Osteoblastic Cells Research Articles

Mimicking Osteoid 3D Porous Dense Microfiber Silk Fibroin Embedded Poly(vinyl alcohol) Scaffold for Alveolar Ridge Preservation

Abstract Alveolar ridge loss presents difficulties for implant placement and stability. To address this, alveolar ridge preservation (ARP) is required to maintain bone and avoid the need for ridge augmentation using socket grafting. In this study, a scaffold for ARP was created by fabricating a 3D porous dense microfiber silk fibroin (mSF) embedded in poly(vinyl alcohol) (PVA), which mimics the osteoid template. The research utilized a freeze-thawing technique to create a mimicked osteoid 3D porous scaffold by incorporating different amounts of mSF into the PVA, namely, 1%, 3%, 5%, and 7%. Subsequently, a 3D profilometer machine and a scanning electron microscope (SEM) were employed to examine the morphology and size of the mSF and the mimicked osteoid 3D porous scaffold in all groups. Thermal characteristics and crystalline structure were analyzed before assessing the water contact angle, swelling behavior, degradation, and mechanical properties. The experiment evaluated the biological performance of the mimicked osteoid 3D porous scaffold by examining the efficacy of osteoblast cell adhesion, proliferation, viability, protein synthesis, alkaline phosphatase (ALP) activity, and calcium synthesis. Finally, the ability of osteoblast cells to regulate the osteoid matrix deposition on the osteoid 3D porous scaffold was assessed by mimicking the dynamic bone environment using rat mesenchymal stem cells (RMSC). The findings suggest that incorporating mSF into PVA enhances the interconnective pore size, crystalline structure, and thermal behavior of the mimicked osteoid 3D porous scaffold. The hydrophilicity of PVA decreased with an increase in the proportion of mSF, while a higher proportion of mSF resulted in increased swelling and mechanical characteristics. Incorporating a greater proportion of mSF, specifically 5% and 7%, led to a reduced rate of degradation. The addition of 5% mSF to the PVA 3D porous scaffold resulted in remarkable biological properties and excellent osteoconductive activity.

Dental Pulp Stem Cell Conditioned Medium Enhance Osteoblastic Differentiation and Bone Regeneration.

Cell-free approaches, utilizing mesenchymal stem cell secretome, have promising prospects in various fields of regenerative medicine. In this study, we examined in vitro and in vivo the potential of dental pulp stem cell-conditioned medium (DPSC-CM) for bone regeneration. The secretome of undifferentiated stem cells from dental pulp were collected, and the effects of this DPSC-CM were assessed for osteodifferentiation of osteoblast-like cells (MG-63) and osteoblasts deriving from DPSC. Cell proliferation, alkaline phosphatase (ALP) activity, gene expression of Runt-related transcription factor 2 (Runx2), Bone Sialoprotein (BSP), Osteocalcin (OCN), and extracellular matrix mineralization were evaluated. The rat caudal vertebrae critical size defect model was to investigate the effect of DPSC-CM in vivo. Results showed that DPSC-CM induced cell growth, and increased ALP activity and the expression of key marker genes at an early stage of osteoblastic differentiation compared to control. A rat bone defect model was used to illustrate the effect of DPSC-CM in vivo. The bone density within the defects were improved using conditioned medium, even though there was no significant difference between the control and DPSC-CM groups. The analysis of DPSC-CM by human growth factor antibody array revealed the presence of several factors involved in osteogenesis. Taken together, these findings indicate that DPSC-CM is a promising therapeutic candidate for bone regenerative therapy, accelerating the maturation of osteoblastic cells. And even though safety and efficiency of DPSC-CM have to be confirmed in preclinical studies, these results represent a first step toward clinical application.

