Human Fetal Osteoblast Research Articles

Assessment of Cytotoxicity of Magnesium Oxide and Magnesium Hydroxide Nanoparticles using the Electric Cell-Substrate Impedance Sensing

Magnesium (Mg)-based alloys have the potential for bone repair due to their properties of biodegradation, biocompatibility, and structural stability, which can eliminate the requirement for a second surgery for the removal of the implant. Nevertheless, uncontrolled degradation rate and possible cytotoxicity of the corrosion products at the implant sites are known current challenges for clinical applications. In this study, we assessed in vitro cytotoxicity of different concentrations (0 to 50 mM) of possible corrosion products in the form of magnesium oxide (MgO) and magnesium hydroxide (Mg(OH)2) nanoparticles (NPs) in human fetal osteoblast (hFOB) 1.19 cells. We measured cell proliferation, adhesion, migration, and cytotoxicity using a real-time, label-free, non-invasive electric cell-substrate impedance sensing (ECIS) system. Our results suggest that 1 mM concentrations of MgO/Mg(OH)2 NPs are tolerable in hFOB 1.19 cells. Based on our findings, we propose the development of innovative biodegradable Mg-based alloys for further in vivo animal testing and clinical trials in orthopedics.

Effect of Different Concentration of Cellulose Nanocrystals Comprising Hydroxyethyl Cellulose / Poly(Vinyl Alcohol) as a Bone Tissue Engineering Scaffold

In this study, biodegradable scaffolds based on hydroxyethyl cellulose (HEC) (5 wt%) and poly (vinyl alcohol) (PVA) (15 wt%) with different percentages of celullose nanocrystal (CNC) (1 and 7 wt%) were fabricated by lyophilization method to get highly porous scaffolds. These scaffolds were made water insoluble by cross-linking via heat treatment. The morphology and thermal properties of HEC/PVA/CNCs scaffolds were characterized by using Scanning Electron Microscope (SEM) and Thermogravimetric Analysis (TGA). The morphological study showed that both prepared scaffold have highly porous structures with good pore interconnected structure. It was observed that thermal properties of scaffolds increased significantly as the concentration of CNCs increased. Cytotoxicity studies on scaffolds were carried out by utilizing human fetal osteoblast (hFOB) cells using DAPI nuclear stain and then confirmed using SEM. hFOB cells were able to attach and spread on all scaffolds. Incorporated CNCs as reinforcing nanofiller on scaffolds promising a superior functionality in bone tissue engineering.

The Affinity of Human Fetal Osteoblast to Laser-Modified Titanium Implant Fixtures

Introduction: Implant surface modification methods have recently involved laser treatment to achieve the desired implant surface characteristics. Meanwhile, surface modification could potentially introduce foreign elements to the implant surface during the manufacturing process. Objectives: The study aimed to investigate the surface chemistry and topography of commercially available laser-modified titanium implants, together with evaluating the cell morphology and cell adhesion of human fetal osteoblast (hFOB) seeded onto the same implants. Method: Six (6) samples of commercially available laser-modified titanium implants were investigated. These implants were manufactured by two different companies. Three (3) implants were made from commercially pure grade 4 Titanium (Brand X); and three were made from grade 5 Ti6Al4V (Brand Y). The surface topography of these implants was analyzed by scanning electron microscope (SEM) and the surface chemistry was evaluated with electron dispersive x-ray spectroscopy(EDS). Human fetal osteoblasts were seeded onto the implant fixtures to investigate the biocompatibility and adhesion. Results & Discussion: Brand X displayed dark areas under SEM while it was rarely found on brand Y. These dark areas were consistent with their organic matter. The hFOB cell experiments revealed cell adhesion with filopodia on Brand X samples which is consistent with cell maturation. The cells on Brand Y were morphologically round and lacked projections, one sample was devoid of any noticeable cells under SEM. Cell adhesion was observed early at 48 hrs in laser-irradiated titanium fixtures from both the brands. Conclusion: The presence of organic impurities in Brand X should not be overlooked because disruption of the osseointegration process may occur due to the rejection of the biomaterial in an in-vivo model. Nevertheless, there was insufficient evidence to link implant failure directly with carbon contaminated implant surfaces. Further studies to determine the toxicity of Vanadium from Ti6Al4V in an in-vivo environment should indicate the reason for different cell maturation.

