Cell Line MG-63 Research Articles

A silver lining in MRSA treatment: The synergistic action of poloxamer-stabilized silver nanoparticles and methicillin against antimicrobial resistance

BackgroundIncreasing antibiotic resistance in bacterial infections, including drug-resistant strains like methicillin-resistant Staphylococcus aureus (MRSA), necessitates innovative therapeutic solutions. Silver nanoparticles are promising for combating infections, but toxicity concerns emphasize the importance of factors like dosage, size, shape, and surface chemistry. Hence, exploring poloxamer as a stabilizing agent to reduce its toxicity and enhance the antibacterial effect on MRSA is investigated. MethodsSilver nanoparticles stabilized with poloxamer (AgNPs@Pol) were synthesized through the chemical reduction method and characterized using UV–visible spectrophotometer, HR-TEM, DLS, and Zeta potential measurements. Subsequently, the antibacterial activity of AgNPs@Pol alone and in combination with methicillin against MRSA and methicillin-susceptible S. aureus (MSSA) was evaluated using the broth microdilution method. ResultsAgNPs@Pol showed significant efficacy against MRSA and MSSA, achieving a 100 % reduction in colony-forming units (CFU) at 9.7 μg/ml. The minimum inhibitory concentration (MIC) against MRSA and MSSA was 8.6 μg/ml and 4.3 μg/ml, respectively. A synergistic effect was observed when AgNPs@Pol was combined with methicillin. Treatment with AgNPs@Pol increased reactive oxygen species (ROS) production in both strains, contributing to its antibacterial activity. Real-time qPCR analysis indicated the downregulation of genes involved in antimicrobial resistance and cell adhesion in both strains. Further, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay demonstrated low cytotoxicity for AgNPs@Pol against MCF-7, MG-63, and NIH-3T3 cell lines. ConclusionThe developed AgNPs@Pol demonstrated extensive colloidal stability, potent antibacterial activity and synergistic effect with methicillin against MRSA and MSSA. Further studies in primary cells and in vivo models may validate its potential for clinical applications.

Multifunctional zinc oxide loaded stearic acid surfaces on biodegradable magnesium WE43 alloy with hydrophobic, self-cleaning and biocompatible attributes

Biodegradable Mg alloys are one of the most promising materials for implant applications, for which the widespread use is hampered by rapid degradation in physiological environment. The present study explores the development of a novel zinc-oxide (ZnO) loaded stearic acid (SA) coating on medical grade WE43 alloy using a facile, and environment-friendly technique for promising biomaterial surface applications. Morphology and chemical analyses reveal the development of a layered coating surface with uniformly dispersed ZnO nanoparticles in SA matrix. The developed coating exhibits hydrophobic performance (contact angle of 135°) along with the retainment of hydrophobicity under varying chemical (acidic pH of 1 and basic pH of 13) and mechanical (post scratch testing and peel-off studies) conditions. The observed surface water repellency facilitates self-cleaning performance pointing towards its efficacy against potential biofilm formation. The developed coating demonstrates superior resistance against surface degradation during immersion studies in phosphate buffer saline solution (pH = 7.4) and better cell viability with MG-63 (human osteosarcoma) cell line. The approach presented here sets a platform for the development of efficient coatings on biodegradable WE43 alloys surfaces for potential biomaterial technologies.

In vitro Preparation and Evaluation of Alendronate-modified Hyaluronic acid-based Nanomicelles as a Bone-Targeted Drug delivery system

