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Keywords = hFOB 1.19

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20 pages, 14247 KiB  
Article
Comparison of Primary Human Osteoblast-like Cells and hFOB 1.19 Cells: Contrasting Effects of Proinflammatory Cytokines
by Juliana Franziska Bousch, Christoph Beyersdorf, Katharina Schultz, Matthis Schnitker, Christoph Viktor Suschek and Uwe Maus
Cells 2025, 14(16), 1264; https://doi.org/10.3390/cells14161264 - 15 Aug 2025
Viewed by 312
Abstract
Proinflammatory cytokines such as IL-1β, IL-6, and TNF-α are key mediators of inflammatory bone loss and are commonly described as inhibitors of osteoblast function. However, their effects on osteogenesis remain controversial, likely due to the differences in the cell models and experimental settings [...] Read more.
Proinflammatory cytokines such as IL-1β, IL-6, and TNF-α are key mediators of inflammatory bone loss and are commonly described as inhibitors of osteoblast function. However, their effects on osteogenesis remain controversial, likely due to the differences in the cell models and experimental settings in in vitro studies. We recently showed that these cytokines significantly enhanced the mineralization of primary human osteoblast-like cells (OBs). Here, we provide the first analysis of cytokine effects on the osteogenesis of the widely used human osteoblastic cell line hFOB 1.19 and compare them to primary OBs. Unexpectedly, all three cytokines significantly inhibited mineralization in hFOB 1.19 cells without affecting the proliferation. IL-1β and TNF-α also suppressed ALP activity, whereas IL-6 acted ALP-independent but increased the osteogenic marker expression despite the reduced mineralization, indicating a possible uncoupled differentiation and mineralization. Morphological and transcriptional analyses indicated that hFOB 1.19 cells represent an earlier osteogenic differentiation stage, while primary OBs show phenotypic heterogeneity and donor-dependent expression profiles. These data demonstrate that proinflammatory cytokines can have severely different effects on the osteogenesis of different cell models, supported by the highly contradictory findings reported in the literature. Nevertheless, elucidating the mechanisms underlying the inhibition of osteogenesis in hFOB 1.19 cells may provide important insights into the cell model and differentiation-stage-specific cytokine effects. Full article
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16 pages, 1210 KiB  
Article
Perilla Seed Meal Extract Enriched with Rosmarinic Acid and Luteolin: Natural Active Pharmaceutical Ingredients (NAPIs) for Osteoprotective Effects
by Thanawat Pattananandecha, Sutasinee Apichai, Treethip Sukkho, Jetsada Ruangsuriya, Fumihiko Ogata, Naohito Kawasaki and Chalermpong Saenjum
Antioxidants 2025, 14(8), 973; https://doi.org/10.3390/antiox14080973 - 8 Aug 2025
Viewed by 402
Abstract
Perilla seed meal (PSM) is a waste biomass of perilla seed extraction that retains flavonoid and phenolic compounds. In this study, we aimed to investigate the potential of PSM extracts (PSMEs) from Perilla frutescens (L.) Britton as a sustainable source of natural active [...] Read more.
Perilla seed meal (PSM) is a waste biomass of perilla seed extraction that retains flavonoid and phenolic compounds. In this study, we aimed to investigate the potential of PSM extracts (PSMEs) from Perilla frutescens (L.) Britton as a sustainable source of natural active pharmaceutical ingredients (NAPIs) containing rosmarinic acid and luteolin for promoting bone health. PSMEs were obtained through shaking incubation and ultrasonic extraction, with 40% ethanol (PS-E40) and 80% ethanol (PS-E80) being found to be the most effective solvents. The effects of PSMEs on bone formation markers were evaluated in human fetal osteoblast cells (hFOB 1.19) using bone formation parameters. The results demonstrated that PS-E40 and PS-E80 extracts significantly increased alkaline phosphatase (ALP) activity, osteocalcin (OC) production, and osteoprotegerin (OPG) levels while concurrently reducing receptor activator of nuclear factor kappa-Β ligand (RANKL) and reactive oxygen species (ROS) production in a dose-dependent manner, particularly at 100 µg/mL on day 7 and 50 and 100 µg/mL on day 14 of the co-incubation period. Moreover, Alizarin Red S staining demonstrated that PS-E40 enhanced calcium deposition in both normal and osteogenic media, further supporting the effect of PSMEs on mineralization and osteoblast differentiation. Our findings suggest that PSMEs rich in rosmarinic acid and luteolin enhance bone health by promoting osteoblast activity and reducing osteoclastogenesis. Full article
(This article belongs to the Special Issue Bioactive Antioxidants from Agri-Food Wastes)
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12 pages, 728 KiB  
Article
Comparison of Microhardness and Depth of Cure of Six Bulk-Fill Resin Composites
by Tomislav Skrinjaric, Kristina Gorseta, Jelena Bagaric, Petra Bucevic Sojcic, Jakov Stojanovic and Luc A. M. Marks
J. Compos. Sci. 2025, 9(8), 418; https://doi.org/10.3390/jcs9080418 - 5 Aug 2025
Viewed by 275
Abstract
Background. Physicomechanical properties and clinical service of bulk-fill composites depend on their adequate polymerization and depth of cure. Some manufacturers claim that these composites can be adequately cured when used in bulks exceeding 4 mm. Objective. The aim of this study was to [...] Read more.