Quantitative Analysis of Deer Bone Hydroethanolic Extract Using Label-Free Proteomics: Investigating Its Safety and Promoting Effect on Mouse Embryonic Osteoblastic Progenitor Cell Proliferation

Background: Deer bone is rich in proteins and free amino acids, offering high nutritional value and benefits such as strengthening bones and antioxidant properties. However, the development and utilization of deer bone resources are limited, and the safety evaluation of health foods is incomplete. Methods: We established a hydrogen ethanol extraction method for deer bone and analyzed the components of the deer bone hydroethanolic extract (DBHE) using liquid chromatography–tandem mass spectrometry (LC-MS/MS), gas chromatography–mass spectrometry (GC-MS), and inductively coupled plasma mass spectrometry (ICP-MS). Results: Using Label-free proteomics technology, we identified 69 proteins and 181 peptides. We also quantified 16 amino acids, 22 fatty acids, and 17 inorganic elements. Finally, we evaluated the safety of DBHE both in vitro and in vivo. The results indicated that DBHE did not exhibit any toxic effects at the doses we tested and can promote the proliferation of mouse embryonic osteoblastic progenitor cells (MC3T3-E1), demonstrating potential efficacy against osteoporosis and arthritis. Conclusions: This study provides a theoretical basis for the quality control, processing, and resource development of deer bone.

Improved visualisation of ACP-engineered osteoblastic spheroids: a comparative study of contrast-enhanced micro-CT and traditional imaging techniques

This study investigates osteoblastic cell spheroid cultivation methods, exploring flat-bottom, U-bottom, and rotary flask techniques with and without amorphous calcium phosphate (ACP) supplementation to replicate the 3D bone tissue microenvironment. ACP particles derived from eggshell waste exhibit enhanced osteogenic activity in 3D models. However, representative imaging of intricate 3D tissue-engineered constructs poses challenges in conventional imaging techniques due to notable scattering and absorption effects in light microscopy, and hence limited penetration depth. We investigated contrast-enhanced micro-CT as a methodological approach for comprehensive morphological 3D-analysis of thein-vitromodel and compared the technique with confocal laser scanning microscopy, scanning electron microscopy and classical histology. Phosphotungstic acid and iodine-based contrast agents were employed for micro-CT imaging in laboratory and synchrotron micro-CT imaging. Results revealed spheroid shape variations and structural integrity influenced by cultivation methods and ACP particles. The study underscores the advantage of 3D spheroid models over traditional 2D cultures in mimicking bone tissue architecture and cellular interactions, emphasising the growing demand for novel imaging techniques to visualise 3D tissue-engineered models. Contrast-enhanced micro-CT emerges as a promising non-invasive imaging method for tissue-engineered constructs containing ACP particles, offering insights into sample morphology, enabling virtual histology before further analysis.

Sonochemical synthesis characterization and biocompatibility studies of polymeric composites for biomedical applications

In this work the in situ synthesis of mixtures of Polymethylmethacrylate (PMMA), High Density Polyethylene (HDPE) and Polypropylene (PP) with biocompatible ceramics was carried out, using high frequency ultrasound as an energy source. Starting from calcium hydroxide and ammonium hydrogen phosphate, two different crystalline phases were obtained: hydroxyapatite (HA) and calcite (CaCO3). FTIR and XRD studies showed that for the materials prepared with HDPE, only CaCO3 was formed during the synthesis process. When Polypropylene used, mixtures of HA with CaCO3 were obtained, while for the materials synthesized with Polymethylmethacrylate, only pure HA was obtained. DSC analyses did not reveal any significant changes in the melting temperatures of HDPE and PP, nor in the glass transition temperature of PMMA with the incorporation of HA and CaCO3, in the polymeric matrices. TGA, showed that the decomposition temperatures of these materials increased with respect to those of the polymers alone, indicating a significant thermal stability. Biocompatibility tests with osteoblastic cells and Scanning Electron Microscopy (SEM) studies showed that the composites of PMMA with HA, PP with HA/CaCO3 and HDPE with CaCO3, are capable of promoting cell adhesion and proliferation mechanisms, revealing potential applications as biomaterials for bone replacement.

Osteoblast Growth in Quaternized Silicon Carbon Nitride Coatings for Dental Implants.