Nanomicelles based on Hyaluronic acid (HA), a linear polysaccharide with high affinity to the CD44 receptor on cancerous cells, offer considerable potential to transport drugs to the target tissue while reducing exposure to normal tissues. The high affinity of alendronate (ALN) to hydroxyapatite crystals, the crucial component of bone, makes it an ideal candidate for bone-targeted delivering therapeutic nanoparticles. Here, we developed targeted and self-assembled nanocarriers by modifying HA nanomicelles with ALN to attach to hydroxyapatite. Curcumin (CUR), as a hydrophobic model drug, was used to evaluate the efficiency of developed nanomicelles. The size of blank and CUR-loaded nanomicelles was below 200 nm and their critical micelle concentrations were in the range of 38 μg/ml, which is suitable for delivery to bone tissue. The CUR-loaded ALN-HA-AC (CUR@ALN-HA-AC) nanomicelles showed a two-step release pattern, a first accelerated release period (12 % in 1 h) followed by a sustained release (65 % up to 50 h). The hydroxyapatite affinity experiment indicated that CUR@ALN-HA-AC nanomicelles exhibited significantly high affinity to the bone. Cytotoxicity studies on MG-63 cell lines demonstrated that CUR-loaded micelles exhibited much higher cytotoxic activity compared to free CUR, indicating that encapsulation in HA-ALN-AC micelles significantly enhanced the tumor cell-killing ability of CUR.

In-vitro and thermal stability study of bioglass synthesized from biogenic sources

In this work, the bioactive glasses are derived from biowaste resources such as corn husk ash, sugarcane leaves ashes, and eggshell powder via the melt quench method. The raw materials were melted in a platinum rhodium crucible at 1550 °C, followed by rapid cooling to obtain a glassy phase. In order to assess the in-vitro bioactivity, glasses were soaked in simulated body fluid at 37 °C for different time durations, i.e., 7,14, 21, and 28 days. Biocompatibility of these glasses was also tested on MG-63 cell lines using a 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) assay test. X-ray diffraction, Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM) with energy dispersive spectroscopy (EDS) were used to analyze the hydroxyapatite (HAp) layer formation and the thermal stability of soaked samples. SEM micrographs showed the cauliflower and flakes type morphology of hydroxyapatite (HAp) on the glass surfaces after 7 days of soaking in SBF. The EDS analysis also confirmed the formation of a hydroxyapatite layer with a Ca/P ratio that varies from 1.6 to 3.9. MTT assay revealed the biocompatibility for different dosages (1, 2, 5, and 10 mg/ml) of the as-prepared glasses with MG-63 cell lines. The heat treatment of soaked glasses converts the amorphous HAp layer into a crystalline HAp layer, followed by its conversion in stable diopside as main and Ca2.589Mg0.41(PO4)2 as minor phase. The tested glasses exhibited good bioactivity and biocompatibility particularly higher CaO content glass, suggesting for use in bone regeneration applications.

Design of Bionanomaterial of Chitosan Carbohydrate Polymer Composited with Broccoli Extract and Zinc Oxide Nanoparticles: Anticancer Activity in Human Osteosarcoma.

In the current research, a chitosan/broccoli extract/ZnO nanoparticle (CH/BE/ZnO) bionanocomposite was created. The physicochemical properties of CH/BE/ZnO bionanocomposite were investigated using a variety of methods, including field emission scanning electron microscopy (FESEM), elemental analysis (CHN-O), X-ray diffraction (XRD), Fourier transform infrared spectrum (FTIR), Brunauer-Emmett-Teller (BET), and transmission electron microscopy (TEM). The CH/BE/ZnO bionanocomposite's biological activity was assessed by examining its cytotoxicity capabilities against a bone cancer cell line (MG63). The total pore volume and specific surface area of CH/BE/ZnO are 0.134 cm3/g and 16.99 m2/g, respectively. The IC50 results for CH/BE/ZnO bionanocomposite in bone cancer investigations using the MTT test against the MG63 cell line was 115μg/mL. The results indicate that the CH/BE/ZnO bionanocomposite is an effective chemotherapeutic agent against human osteosarcoma. The CH/BE/ZnO bionanocomposite showed high performance and structure, which means innovating nanomaterial agents for biological applications in the future.