Type I Collagen-chitosan Membranes Crosslinked Chemically with N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide Hydrochloride for Guided Bone Regeneration: A Comparative Study

In this work, chemical crosslinking with N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC) was studied in type I collagen-chitosan (Col I-CS) membranes for their use in guided bone regeneration (GBR). The membranes were prepared in a wide range of Col I-CS proportions, produced in the form of films by the solvent evaporation technique and crosslinked with EDC. The membranes presented a rough surface that is adequate for cell adhesion and a compact cross-section to prevent some infiltration of unwanted cell tissues, according to scanning electron microscopy. The physicochemical characterization (thermogravimetric analysis, differential scanning calorimetry, uniaxial tensile test and in vitro degradation) showed that the crosslinked membranes are more resistant to dehydration, tensile strength and enzymatic digestion. The biological assays (adhesion, viability and morphology of cells) using human fetal osteoblasts (hFOB) indicated that the crosslinked membranes are not cytotoxic. Therefore, the chemical crosslinking with EDC improves the physicochemical properties of the membranes without affecting their biocompatibility, indicating that this type of membranes is a potential alternative to be used in GBR. In addition, this study also allowed to know the best type of crosslinked membranes, as a proposal for its application in GBR strategies.

Cytotoxic and osteogenic effects of crocin and bicarbonate from calcium phosphates for potential chemopreventative and anti-inflammatory applications in vitro and in vivo.

Delayed healing and nonhealing of bone defects or resected bone sites remains an important clinical concern in the biomedical field. Osteosarcoma is one of the most common types of primary bone cancers. Among calcium phosphates, hydroxyapatite (HA) and tricalcium phosphate (TCP) are the most widely used in various biomedical applications for bone reconstruction and replacement. In this study, crocin, saffron's natural bioactive and anti-inflammatory molecule, and bicarbonate, a neutralizing agent, were directly loaded onto HA disks to evaluate their in vitro release and effect on human osteoblast and osteosarcoma cell lines. This was assessed through release, initial toxicity, drug optimization, final toxicity studies and in vivo anti-inflammatory assessment through H&E indexing. It is hypothesized that the release of crocin, bicarbonate, and the dual release of both agents will decrease osteosarcoma cellular viability with no effect on osteoblast cells. A plateaued release of crocin and bicarbonate was achieved over seven weeks in physiological and acidic environments, where bicarbonate was shown to modulate the release of crocin. Through morphological characterization and MTT assay analysis, bicarbonate showed no toxicity to human fetal osteoblast (hFOB) cells and crocin significantly enhanced osteoblast proliferation. Through drug concentration optimization, all drug loaded samples decreased human osteosarcoma (MG-63) viability by 50% compared to control samples by Day 11, with clear changes in cell spreading and morphology. Moreover, 3D printed TCP scaffolds loaded with crocin and bicarbonate were tested in vivo in order to assess their preliminary effects on inflammation in a rat distal femur model at 4 days. Lower inflammatory cellular recruitment was achieved in the presence of crocin and bicarbonate, compared to the control. These results suggest a pro-apoptotic mechanism against osteosarcoma as well as anti-inflammatory properties of crocin and bicarbonate, elucidating a potential application for osteosarcoma regulation and wound healing for bone tissue regeneration applications.

Short Communication: Fructose-Enhanced Antibacterial Activity of Self-Assembled Nano-Peptide Amphiphiles for Treating Antibiotic-Resistant Bacteria.

BackgroundIn recent years, numerous bacteria have become resistant to conventional antibiotics. Fortunately, an increasing body of research indicates that through the addition of specific metabolites (like sugars), the antibacterial activity of certain drugs can be enhanced. A new type of self-assembled nano-peptide amphiphile (SANPA) was designed in this study to treat antibiotic-resistant bacterial infections and to reduce the use of antibiotics.MethodsHere, SANPAs were self-assembled into nanorod structures with a diameter of ca. 10.5 nm at concentrations greater than the critical micelle concentration (CMC) of 44.67 μM. Both Gram-positive and Gram-negative bacteria were treated with SANPAs with fructose supplementation.ResultsAfter a 30-min fructose pre-incubation, SANPAs reduced bacteria growth relative to non-fructose treatments at all concentrations. Cytotoxicity assays indicated that the presence of fructose seemed to slightly ameliorate the cytotoxic effect of the treatment on model human fetal osteoblasts (or bone-forming cells) and human dermal fibroblasts.ConclusionWe demonstrated here that SANPAs-like nanomaterials have a promising potential to treat antibiotic-resistant bacteria, especially when added to fructose, potentially limiting their associated infections.

