Human Osteoblastic Cells Research Articles

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.

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.

Low toxicity of dissolved silver from silver-coated titanium dental implants to human primary osteoblast cells

Objectivesthe controlled release of silver as a biocide from Ag-coated medical implants is desirable. However, the biocompatibility of Ag leachates is poorly understood. This study investigated the toxicity of silver released from the silver plated titanium implants to human primary osteoblast cells; and the effect of cell culture medium on the silver speciation and bioavailability. Methods: Ti6Al4V discs were coated with Ag nanoparticles (NPs), silver plus hydroxyapatite (HA) nanoparticles (Ag+nHA), or Ag NPs plus microparticles (Ag+mHA). Primary human osteoblast cells were exposed to the leachates from the various discs for up to 7 days. Results: the total Ag concentrations released as leachate from the silver-plated titanium discs were 0.7–1.6 mg L−1, regardless of treatment. Cumulative silver release over 7 days was approximately 3 mg L−1 in all treatments. The concentration of total Ag in the cell homogenates from all the Ag-containing treatments was modest, ∼ 0.1 µg mg protein−1 or less at day 7. Cells showed normal healthy morphology with no appreciable leak of LDH or ALP activity into the external media compared to the reference control. Similarly, there was no significant differences (Kruskal Wallis, p > 0.05) in the LDH or ALP activity in the cell homogenate between treatments. Conclusions: overall, there was a controlled release of Ag into the external media, but this remained biocompatible with no deleterious effects on the osteoblast cells, which means that the released silver to the peri-implant environment is not toxic making the coating potential for clinical use

Development of Novel Biocomposites with Antimicrobial-Activity-Based Magnesium-Doped Hydroxyapatite with Amoxicillin.

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.

Customized bioresin formulation for stereolithography in tissue engineering

Recently, advancements in fabrication technology have brought a new aspect to the field of tissue engineering. By utilizing advanced techniques in 3D manufacturing and biomaterials, scientists have successfully created tissue engineering scaffolds with complex three-dimensional structures and customized chemical compositions that closely mimic the natural environment of living tissues. These methodologies show potential not only for developing therapies that restore lost tissue function but also for creating in vitro models that replicate living tissue. The current investigation involved the synthesis of methacrylated polycaprolactone (PCLMA) by incorporating methacryloyl chloride (Meth-Cl) into polycaprolactone (PCL) with a molecular weight of 80,000 Da. Afterwards, PCLMA was subjected to crosslinking with glycerol acrylate (GA) and, by utilizing Diphenyl(2,4,6-trimethylbenzoyl) phosphine oxide (TPO) as a photoinitiator, achieved the three-dimensional (3D) printing of tissue materials using Stereolithography (SLA). Analytical techniques included nuclear magnetic resonance (NMR), Fourier-transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). Cell viability was investigated using Human Osteoblast (HOB) cells. The biocompatibility of glycerol acrylate (GA) crosslinked polymethacrylated polycaprolactone (PCLMA) was confirmed using cell viability experiments. Overall, the GA-crosslinked PCLMA bioresin, particularly PCLMA-8, shows promise for further use in tissue engineering applications.

Phage-Loaded Biomimetic Apatite Powder With Antibiofilm Activity to Treat Bone-Associated Infections.

For decades, calcium phosphate (CaP)-based ceramics have been used for coating of bone and joint substitutes after arthroplasty due to their biocompatible properties. Infections following orthopedic replacement occur in 1%-5% of cases, causing serious complications. Biofilm formation either on the biomaterial's surface or on patient's tissues greatly enhances the resistance against antibiotic treatments and can induce a chronic infection, emphasizing the need for novel antimicrobial delivery systems. In this study, we established a protocol enabling bacteriophage loading during the synthesis of a CaP-based powder. The resulting biomaterial proved to be noncytotoxic against human osteoblastic cells and able to significantly inhibit 24-h matured S. aureus biofilm cultures or even completely eradicate it after 5 days of contact. Additional S. aureus biofilm assays with a freeze-dried material using two different excipients showed that sucrose had a protective role against Remus bacteriophage treatment of S. aureus biofilms, whereas lactose-freeze-dried powder maintained the antibiofilm activity.