Targeted Drug Delivery Using 5-Fluorouracil Loaded Porous Silica Nanoparticles for Enhanced Osteosarcoma Treatment: An In Vitro Study on the MG-63 Cell Line

Targeted Drug Delivery Using 5-Fluorouracil Loaded Porous Silica Nanoparticles for Enhanced Osteosarcoma Treatment: An In Vitro Study on the MG-63 Cell Line

Effects of Three Retrograde Filling Materials on Production of Inflammatory Cytokines and Resorbing Mediators

: Root-end filling materials are typically used following endodontic surgeries, and due to their different chemical compositions, they have varying effects on the survival and differentiation of tissues around the root apex. The purpose of this in vitro study was to evaluate the effects of three retrograde filling materials—ProRoot MTA, Super ethoxy benzoic acid (EBA), and Geristore—on the production of pro-inflammatory and resorptive cytokines (RANK, RANK-L, OPG, IL-1β, and TNF-α). After mixing and setting, the experimental materials were exposed to UV rays and then applied to the prepared cells. This study used osteoblast (MG-63) and THP-1 monocyte cell lines, as well as peripheral blood monocytes (PBM). The evaluation times for RANK-L and OPG were 24 and 48 hours, while IL-1β and tumor necrosis factor (TNF-α) were evaluated at 24 hours. The RANK levels were measured using flow cytometry, and monocyte survival was assessed at different times using the MTT assay. The results were analyzed using Minitab statistical software, which indicated that the type of material significantly affected cytokine production. Exposure of monocytes to EBA resulted in a decrease in TNF-α and IL-1β secretion compared to the other groups. However, in the THP-1 cell line, there was no significant difference in the release of pro-inflammatory cytokines between the groups. Conversely, Geristore induced the highest level of RANK-L and the lowest level of OPG in cultures with the MG-63 cell line. According to the MTT assay, the viability order of the materials from early to late was Geristore < EBA < MTA.

Fabrication and characterization of multifunctional, asymmetric bilayer films based on chitosan/gelatin/mesoporous bioactive glass nanoparticles for guided bone regeneration

AbstractTwo-faced GBR membranes were fabricated by electrophoretic deposition (EPD) using a combination of biopolymers and mesoporous bioactive glass nanoparticles (MBGNs). The membrane design was aimed at leveraging the advantageous properties of both biopolymers and MBGNs. The dense composite layer consisted of chitosan (CS) incorporating MBGNs and it was functionalized with a phytotherapeutic drug, naringin (Nar). The porous layer consisted of CS-gelatin (Gel)- MBGNs as well as copper chelated chitosan (Cu(II)-CS)-Gel-MBGNs composites. EPD was conducted in direct current mode. The antibacterial activity of the membranes as a result of the presence of Cu(II) and Nar was confirmed. The films were cytocompatible when tested with MC3T3-E1 (pre-osteoblastic) and MG-63 (osteoblast like) cell lines. However, a slight cytotoxic effect of the releasing Cu(II) ions was determined. In contrast, Nar-loaded films revealed improved cell viability. The results indicate the high potential of EPD to fabricate bilayer structures for GBR applications. Graphical abstract

Chalcones induce apoptosis, autophagy and reduce spreading in osteosarcoma 3D models

Osteosarcoma is the most common primary bone malignancy with a challenging prognosis marked by a high rate of metastasis. The limited success of current treatments may be partially attributed to an incomplete understanding of osteosarcoma pathophysiology and to the absence of reliable in vitro models to select the best molecules for in vivo studies. Among the natural compounds relevant for osteosarcoma treatment, Licochalcone A (Lic-A) and chalcone derivatives are particularly interesting. Here, Lic-A and selected derivatives have been evaluated for their anticancer effect on multicellular tumor spheroids from MG63 and 143B osteosarcoma cell lines. A metabolic activity assay revealed Lic-A, 1i, and 1k derivatives as the most promising candidates. To delve into their mechanism of action, caspase activity assay was conducted in 2D and 3D in vitro models. Notably, apoptosis and autophagic induction was generally observed for Lic-A and 1k. The invasion assay demonstrated that Lic-A and 1k possess the ability to mitigate the spread of osteosarcoma cells within a matrix. The effectiveness of chalcone as a natural scaffold for generating potential antiproliferative agents against osteosarcoma has been demonstrated. In particular, chalcones exert their antiproliferative activity by inducing apoptosis and autophagy, and in addition they are capable of reducing cell invasion. These findings suggest Lic-A and 1k as promising antitumor agents against osteosarcoma cells.