Development of label-free biophysical markers in osteogenic maturation

The spatial and temporal changes of morphological and mechanical properties of living cells reflect complex functionally-associated processes. Monitoring these modifications could provide a direct information on the cellular functional state. Here we present an integrated biophysical approach to the quantification of the morphological and mechanical phenotype of single cells along a maturation pathway. Specifically, quantitative phase microscopy and single cell biomechanical testing were applied to the characterization of the maturation of human foetal osteoblasts, demonstrating the ability to identify effective label-free biomarkers along this fundamental biological process.

In Vitro Cytotoxicity of Possible Corrosion Products from Mg-Based Biodegradable Metals: Magnesium Oxide and Magnesium Hydroxide Nanoparticles

Biodegradable magnesium (Mg) alloys have potential applications in orthopedic implants due to their mechanical and osseointegration properties. However, the surface characteristics, biocompatibility, and toxicity of the released corrosion products in the form of magnesium oxide (MgO) and magnesium hydroxide (Mg(OH)2) nanoparticles (NPs) at the junction of implants and in the surrounding tissue are not completely understood. Here, we investigated in vitro cytotoxicity and morphological changes in human fetal osteoblast (hFOB) 1.19 cells in response to various concentrations (1 mM, 5 mM, 10 mM, and 50 mM) of MgO/Mg(OH)2 NPs by live/dead assay and scanning electron microscopy (SEM). In this study, we performed a surface characterization of MgO/Mg(OH)2 NPs to evaluate the size of the NPs. Further, an immersion test was performed in Dulbecco’s Modified Eagle’s Medium (DMEM) with randomly selected various concentrations (1 mM, 5 mM, 10 mM, 50 mM, and 100 mM) of MgO/Mg(OH)2 NPs to understand the degradation behavior of the NPs, and the change in the pH values from days 1 to 7 was measured. After conducting an immersion test for seven days, the highest concentration (100 mM) of MgO/Mg(OH)2 NPs was selected to study the element depositions on nanoparticles through scanning electron microscopy–energy-dispersive X-ray spectroscopy (SEM–EDX) mapping. The results from this in vitro cytotoxicity study suggest that less than or equal to 5-mM concentrations of MgO/Mg(OH)2 NPs are tolerable concentrations for hFOB 1.19 cells. This study provides a foundational knowledge of MgO/Mg(OH)2 NP cytotoxicity in hFOB 1.19 cells that can help to develop future sustainable biodegradable magnesium-based alloys for orthopedic applications.

Preparation and characterization of silk fibroin from four different species of Thai-local silk cocoon for Bone implanted applications

The metal interlocking nail is normally used in diaphyseal fractures. The bio-composite interlocking nail will be fabricated in the near future by using local silk fibroin reinforce Polylactic acid filament for 3D printing. Four species of local Thai Bombyx mori silk cocoons were selected to extract silk fibroin. The silk cocoon species consist of 1) Nangnoi Srisaket-I (NN), 2) Nanglai (NL), 3) Luang Saraburi (LS), and 4) J108. After the extraction, fibroins of each silk cocoon species were characterized and compared the physical property by using Scanning Electron Microscopy (SEM), Energy Dispersive X-ray (EDS) and Fourier Transform Infrared Spectroscopy (FT-IR). Then, the biological test was performed on cell viability and cytotoxicity with human fetal osteoblast cell line. The result presents that all of local silk cocoons species presented non-cytotoxicity ability which can be used in human or animal body without endangerment.

Proliferation study and microscopy evaluation on the effects of tannic acid in human fetal osteoblast cell line (hFOB 1.19).