The demand for dental implants has increased, establishing them as the standard of care for replacing missing teeth. Several factors contribute to the success or failure of an implant post-placement. Modifications to implant surfaces can enhance the biological interactions between bone cells and the implant, promoting better outcomes. Surface coatings have been developed to electrochemically alter implant surfaces, aiming to reduce healing time, enhance bone growth, and prevent bacterial adhesion. Quaternized silicon carbon nitride (QSiCN) is a novel material with unique electrochemical and biological properties. This study aimed to assess the influence of QSiCN, silicon carbide nitride (SiCN), and silicon carbide (SiC) coatings on the viability of osteoblast cells on nanostructured titanium surfaces. The experiment utilized thirty-two titanium sheets with anodized TiO2 nanotubes featuring nanotube diameters of 50 nm and 150 nm. These sheets were divided into eight groups (n = 4): QSiCN-coated 50 nm, QSiCN-coated 150 nm, SiCN-coated 50 nm, SiCN-coated 150 nm, SiC-coated 50 nm, SiC-coated 150 nm, non-coated 50 nm, and non-coated 150 nm. Preosteoblast MC3T3-E1 Subclone 4 cells (ATCC, USA) were used to evaluate osteoblast viability. After three days of cell growth, samples were assessed using scanning electron microscopy (SEM). The results indicated that QSiCN coatings significantly increased osteoblast proliferation (p < 0.005) compared to other groups. The enhanced cell adhesion observed with QSiCN coatings is likely due to the positive surface charge imparted by N+.

Osteoprotegerin secretion and its inhibition by RANKL in osteoblastic cells visualized using bioluminescence imaging

Bone remodeling is regulated by the interaction between receptor activator of nuclear factor kappa-B ligand (RANKL) and its receptor RANK on osteoblasts and osteoclasts, respectively. Osteoprotegerin (OPG) is secreted from osteoblasts and inhibits osteoclast differentiation by acting as a decoy receptor for RANKL. Despite its importance, the mechanism underlying the secretion of OPG remains poorly understood. Here, we applied a method of video-rate bioluminescence imaging using a fusion protein with Gaussia luciferase (GLase) and visualized the secretion of OPG from living mouse osteoblastic MC3T3-E1 cells. The bioluminescence imaging revealed that the secretion of OPG fused to GLase (OPG-GLase) occurred frequently and widely across the cell surface. Notably, co-expression of RANKL significantly reduced the secretion of OPG-GLase, indicating an inhibitory role of RANKL on OPG secretion within cells. Further imaging and biochemical analyses using deletion mutants of OPG and RANKL, as well as RANKL mutants that cause autosomal recessive osteopetrosis, demonstrated the essential role of protein-protein interaction between OPG and RANKL in the inhibition of OPG secretion. Treatment with proteasome inhibitors resulted in increased levels of OPG in both culture medium and cell lysates. However, the fold-increase of OPG was similar regardless of the presence or absence of RANKL, suggesting that the regulation of OPG secretion by RANKL is independent of proteasome activity. This report visualized the secretion of OPG from living cells and provided evidence for a novel intracellular inhibitory effect of RANKL on OPG secretion.

Dhvar5- and MSI78-coated titanium are bactericidal against methicillin-resistant Staphylococcus aureus, immunomodulatory and osteogenic

Infection is one of the major issues associated with the failure of orthopedic devices, mainly due to implant bacterial colonization, biofilm formation, and associated antibiotic resistance. Antimicrobial peptides (AMP) are a promising alternative to conventional antibiotics given their broad-spectrum of activity, low propensity to induce bacterial resistance, and ability to modulate host immune responses. Dhvar5 (LLLFLLKKRKKRKY) and MSI78 (GIGKFLKKAKKFGKAFVKILKK) are two AMP with broad-spectrum activity against bacteria, including methicillin-resistant Staphylococcus aureus (MRSA), one of the most problematic etiologic agents in Orthopedic Devices-Related Infections (ODRI). This work aims to evaluate the bactericidal, immunomodulatory and osteogenic potential of Dhvar5- and MSI78-coated titanium surfaces (AMP-Ti). Two AMP-Ti surfaces were successfully obtained by grafting Dhvar5 (0.8 ± 0.1 µM/mm2) or MSI78 (0.5 ± 0.3 µM/mm2) onto titanium substrates through a polydopamine layer. Both AMP-Ti were bactericidal against MRSA, eradicating bacteria upon contact for 6 h in a culture medium supplemented with human plasma proteins. The AMP-Ti immunomodulatory potential was evaluated using human primary macrophages, by assessing surfaces capacity to induce pro-/anti-inflammatory (M1/M2) markers and cytokines. While in naïve conditions both AMP-Ti surfaces slightly promoted the M2 marker CD163, in response to LPS and IFN-γ (simulating a bacterial infection), both AMP increased the M1 marker CCR7 and enhanced macrophage secretion of pro-inflammatory IL-6 and TNF-α cytokines, particularly for Ti-MSI78 surfaces. Additionally, both AMP-Ti surfaces were cytocompatible and promoted osteoblastic cell differentiation. This proof-of-concept study demonstrated the high potential of Dhvar5- and MSI78-Ti as antimicrobial coatings for ODRI prevention. STATEMENT OF SIGNIFICANCE: This study investigates the bactericidal effects of the antimicrobial peptides Dhvar5 and MSI78, immobilized on titanium (Ti) surfaces through a polydopamine coating, aiming at the prevention of Orthopedic-Device Related Infections (ODRIs). The developed coatings displayed MRSA-sterilizing activity, while revealing an immunomodulatory potential towards macrophages and promoting osteoblastic cell differentiation. This strategy allows a quick and easy immobilization of high quantities of AMP, unlike most other approaches, thus favoring its clinical translation.