Background. Physicomechanical properties and clinical service of bulk-fill composites depend on their adequate polymerization and depth of cure. Some manufacturers claim that these composites can be adequately cured when used in bulks exceeding 4 mm. Objective. The aim of this study was to compare Vickers microhardness (VMH) and depth of cure (DOC) of six contemporary bulk-fill resin composites at depths of 4 mm and 6 mm. Material and methods. Six bulk-fill composites were evaluated in this study: 1. Tetric EvoCeram Bulk (Ivoclar Vivadent, Schaan, Liechtenstein), (TEC); 2. Filtek Bulk Fill Posterior (3M ESPE Dental Products Division, St. Paul, MN, USA), (FBF); 3. Filtek One Bulk Fill (3M ESPE Dental Products Division, St. Paul, MN, USA, (FOB); 4. SonicFill 2 (Kerr, Orange, CA, USA), (SF2); 5. Admira Fusion X-tra (Voco, GmbH, Cuxhaven, Germany), (AFX); 6. GrandioSO X-tra (Voco, GmbH, Cuxhaven, Germany), (GSX). The 18 specimens (3 of each composite) were prepared in split Teflon moulds of 4 mm diameter and 6 mm thickness. All composites were cured in standard mode for 20 s using LED LCU (D-Light Duo, RF-Pharmaceuticals Sarl, Geneva, Switzerland; 1200–1300 mW/cm). The VMH was measured using a digital Micro Hardness Tester Shimadzu (HMV-2T E, Shimadzu Corporation, Kyoto, Japan). A 50 g (0.5 N) load force was applied for 30 s. Each specimen was measured at five places selected by chance at each level (N = 15). The hardness ratio or DOC was calculated for all samples as the ratio of bottom and surface microhardness at levels of 4 and 6 mm. Data were analysed using one-way ANOVA and Tukey’s post hoc test. Results. Significant reduction in VMH was observed for all tested materials when comparing top surface and bottom (p < 0.01). The highest VMH was obtained for GSX and AFX, and the lowest for TEC. The results show that the degree of polymerization was adequate for all tested materials at a depth of 6 mm, since the hardness ratio exceeded 0.80 in all cases. The hardness ratio at 4 mm was high for all tested composites ranging from 0.91 for TEC to 0.98 for GSX. All composites showed adequate DOC at the bottom of the 6 mm bulk samples. However, the hardness ratio was the highest for Admira Fusion X-tra (0.96) and GrandioSO X-tra (0.97). Conclusions. All tested materials showed a significant decrease in microhardness from the top surface to the bottom. The DOC was adequate for all bulk-fill composites at a depth of 6 mm cured under standard mode for 20 s. All bulk-fill resin composites evaluated in this study can be used in bulk, up to 6 mm. Full article
(This article belongs to the Special Issue Innovations in Direct and Indirect Dental Composite Restorations)
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13 pages, 2115 KiB  
Article
Residual-Free Micro–Nano Titanium Surfaces via Titanium Blasting and Single Acid-Etching: A Cleaner Alternative
by Artiom Lijnev, José Eduardo Maté Sánchez de Val, Jeevithan Elango, Carlos Pérez-Albacete Martínez, José Manuel Granero Marín, Antonio Scarano and Sergio Alexandre Gehrke
Bioengineering 2025, 12(7), 735; https://doi.org/10.3390/bioengineering12070735 - 5 Jul 2025
Viewed by 2230
Abstract
Background: Traditional sandblasted large-grit acid-etched (SLA) surface treatments frequently utilize alumina (Al2O3) blasting, which may leave residual particles embedded in implant surfaces, potentially compromising biocompatibility and osseointegration. This study investigates a contamination-free alternative: titanium dioxide particle (TiO2) [...] Read more.
