Biomaterial scientists design organic bone substitutes based on the biochemical properties of the mimicked tissue to achieve near native functionality. Several non-collagenous proteins in bone are known as intrinsically disordered proteins (IDPs), as they lack detectible ordered domains and a fixed 3D structure under physiological conditions. Many IDPs perform regulatory roles in a range of cellular functions, which motivated us to design two proline-rich disordered peptides (P2 and P6) and augmented them into the SmartBone® (SBN) biohybrid substitute. Recently we reported an improved proliferation and osteogensis of human osteoblasts and mesenchymal stem cells in the composite groups containing peptides (named here as SBN + P2 and SBN + P6) in vitro. To address the effects of these composites on bone formation and biomineralization, this in vivo study investigated their functions in critical size craniotomy defects in 16 domestic pigs after 8 and 16 weeks of healing. For this purpose, we used cone beam computed tomography (CBCT), microCT (µCT), histology, immunohistochemistry, fluorescent labeling of abundant reactive entities (FLARE), synchrotron SAXS/XRD, optical photothermal IR (O-PTIR) microscopy and nanoscale atomic force microscopy-infrared (AFM-IR) analyses. Our results represent new synthetic IDPs as potential candidates for directing bone formation and biomineralization. The SBN + P6 stimulated significantly higher bone formation and biomineralization after 8 weeks of healing compared to other groups indicating its potential in stimulating early biomineralization. After 16 weeks of healing, the SBN + P2 induced significantly higher bone formation and biomineralization compared to other groups indicating its effects on later bone formation and biomineralization processes. The vigorous stretching of amide primary and secondary IR absorbance peaks at 1660 and 1546 cm−1 in the SBN + P2 group verified that this peptide experienced more conformational changes after 16 weeks of implantation with a higher phosphate intensity at 1037 cm−1 compared to peptide 6. Overall, P2 and P6 are promising candidates for bone augmentation strategies in critical clinical applications. We concluded that FLARE and O-PTIR are promising tools in evaluating and diagnosing the biochemical structure of bone tissue and the bone-biomaterial interface.
Today, substantial attention is given to biomaterial strategies for bone regeneration, and among them, there is a growing interest in using immunomodulatory biomaterials. The ability of a biomaterial to induce neo vascularization and macrophage polarization is a major factor in defining its success. Magnesium (Mg)-based degradable alloys have attracted significant attention for bone regeneration owing to their biodegradability and potential for avoiding secondary removal surgeries. However, there is insufficient evidence in the literature regarding the early inflammatory responses to these alloys in vivo. In this study, we investigated the early body responses to Mg-0.45wt%Zn-0.45wt%Ca pin-shaped alloy (known as ZX00 alloy) in rat femora 2, 5, and 10 days after implantation. We used 3D micro computed tomography (µCT), histological, immunohistochemical, histomorphometrical, and small angle X-ray scattering (SAXS) analyses to study new bone formation, early macrophage polarization, neo vascularization, and bone quality at the implant bone interface. The expression of macrophage type 2 biological markers increased significantly after 10 days of Mg alloy implantation, indicating its potential in stimulating macrophage polarization. Our biomineralization results using µCT as well as histological stained sections did not indicate any statistically significant differences between different time points for both groups. The activity of alkaline phosphatase (ALP) and Runt-related transcription factor 2 (Runx 2) biological markers decreased significantly for Mg group, indicating less osteoblast activity. Generally, our results supported the potential of ZX00 alloy to enhance the expression of macrophage polarization in vivo; however, we could not observe any statistically significant changes regarding biomineralization.