9263 Characterization Of A Novel Oral Small Molecule PTH1R Agonist: Proof Of Concept For An Alternative To Injectable Peptide-Based Therapy For Hypoparathyroidism

Abstract Disclosure: J. Zhang: Employee; Self; Septerna Inc.. I. Gomez: None. C. He: Employee; Self; Septerna Inc. J. Wang: Employee; Self; Septerna Inc. X. Du: Employee; Self; Septerna Inc. K. Nguyen: Employee; Self; Septerna Inc. K. Alvarez: Employee; Self; Septerna Inc. A.M. Mirza: Employee; Self; Septerna Inc.. Hypoparathyroidism (HP), a rare endocrine disorder characterized by insufficient levels of parathyroid hormone (PTH), leads to hypocalcemia and hyperphosphatemia. Most HP cases arise due to damage of the parathyroid glands during thyroid or neck surgery. HP symptoms include severe muscle cramps, tingling, burning and numbness, memory loss, and headaches, all leading to a decreased quality of life. Calcium and vitamin D supplementation do not fully ameliorate the disease and may contribute to renal disease. Chronic HP increases the risk of major complications, such as calcium depositions in the brain, eye, and kidneys. Similarly, it can lead to low bone turnover and increased bone mineral density with associated increased bone mineralization. In comparison to standard of care and traditional injectable peptide-based therapies, oral small molecules may have multiple advantages clinically. However, there are no oral small molecules currently in development. PTH functions through activation of the parathyroid hormone 1 receptor (PTH1R), a class B G protein-coupled receptor. The main target tissues for PTH1R activation include bone and kidney. In the kidney, PTH increases tubular reabsorption of calcium and promotes renal excretion of phosphate. In bone, PTH stimulates bone turnover and mobilizes calcium into the circulation to maintain serum calcium. Here we describe the characterization of SP-1462, a novel, potent, and selective oral small molecule, and show that, like the PTH peptides, it acts on PTH1R as an agonist and elicits similar downstream functions in vitro and in vivo. To assess PTH1R engagement in the major targeted organs, kidney and bone, primary human renal proximal epithelial cells (RPTEC) and a human osteoblast cell line (Saos-2) were used, respectively. RPTECs treated with PTH and SP-1462 showed significant changes in transporter genes (SLC22A2 and SLC13A1), immune related genes (CCL2 & DPP4), and matrix reorganization genes (ITGA5 and COL1A1). Saos-2 cells treated with PTH and SP-1462 both significantly affected the expression of Wnt signaling genes (DKK1 and TCF7) and the expression of coupling genes that bridge anabolic/catabolic activities (M-CSF, RANKL, HDAC4, and OPG). Importantly, oral administration of a single dose of SP-1805 in a rat surgical thyroparathyroidectomy (TPTx) model resulted in significant upregulation of serum calcium levels for a period of 24 hours in a dose dependent manner at 3, 10 and 30 mg/kg. The window of calcium upregulation is comparable to injectable PTH peptides currently in development to treat hypoparathyroidism. These data suggest that oral small molecule PTH1R agonists engage PTH pathways similar to native PTH and have the potential to replace injectable PTH peptides to treat PTH-related disorders, including hypoparathyroidism. Presentation: 6/3/2024

6821 Characterization Of A Novel Oral Small Molecule PTH1R Agonist: Proof Of Concept For An Alternative To Injectable Peptide-Based Therapy For Hypoparathyroidism