The effect of resveratrol on the viability of rat bone marrow mesenchymal stem cells

Background and aimResveratrol is a plant polyphenol that has antioxidant properties. In this research, in order to better understand the role of resveratrol on bone, the effect of different doses of resveratrol on the viability, proliferation, differentiation and biochemical factors of rat bone marrow mesenchyme stem cells (MSCs) was investigated. MethodsTo evaluate the ability of proliferation and differentiation, the MSCs were cultured up to three passages and were placed in culture media without osteogenic stimulation. At first to investigation the proliferation ability, the viability was evaluated by MTT test and trypan blue staining with 7.5, 15 and 30 mg/L treatment doses after 12, 24 and 36 hours. Then, to investigation the differentiation ability, the effect of 7.5, 15, and 30 mg/L treatment doses was assessed on the viability, electrolyte levels, and the activity of LDH, AST, ALP and ALT enzymes in differentiated cells after 5, 10, 15, and 21 days. To investigation the differentiation ability, the effect of 7.5, 15, and 30 mg/L treatment doses was assessed on the viability, electrolyte levels, and the activity of Lactate dehydrogenase (LDH), Aspartate aminotransferase (AST), Alkaline phosphatase (ALP) and alanine aminotransferase (ALT) enzymes in differentiated cells after 5, 10, 15, and 21 days. The data were analyzed by ANOVA and Tukey's test, and p<0.05 was considered as a significant level. ResultsThe obtained results showed that in mesenchyme cells, the level of viability does not change in low dose of resveratrol, however increase in the time followed by decrease in the activity of metabolic enzymes (ALP, ALT and AST P<0.5) and subsequently viability (p<0.05), and this trend was reversed with increasing resveratrol concentration (significant differences among 5 days and others). ConclusionThe effect of resveratrol depends on the type of cell and the treatment conditions, so it was shown in this study that the sensitivity of mesenchymal cells is higher than that of osteoblast cells. In addition, high doses of resveratrol had a positive effect on bone differentiation, so high doses of resveratrol can have a beneficial role on the health of mesenchymal cells and their differentiation into osteoblasts.

Uniaxial static strain enhances osteogenic and angiogenic potential under hypoxic conditions in distraction osteogenesis