Background: Traditional sandblasted large-grit acid-etched (SLA) surface treatments frequently utilize alumina (Al2O3) blasting, which may leave residual particles embedded in implant surfaces, potentially compromising biocompatibility and osseointegration. This study investigates a contamination-free alternative: titanium dioxide particle (TiO2) blasting followed by hydrochloric acid (HCl) etching, aimed at generating a cleaner, hierarchical micro–nano-textured surface. Methods: Grade IV titanium disks were treated either with TiO2 sandblasting alone or with an additional HCl etching step. Surfaces were analyzed via atomic force microscopy (AFM), scanning electron microscopy (SEM), contact angle measurements, and profilometry. hFOB osteoblasts were cultured to assess adhesion, proliferation, metabolic activity, and morphology. Results: The combination treatment produced a more homogeneous micro–nano structure with significantly increased roughness and a cleaner surface chemistry. Osteoblast proliferation and metabolic activity were notably improved in the TiO2 and HCl group. SEM imaging showed a more organized cytoskeletal structure and pronounced filopodia at 72 h. Conclusions: Titanium blasting combined with HCl etching yields a cost-effective, contamination-free surface modification with promising early-stage cellular responses. This approach represents a safer and effective alternative to conventional SLA treatment. Full article
(This article belongs to the Special Issue Periodontics and Implant Dentistry)
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11 pages, 2562 KiB  
Article
Biocompatibility of Titanium Oxide Nanotubes Layer Formed on a Ti-6Al-4V Dental Implant Screw in hFOB Cells In Vitro
by José Luis Castrejón Flores, Ángel Daniel Campos Juarez, Alexis Chino Ulloa, Fernando Nava Palafox, David Cruz Ortiz and Itzel Pamela Torres Avila
Coatings 2025, 15(6), 715; https://doi.org/10.3390/coatings15060715 - 13 Jun 2025
Viewed by 569
Abstract
The surface modification of dental implants with nanostructured films enables the development of the next generation of biomaterials that promote osseointegration. In this study, a uniform layer of titanium oxide nanotubes (TNTs) was successfully formed on a Ti-6Al-4V dental implant screw through anodic [...] Read more.
The surface modification of dental implants with nanostructured films enables the development of the next generation of biomaterials that promote osseointegration. In this study, a uniform layer of titanium oxide nanotubes (TNTs) was successfully formed on a Ti-6Al-4V dental implant screw through anodic oxidation. TNTs were morphologically characterized by Scanning Electron Microscopy (SEM), obtaining dimensions of 64.88 ± 10 nm in diameter and 5.34 ± 5 µm in length. Additionally, a crystal size of 23.45 nm was determined by X-ray diffraction (XRD) analysis. The TNT layer on the dental implant screw was evaluated in an in vitro system in direct contact with human osteoblast cells (hFOB) for 24 h and 48 h, finding cell growth near to the screw threads. Further, the biocompatibility of the dental screw coated with TNTs was evaluated using a flow cytometric assay with 7-AAD, demonstrating that cell viability was not affected at 24 h and 48 h. This study opens the perspective of the study of inflammation and osseointegration induced by implants coated with TNTs. Full article
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17 pages, 4881 KiB  
Article
Functionalization-Dependent Cytotoxicity of Silver Nanoparticles: A Comparative Study of Chlorhexidine and Metronidazole Conjugates
by Karol P. Steckiewicz, Monika Dmochowska, Elżbieta Megiel, Ewelina Barcińska and Iwona Inkielewicz-Stępniak
Biomolecules 2025, 15(6), 850; https://doi.org/10.3390/biom15060850 - 10 Jun 2025
Viewed by 848
Abstract
This study examines the cytotoxicity of two silver nanoparticle formulations—AgNPs conjugated with chlorhexidine (AgNPs-CHL) and AgNPs conjugated with polyethylene glycol and metronidazole (AgNPs-PEG-MET)—as examples of the surface functionalization of silver nanoparticles with drugs via sulfur–silver bonds and nitrogen–silver interactions. We previously reported the [...] Read more.