Abstract Disclosure: J. Zhang: None. I. Gomez: None. C. He: None. J. Wang: None. X. Du: None. K. Nguyen: None. K. Alvarez: None. A.M. Mirza: None. Hypoparathyroidism (HP), a rare endocrine disorder characterized by insufficient levels of parathyroid hormone (PTH), leads to hypocalcemia and hyperphosphatemia. Most HP cases arise due to damage of the parathyroid glands during thyroid or neck surgery. HP symptoms include severe muscle cramps, tingling, burning and numbness, memory loss, and headaches, all leading to a decreased quality of life. Calcium and vitamin D supplementation do not fully ameliorate the disease and may contribute to renal disease. Chronic HP increases the risk of major complications, such as calcium depositions in the brain, eye, and kidneys. Similarly, it can lead to low bone turnover and increased bone mineral density with associated increased bone mineralization. In comparison to standard of care and traditional injectable peptide-based therapies, oral small molecules may have multiple advantages clinically. However, there are no oral small molecules currently in development. PTH functions through activation of the parathyroid hormone 1 receptor (PTH1R), a class B G protein-coupled receptor. The main target tissues for PTH1R activation include bone and kidney. In the kidney, PTH increases tubular reabsorption of calcium and promotes renal excretion of phosphate. In bone, PTH stimulates bone turnover and mobilizes calcium into the circulation to maintain serum calcium. Here we describe the characterization of SP-1462, a novel, potent, and selective oral small molecule, and show that, like the PTH peptides, it acts on PTH1R as an agonist and elicits similar downstream functions in vitro and in vivo. To assess PTH1R engagement in the major targeted organs, kidney and bone, primary human renal proximal epithelial cells (RPTEC) and a human osteoblast cell line (Saos-2) were used, respectively. RPTECs treated with PTH and SP-1462 showed significant changes in transporter genes (SLC22A2 and SLC13A1), immune related genes (CCL2 & DPP4), and matrix reorganization genes (ITGA5 and COL1A1). Saos-2 cells treated with PTH and SP-1462 both significantly affected the expression of Wnt signaling genes (DKK1 and TCF7) and the expression of coupling genes that bridge anabolic/catabolic activities (M-CSF, RANKL, HDAC4, and OPG). Importantly, oral administration of a single dose of SP-1805 in a rat surgical thyroparathyroidectomy (TPTx) model resulted in significant upregulation of serum calcium levels for a period of 24 hours in a dose dependent manner at 3, 10 and 30 mg/kg. The window of calcium upregulation is comparable to injectable PTH peptides currently in development to treat hypoparathyroidism. These data suggest that oral small molecule PTH1R agonists engage PTH pathways similar to native PTH and have the potential to replace injectable PTH peptides to treat PTH-related disorders, including hypoparathyroidism. Presentation: 6/3/2024

Formulation and evaluation of Phaseolus lunatus seed coat mediated silver nanoparticles mouthwash: A comprehensive study on biomedical properties and toxicological assessment

Silver nanoparticles have promising therapeutic potential in the field of dentistry, as newly emerging oral therapeutics, in the form of mouthwashes based on silver nanoparticles, demonstrate significant potential for enhancing oral management thus the present investigation aims to formulate silver nanoparticles-based mouthwash from an aqueous extract of Phaseolus lunatus seed coat (PLSC) and evaluate its biomedical properties. The green synthesized AgNPs in the mouthwash were characterized using UV–visible spectroscopy, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and x-ray diffraction (XRD). The formulated mouthwash was assessed for its anti-microbial activity using the agar well diffusion technique and time-kill curve assay. Its anti-inflammatory and anti-oxidant activities are assessed through egg albumin assay and hydrogen peroxide assays. The Cytotoxic effect of formulated mouthwash was assessed through a brine shrimp lethality assay and MTT assay over the human osteoblast cell line (MG-63). Furthermore, the study also assessed the toxicity effect of formulated mouthwash through zebrafish embryos. The results suggest that the green synthesized AgNPs were spherical and had an average size of 42 nm and the formulated mouthwash exhibited significant anti-microbial activity, anti-inflammatory, and antioxidant properties, making them a better candidate for oral health therapeutics. The Zebrafish embryo toxicology studies of mouthwash revealed a consistent lack of abnormalities and a good viability and hatchability rate and the cytotoxic effect shows less toxicity over brine shrimp and human osteoblast cells. In conclusion, PLSC-AgNPs mouthwash is a potential oral therapeutic option with minimal toxicity for better oral management.

The apatite-forming ability of bioactive glasses – A comparative study in human serum and Kokubo's simulated body fluid

Currently, the apatite-forming ability evaluation of a biomaterial employs Kokubo's simulated body fluid (SBF) as a testing medium. As the conventional SBF simulates only the ionic composition and pH of human serum (HS), this study aimed to compare the apatite formation in SBF and HS on the sol-gel-derived bioglass (BG) 83S2 and commercial 45S5 as well as to verify the influence of preincubation of BG in both fluids on cytocompatibility towards human fetal osteoblast cell line hFOB 1.19. The apatite formation in HS was hindered, regardless of the type of used BG. Relatively higher apatite-forming ability was noticed for BG83S2. In direct contact test, both BGs were cytocompatible towards osteoblasts, regardless of the type of used fluid. In the case of extracts; however, only the extract obtained from BG83S2 preincubated in HS did not show the cytotoxic effect. The evaluation of apatite-forming ability in SBF appears to be a fast screening method for comparing the apatite-forming ability of various bioglasses. Nonetheless, SBF is unable to precisely imitate the morphology and crystallinity of apatite observed during the assay in HS, as well as it does not mimic the changes in profiles of osteogenic ions.

