution combustion method using urea as a fuel source. Furthermore, CaTiO3 and MgTiO3 were synthesized through an easy and simple solid-state method by mixing thoroughly the synthesized CaO or MgO with TiO2 before calcination at 900 °C. XRD and EDX investigations confirmed the phase formation of the materials. Moreover, FTIR spectra revealed the existence of Ca-Ti-O, Mg-Ti-O, and Ti-O which resembles the chemical composition of the proposed materials. SEM micrographs revealed that the surface of CaTiO3 is rougher with relatively dispersed particles compared to MgTiO3, reflecting a higher surface area of CaTiO3. Diffuse reflectance spectroscopy investigations indicated that the synthesized materials can act as photocatalysts under UV illumination. Accordingly, CaO and CaTiO3 effectively degraded rhodamine B dye within 120 min with a photodegradation activity of 63% and 72%, respectively. In contrast, the photocatalytic degradation activity of MgO and MgTiO3 was much lower, since only 21.39 and 29.44% of the dye were degraded, respectively after 120 min of irradiation. Furtheremore, the photocatalytic activity of the mixture from both Ca and Mg titanates was 64.63%. These findings might be valuable for designing potential and affordable photocatalysts for wastewater purification.

Calcium-Silicate-Incorporated Gellan-Chitosan Induced Osteogenic Differentiation in Mesenchymal Stromal Cells.

Gellan-chitosan (GC) incorporated with CS: 0% (GC-0 CS), 10% (GC-10 CS), 20% (GC-20 CS) or 40% (GC-40 CS) w/w was prepared using freeze-drying method to investigate its physicochemical, biocompatible, and osteoinductive properties in human bone-marrow mesenchymal stromal cells (hBMSCs). The composition of different groups was reflected in physicochemical analyses performed using BET, FTIR, and XRD. The SEM micrographs revealed excellent hBMSCs attachment in GC-40 CS. The Alamar Blue assay indicated an increased proliferation and viability of seeded hBMSCs in all groups on day 21 as compared with day 0. The hBMSCs seeded in GC-40 CS indicated osteogenic differentiation based on an amplified alkaline-phosphatase release on day 7 and 14 as compared with day 0. These cells supported bone mineralization on GC-40 CS based on Alizarin-Red assay on day 21 as compared with day 7 and increased their osteogenic gene expression (RUNX2, ALP, BGLAP, BMP, and Osteonectin) on day 21. The GC-40 CS-seeded hBMSCs initiated their osteogenic differentiation on day 7 as compared with counterparts based on an increased expression of type-1 collagen and BMP2 in immunocytochemistry analysis. In conclusion, the incorporation of 40% (w/w) calcium silicate in gellan-chitosan showed osteoinduction potential in hBMSCs, making it a potential biomaterial to treat critical bone defects.

Biomineralization, dissolution and cellular studies of silicate bioceramics prepared from eggshell and rice husk.

The current investigation aims to replace the synthetic starting materials with biowaste to synthesize and explore three different silicate bioceramics. Pure silica from rice husk was extracted by decomposition of rice husk in muffle furnace followed by alkali treatment and acid precipitation. Raw eggshell and extracted silica were utilized for the preparation of wollastonite, diopside and forsterite by the solid-state method. The TG-DSC analysis shows that the crystallization temperature of wollastonite, diopside and forsterite was found to be 883 °C, 870 °C and 980 °C, respectively. The phase purity of wollastonite was attained at 1100 °C whereas diopside and forsterite were composed of secondary phases even after calcination at 1250 °C and 1300 °C respectively. All three materials behaved differently when exposed to the physiological environment, as wollastonite exhibited remarkable apatite deposition within 3 days whereas a distinct apatite phase was noticed on the surface of diopside after 2 weeks and forsterite shows the formation of apatite phase after five weeks of immersion. The rapid dissolution of Mg2+ ion from forsterite lowered the leaching of silicate ions into the simulated body fluid leading to poor apatite deposition over its surface. Chemical composition was found to plays a key role in the biomineralization ability of these bioceramics. Hemolysis and Lactate Dehydrogenase (LDH) release assays were performed to evaluate the hemocompatibility of silicate ceramics cultured at different concentrations (62.5, 125, and 250 µg/mL) with red blood cells and mononuclear leucocytes (MLs) of mice. The hemolytic activity of all the tested bioceramics was insignificant (less than 1%). The interaction between diopside and mouse multipotent mesenchymal stromal cells (MMSCs) caused a negligible increase in the number of apoptosis-associated Annexin V-binding cells whereas forsterite and wollastonite induced an increase in the number of the apoptotic cells only at the concentration of 250 µg/mL. The LDH assay did not show statistically significant changes in the proliferation of MMSCs after treatment with the bioceramics at the tested concentrations when compared to control (p > 0.05). This finding showed that the death of a part of cells during the first 24 h of incubation did not prevent the proliferation of MMSCs incubated with diopside, forsterite and wollastonite for 72 h.