ObjectiveBone incision leads to interrupted and sluggish blood flow in the process of distraction osteogenesis (DO), creating a hypoxia (0–2% oxygen tension) at the center of the bone callus. This hypoxia is critical in the coupling of osteogenesis and angiogenesis during DO. This study aimed to investigate the effect of Uniaxial Static Strain (USS) on osteogenesis in osteoblasts under hypoxic conditions, with a focus on the expression of osteogenic markers and angiogenic factors.MethodsThe USS was made by a multi-unit tension compression device.Osteoblasts were subjected to 10% USS made under hypoxic conditions to mimic the process of DO in vitro. The cell proliferation, alkaline phosphatase (ALP) activity, mineralized nodule formation, and expression of osteogenic and angiogenic markers were evaluated by using a CCK-8 assay, alkaline phosphatase (ALP) staining, ALP activity assay, alizarin red S staining, qRT-PCR, Western blotting and ELISA.ResultsHypoxia inhibited osteoblast cell proliferation, ALP activity, mineralized nodule formation, and the expression of runt-related transcription factor 2 (Runx- 2), osteopontin(OPN), osteocalcin (OCN), collagen type I (Col1a1). Conversely, hypoxia upregulated the expression of hypoxia-inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor (VEGF), which are associated with angiogenesis. However, the application of USS enhanced osteoblasts’ osteogenic capacity and upregulated angiogenic factors under hypoxic conditions.ConclusionUSS can enhance osteogenesis in osteoblasts under hypoxic conditions. Moreover, it may stimulate angiogenesis by promoting the expression of VEGF, which further contributes to bone formation. This finding provides important implications for understanding the mechanisms involved in bone regeneration and may have clinical applications in optimizing the effectiveness of DO techniques.

Synergistic integration of plant derived galactomannan and MXene to produce multifunctional nanocomposites with antibacterial and osteogenic properties

This study proposes to combine plant-derived galactomannan and MXene to form an organic-inorganic hydrogel network with integrative antibacterial and osteogenic properties. Oxidized galactomannan (OxGa) was blended with N-succinyl chitosan (NSC) and varying concentrations of Mxene, owing to its high reactivity and biocompatibility. The results indicated that Mxene-loaded OxGa/NSC scaffolds exhibited biomimetic topography, adequate mechanical features and excellent physicochemical properties with strong antibacterial properties against pathogenic microbes. The cell culture experiments showed dose-dependent cytotoxic behaviour. An optimized MXene content of ≤ 4 mg/ml revealed no cytotoxicity and augmented the proliferation of osteoblast cells. Similar results were obtained in gene expression analysis where ALP, COL1A1, RUNX2, and SOX2 genes showed up-regulation. In addition, the chorioallantoic membrane (CAM) assay also illustrated augmentation of angiogenesis without any toxicity. The overall results highlight the dual performance of OxGa/NSC-MXene scaffolds to exhibit anti-infection with simultaneous osteogenic properties, which is purely determined by the concentration of MXene. Therefore, regulating MXene concentration can serve as a chemical switch in imparting adequate antibacterial functions with the ability to allow new osseous generation. Our findings on the newly designed Mxene-loaded OxGa/NSC scaffolds can be a promising biomaterial for bone-related infections with a concurrent ability to promote osteogenesis.

Mimicking the Architecture and Dissolution Chemistry of Cancellous Bone Tissue to Optimize the Biocompatibility of Bioactive Scaffolds.

Synthesis of mechanically stable porous scaffolds with an architecture analogous to cancellous bone tissue poses significant challenges to bioactive glass (BG) based scaffolds. This is primarily due to densification and crystallization of the BG's during heat treatment. This study presents a modified BG series (42SiO2-xTiO2-24Na2O-21CaO-13P2O5, where x = 8 and 16 TiO2). TiO2 replaced the SiO2 concentration in the glass and was incorporated due to its biocompatibility and influence on glass structure. Material characterization determined that TiO2 did not induce crystallization within the glass but did increase the glass transition temperature (Tg) from 520°C to 600°C thereby indicating a more stable network connectivity. Scaffolds were synthesized using the foam replication method, resulting in scaffolds with a pore size of approximately 500 μm with the BG-4 composition (30SiO2-12TiO2-24Na2O-21CaO-13P2O5) retaining its amorphous character post-heat treatment. Scaffold ion release was monitored over 5-60 days in simulated body fluid (SBF). Si4+ release was found to decrease, while Ca2+ levels increased in SBF as TiO2 replaced SiO2 within the glass series. Cytocompatibility studies revealed that MC3T3 Osteoblast cells proliferated on the BG-4 scaffold surface and at its interface within culture media, and cell numbers were not significantly reduced.