This study examines the cytotoxicity of two silver nanoparticle formulations—AgNPs conjugated with chlorhexidine (AgNPs-CHL) and AgNPs conjugated with polyethylene glycol and metronidazole (AgNPs-PEG-MET)—as examples of the surface functionalization of silver nanoparticles with drugs via sulfur–silver bonds and nitrogen–silver interactions. We previously reported the synthesis of these NPs and their efficiency in periodontitis treatment. Here, we analyze the relationships between the cytotoxic mechanisms of AgNPs and their surface chemistry. UV–Vis spectroscopy, dynamic light scattering (DLS), and scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy (EDX) were used for physicochemical studies of the conjugates in two environments: aqueous solutions and commonly used cell culture media. Cytotoxicity was assessed in human fetal osteoblasts (hFOB 1.19) and human gingival fibroblasts (HGF-1) through BrdU and LDH assays, ROS detection, cell cycle analysis, apoptosis assays, and protein expression studies. AgNPs-CHL showed aggregation and increased hydrodynamic diameters in the culture medium, while AgNPs-PEG-MET remained stable. Both exhibited concentration-dependent cytotoxicity: AgNPs-CHL at 0.4–10 μg/mL and AgNPs-PEG-MET at 0.75–10 μg/mL. AgNPs-CHL, in which silver surface functionalization was realized via nitrogen–silver interactions, induced significant ROS generation, LDH release, and necroptosis, marked by increased RIP1, RIP3, and MLKL proteins. In the case of AgNPs-PEG-MET, where sulfur–silver bonds combined the drug via a PEG linker, they triggered apoptosis, as evidenced by elevated caspase-2 levels and flow cytometry. These findings highlight that the type of surface functionalization of silver nanoparticles significantly influences their physicochemical behavior and biological effects. Understanding these mechanisms is crucial in designing safer, more effective nanoparticle-based therapies for periodontal and other inflammatory conditions. Full article
(This article belongs to the Special Issue Metallic Nanoparticles: Biosynthesis and Therapeutic Potential)
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16 pages, 5061 KiB  
Article
Bioactive Hydroxyapatite–Carboplatin–Quercetin Coatings for Enhanced Osteointegration and Antitumoral Protection in Hip Endoprostheses
by Gheorghe Iosub, Dana-Ionela Tudorache (Trifa), Ionuț Marinel Iova, Liviu Duta, Valentina Grumezescu, Alexandra Cătălina Bîrcă, Adelina-Gabriela Niculescu, Paul Cătălin Balaure, Ionela Cristina Voinea, Miruna S. Stan, Dragoș Mihai Rădulescu, Adrian Emilian Bădilă, Bogdan Ștefan Vasile, Alexandru Mihai Grumezescu and Adrian Radu Rădulescu
Coatings 2025, 15(4), 489; https://doi.org/10.3390/coatings15040489 - 20 Apr 2025
Cited by 1 | Viewed by 677
Abstract
The recurrence of bone cancer poses severe complications, particularly after orthopedic surgery, necessitating advanced biomaterials with dual functionality. This study develops nanostructured coatings composed of hydroxyapatite, carboplatin, and quercetin, designed to enhance bone regeneration while delivering localized cancer therapy. These coatings present a [...] Read more.
The recurrence of bone cancer poses severe complications, particularly after orthopedic surgery, necessitating advanced biomaterials with dual functionality. This study develops nanostructured coatings composed of hydroxyapatite, carboplatin, and quercetin, designed to enhance bone regeneration while delivering localized cancer therapy. These coatings present a promising solution for hip endoprostheses, addressing osteointegration and tumor recurrence prevention simultaneously. Hydroxyapatite was synthesized and characterized using XRD, TEM, SAED, FTIR, and SEM to assess crystallinity, surface morphology, and functional groups. The coatings were obtained by MAPLE. In vitro biocompatibility tests showed that HAp@CPT and HAp@CPT/QUE coatings supported osteoblast viability and adhesion while exhibiting selective cytotoxic effects on osteosarcoma cells. The Griess assay indicated that nitric oxide (NO) levels remained unchanged in hFOB osteoblasts, confirming that neither coating induced inflammatory responses in healthy cells. In contrast, MG63 osteosarcoma cells exhibited significantly elevated NO levels (p < 0.05) in response to HAp@CPT/QUE, suggesting increased oxidative stress. MTT assay results showed a 12% and 28% reduction in osteosarcoma cell viability for HAp@CPT and HAp@CPT/QUE, respectively. Phase-contrast microscopy further confirmed strong osteoblast adhesion and reduced osteosarcoma attachment, particularly on HAp@CPT/QUE surfaces. These findings highlight the dual functionality of hydroxyapatite–carboplatin–quercetin coatings, promoting osteointegration while exerting localized anticancer effects. Their bone-regenerative and selective cytotoxic properties make them a promising material for hip endoprostheses in oncological orthopedic applications. Full article
(This article belongs to the Special Issue Synthesis and Applications of Bioactive Coatings)
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19 pages, 6443 KiB  
Article
Biocompatible and Antibacterial Chemical Coatings on TiZr Dental Implants
by Vlad Gabriel Vasilescu, Toma Lucian Ciocan, Andreea Mihaela Custura, Florin Miculescu, Miruna Stan, Ionela Cristina Voinea, Dumitru Dima, Florentina Ionela Bucur, Andreea Veronica Dediu-Botezatu, Marian Iulian Neacșu, Elisabeta Vasilescu and Marina Imre
J. Funct. Biomater. 2025, 16(3), 112; https://doi.org/10.3390/jfb16030112 - 20 Mar 2025
Viewed by 1053
Abstract
This research aims to study the antibacterial coatings of invasive surgical medical devices, including dental implants, to reduce superficial and deep local infections over the long term. To obtain the coating without altering the initial properties of the substrate (dental implant made of [...] Read more.