Formulation and characterization of a novel anti-human bone cancer supplement from silver nanoparticles green-synthesized using Allium sativum l. Leaf aqueous extract

Several studies have recently uncovered the ability of plants to enhance the therapeutic efficacies of Ag NPs on various carcinoma cells, particularly those associated with human bone tumors. Developing and formulating novel chemotherapy supplements or medications for the management of bone tumors in humans is a top research focus for both developing and developed nations. Our current study involved the development of a contemporary chemotherapeutic medication utilizing silver nanoparticles (AgNPs) combined with an aqueous extract of Allium sativum L. for the purpose of treating bone tumors in humans. AgNPs were characterized using XRD, TEM, UV–Vis, and FE-SEM. The peaks detected at 2θ values of 77.4, 65.1, and 37.8 were associated with the crystal planes 311, 220, and 111, respectively. To examine the antioxidant characteristics of AgNPs, the DPPH assay was employed with BHT serving as the positive control. The MTT assay was employed to examine the anti-human bone tumor and cytotoxicity properties of AgNPs on bone cancer cells, including A-673 and HS-729 (rhabdomyosarcoma), MHH-ES1 and CADO-ES1 (Ewing’s sarcoma), MG-63 and HOS (osteosarcoma), as well as CH-3573 and SW-1353 (chondrosarcoma). The DPPH assay demonstrated comparable antioxidant capacities among AgNPs, and butylated hydroxytoluene. The anti-human bone tumor properties and cell viability of silver nanoparticles exhibited a dose-dependent decrease against CH-3573, SW-1353, MG-63, HOS, MHH-ES1, CADO-ES1, HS-729, and A-673 cells, while showing no cytotoxic effects on the normal cell line. The viability of the cells and their response to the nanoparticles varied based on the dosage administered. The silver nanoparticles exhibited IC50 values of 154, 67, 129, 110, 124, 107, 70, and 81 µg/ml against H-3573, SW-1353, MG-63, HOS, MHH-ES1, CADO-ES1, HS-729, and A-673 cell lines, respectively. The HOS revealed the most effective outcome regarding anti-human bone tumor efficacies when treated with AgNPs, surpassing the results observed in other cell lines. Based on the aforementioned results, it is possible to utilize the silver nanoparticles with A. sativum extract for the management of various human bone tumors, particularly human bone rhabdomyosarcoma, human bone chondrosarcoma, human bone osteosarcoma, and human bone Ewing’s sarcoma.

Fibronectin Functionalization: A Way to Enhance Dynamic Cell Culture on Alginate/Hydroxyapatite Scaffolds.

Bone defects are a global health concern; bone tissue engineering (BTE) is the most promising alternative to reduce patient morbidity and overcome the inherent drawbacks of autograft and allograft bone. Three-dimensional scaffolds are pivotal in this field due to their potential to provide structural support and mimic the natural bone microenvironment. Following an already published protocol, a 3D porous structure consisting of alginate and hydroxyapatite was prepared after a gelation step and a freezing-drying step. Despite the frequent use of alginate in tissue regeneration, the biological inertness of this polysaccharide hampers proper cell colonization and proliferation. Therefore, the purpose of this work was to enhance the biological properties by promoting the interaction and adhesion between cells and biomaterial with the use of Fibronectin. This extracellular matrix protein was physically adsorbed on the scaffold, and its presence was evaluated with environmental scanning electron microscopy (eSEM) and the Micro-Bicinchoninic Acid (μBCA) protein assay. The MG-63 cell line was used for both static and dynamic (i.e., in bioreactor) 3D cell culturing on the scaffolds. The use of the bioreactor allowed for a better exchange of nutrients and oxygen and a better removal of cell catabolites from the inner portion of the construct, mimicking the physiological environment. The functionalized scaffolds showed an improvement in cell proliferation and colonization compared to non-functionalized ones; the effect of the addition of Fibronectin was more evident in the dynamic culturing conditions, where the cells clearly adhered on the surface of functionalized scaffolds.