Frequency and amplitude dependence of nuclear displacement and phase delay in mechanical vibrations for determining cellular natural frequency

Cultured cells biochemically respond to mechanical vibrations. However, the mechanisms of sensing mechanical vibrations and transducing biochemical responses remain unclear. A previous study reported that the expression of the alkaline phosphatase gene of osteoblastic cell under mechanical vibrations peaks at 50 Hz, which seems like a resonance curve in the mechanical vibration theory. Since forced displacement excitation is a dynamic mechanical stimulus that differs from other static mechanical stimuli in that an external force is equivalent to inertia, force is apparently exerted on the mass element by considering the equation of motion. In this study, the method for obtaining the change of a nucleus’s relative displacement to an excited dish was refined, and the frequency and acceleration amplitude dependence of the nucleus’s relative displacement and phase delay under mechanical vibrations were demonstrated by regarding a cell model as a vibration system. The change of the relative displacement of a HeLa nucleus to an excited dish decreases with increasing frequency in the 12.5–100 Hz range at 0.5 G and increases with increasing acceleration amplitude in the 0.5–2.0 G range at 50 Hz. Phase reversal occurs between 12.5 Hz and 50 Hz, which suggests the existence of the natural frequency of the cell between 12.5 Hz and 50 Hz. The tension estimated from the nucleus’s relative displacement change in this method was 2.3–10 pN and can be a biochemical response of the mechanotransducer. These findings can contribute to clarifying the mechanism of cell mechanotransduction in dynamic mechanical stimuli.

Green Synthesis and Characterization of Hydroxyapatite Nanorods with Enhanced Antibacterial and Anticancer Properties

This study reports the microwave synthesis of hydroxyapatite nanorods using Lantana camara L. flower extract (LCF-HA) and characterized by FTIR, XRD, TEM and SEM-EDX techniques. The extract coating on the hydroxyapatite nanorods improves the antibacterial efficacy against Gram-positive bacteria. The study also evaluates the mitochondrial membrane potential (MMP), generation of reactive oxygen species (ROS) and capacity to induce apoptosis of LCF-HA. It shows that LCF-HA efficiently inhibits the growth of osteosarcoma cells (MG-63) in a dose-dependent manner. The efficacy of LCF-HA in inducing apoptosis in osteosarcoma cells is established by dual acridine orange/ethidium bromide (AO/EB) fluorescence labelling. Additionally, LCF-HA induces an increase in reactive oxygen species (ROS) generation and a reduction in mitochondrial membrane potential (MMP) levels. The findings indicate that derived LCF-HA has significant anticancer efficacy by inducing apoptosis in osteosarcoma cells through enhanced ROS generation, suggesting that derived LCF-HA may serve as a promising therapeutic target for bone cancer cell lines. Furthermore, the study demostrates excellent compatibility with osteoblast cells, highlighting its potential for bone regeneration applications.

KEFIRAN in vitro biological activity on enterocytes and mesodermal origin cell lines: Focus on adenocarcinoma cells HT29

Kefiran, the exopolysaccharide found in the fermented milk named “kefir” has been receiving attention over the last years due to its interesting biological and functional properties. It is a non-digestible branched glucogalactan with a molecular weight higher than 1 106 Da. Kefiran was isolated from kefir grains and its biological activity was assessed on different cell lines. Proliferation (trypan blue exclusion assay), mitochondrial activity (MTT assay), viability (Alive-Vulnerable-Dead model, AVD) and cell death were studied. It was found that proliferation was not inhibited in mesodermal cell line MC3 (osteoblast cell line) or in not-invasive colonocytes (Caco-2/TC7 cell line); on the contrary, the high invasive HT29 cell line and the UMR106 osteosarcoma cell line were sensitive to kefiran 0.5 mg/mL, and a significant diminishment on cell proliferation was found by trypan blue dye exclusion assay. Kefiran maintained mitochondrial activity while increased cell death processes in a dose-dependent way in HT29 cell line. When viability was assessed by AVD model, it was found that kefiran increased vulnerability status of neoplasic cell line HT29. In summary, in the present work, we shown that kefiran has a different effect on tumor and non-tumor cell lines. Our results contribute to the understanding of kefiran effect on the empirical antitumoral effect attributed to kefir consumption.

Bone Healing via Carvacrol and Curcumin Nanoparticle on 3D Printed Scaffolds.