This research aims to study the antibacterial coatings of invasive surgical medical devices, including dental implants, to reduce superficial and deep local infections over the long term. To obtain the coating without altering the initial properties of the substrate (dental implant made of TiZr bioalloy), simple, cost-effective, and efficient methods were employed, such as chemical deposition of silver (Ag). The deposition characteristics were analyzed using scanning electron microscopy (SEM), EDX analysis, and FT-IR infrared analysis. The in vitro testing of antimicrobial activity was conducted using the diffusion method by cultivating the bacterial strains Escherichia coli (E. coli) ATCC25922 and Staphylococcus aureus (S. aureus) ATCC25923 and measuring the diameter of the bacterial inhibition zone. Investigations and biocompatibility evaluations were performed on both uncoated and silver-coated (Ag) samples by analyzing cell viability and morphology in the presence of human fetal osteoblasts (hFOB cell line) and human gingival fibroblasts (HFIB-G cells) after 8 days of incubation. The research results confirm the biocompatibility of the coating, demonstrated by the lack of significant differences in cell density between the Ag-coated samples and the control group, as well as by the fact that the silver-coated surface effectively supports actin cytoskeleton organization, adhesion, and migration of both human osteoblasts and gingival fibroblasts. The results regarding the antibacterial efficiency of the silver implant coating indicated that the E. coli bacterial strain is more resistant than S. aureus. The resistance difference between the two bacterial strains was attributed to differences in the structure of their cell envelopes. Full article
(This article belongs to the Special Issue The Development and Future of Dental Implants)
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17 pages, 5006 KiB  
Article
Red Algae Alters Expression of Inflammatory Pathways in an Osteoarthritis In Vitro Co-Culture
by Shane M. Heffernan, Mark Waldron, Kirsty Meldrum, Stephen J. Evans and Gillian E. Conway
Pharmaceuticals 2025, 18(3), 315; https://doi.org/10.3390/ph18030315 - 24 Feb 2025
Viewed by 713
Abstract
Background/Objectives: Osteoarthritis (OA) is one of the most prevalent chronic conditions and significantly contributes to local and global disease burden. Common pharmaceuticals that are used to treat OA cause significant side effects, thus non-pharmaceutical bioactive alternatives have been developed that can impact OA [...] Read more.
Background/Objectives: Osteoarthritis (OA) is one of the most prevalent chronic conditions and significantly contributes to local and global disease burden. Common pharmaceuticals that are used to treat OA cause significant side effects, thus non-pharmaceutical bioactive alternatives have been developed that can impact OA symptoms without severe side-effects. One such alternative is the Red Algae Lithothamnion species (Litho). However, there is little mechanistic knowledge of its potential to effect OA gene expression, and a human in vitro model using commercially available cell lines to test its effectiveness has yet to be developed. Methods: Human osteoblast (hFOB 1.19. CRL-11372) and chondrocyte (C28/I2) cell lines were co-cultured indirectly using transwells. IL1-β was used to induce an inflammatory state and gene expression profiles following treatment were the primary outcome. Conclusions: Results indicated that the model was physiologically relevant, remained viable over at least seven days, untreated or following induction of an inflammatory state while maintaining hFOB 1.19. and C28/I2 cell phenotypic characteristics. Following treatment, Litho reduced the expression of inflammatory and pain associated genes, most notably IL-1β, IL-6, PTGS2 (COX-2) and C1qTNF2 (CTRP2). Confirmatory analysis with droplet digital PCR (ddPCR) revealed that Il-1β induced a significant reduction in C1qTNF2 at 7 days which was ameliorated with Litho treatment. These data present a novel and replicable co-culture model of inflammatory OA that can be used to investigate bioactive nutraceuticals. For the first time, this model demonstrated a reduction in C1qTNF2 expression that was mitigated by Red Algae Lithothamnion species. Full article
(This article belongs to the Section Natural Products)
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13 pages, 6696 KiB  
Article
Effect of Different Forms of Human Platelet Lysate on the Proliferation and Phenotype of Human Osteoblasts
by Mohamad Raihan Kamaruddin, Bahiratuz Zulfa Baharuddin, Nahgeshwarie Ratha Manaalan, Yi Lyn Wong, Muhammad Najib Fathi Hassan, Suria Abdul Aziz, Barathan Muttiah and Jia Xian Law
Appl. Sci. 2025, 15(4), 2074; https://doi.org/10.3390/app15042074 - 16 Feb 2025
Viewed by 1031
Abstract
Background and aims: Enhanced cell proliferation is crucial for reducing production time and cost in cell therapy, and human platelet lysate (HPL) is often used to boost cell proliferation due to its favorable safety profile. Understanding the roles of different HPL components and [...] Read more.