Physico-Chemical Properties of Copper-Doped Hydroxyapatite Coatings Obtained by Vacuum Deposition Technique.

The hydroxyapatite and copper-doped hydroxyapatite coatings (Ca10-xCux(PO4)6(OH)2; xCu = 0, 0.03; HAp and 3CuHAp) were obtained by the vacuum deposition technique. Then, both coatings were analyzed by the X-ray diffraction (XRD), scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR) and water contact angle techniques. Information regarding the in vitro antibacterial activity and biological evaluation were obtained. The XRD studies confirmed that the obtained thin films consist of a single phase associated with hydroxyapatite (HAp). The obtained 2D and 3D SEM images did not show cracks or other types of surface defects. The FTIR studies' results proved the presence of vibrational bands characteristic of the hydroxyapatite structure in the studied coating. Moreover, information regarding the HAp and 3CuHAp surface wettability was obtained by water contact angle measurements. The biocompatibility of the HAp and 3CuHAp coatings was evaluated using the HeLa and MG63 cell lines. The cytotoxicity evaluation of the coatings was performed by assessing the cell viability through the MTT assay after incubation with the HAp and 3CuHAp coatings for 24, 48, and 72 h. The results proved that the 3CuHAp coatings exhibited good biocompatible activity for all the tested intervals. The ability of Pseudomonas aeruginosa 27853 ATCC (P. aeruginosa) cells to adhere to and develop on the surface of the HAp and 3CuHAp coatings was investigated using AFM studies. The AFM studies revealed that the 3CuHAp coatings inhibited the formation of P. aeruginosa biofilms. The AFM data indicated that P. aeruginosa's attachment and development on the 3CuHAp coatings were significantly inhibited within the first 24 h. Both the 2D and 3D topographies showed a rapid decrease in attached bacterial cells over time, with a significant reduction observed after 72 h of exposure. Our studies suggest that 3CuHAp coatings could be suitable candidates for biomedical uses such as the development of new antimicrobial agents.

Benzimidazole-oxindole hybrids: A novel class of selective dual CDK2 and GSK-3β inhibitors of potent anticancer activity.

A new series of benzimidazole-oxindole hybrids 8a-x was discovered as dual cyclin-dependent kinase (CDK2) and glycogen synthase kinase-3-beta (GSK-3β) inhibitors with potent anticancer activity. The synthesized hits displayed potent anticancer activity against national cancer institute cancer cell lines in single-dose and five-dose assays. Moreover, the derivatives 8k, 8l, 8n, 8o, and 8p demonstrated potent cytotoxic activity against PANC-1 cells with IC50 = 1.88-2.79 µM. In addition, the hybrids 8l, 8n, 8o, and 8p displayed potent antiproliferative activity on the MG-63 cell line (IC50 = 0.99-1.90 µM). Concurrently, the benzimidazole-oxindole hybrid 8v exhibited potent dual CDK2/GSK-3β inhibitory activity with IC50 values of 0.04 and 0.021 µM, respectively. In addition, 8v displayed more than 10-fold higher selectivity toward CDK2 and GSK-3 β over CDK1, CDK5, GSK-3α, vascular endothelial growth factor receptor-2, and B-rapidly accelerated fibrosarcoma. Screening of the effect of 8n and 8v on the cell cycle and apoptosis of PANC-1 and MG-63 cells displayed their ability to arrest their cell cycle at the G2-M phase and to potentiate the apoptosis of both cell lines. In silico docking of the benzimidazole-oxindole hybrid 8v into the catalytic pocket of both CDK2 and GSK-3β revealed its perfect fitting through the formation of hydrogen bonding and hydrophobic interactions with the key amino acids in the binding sites. In addition, in silico absorption, distribution, metabolism, excretion studies proved that 8a-x exhibit satisfactory drug-likeness properties for drug development.

Apoptotic Induction by Biosynthesized Gold Nanoparticles Using Phormidesmis communis Strain AB_11_10 against Osteosarcoma Cancer.