One-step synthesis of a piezoelectric hybrid BNNT/BaTiO3 composite and its application in bone tissue engineering

Nanomaterials with piezoelectric properties can significantly improve the applicability of polymers used in tissue engineering applications. In this study, we report the one-step synthesis of a novel hybrid piezoelectric composite comprising barium titanates and boron nitride nanotubes. This composite is distinguished by its unique microstructures, including nanoflakes, triangular boron nitride structures, and fiber-like boron nitride nanotube configurations, which contribute to its enhanced piezoelectric properties. The composite was incorporated into a chitosan-based tissue scaffold and evaluated in vitro. Electric-responsive Human Osteoblast cells cultured on the scaffolds are exposed to low-frequency ultrasound stimulation during cell growth. The biocompatibility, cell adhesion, alkaline phosphatase activities, and mineralization of osteoblast cells on the piezo-composite scaffolds were evaluated. The results show that the hybrid piezoelectric composite significantly enhances the properties of chitosan-based scaffold.

The Effect of Water Fraction Derived from Green Clover (Marsilea crenata C. Presl.) Leaves on Differentiation and Maturation of Human Osteoblast Cell

Women over the age of 40 often experience a decrease in bone mass due to a lack of estrogen, which can increase the risk of osteoporosis. Previous studies have shown that the leaves of Green Clover (Marsilea crenata C. Presl.) contains phytoestrogen compounds which can potentially replace the function of estrogen in the body. In this context, this study aims to evaluate the ability of water fractions from M. crenata leaves in increasing the expression of osterix (Osx) and osteocalcin (Ocn), which are important indicators of increased bone formation, specificially on differentiation and maturation of osteoblast cell. In this study, the water fraction with a dose of 62.5; 125; and 250 µg/L, as well as Genistein 2.5 µg/ml, were given to human fetal osteoblast (hFOB) 1.19 cells that had reached the confluence point. The analysis method uses immunocytochemical techniques with confocal laser scanning microscopy (CLSM) to identify changes in the bone formation process. All dosages of water fraction considerably enhanced the bone formation in hFOB 1.19 cells, as indicated by the results. The results of this study provide confirmation that the water fraction derived from M. crenata leaves has promise as a possible agent for combating osteoporosis. This is achieved by enhancing differentiation and maturation of osteoblast cell through the upregulation of Osx and Ocn genes.

RGD-functionalised self-assembling peptide hydrogel induces a proliferative profile in human osteoblasts in vitro.

Self-assembling peptide hydrogels (SAPHs) have been used in the past decade as reliable three-dimensional (3D) synthetic scaffolds for the culture of a variety of mammalian cells in vitro. Thanks to their versatile physicochemical properties, they allow researchers to tailor the hydrogel properties, including stiffness and functionality to the targeted cells and cells' behaviour. One of the advantages of using SAPH scaffolds is the ease of functionalisation. In the present work, we discuss the effect that functionalising the FEFEFKFK (F, phenylalanine; K, lysine; and E, glutamic acid) hydrogel scaffold using the cell-binding RGDS (fibronectin - R, arginine; G, glycine; D, aspartic acid; S, serine) epitope affects the material properties as well as the function of encapsulated human osteoblast cells. RGDS functionalisation resulted in cells adopting an elongated morphology, suggesting attachment and increased proliferation. While this led to higher cell viability, it also resulted in a decrease in extra-cellular matrix (ECM) protein production as well as a decrease in calcium ion deposition, suggesting lower mineralisation capabilities. The work clearly shows that SAPHs are a flexible platform that allow the modification of scaffolds in a controlled manner to investigate cell-material interactions.