Background and aims: Enhanced cell proliferation is crucial for reducing production time and cost in cell therapy, and human platelet lysate (HPL) is often used to boost cell proliferation due to its favorable safety profile. Understanding the roles of different HPL components and their effects on cell culture can lead to more informed choices in medium formulation, which in turn can influence cell behavior and outcomes. Therefore, this study aimed to investigate the effects of two types of HPL, i.e., heparin-supplemented HPL (He-HPL) and fibrinogen-depleted HPL without heparin (Fd-HPL), on human osteoblasts. Materials and Methods: He-HPL and Fd-HPL were prepared from expired platelet concentrates. The presence of growth factors, i.e., brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF), and cytokines, i.e., interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α), in HPL was evaluated. Human fetal osteoblast (hFOB) cells were cultured in Dulbecco’s Modified Eagle Medium supplemented with either He-HPL or Fd-HPL. The cell morphology, viability, calcium deposition, and expression of osteogenic genes were assessed. Results: Comparable levels of BDNF (p > 0.05), VEGF (p > 0.05), and IL-6 (p > 0.05) were detected in both types of HPL, whereas He-HPL exhibited significantly higher levels of TNF-α (p < 0.05). However, there were no notable differences in cell morphology, viability, population doubling time, or total cell yield between the two HPL types. Similarly, no differences were observed in the mineralization of cells treated with He-HPL compared to Fd-HPL. Nonetheless, hFOB cells cultured with He-HPL demonstrated significantly higher expression of osteogenic markers Runx2 and ALP (p < 0.05) compared to those cultured with Fd-HPL. Conclusions: He-HPL and Fd-HPL demonstrate comparable performance in promoting osteoblast proliferation and mineralization, making both usable for bone tissue engineering. However, He-HPL might have a slight edge as it enhances osteogenic gene expression. Full article
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14 pages, 17159 KiB  
Article
Eicosapentaenoic Acid and Docosahexaenoic Acid as an Antimicrobial Agent in Orthopedics—An In Vitro Study About the Race for Surface
by Christopher Spiegel, Burak Ünalan, Andreas Kaserbacher, Rohit Arora and Débora C. Coraça-Huber
Pathogens 2025, 14(1), 57; https://doi.org/10.3390/pathogens14010057 - 10 Jan 2025
Cited by 1 | Viewed by 1049
Abstract
Background: The burden of prosthetic joint infection in combination with antibiotic-resistant bacterial strains is a rising dilemma for patients experiencing total joint replacements. Around 0.8–2% of patients experience prosthetic joint infections, while up to 21% of patients are considered fatal cases after 5 [...] Read more.
Background: The burden of prosthetic joint infection in combination with antibiotic-resistant bacterial strains is a rising dilemma for patients experiencing total joint replacements. Around 0.8–2% of patients experience prosthetic joint infections, while up to 21% of patients are considered fatal cases after 5 years. Staphylococcus aureus is one of the main reasons for prosthetic joint infections. Its capability of forming biofilms and developing mechanisms against antibiotics is one of the most dangerous clinical topics being currently discussed. Previous studies have shown the promising results of omega-3 fatty acids as an antimicrobial agent against Staphylococcus aureus. Though an antimicrobial effect has been examined, the influence of polyunsaturated fatty acids on Staphylococcus aureus in the presence of human osteoblasts has not been reported yet. In this study, we aimed to investigate the influence of omega-3 fatty acids on the biofilm formation of Staphylococcus aureus ATCC 29213 in the presence of hFOB 1.19 cells. The co-culture setup helped to examine the influence of omega-3 fatty acids on the race for surface to simulate prosthetic joint infections. Methods: In this study, we tested Staphylococcus aureus ATCC 29213 co-cultured with human fetal osteoblasts hFOB 1.19 in the presence of sub-MIC and MIC concentrations of docosahexaenoic acid (1.25 mg/L, 2.5 mg/L) and eicosapentaenoic acid (0.15 mg/L, 0.3 mg/L) after 1, 6 and 24 h of incubation. After establishing the co-culture, cell culture and biofilm, we performed colony-forming unit counting and cell counting to examine cell survivability. In addition, we carried out scanning electron microscopy to study the race for surface behaviour of the cells. Results: We found a protective influence of omega-3 fatty acids on osteoblasts when present in co-culture with Staphylococcus aureus after 6 h of incubation. Omega-3 fatty acids increase the cell survival of osteoblasts after 6 h in a co-culture with bacteria and are able to influence the race for surface. In this study, the strain of Staphylcoccus aureus ATCC 29213 showed signs of growth inhibition within the first 6 h. Conclusions: Omega-3 fatty acids can be a valuable antimicrobial agent in terms of decreasing the risk of on-site infection during surgery. Omega-3 fatty acids were shown to decrease the bacterial load within the first 6 h of incubation and increase the survivability of osteoblasts. Full article
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25 pages, 3859 KiB  
Article
Polydatin-Induced Shift of Redox Balance and Its Anti-Cancer Impact on Human Osteosarcoma Cells
by Alessio Cimmino, Magda Gioia, Maria Elisabetta Clementi, Isabella Faraoni, Stefano Marini and Chiara Ciaccio
Curr. Issues Mol. Biol. 2025, 47(1), 21; https://doi.org/10.3390/cimb47010021 - 31 Dec 2024
Viewed by 1397
Abstract
Cancer cells demonstrate remarkable resilience by adapting to oxidative stress and undergoing metabolic reprogramming, making oxidative stress a critical target for cancer therapy. This study explores, for the first time, the redox-dependent anticancer effects of Polydatin (PD), a glucoside derivative of resveratrol, on [...] Read more.