Phormidesmis communis strain AB_11_10 was isolated and identified using microscopy and 16s rRNA sequencing, and its phytochemical constituents were determined using liquid chromatography-quadrupole time-of-flight mass spectrometry. The isolate had a segmented filamentous shape with a blue-green color. Many biomolecules, including organic compounds, amino acids, and fatty acids, were detected. P. communis strain AB_11_10 was used to synthesize gold nanoparticles (Ph-AuNPs) by adjusting the optimum reaction conditions. The concentration, algal/precursor ratio, temperature, reaction time, and pH significantly influenced the synthesis of the Ph-AuNPs. Mixing 1 mL of 0.5 mM of HAuCl4 with 1 mL of algal extract and exposing the mixture to 100 °C for 30 min at pH 5.6 were the optimum conditions for the biosynthesis of Ph-AuNPs at a wavelength of 524.5 nm. The Ph-AuNPs were characterized using TEM, SEM, EDX, and mapping Zeta sizer and FTIR. The Ph-AuNPs had quasi-spherical to triangular shapes with an average diameter of 9.6 ± 4.3 nm. Ph-AuNPs composed of 76.10 ± 3.14% of Au and trace amounts of carbon and oxygen were detected, indicating that the P. communis strain AB_11_10 successfully synthesized Ph-AuNPs. The hydrodynamic diameter of the Ph-AuNPs was 28.5 nm, and their potential charge was -17.7 mV. O-H, N-H, C=C, N-O, C-H, and C-O were coated onto the surfaces of the Ph-AuNPs. These groups correspond to algal phytochemicals, which may have been the main reducing and stabilizing substances during the Ph-AuNP synthesis. The therapeutic activity of the Ph-AuNPs against osteosarcoma cancers was examined in MG-63 and SAOS-2 cell lines, while their biocompatibility was tested against Vero cell lines using a sulforhodamine B assay. The Ph-AuNPs had potent antitumor activity against the MG-63 and SAOS-2 cells, with a low toxicity toward Vero cells. Flow cytometry and cell cycle arrest analyses revealed that the Ph-AuNPs enhanced the apoptotic pathway and arrested the cell cycle in the MG-63 and SAOS-2 cells. P. communis strain AB_11_10 provides a new source to synthesize small, stable, and biocompatible AuNPs that act as apoptotic enhancers in osteosarcoma.

Poly(styrene-co-methyl methacrylate)-silver/reduced graphene oxide-nano hydroxyapatite nanocomposites for bone cement applications

Herein, we report (polystyrene-co-polymethylmethacrylate/silver/reduced graphene oxide)-(nano-hydroxyapatite) ((PS-PMMA/Ag/RGO)-(nHA)) nanocomposites as potential bone cement. The polymer nanocomposites were characterized using Fourier Transform Infrared spectroscopy (FT-IR), X-ray diffraction (XRD), nano-indentation, thermal, and electron microscopic methods. The FT-IR and XRD results confirmed the successful formation of the nanocomposites. The elastic modulus (E) was found to be 6.85 GPa, and hardness (H) was 0.181 GPa for the nanocomposite bone cement samples, while the E and H values of commercial counterparts are 7.22 and 0.218 GPa, respectively. The thermogravimetric analysis (TGA) has revealed that the nanocomposite bone cement are stable up to 371 °C. The addition of nHA enhances the biocompatibility of bone cement. Cytotoxicity studies were conducted using human osteosarcoma MG63 cell lines. In-vitro studies showed that the material does not display any toxicity on the MG63 cell line, and increased concentration of nHA did not significantly impact the material’s toxicity, only impacting the curing kinetics of the cement.