Biomineralization, mechanical, antibacterial and biological investigation of larnite and rankinite bioceramics.

This work is aimed to develop a biocompatible, bactericidal and mechanically stable biomaterial to overcome the challenges associated with calcium phosphate bioceramics. The influence of chemical composition on synthesis temperature, bioactivity, antibacterial activity and mechanical stability of least explored calcium silicate bioceramics was studied. The current study also investigates the biomedical applications of rankinite (Ca3Si2O7) for the first time. Sol-gel combustion method was employed for their preparation using citric acid as a fuel. Differential thermal analysis indicated that the crystallization of larnite and rankinite occurred at 795 °C and 1000 °C respectively. The transformation of secondary phases into the desired product was confirmed by XRD and FT-IR. TEM micrographs showed the particle size of larnite in the range of 100-200 nm. The surface of the samples was entirely covered by the dominant apatite phase within one week of immersion. Moreover, the compressive strength of larnite and rankinite was found to be 143 MPa and 233 MPa even after 28 days of soaking in SBF. Both samples prevented the growth of clinical pathogens at a concentration of 2 mg/mL. Larnite and rankinite supported the adhesion, proliferation and osteogenic differentiation of hBMSCs. The variation in chemical composition was found to influence the properties of larnite and rankinite. The results observed in this work signify that these materials not only exhibit faster biomineralization ability, excellent cytocompatibility but also enhanced mechanical stability and antibacterial properties.

Review on calcium- and magnesium-based silicates for bone tissue engineering applications.

Bone is a self-engineered structural component of the human body with multifaceted mechanical strength, which provides indomitable support to the effective functioning of the human body. It is indispensable to find a suitable biomaterial for substituting the bone as the bone substitute material requirement is very high due to the rate of bone fracture and infection lead to osteoporosis in human beings increases rapidly. It is not an easy task to design a material with good apatite deposition ability, a faster rate of dissolution, superior resorbability, high mechanical strength, and significant bactericidal activity. Since the synthetic hydroxyapatite was not able to achieve the dahlite phase of hydroxyapatite (natural bone mineral phase), silicates emerged as an alternate biomaterial to meet the need for bone graft substitutes. All silicates do not exhibit the properties required for bone graft substitutes, as their composition and methodology adopted for the synthesis are different. Calcium, magnesium, and silicon play a major role in the formation of bone mineral and their metabolism during bone formation. In this review, the relationship between composition and activity of calcium, magnesium-based silicates have been discussed along with the future scope of these materials for hard tissue engineering applications.

Antibacterial forsterite (Mg2SiO4) scaffold: A promising bioceramic for load bearing applications.

In the current work, forsterite samples with different surface area were investigated for its antibacterial activity. Dissolution studies show that the lower degradation of forsterite compared to other silicate bioceramics, which is a desirable property for repairing bone defects. Forsterite scaffold shows superior compressive strength than the cortical bone after immersion in simulated body fluid. Bactericidal tests indicate that the forsterite had inhibition effect on the growth of clinical bacterial isolates. Forsterite may be a suitable candidate material for load bearing applications with enhanced mechanical properties and lower degradation rate.