Tannic acid (TA) is a phenolic compound that might act directly on osteoblast metabolism. The study was performed to investigate the effects of TA on the proliferation, mineralization, and morphology of human fetal osteoblast cells (hFOB 1.19). The cells were divided into TA-treated, untreated, and pamidronate-treated (control drug) groups. Half maximal effective concentration (EC50 ) values for TA and pamidronate were measured using MTT assay. The EC50 of hFOB 1.19 cells treated with TA was 2.94 M. This concentration was more effective compared to the pamidronate (15.27 M). Cell proliferation assay was performed to compare cell viability from Day 1 until Day 14. The morphology of hFOB 1.19 was observed via inverted microscope and scanning electron microscope. Calcium (Ca) and phosphate (P) were assessed using energy-dispersive X-ray (EDX) analysis. Furthermore, the mineralization of hFOB 1.19 was determined by von Kossa staining (P depositions) and Alizarin Red S staining (Ca depositions). The number of cells treated with TA was significantly higher than the two control groups at Day 10 and Day 14. The morphology of cells treated with TA was uniformly fusiform-shaped with filopodia extensions. Besides, globular-like structures of deposited minerals were observed in the TA-treated group. In line with other findings, EDX spectrum analysis confirmed the presence of Ca and P. The cells treated with TA had significantly higher percentage of both minerals at Day 3 and Day 10 compared to the two control groups. In conclusion, TA enhances cell proliferation and causes cell morphology changes, as well as improved mineralization.

Sustained release of vitamin C from PCL coated TCP induces proliferation and differentiation of osteoblast cells and suppresses osteosarcoma cell growth.

The objective of this study is to understand the effect of sustained release of vitamin C from β-tricalcium phosphate (β-TCP) scaffold on proliferation, viability and differentiation of human fetal osteoblast cells (hFOB). The influence of pH, drug concentration, and presence of polymer on the sustained release of vitamin C from polycaprolactone (PCL) coated β-TCP scaffolds are studied. Prolonged and sustained release of vitamin C, over 60 days is observed in PCL coated β-TCP scaffolds compared to uncoated scaffolds. Presence of PCL helps to minimize the burst release of vitamin C from β-TCP scaffolds in the initial 24 h of release. To evaluate the osteogenic potential of vitamin C incorporated β-TCP scaffolds, osteoblast cells are cultured and cell morphology, proliferation, viability, and differentiation are assessed. Morphological characterization shows layer like osteoblast cell attachment in the presence of vitamin C compared to the control. MTT cell viability assay shows 2 folds increase in osteoblast cell density in the presence of vitamin C after 3,7 and 11 days of culture. Furthermore, increased ALP activity at 11 days of culture indicates the possible role of vitamin C on osteoblast differentiation. Additionally, a preliminary study shows vitamin C loaded scaffolds suppress osteosarcoma (MG-63) cell proliferation to 4 folds after 3 days compared to control. These results show a sustained release of vitamin C from PCL coated β-TCP scaffolds improve proliferation, viability, and differentiation of osteoblasts cell as well as mitigate osteosarcoma cell proliferation, suggesting its potential application as synthetic bone graft substitutes in tissue engineering application.

Modulation of biological properties by grain refinement and surface modification on titanium surfaces for implant-related infections

The nanostructured titanium (Ti) obtained by the equal-channel angular pressing (ECAP) has shown great promise as a biomedical implant material over the past few decades. The present work aims to investigate the effect of topographical changes caused by ECAP and piranha treatment (Tr) on the surface performance and biological properties of Ti for bone tissue engineering applications. The effects of dual treatments, i.e., ECAP and Tr, on Ti were systematically investigated by multiple characterization techniques, surface wettability, apatite-forming ability, cellular behavior, and antibacterial studies. We demonstrate that both ECAP and ECApTr samples possess desirable mechanical and physical properties and are biocompatible to cultured human fetal osteoblast (hFOB) cells. The potential of adhesion and proliferation of hFOB cells on ECAP and ECApTr samples was found to be superior to that of control unprocessed sample (annealed). Ti samples prepared by both methods showed excellent antimicrobial properties against clinical strains of the most common pathogenic bacteria causing orthopedic implant infections, Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa). This study supports the established claim about mechanical properties improvement by ultrafine refinement and further enhances the antibacterial properties when chemically etched with a piranha solution.