Cancer cells demonstrate remarkable resilience by adapting to oxidative stress and undergoing metabolic reprogramming, making oxidative stress a critical target for cancer therapy. This study explores, for the first time, the redox-dependent anticancer effects of Polydatin (PD), a glucoside derivative of resveratrol, on the human Osteosarcoma (OS) cells SAOS-2 and U2OS. Using cell-based biochemical assays, we found that cytotoxic doses of PD (100–200 µM) promote ROS production, deplete glutathione (GSH), and elevate levels of both total iron and intracellular malondialdehyde (MDA), which are key markers of ferroptosis. Notably, the ROS scavenger N-acetylcysteine (NAC) and the ferroptosis inhibitor ferrostatin-1 (Fer-1) partially reverse PD’s cytotoxic effects. Interestingly, PD’s ability to hinder cell adhesion and migration appears independent of its pro-oxidant effect. Analysis of the oxidative stress regulators SIRT1 and Nrf2 at the gene and protein levels using real-time PCR and Western blot indicates an early oxidative response to PD treatment. PD remains effective under tumor-like conditions of hypoxia and serum starvation, and sensitizes OS cells to ROS-inducing chemotherapeutics like doxorubicin (DOX) and cisplatin (CIS). Importantly, PD exhibits minimal toxicity to non-tumorigenic cells (hFOB), suggesting a favorable therapeutic profile. Overall, our findings underscore that PD-induced redox imbalance plays a crucial role in its anti-OS effects, warranting further exploration into the molecular mechanisms behind its pro-oxidant activity. Full article
(This article belongs to the Special Issue Phytochemicals and Cancer, 2nd Edition)
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11 pages, 1775 KiB  
Article
The Role of Formononetin in Osteoblast Function and Mineralization Potential with Deproteinized Bovine Bone Material
by Ebru Haciosmanoglu Aldogan, Deniz Başaran, Bilgin Öner and Başak Günçer
Curr. Issues Mol. Biol. 2024, 46(12), 14215-14225; https://doi.org/10.3390/cimb46120851 - 17 Dec 2024
Viewed by 887
Abstract
Objectives: Dental bone formation involves various cellular and molecular mechanisms, and phytoestrogens such as formononetin (FORM) are promising because of their estrogenic, anti-inflammatory, and antioxidant effects. This study investigated the effect of FORM on osteoblast proliferation, differentiation, and mineralization in combination with spongiosa [...] Read more.
Objectives: Dental bone formation involves various cellular and molecular mechanisms, and phytoestrogens such as formononetin (FORM) are promising because of their estrogenic, anti-inflammatory, and antioxidant effects. This study investigated the effect of FORM on osteoblast proliferation, differentiation, and mineralization in combination with spongiosa granulates (BO) in vitro. Materials and Methods: Human fetal osteoblast cells (hFOB1.19) were treated with increasing concentrations of FORM (1, 10, and 100 µg/mL), BO, or their combination. Cell proliferation was assessed using a MTT assay. Alkaline phosphatase (ALP) activity, intracellular Ca2+, and Pi levels were measured using ELISA. Vascular endothelial growth factor (VEGF) and osteocalcin expression levels were analyzed by western blotting. Results: Cell proliferation increased with FORM, with or without BO, after 6 days (p < 0.001). FORM and BO had a synergistic effect on ALP activity (p < 0.001). Intracellular Ca2+ and Pi levels were highest in the BO-FORM group, suggesting superior mineralization (p < 0.05). VEGF and osteocalcin expression was significantly upregulated with FORM, alone and with BO (p < 0.05), indicating improved angiogenesis and bone maturation over 9 days. Conclusions: FORM enhances osteoblast proliferation, differentiation, and mineralization potential, particularly in BO spongiosa granulates. These data support the in vitro potential of formononetin-phytoestrogen in promoting osteoblast differentiation and mineralization potential with BO. These findings suggest that FORM, combined with BO, could improve bone augmentation in clinical applications such as maxillofacial surgery. FORM shows valuable potential for clinical applications, such as maxillofacial surgery, by promoting faster and more effective healing. Full article
(This article belongs to the Section Molecular Medicine)
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11 pages, 3982 KiB  
Communication
Bioactive Agrocomposite for Tissue Engineering and Bone Regeneration
by Miguel Suffo, Celia Pérez-Muñoz, Daniel Goma-Jiménez, Carlos Revenga, Pablo Andrés-Cano and Miguel Ángel Cauqui-López
Inventions 2024, 9(6), 123; https://doi.org/10.3390/inventions9060123 - 9 Dec 2024
Viewed by 1380
Abstract
Background: This study describes a novel biomaterial consisting of a mixture of biphasic bioceramic obtained from waste generated by the sugar industry (Carbocal) and a medical-grade epoxy resin adhesive called LOCTITE® M31 CLTM. The objective was to demonstrate the possibility of coating [...] Read more.