Plasma electrolytic oxidation coatings were prepared in aluminate, phosphate, and silicate-based electrolytic solutions using a soft-sparking regime in a multi-frequency stepped process to compare the structure, corrosion, and wear characteristics of the obtained coatings on AZ31 magnesium alloy. The XRD results indicated that all coatings consist of MgO and MgF2, while specific products such as Mg2SiO4, MgSiO3, Mg2P2O7, and MgAl2O4 were also present in specimens based on the selected solution. Surface morphology of the obtained coatings was strongly affected by the electrolyte composition. Aluminate-containing coating showed volcano-like, nodular particles and craters distributed over the surface. Phosphate-containing coating presented a sintering-crater structure, with non-uniform distributions of micro-pores and micro-cracks. Silicate-containing coating exhibited a scaffold surface involving a network of numerous micro-pores and oxide granules. The aluminate-treated sample offered the highest corrosion resistance and the minimum wear rate (5 × 10−5 mm3 N−1 m−1), owing to its compact structure containing solely 1.75% relative porosity, which is the lowest value in comparison with other samples. The silicate-treated sample was degraded faster in long-term corrosion and wear tests due to its porous structure, and with more delay in the phosphate-containing coating due to its larger thickness (30 µm).
Thin films of barium fluoride (BaF2 ), used as a buffer layer between the substrate and the superconducting films, dramatically improve the properties of Y-Ba-Cu-O (YBCO) superconducting films deposited by metalorganic chemical vapor deposition on yttrium-stabilized zirconia (YSZ) substrates. For the as-deposited films at a substrate temperature of 780 °C, the observed transition temperature Tc of 80 K on the BaF2 /YSZ structure represents the highest value reported to date. By using BaF2 buffer layers, superconducting films formed by using a paste of the YBCO powder and a binder, on BaF2 /Si provided a Tc of 80 K.
The purpose of the present study was to provide insights into the interactive effects of β-glycerophosphate (β-GP) molecules on thermosensitive chitosan-based hydrogels, which experience sol–gel transformation around the physiological temperature. This study also investigated the influence of β-GP as a physical cross-linker and pH modifier on chitosan-based hydrogels by Fourier transform infrared spectroscopy with the aim of achieving a better understanding of the effect of β-GP action mechanism. For this purpose, some solutions with 2% (w/v) chitosan were mixed with different amounts of β-GP in the range of 0–20% (w/v) concentrations. It was shown that by further addition of β-GP, the anionic phosphate units of β-GP in combination with the cationic amine units of chitosan could provide electrostatic cross-linking for making chitosan-based hydrogels. All the β-GP–chitosan solutions remained at the physiological pH after complete mixing. In addition, the viability of 3T3 fibroblast cells determined by MTT assay and scanning suggested that the samples containing 12% β-GP would show optimum biological characteristics. This study demonstrated that the synthesized chitosan–β-GP is a biocompatible thermosensitive system, which can be potentially used for tissue engineering applications in different areas, such as cartilage injections, and that injectability and appropriate gelation behavior are important factors.
This study investigated the elicitation effects of several methyl jasmonate (MeJ) concentrations (0, 25, 50, and 100 µM) on various biochemical traits of caraway (Carum carvi L.) callus cultures. The 25 μM MeJ concentration yielded the highest callus growth rate (0.57 mm day−1), total flavonols content (2.58 mg QE g−1 FW) and total carotenoids content (0.04 μg g−1 FW), whereas the highest relative fresh weight (75.72%), total phenolics content (76.90 mg GAE g−1 FW), total flavonoids content (58.49 mg QE g−1 FW) and phenylalanine ammonia lyase activity (3.40 nmol cinnamic acid mg−1 h−1 FW) were obtained with the 50 μM MeJ concentration. The highest antioxidant activity through DPPH assay (8.37%) and malondialdehyde content (7.82 μmol g−1 FW) were observed at 100 μM MeJ. The HPLC conducted 21 days post-elicitation revealed a 5.55-fold increase of carvone accumulation (1.83 μg g−1 DW) at 50 μM MeJ and a 2.7-fold increase (0.62 μg g−1 DW) of limonene at 50 μM MeJ. The optimal dosages applied for carvone and limonene accumulation under MeJ elicitation could be used to scale up the economic production of these elite medicinal compounds through caraway callus cultures.