3-D printed Ti-6Al-4V scaffolds for supporting osteoblast and restricting bacterial functions without using drugs: Predictive equations and experiments

Conditions resulting from musculoskeletal deficiencies (MSDs) are wide-ranging and retain the likelihood for restricting motion or producing pain, especially in the lower back, neck, and upper limbs. Engineered scaffold devices are being produced to replace antiquated modalities that suffer from structural and mechanical deficiencies in the treatment of MSDs. Here, as-fabricated Ti-6Al-4V-based Hive™ interbody fusion scaffolds, commercialized by HD Lifesciences LLC, were assayed for their osteogenicity and antibacterial potential using a series of characterization and in vitro tests, as well as by quantitative analyses. A topographical assessment of the Hive™ meshes indicated that the elementally pure substrates are microscopically porous and rough, in addition to displaying structural heterogeneity. Roughness estimations and static contact angle measurements recommended the use of the as-fabricated Ti-6Al-4V substrates for supporting osteoblast attachment, especially, due to the improved surface roughness and wettability values of these scaffolds relative to the unembellished Ti-6Al-4V surfaces. Quantitative correlations relating the surface properties of roughness and energy were applied to predict cellular behaviors. Cell growth suppositions were experimentally corroborated. Critical in vitro data indicated the competencies of the Hive™ scaffolds for promoting the adhesion and proliferation of human fetal osteoblasts (hFOBs), accumulating substantial calcium deposition from metabolizing hFOBs, and restricting the attachment of bacteria. The model system that investigated the pre-adsorption of casein proteins along the Hive™ test substrates additionally furthered the notion that bacterial attachment may be restricted, with short-scale adhesion dynamics serving as the theoretical basis for this hypothesis. In this manner, this study showed that through predictive models and experiments, these novel 3D printed Ti-based scaffolds can increase bone cell while decreasing bacteria functions without using drugs. Statement of SignificanceSintered Ti-6Al-4V spinal fusion devices (Hive™) manufactured and marketed by HD Lifesciences LLC were assessed for their biocompatibility and antibacterial performance. A mixed methods approach was employed, whereby quantitative measures were used to predict the ability for Hive™ substrates to adsorb specialized proteins and to restrict bacterial surface colonization. In vitro tests that evaluated bone cell and bacterial adhesion, calcium deposition, and protein adsorption supported quantitative predictions. The data herein presented demonstrate the following: (1) surface energy is an important predictor of implant-cell interactions, (2) strong correlations exist between surface energy and surface roughness, (3) mathematical models can be used to improve and predict implant device perofrmance, and (4) porous, rough, 3D-printed materials perform well in terms of biocompatibility and antimicrobial efficacy.

Drug loaded electrospun polymer/ceramic composite nanofibrous coatings on titanium for implant related infections

Abstract Developing an effective antibacterial surface with the help of drugs that prevent bacterial adhesion, colonization, and proliferation into the surrounding tissues is of great demand. Rifampicin (Rf) is effective antibiotic drug proved has proved its potential in treating bacteria in biofilms, especially against the microbes causing bone infections. Hydroxyapatite (HA), a biocompatible osteoconductive ceramic, has been verified to be a significant material for bioactivity enhancement. Electrospinning is an effective inexpensive method for incorporating nanoparticles into nanofibers with uniform distribution for the drug delivery system for tissue engineering applications. In the current study, for improving bioactivity and antibacterial properties, novel functional polycaprolactone (PCL) composite nanofibers loaded HA and Rf was developed and coated on titanium (Ti). Different characterization techniques such as SEM, EDS, XRD, FITR were used to analyze these PCL/Rf/HA nanocomposites. The results showed that the bioactivity and tensile strength of the composite scaffold increased with the addition of HA nanoparticles. In vitro bioactivity demonstrated that the PCL/HA/Rf composite nanofibers possess enhanced calcium deposition when compared to the pure sample. Cellular interactive responses such as adhesive and proliferation were evaluated using hFOB human fetal osteoblast cell lines. After 6 days of culturing, the cellular properties on Ti sample coated with PCL/HA/Rf was significantly improved. Antibacterial evaluations on the substrates showed that Rf-loaded PCL/HA fibers displayed >3 log reduction against S.aureus MRSA, and S.epidermidis bacterial strain and >2 log reduction against P.aeruginosa bacteria. In vitro drug release study shows initial burst release of Rf, followed by sustained released of 62% at the end of 32 days. The cell viability, adhesion, and proliferation evaluation suggest that the PCL/HA/Rf coated substrate possess good cytocompatibility. Further incorporation of Rf enhanced the antibacterial property of this nanofibrous scaffold.

Nanocrystalline Zn2+ and SO42- binary doped fluorohydroxyapatite: A novel biomaterial with enhanced osteoconductive and osteoinconductive properties.