Background: This study describes a novel biomaterial consisting of a mixture of biphasic bioceramic obtained from waste generated by the sugar industry (Carbocal) and a medical-grade epoxy resin adhesive called LOCTITE® M31 CLTM. The objective was to demonstrate the possibility of coating non-bioactive and non-biodegradable metallic surfaces on implantable elements. Methods: After preparation, the mixture was applied to the surfaces of hip prostheses composed of two distinct materials: polyetherimide and grade 5 titanium. In both cases, adhesion tests produced favourable results. Additionally, cell cultures were conducted using human foetal osteoblastic cell lines (hFOB 1.19). Results: It was observed that the mixture did not affect the proliferation of bone cells. Conclusions: This composite material was found to promote the growth of bone cells, suggesting its potential for fostering bone tissue development. Full article
(This article belongs to the Section Inventions and Innovation in Biotechnology and Materials)
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32 pages, 26302 KiB  
Article
Development of Novel Biocomposites with Antimicrobial-Activity-Based Magnesium-Doped Hydroxyapatite with Amoxicillin
by Carmen Cimpeanu, Daniela Predoi, Carmen Steluta Ciobanu, Simona Liliana Iconaru, Krzysztof Rokosz, Mihai Valentin Predoi, Steinar Raaen and Monica Luminita Badea
Antibiotics 2024, 13(10), 963; https://doi.org/10.3390/antibiotics13100963 - 12 Oct 2024
Cited by 6 | Viewed by 2120
Abstract
Background/Objectives: A biocomposite based on magnesium-doped hydroxyapatite and enriched with amoxicillin (MgHApOx) was synthesized using the coprecipitation method and is presented here for the first time. Methods: The stability of MgHAp and MgHApOx suspensions was evaluated by ultrasound measurements. The structure [...] Read more.
Background/Objectives: A biocomposite based on magnesium-doped hydroxyapatite and enriched with amoxicillin (MgHApOx) was synthesized using the coprecipitation method and is presented here for the first time. Methods: The stability of MgHAp and MgHApOx suspensions was evaluated by ultrasound measurements. The structure of the synthesized MgHAp and MgHApOx was examined with X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy and X-ray photoelectron spectroscopy (XPS). The crystalline structure was determined by X-ray diffraction. The FTIR data were collected in the range of 4000–400 cm−1. The morphology of the nanoparticles was evaluated by scanning electron microscopy (SEM). Furthermore, the biocompatible properties of MgHAp, MgHApOx and amoxicillin (Ox) suspensions were assessed using human fetal osteoblastic cells (hFOB 1.19 cell line). The antimicrobial properties of the MgHAp, MgHApOx and Ox suspension nanoparticles were assessed using the standard reference microbial strains Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Candida albicans ATCC 10231. Results: X-ray studies have shown that the biocomposite retains the characteristics of HAp and amoxicillin. The SEM assessment exhibited that the apatite contains particles at nanometric scale with acicular flakes morphology. The XRD and SEM results exhibited crystalline nanoparticles. The average crystallite size calculated from XRD analysis increased from 15.31 nm for MgHAp to 17.79 nm in the case of the MgHApOx sample. The energy-dispersive X-ray spectroscopy (EDS) and X-ray photoelectron spectroscopy (XPS) analysis highlighted the presence of the constituent elements of MgHAp and amoxicillin. Moreover, XPS confirmed the substitution of Ca2+ ions with Mg2+ and the presence of amoxicillin constituents in the MgHAp lattice. The results of the in vitro antimicrobial assay demonstrated that MgHAp, MgHApOx and Ox suspensions exhibited good antimicrobial activity against the tested microbial strains. The results showed that the antimicrobial activity of the samples was influenced by the presence of the antibiotic and also by the incubation time. Conclusions: The findings from the biological assays indicate that MgHAp and MgHApOx are promising candidates for the development of new biocompatible and antimicrobial agents for biomedical applications. Full article
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