Additively manufactured 17–4 PH stainless steels for fracture management devices

ABSTRACT Stainless steel 316L (SS316L) is widely used in fracture management devices. However, SS316L does not offer any bacterial infection resistance and can cause metal-ion sensitivity due to Ni-ions’ presence. 17-4PH can emerge as a promising substitute due to the intrinsic antibacterial properties of copper, a 75% reduction in nickel content, and superior mechanical properties. SS316L and 17–4 PH were manufactured using laser-directed energy deposition (LDED). 17-4PH specimens surpassed the compressive strength of SS316L by over 150%. A static magnetic field was generated in 17–4 PH specimens to understand in vitro bone cell–material interactions. In vitro human fetal osteoblast cell culture and bacterial inhibition study using Staphylococcus aureus and Pseudomonas aeruginosa were carried out on these specimens with SS316L as control and as-processed and magnetised 17–4 PH as treatments. Results demonstrated that magnetised 17–4 PH exhibited 25% enhancement in hFOB proliferation and 70% reduction in bacterial colonisation compared to SS316L.

In Vitro Proliferation of MG-63 Cells in Additively Manufactured Ti-6Al-4V Biomimetic Lattice Structures with Varying Strut Geometry and Porosity.

Lattice structures have demonstrated the ability to provide secondary stability in orthopedic implants by promoting internal bone growth. In response to the growing prevalence of lattices in orthopedic design, we investigated the effects of porosity and unit cell geometry in additively manufactured Ti-6Al-4V biomimetic lattice structures on the osteogenesis of human MG-63 osteoblastic cell lines in vitro. We analyzed glucose consumption, alkaline phosphatase (ALP) concentration, and end-of-culture cell count as markers for osteogenic growth. Two different strut geometries were utilized (cubic and body-centered cubic), along with four different pore sizes (400, 500, 600, and 900 µm, representing 40-90% porosity in a 10 mm cube), in addition to a solid specimen. Structural characterization was performed using scanning electron microscopy. The results indicated that lattices with a 900 µm pore size exhibited the highest glucose consumption, the greatest change in ALP activity, and the highest cell count when compared to other pore sizes. Cubic 900 µm lattice structures outperformed other specimens in facilitating the maturation of viable MG-63 cells from the formation to the mineralization phase of bone remodeling, offering the most promise for osseointegration in additively manufactured titanium implants in the future. However, irrespective of a particular pore size or unit cell geometry, it was found that all the lattices were capable of promoting osteogenic growth due to surface roughness in the printed parts.

Photobiomodulation Modulates Proliferation and Gene Expression Related to Calcium Signaling in Human Osteoblast Cells.

Introduction: Photobiomodulation with low-level laser treatment can enhance bone formation by stimulating the cell division of osteoblasts and increasing the amount of protein deposition, thus encouraging the formation of new bone. The aim of this study was to evaluate the effects of photobiomodulation with a low-level laser on proliferation and gene expression related to calcium signaling in human osteoblasts. Methods: Osteoblastic cell lines of the hFOB1.19 lineage, human osteoblasts, were grown and assigned into two groups, control (C; n=78 cultured wells) and photobiomodulation (L; n=78 cultured wells) with n=6 per day of the experimental period. Cells were cultured (immature at 34 ºC), and after maturation at 37 ºC, group L cells were exposed to laser irradiation with a low-level laser device (gallium and aluminum arsenide), at a wavelength of 808 nm, a power output of 200 mW, and a power density of 200 mW/cm2. The energy delivered to the cells was 37 J/cm2, with a beam area of 0.02 mm2 and an exposure time of 5 seconds. This treatment was applied daily for a period of 13 days. Following this, the number of cells was counted, and RNA was isolated, measured, and then converted into cDNA for further quantification using a comparative Ct method with real-time polymerase chain reaction. The results were then subjected to statistical analysis through a Mann-Whitney test, with a significance level of P<0.05. Results: The cell count in the L group (37.25x10±4±22.02) was statistically higher compared to the control group (22.75x10±4±7.660) with a P value of 0.0259. The values of 2-ΔΔCt for S100A6, plasma membrane calcium ATPase (PMCA), and calmodulin genes indicated hyper-expression on the thirteenth day, while the osteocalcin gene showed hypo-expression. Conclusion: The study suggests that the photobiomodulation mechanism with a low-level laser may regulate gene expression in human osteoblasts in a dose-dependent and cumulative manner.

The Effect of Marginal Zn2+ Excess Released from Titanium Coating on Differentiation of Human Osteoblastic Cells.