In this study, we have successfully doped hydroxyapatite (HA) with zinc (Zn2+), sulphate (SO42-) and fluoride (F-) ions to develop a new composition of bioceramic, Ca10-x Znx(PO4)6-y(SO4)y(OH)2-z-yFz(SO4)y, (x = 0, 0.2, 0.6, 1.0, y = 0, 0.5 and z = 0,1.0 mol), using wet precipitation method. The obtained materials were analysed using XRD, FTIR, FESEM, and XPS techniques to investigate the phase purity, particle morphology and elemental composition, respectively. A model anticancer drug (Doxorubicin, DOX) was loaded onto the surface of the Zn/SO4-FHA materials. About 100% loading of DOX with a controlled release profile was obtained. Degradation of materials in Simulated body fluid (SBF) was greatly improved with the incorporation of Zn2+/SO42- ions in comparison to HA/FHA, which makes it highly bioactive materials. In vitro cell viability and adhesion of Human fetal osteoblast (hFOB) cell were investigated. Cell viability has demonstrated that the hFOB cells proliferated at a high rate on Zn/SO4-FHA materials, confirming the in vitro biocompatibility of the materials. Alkaline phosphatase (ALP) activity and intracellular calcium deposition of hFOB cells seeded on 1ZnSO4-FHA disc surface was statistically higher than observed on pure HA and FHA discs, indicating that hFOB cells differentiated into mature osteoblasts on 1Zn/SO4-FHA disc surfaces. Taken together, our results suggest that HA substituted by (Zn2+, 0.2 mol), (SO42-, 0.5 mol) and (F-, 1 mol) (1Zn/SO4-FHA) material was a promising material for hard tissue scaffolds.

Biocompatibility of Developing 3D-Printed Tubular Scaffold Coated with Nanofibers for Bone Applications

3D printing with controlled microarchitectures has gained traction in a wide variety of fields, including bone tissue engineering, because it represents an exciting alternative for the synthesis of new scaffolds due to its rapid manufacturing process, high precision, cost-effectiveness, and ease of use. Thus, this study is aimed at evaluating the biocompatibility response of a 3D-printed tubular scaffold coated by a layer of 7% PLA nanofibers. The morphology, structure, and chemical composition of the 3D-printed tubular scaffold were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier Transform Infrared (FTIR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and surface property analysis by profilometry. The biocompatibility response of the scaffold was assessed by cell adhesion, proliferation, and cell-material interactions of human fetal osteoblasts. Our results showed that 3D printing allowed obtaining similar and reproducible structures and the biocompatibility assays showed that nanofiber coating of the surface of the 3D tubular scaffold promoted an improvement on cell attachment, proliferation, and the morphology of osteoblast cells when compared with a noncoated scaffold. In conclusion, the surface of the 3D-printed tubular scaffold could be improved by the deposition of a nanofiber layer to render a more mimetic and active topography with excellent cellular biocompatibility for bone tissue applications.

Characterization and In Vitro evaluation of a novel coated nanocomposite porous 3D scaffold for bone repair

The aim of this study is to tissue engineer a 3D scaffold that can be used for load bearing segmental bone defects (SBDs) repair. Three different scaffolds were fabricated using cockle shell-derived CaCO3 aragonite nanoparticles (CCAN), gelatin, dextran and dextrin with coated framework via Freeze-Drying Method (FDM) labeled as 5211, 5211GTA+Alginate, 5211PLA. Scaffolds were assessed using Scanning Electron Microscopy (SEM). The cytocompatibility of the organized scaffolds was assessed using cells multiplication and alkaline phosphatase (ALP) concentration via In Vitro cell culture using human Fetal OsteoBlast cells line (hFOB). The results showed a substantial difference in ALP concentrations between the cultures of different scaffolds leachable medium during the study period. The biological evaluation also showed that three scaffolds did enhanced the osteoblast proliferation rate and improved the osteoblast function as demonstrated by the significant increase in ALP concentration. Engineering analyses showed that scaffolds possessed 3D interconnected homogenous porous structure with a porosity ranging 6%-49%, pore sizes ranging 8-345 µm, mechanical strength ranging 20-65 MPa, young’s modulus ranging 166-296 MPa and enzymatic degradation rate between 16%-38% within 2-10 weeks. The in vitro evaluation revealed that the scaffold 5211, 5211GTA+Alginate and 5211PLA fulfill all the main requirements to be considered as an ideal bone replacement.

Liposome-Encapsulated Curcumin-Loaded 3D Printed Scaffold for Bone Tissue Engineering.