A comprehensive investigation of strontium doped hydroxyapatite-merwinite composite: In-vitro biomineralization, mechanical testing, and cytotoxicity analysis

The advancement of biomaterials for reconstruction and replacement of damaged tissues is essential, which would possess biocompatibility, better mechanical, biochemical, and physical properties. Merwinite (Ca3MgSi2O8) has emerged as a promising material for hard tissue engineering, particularly in composite with hydroxyapatite (HAp). Strontium-doped hydroxyapatite (SrHAp) was prepared with varying dopant concentrations (1%, 3%, and 5% Sr) on the calcium site of HAp. The materials were analyzed by XRD, FTIR and SEM/EDX techniques. XRD analysis revealed average crystallite sizes ranging from 46 to 50 nm for SrHAp. It was found that the Merwinite-SrHAp composite doped with 5% strontium had a higher rate of apatite nucleation compared to pure composite, indicating that strontium doping enhances the bioactivity of the composite. The composite scaffolds, prepared in an 80:20 ratio of SrHAp to merwinite, exhibited compressive strengths ranging from 63.43 MPa (1% Sr doping) to 80.68 MPa (5% Sr doping). Cytotoxicity assessment against MG63 cell line via MTT assay demonstrated cell inhibition rates ranging from 11.71% (at 6.5 μg/mL) to 68.35% (at 100 μg/mL) for the 5% Sr-doped composite. Alkaline phosphatase (ALP) investigation revealed enhanced ALP activity for cells cultured on the 5% Sr-doped composite compared to control samples, indicating increased cell differentiation and mineralization potential.

In Vitro and In Vivo Analysis of the Mg-Ca-Zn Biodegradable Alloys.

The objective of this work was to analyze the in vitro and in vivo tests of a novel Mg-based biodegradable alloy-Mg-0.5%Ca-with various amounts of Zn (0.5, 1, 1.5, 2.0, and 3.0 wt.%). In terms of in vitro biocompatibility, MTT and Calcein-AM cell viability assays, performed on the MG-63 cell line through the extract method, revealed that all five alloy extracts are non-cytotoxic at an extraction ratio of 0.025 g alloy per mL of cell culture medium. In the in vivo histological analysis, Mg-0.5Ca-1.5Zn demonstrated exceptional potential for stimulating bone remodeling and showed excellent biocompatibility. It was observed that Mg-0.5Ca-0.5Zn, Mg-0.5Ca-1.5Zn, and Mg-0.5Ca-3Zn displayed good biocompatibility. Furthermore, the histological examination highlighted the differentiation of periosteal cells into chondrocytes and subsequent bone tissue replacement through endochondral ossification. This process highlighted the importance of the initial implant's integrity and the role of the periosteum. In summary, Mg-0.5Ca-1.5Zn stands out as a promising candidate for bone regeneration and osseointegration, supported by both in vitro and in vivo findings.

Flexible, electrospun boron nitride nanosheets (BNNS)/hydroxyapatite –PVDF nanofibers with superior piezoelectric/ferroelectric, biocompatible features for effective bone tissue regeneration

Piezoelectric/ferroelectric scaffolds are emerging tools in tissue engineering that offer new platforms for the repair and regeneration of damaged soft and hard tissues by means of electrical stimulation. In line with this, boron nitride nanosheets (BNNS)/hydroxyapatite (H)-polyvinylidene fluoride (PVDF) based piezoelectric electrospun nanofibers were developed for effective bone tissue regeneration. BNNS were prepared through a sequential process involving lithium intercalation, followed by hydrothermal exfoliation. Four distinct sets of nanofibers were prepared by altering the concentration of BNNS (2.5 wt%, 5 wt%, 10 wt%) while maintaining a constant concentration of HAP nanoparticles (2.5 wt%) in the PVDF polymer matrix. The piezoelectric/ferroelectric performance of the PVDF-H-BNNS nanofibers were found to be enhanced with the addition of BNNS. The PVDF-H-BNNS nanofibers, with an effective contact area of 1 cm2, exhibited remarkable piezoelectric characteristics, exhibiting a peak voltage of 13 V and a current of 1.63 µA under the influence of 10 N force at a frequency of 10 Hz, showcasing their exceptional electromechanical response. The cytocompatibility studies of PVDF-H-BNNS nanofibers on MG-63 and HOS cell lines reveals excellent viability of nanofibers, akin to that control. Cell proliferation investigations conducted with MG-63 and HOS cell lines on PVDF-H-BNNS showcased the remarkable biocompatibility of the engineered membranes. The PVDF-H-BNNS piezoelectric nanofibers, having exceptional piezo-response and enhanced cytocompatibility, hold promise as ideal candidates for applications in the field of bone tissue engineering.