Composite coatings based on chitosan and zinc nanoparticles (ZnNPs) were successfully produced on Ti13Zr13Nb substrates by cathodic electrophoretic deposition (EPD). The unfavorable phenomenon of water electrolysis-induced nonuniformity was reduced by applying a low voltage (20 V) and a short deposition time (1 min). Surface analysis (roughness and hydrophilicity) reveals the potential of these coatings for enhancing cell attachment and bone-implant integration. However, there is a concern about adhesion and strength; therefore, incorporating ZnNPs shows promise for enhancing mechanical properties, suggesting opportunities for further optimization of the process. The aim of this work was to investigate whether Zn2+ released from coating yields overt cellular impairment. hFOB1.19 osteoblastic cells were used as a model in this study. A subtoxic, 0.125 mmol/L, Zn concentration did not cause significant negative changes in cultured osteoblastic cells, as there was no significant change in their viability and their mitochondrial metabolism. Moreover, the alkaline phosphatase and lactate dehydrogenase activities were aggravated. However, a high, over 0.175 mmol/L, Zn2+ concentration caused total cell death. This was caused by the inhibition of mitochondrial enzymes' activities. Our data indicate that composite coatings releasing Zn2+ may be used as the differentiating factor toward osteoblastic cells.

Regulation of Osteogenic and Angiogenic Markers in Alkali Treated Titanium for Hard Tissue Engineering Applications.

Titanium (Ti) and Ti alloys are of great interest in bone and dental tissue engineering applications due to their biocompatibility, corrosion resistance, and close mechanical properties to natural bone. However, the formation of fibrous tissue prevents osteointegration and results in implant loosening. Thus, physical and chemical methods are used to improve the surface properties of Ti. This study aimed to understand the role of alkali treatment conditions, including alkali medium concentration, temperature, rotation speed, and post-heat treatment. Our results showed that alkali treatment using 5 and 10 molar sodium hydroxide solution allows the formation of web-like microstructure. However, a higher concentration of 15 molar resulted in cracks along the surface. Interaction between the human fetal osteoblast cells (hfOBs) and Ti samples showed that heat treatment is necessary for increased cellular proliferation, which was not significantly different at later time points compared to the polished Ti. Alkali heat treatment did not induce inflammatory reactions at later time points. It showed an increase in vascular endothelial growth factor, osteoprotegerin/nuclear factor kappa-В ligand ratio, and osteocalcin expression, which is evidence for accelerated osteoblast cell maturation and bone remodeling in surface-modified samples. Together, these data show that alkali treatment using 5 or 10 molar of NaOH followed by heat treatment may have therapeutic effect and assist with bone tissue integration with Ti implant.

Potential effect of Yemeni Sidr Honey on the Viability of Gingival Fibroblasts and Osteoblast Cells

Background/aim: The aim of this research is to investigate the potential beneficial effects of Yemen Sidr honey on human gingival fibroblast and osteoblast cell viability and proliferation. Materials and methods: MTT assays were conducted, and the cells were analyzed using an ELISA reader at 590nm. Cell viability was determined based on the optical density readings. Results: At a lower concentration 5μL, the fibroblast and osteoblast cells exhibited a cell viability percentage of over 50%. Conclusion: Yemen Sidr honey is biocompatible and non-toxic to the cultured fibroblast and osteoblast cells.

Novel Antimicrobial Agents Based on Zinc-Doped Hydroxyapatite Loaded with Tetracycline.

In this paper, we present for the first time the development of zinc-doped hydroxyapatite enriched with tetracycline (ZnHApTe) powders and provide a comprehensive evaluation of their physico-chemical and biological properties. Various techniques such as X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR) were used for the sample's complex evaluation. Moreover, the biocompatibility of zinc-doped hydroxyapatite (ZnHAp) and ZnHApTe nanoparticles was evaluated with the aid of human fetal osteoblastic cells (hFOB 1.19 cell line). The results of the biological assays suggested that these nanoparticles hold great promise as potential candidates for the future development of novel biocompatible and antimicrobial agents for biomedical applications. The antimicrobial properties of the ZnHAp and ZnHApTe nanoparticles were assessed using the standard reference microbial strains Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Candida albicans ATCC 10231. The results of the in vitro antimicrobial assay demonstrated that both tested materials exhibited good antimicrobial activity. Additionally, these data also indicated that the antimicrobial effects of the ZnHAp nanoparticles were intensified by the presence of tetracycline (Te). Furthermore, the results also suggested that the antimicrobial activity of the samples increased with the incubation time.