Curcumin, the active constituent for turmeric, is known for its antioxidant, anti-inflammatory, anticancer, and osteogenic activities. However, it shows extremely poor bioavailability, rapid metabolism, and rapid systemic elimination. In this study, we have increased the bioavailability of curcumin by encapsulating it in a liposome, followed by the incorporation onto 3D printed (3DP) calcium phosphate (CaP) scaffolds with designed porosity. 3DP scaffolds with a designed shape and interconnected porosity allow for the fabrication of patient-specific implants, providing new tissue ingrowth by mechanical interlocking between the surrounding host tissue and the scaffold. Upon successful encapsulation of curcumin into the liposomes, we have investigated the effect of liposomal curcumin released from the 3DP scaffolds on both human fetal osteoblast cells (hFOB) and human osteosarcoma (MG-63) cells. Interestingly, liposomal curcumin released from the 3DP scaffold showed significant cytotoxicity toward in vitro osteosarcoma (bone cancer) cells, whereas it promoted osteoblast (healthy bone cell) cell viability and proliferation. These results reveal a novel approach toward the fabrication of tissue engineering scaffolds, which couples the advanced additive manufacturing technology with the wisdom of alternative medicine. These bifunctional scaffolds eradicate the osteosarcoma cells and also promote osteoblast proliferation, offering new opportunities to treat bone defects after tumor resection.

Changes in Human Foetal Osteoblasts Exposed to the Random Positioning Machine and Bone Construct Tissue Engineering.

Human cells, when exposed to both real and simulated microgravity (s-µg), form 3D tissue constructs mirroring in vivo architectures (e.g., cartilage, intima constructs, cancer spheroids and others). In this study, we exposed human foetal osteoblast (hFOB 1.19) cells to a Random Positioning Machine (RPM) for 7 days and 14 days, with the purpose of investigating the effects of s-µg on biological processes and to engineer 3D bone constructs. RPM exposure of the hFOB 1.19 cells induces alterations in the cytoskeleton, cell adhesion, extra cellular matrix (ECM) and the 3D multicellular spheroid (MCS) formation. In addition, after 7 days, it influences the morphological appearance of these cells, as it forces adherent cells to detach from the surface and assemble into 3D structures. The RPM-exposed hFOB 1.19 cells exhibited a differential gene expression of the following genes: transforming growth factor beta 1 (TGFB1, bone morphogenic protein 2 (BMP2), SRY-Box 9 (SOX9), actin beta (ACTB), beta tubulin (TUBB), vimentin (VIM), laminin subunit alpha 1 (LAMA1), collagen type 1 alpha 1 (COL1A1), phosphoprotein 1 (SPP1) and fibronectin 1 (FN1). RPM exposure also induced a significantly altered release of the cytokines and bone biomarkers sclerostin (SOST), osteocalcin (OC), osteoprotegerin (OPG), osteopontin (OPN), interleukin 1 beta (IL-1β) and tumour necrosis factor 1 alpha (TNF-1α). After the two-week RPM exposure, the spheroids presented a bone-specific morphology. In conclusion, culturing cells in s-µg under gravitational unloading represents a novel technology for tissue-engineering of bone constructs and it can be used for investigating the mechanisms behind spaceflight-related bone loss as well as bone diseases such as osteonecrosis or bone injuries.

The Osteogenic Potential of Brown Seaweed Extracts.

Marine drugs hold significantly more promise than their terrestrial counterparts, which could help to solve the current shortfall in treatments for osteoporosis and other bone related diseases. Fucoxanthin is the main carotenoid found in brown seaweed, and has many perceived health benefits, including potential bone therapeutic properties. This study assessed the osteogenic potential of pure fucoxanthin and crude extracts containing both fucoxanthin and phenolic fractions (also cited to have osteogenic potential) isolated from two intertidal species of brown seaweed, Laminaria digitata and Ascophyllum nodosum. In vitro studies were performed using a human foetal osteoblast cell line (hFOBs) and primary human bone marrow stromal cells (hBMSCs). The results found pure fucoxanthin inhibitory to cell proliferation in hFOBs at higher concentrations, whereas, the crude extracts containing both polyphenols and fucoxanthin showed the ability to scavenge free radicals, which masked this effect. None of the extracts tested showed strong pro-osteogenic effects in either cell type tested, failing to support previously reported positive effects.