Preparation of nanocrystalline forsterite by combustion of different fuels and their comparative in-vitro bioactivity, dissolution behaviour and antibacterial studies.

This study presents different fuels (Glycine and Urea) that can be used to synthesize nanocrystalline forsterite by the sol-gel combustion method. The weight change of precursor during thermal treatment was studied by thermo-gravimetric analysis (TGA). Pure forsterite was characterized by heating microscopy, Fourier transform infrared spectroscopy, X-ray Diffraction, Brunauer-Emmett-Teller, Scanning Electron Microscopy, and Energy dispersive X-ray spectroscopy. The HAP (hydroxyapatite) deposition ability, degradation and dissolution behaviour of forsterite was examined in simulated body fluid (SBF). The combusted forsterite precursor showed distinct thermal behaviour for each fuel when analyzed by heating microscopy. BET analysis showed that the particle size of forsterite synthesized using glycine was 28nm, specific surface area 65.11m2/g and average pore diameter 16.4nm while using urea 1.951µm, 0.939m2/g, and 30.5nm are the respective parameters. The dissolution of forsterite pointed to the consumption of Ca and P ions from SBF, the negligible release of Si ion into the SBF and these ionic interactions with SBF can be altered as per the material properties. The forsterite showed good antibacterial activity against S. aureus but lower activity against E. coli. The bactericidal activity of forsterite indicated that it can be used to inhibit biofilm formation in dental, bone implants and bacterial infection during surgical operations.

Sodium Alginate with PEG/PEO Blends as a Floating Drug Delivery Carrier - In vitro Evaluation.

Purpose: Floating drug delivery system reduces the quantity of drug intake and the risk of overloading the organs with excess drug. Methods: In the present study, we prepared the blends of sodium alginate with polyethylene glycol (PEG) and polyethylene oxide (PEO) as a matrix, sodium hydrogen carbonate as a pore forming agent, methyl cellulose as a binder and barium chloride containing 10% acetic acid as a hardening agent. Different ratios of pore forming agent to the polymer blend was used to prepare the floating beads with different porosity and morphology. Ciprofloxacin hydrochloride was used as a model drug for the release kinetics studies. Results: The beads were characterized by optical and FESEM microscopy to study the morphology and pore dimensions. The results obtained shows decrease in beads size with increase in the concentration of the pore forming agent. The swelling properties of the beads were found to be in the range of 80% to 125%. The release kinetics of the ciprofloxacin from the beads was measured by UV-Visible spectroscopy at λmax of 278nm and the results shows for highly porous beads. Conclusion: By varying the amount of alginate and pore forming agent the release kinetics is found to get altered. As a result, ciprofloxacin hydrochloride release is found to be sustained from the blended beads.

In-vitro bioactivity, biocompatibility and dissolution studies of diopside prepared from biowaste by using sol-gel combustion method.

Diopside was synthesized from biowaste (Eggshell) by sol-gel combustion method at low calcination temperature and the influence of two different fuels (urea, l-alanine) on the phase formation temperature, physical and biological properties of the resultant diopside was studied. The synthesized materials were characterized by heating microscopy, FTIR, XRD, BET, SEM and EDAX techniques. BET analysis reveals particles were of submicron size with porosity in the nanometer range. Bone-like apatite deposition ability of diopside scaffolds was examined under static and circulation mode of SBF (Simulated Body Fluid). It was noticed that diopside has the capability to deposit HAP (hydroxyapatite) within the early stages of immersion. ICP-OES analysis indicates release of Ca, Mg, Si ions and removal of P ions from the SBF, but in different quantities from diopside scaffolds. Cytocompatability studies on human bone marrow stromal cells (hBMSCs) revealed good cellular attachment on the surface of diopside scaffolds and formation of extracellular matrix (ECM). This study suggests that the usage of eggshell biowaste as calcium source provides an effective substitute for synthetic starting materials to fabricate bioproducts for biomedical applications.