Thiolated eudragit/β-cyclodextrin/centella asiatica nanomaterial for bone tissue engineering construct

Polymeric scaffolds and nanomaterials enriched with herbal extracts have been extensively studied for bone tissue regeneration due to their well-organized targeting ability and site-specific action at the intended locus. The use of polymer hybrids functionalized with plant extract in developing bone-specific material is a significant approach. Therefore, the current study aimed to develop a nanomaterial using a combination of thiolated eudragit, β-cyclodextrin, and Centella Asiatica to promote bone tissue regeneration. Initially, a composite material was developed by freeze-thawing technique and subsequently, the composite was developed into a nanocomposite by combining ionic gelation and solvent precipitation techniques. The nanocomposite was characterized by dynamic light scattering, scanning electron microscopy, atomic force microscopy, XRD, and FTIR studies. To assess the cytotoxicity and cell differentiation efficiency of the composite, in vitro cell line studies such as MTT assay and cell proliferation studies were performed using MG 63 (human bone fibroblasts) cell lines. In silico studies were conducted to investigate the bone-targeting efficacy of the developed nanocomposite. The results of FTIR and XRD analysis confirmed the successful thiolation of eudragit. SEM analysis of the composite showed a porous structure with an average pore diameter of 1 μm. DLS and AFM characterization indicated the zeta potential value of −37.06 mV and the average size of the nanocomposite was within 100–300 nm. The gel fraction and swelling degree of the developed nanocomposite were observed to be 80.08 ± 4.47% and 2.313 ± 0.04 respectively. Viscosity was determined to be 133.6 ± 26.008 cps. Optical profilometry data of the composite indicated mean roughness (Ra) and root mean square roughness (Rq) of 4.727 μm and 6.075 μm respectively. MTT assay inferred the cytotoxic nature and tolerance of the nanoparticles against the MG-63 cell lines. Cell proliferation studies on the MG-63 cell line exhibited a characteristic growth with the developed nanoparticles. In vitro studies in corroboration with the in silico method of molecular docking confirmed that the composite has the bone targeting efficacy and the osteogenic property where it plays a key role in the isoprenoid biosynthetic pathway by being the inhibitor of Farnesyl diphosphate synthase (FDPS) which in turn results in bone tissue regeneration.

Exploring the biocompatibility and healing activity of actinobacterial-enhanced reduced nano-graphene oxide in in vitro and in vivo model and induce bone regeneration through modulation of OPG/RANKL/RUNX2/ALP pathways.

The development of cost-effective, simple, environment-friendly biographene is an area of interest. To accomplish environmentally safe, benign culturing that has advantages over other methods to reduce the graphene oxide (GO), extracellular metabolites from actinobacteria associated with mushrooms were used for the first time. Bactericidal effect of GO against methicillin-resistant Staphylococcus aureus, antioxidant activity, and hydroxyapatite-like bone layer formation, gene expression analysis and appropriate biodegradation of the microbe-mediated synthesis of graphene was studied. Isolated extracellular contents Streptomyces achromogenes sub sp rubradiris reduced nano-GO to graphene (rGO), which was further examined by spectrometry and suggested an efficient conversion and significant reduction in the intensity of all oxygen-containing moieties and shifted crystalline peaks. Electron microscopic results also suggested the reduction of GO layer. In addition, absence of significant toxicity in MG-63 cell line, intentional free radical scavenging prowess, liver and kidney histopathology, and Wistar rat bone regeneration through modulation of OPG/RANKL/RUNX2/ALP pathways show the feasibility of the prepared nano GO. The study demonstrates the successful synthesis of biographene from actinobacterial extracellular metabolites, its potential biomedical applications, and its promising role in addressing health and environmental concerns.