Carvacrol is a potent antimicrobial and anti-inflammatory agent, while curcumin possesses antioxidant, anti-inflammatory, and anticancer properties. These phytochemicals have poor solubility, bioavailability, and stability in their free form. Nanoencapsulation can reduce these limitations with enhanced translational capability. Integrating nanocarriers with 3D-printedcalcium phosphate (CaP)scaffolds presents a novel strategy for bone regeneration. Carvacrol and curcumin-loaded nanoparticles (CC-NP) synthesized with melt emulsification produced negatively charged, monodispersed particles with a hydrodynamic diameter of ≈127nm. Their release from the scaffold shows a biphasic release under physiological and acidic conditions. At pH 5.0, the CC-NP exhibits a 53% release of curcumin and nearly 100% release of carvacrol, compared to 19% and 36% from their respective drug solutions. At pH 7.4, ≈40% of curcumin and 76% of carvacrol releases, highlighting their pH-sensitive release mechanism. In vitro studies demonstrate a 1.4-fold increase in osteoblast cell viability with CC-NP treatment. CC-NP exhibit cytotoxic effects against osteosarcoma cells, reducing cell viability by ≈2.9-fold. The antibacterial efficacy of CC-NP evaluated against Staphylococcus aureus (SA) and Pseudomonas aeruginosa (PA) exhibiting 98% antibacterial efficacy. This approach enhances therapeutic outcomes and minimizes the potential side effects associated with conventional treatments, paving the way for innovative applications in regenerative medicine.

Screening of active components of melastoma dodecandrum lour. against diabetic osteoporosis using cell membrane chromatography-mass spectrometry.

Melastoma dodecandrum Lour. (MD), a traditional botanical drug known for its hypoglycemic, antioxidant, and anti-inflammatory properties, is commonly used to treat diabetes, osteoarthritis, and osteoporosis. However, its specific active components against diabetic osteoporosis remain unclear. This study aimed to identify the key active components in MD using cell membrane chromatography coupled with mass spectrometry and validate their effects in vitro. An AGEs-induced osteoblast injury model was established. MTT assays measured cell viability, and ALP activity was assessed using a biochemical kit. Western blotting was employed to detect the expression levels of osteoblast-related proteins OCN and RUNX2 and the AGE receptor protein RAGE. ELISA was used to determine the levels of SOD, MDA, CAT, and GPx. PCR quantified TNF-α expression to evaluate the protective effects and potential mechanisms of MD. The AGEs-induced osteoblast cell membrane chromatography-mass spectrometry method facilitated the rapid identification of potentially active compounds based on their affinity for the osteoblast cell membrane. Cell experiments further validated the activity of the characteristic component isovitexin. MD significantly improved cell viability in AGEs-damaged osteoblasts, enhanced ALP, SOD, CAT, and GPx activities, reduced MDA levels, increased OCN and RUNX2 protein expression, and decreased TNF-α mRNA and RAGE protein expression. Cell membrane chromatography identified 20 chemical constituents, including 13 flavonoids, 4 organic acids, 1 phenylpropanoids, 1 terpenoids, and 1 alkaloid. Cell experiments have confirmed that isovitexin has significant protective activity against osteoblasts and can inhibit the expression of specific receptor RAGE on the osteoblast membrane, consistent with the effect of MD. MD and its active component, isovitexin, provide protective effects against AGEs-induced osteoblast injury, offering a basis for subsequent drug development.

Zibibbo Grape Seeds’ Polyphenolic Profile: Effects on Bone Turnover and Metabolism

Background: The consumption of seeds as food has become increasingly common due to their numerous health benefits. Among these, the seeds of the Zibibbo grape from Pantelleria, a native species of southern Italy, remain largely unexplored and are usually considered waste material from viticulture. Nevertheless, Zibibbo grape seeds may offer health benefits, particularly for the elderly and people with metabolic disorders, due to their potential content of beneficial compounds such as polyphenols. Methods: The Zibibbo grape seeds extract (ZSE) was characterized using UV-visible spectrophotometry and HPLC chromatography. The antioxidant activity of ZSE was measured by different colorimetric assays and Electronic Paramagnetic Resonance (EPR). Additionally, specific in vitro tests were conducted on human osteoblast cell lines (Saos-2 and MG63) aiming to evaluate the ZSE’s effects on bone turnover and metabolism. Western blotting was used to assess the impact on specific proteins and pathways related to bone health. Results: The ZSE contained almost ~3 mg/mL of carbohydrates and phenolic compounds, including rutin (~6.4 ppm) and hesperidin (~44.6 ppm). The extracts exhibited an antioxidant activity greater than 90% across all tests performed. Moreover, the Zibibbo seed extracts exerted a significant proliferative effect on the Saos-2 cell human osteoblast-like cell line, also modulating the phosphorylation of specific kinases involved in cell health and metabolism. Conclusions: Zibibbo grape seeds are rich in phenolic compounds, especially flavonoids with strong antioxidant and free radical scavenging properties. ZSE demonstrated beneficial effects on bone metabolism and osteoblast proliferation, suggesting potential for counteracting osteodegenerative conditions like osteoporosis. If confirmed through further studies, Zibibbo grape seed phenolic compounds could serve as an adjunctive therapy for osteoporosis, helping to slow aging and bone degeneration.

Enhanced bipolar electrode electrochemiluminescence biosensor for ultrasensitive monitoring of alkaline phosphatase activity during osteoblast differentiation by integrating electrocatalytic and enzymatic strategies

Alkaline phosphatase (ALP) is an important biomarker that reflects osteoblast activity and bone growth. Accurately detecting ALP activity is of pivotal clinical significance for the diagnosis and differentiation of bone system diseases. In this work, an enhanced bipolar electrode electrochemiluminescence (BPE-ECL) biosensor based on enzyme catalysis and electrocatalysis was proposed for ultrasensitive detection of ALP in osteoblasts. Carbon nitride nanosheets (CNNS) were utilized as electrocatalysts to facilitate the electrochemical reaction of bipyridine ruthenium (Ru(bpy)32+) and tripropylamine (TPrA) in the reporting cell. Meanwhile, in the sensing cell, the target ALP catalyzed the conversion of p-aminophenyl phosphate monohydrate into p-aminophenol (p-AP). The p-AP was subsequently oxidized at the anode of the driving electrode, producing electrons and increasing the Faradaic current of BPE. The dual-signal amplification strategy, combining electrocatalysis and enzyme catalysis, resulted in a 10-fold enhancement of the ECL intensity of Ru(bpy)32+/TPrA. The BPE-ECL biosensor achieved ultrasensitive detection of ALP with a detection limit as low as 1.9×10−6 U/L, and was successfully applied to monitor ALP activity in osteoblast cells. Additionally, a portable smartphone imaging was developed for the visual detection of ALP. This research introduces an innovative BPE-ECL biosensor for the monitoring of ALP activity and point-of-care testing (POCT) in osteoblasts in clinical settings.

Fabrication and comprehensive evaluation of Zr-based bulk metallic glass matrix composites for biomedical applications

AbstractThe aim of this study is to fabricate Zr-based bulk metallic glass matrix composites (BMG-MCs) for biomedical usage and subject them to a comprehensive and farreaching analysis with respect to their mechanical properties, biocorrosion resistance, biocompatibility, and interactions with biofilms that all may arise from their chemical compositions and unusual disordered internal structure. In this study, we fabricate Zr40Ti15Cu10Ni10Be25, Zr50Ti15Cu10Ni10Be25, and Zr40Ti15Cu10Ni5Si5Be25 alloys and confirm their glassy matrix nature through differential scanning calorimetry (DSC) and scanning electron microscopy (SEM) analyses. The mechanical properties, assessed via nanoindentation, demonstrate the high hardness, strength, and elasticity of the produced materials. Corrosion resistance is investigated in simulated body fluid, with Zr-based BMG-MCs exhibiting superior performance compared to conventional biomedical materials, including 316L stainless steel and Ti6Al4V alloy. Biocompatibility is assessed using human fetal osteoblastic cell line hFOB 1.19, revealing low levels of cytotoxicity. The study also examines the potential for biofilm formation, a critical factor in the success of biomedical implantation, where bacterial infection is a major concern. Our findings suggest, as never reported before, that Zr-based BMG-MCs, with their unique composite glassy structure and excellent physicochemical properties, are promising candidates for various biomedical applications, potentially offering improved performance over traditional metallic biomaterials.