Human Osteoblastic Cells Research Articles

Biological and antibacterial properties of chitosan-based coatings with AgNPs and CuNPs obtained on oxidized Ti13Zr13Nb titanium alloy

Despite numerous studies, the antibacterial efficiency and cytotoxicity of chitosan-based coatings with nanometals need further studies. The purpose of this research was to determine the biological properties of chitosan coatings implemented with either silver nanoparticles (AgNPs) or copper nanoparticles (CuNPs), that were electrophoretically deposited on electrochemically oxidized Ti13Zr13Nb alloy. The SEM tests were used to examine the surface uniformity. The biological tests against human osteoblast cells hFOB 1.19 and antibacterial activity against Staphylococcus aureus and Escherichia coli bacteria were performed. The release of Cu2+ from coating dependent on the pH of medium. The highest was at pH 3.0 and it achieved 47.73 mg/ml. No cytotoxic effects of the chitosan-AgNPs coatings were observed in direct and indirect contact studies, as there was 20 % inhibition of cell growth but no LDH release which indicate the cell breakdown. Antibacterial properties were proven for all modifications, but this effect was stronger for coatings with implemented AgNPs (log degree of bacteria number reduction >3.75 for E. coli and S. aureus).

Targeting POLRMT by a first-in-class inhibitor IMT1 inhibits osteosarcoma cell growth in vitro and in vivo

Osteosarcoma (OS) is a highly aggressive form of bone cancer that predominantly affects adolescents and young adults. In this study, we have undertaken an investigation into the potential anti-OS cell activity of IMT1 (inhibitor of mitochondrial transcription 1), a first-in-class inhibitor of RNA polymerase mitochondrial (POLRMT). IMT1 exhibited a profound inhibitory effect on cell survival, proliferation, cell cycle progression, and migration in primary and immortalized OS cells. Furthermore, this POLRMT inhibitor elicited apoptosis in the OS cells, without, however, inducing cytotoxicity in human osteoblasts or osteoblastic cells. IMT1 disrupted mitochondrial functions in OS cells, resulting in mitochondrial depolarization, oxidative injury, lipid peroxidation, and ATP reduction in OS cells. Silencing POLRMT using targeted shRNA closely mimicked the actions of IMT1 and exerted potent anti-OS cell activity. Importantly, IMT1’s effectiveness was diminished in POLRMT-silenced OS cells. Subsequent investigations revealed that IMT1 suppressed the activation of the Akt-mammalian target of rapamycin (mTOR) cascade in OS cells. IMT1 treatment or POLRMT silencing in primary OS cells led to a significant reduction in Akt1-S6K-S6 phosphorylation. Conversely, it was enhanced upon POLRMT overexpression. The restoration of Akt-mTOR activation through the introduction of a constitutively active S473D mutant Akt1 (caAkt1) mitigated IMT1-induced cytotoxicity in OS cells. In vivo, oral administration of IMT1 robustly curtailed the growth of OS xenografts in nude mice. Furthermore, IMT1 suppressed POLRMT activity, impaired mitochondrial function, repressed Akt-mTOR activation, and induced apoptosis within xenograft tissues. Collectively, these findings underscore the potent growth-inhibitory effects attributed to IMT1 via targeted POLRMT inhibition. The utilization of this POLRMT inhibitor carries substantial therapeutic promise in the context of OS treatment.

Ultrasound stimulated piezoelectric barium titanate and boron nitride nanotubes in nonconductive poly-ε-caprolactone nanofibrous scaffold for bone tissue engineering

Nanomaterials can provide unique solutions for the problems experienced in tissue engineering by improving a scaffold’s physico-bio-chemical properties. With its piezoelectric property, bone is an active tissue with easy adaptation and remodeling through complicated mechanisms of electromechanical operations. Although poly(ε-caprolactone) (PCL) is an excellent polymer for bone tissue engineering, it is lack of conductivity. In this study, piezoelectric barium titanates (BaTiO3) and boron nitride nanotubes (BNNTs) are used as ultrasound (US) stimulated piezoelectric components in PCL to mimic piezoelectric nature of bone tissue. Electric-responsive Human Osteoblast cells on the scaffolds were stimulated by applying low-frequency US during cell growth. Biocompatibility, cell adhesion, alkaline phosphatase activities and mineralization of osteoblast cells on piezo-composite scaffolds were investigated. BaTiO3 or BNNTs as reinforcement agents improved physical and mechanical properties of PCL scaffolds. In vitro studies show that the use of BaTiO3 or BNNTs as additives in non-conductive scaffolds significantly induces and increases the osteogenic activities even without US stimulation. Although BaTiO3 is one of the best piezoelectric materials, the improvement is more dramatic in the case of BNNTs with the increased mineralization, and excellent chemical and mechanical properties.

Investigating the interplay between environmental conditioning and nanotopographical cueing on the response of human MG63 osteoblastic cells to titanium nanotubes.

Titanium nanotubular surfaces have been extensively studied for their potential use in biomedical implants due to their ability to promote relevant phenomena associated with osseointegration, among other functions. However, despite the large body of literature on the subject, potential synergistic/antagonistic effects resulting from the combined influence of environmental variables and nanotopographical cues remain poorly investigated. Specifically, it is still unclear whether the nanotube-induced variations in cellular activity are preserved across different biochemical contexts. To bridge this gap, this study systematically evaluates the combined influence of nanotopographical cues and environmental factors on human MG63 osteoblastic cells. To this end, we capitalized on a triphasic anodization protocol to create nanostructured surfaces characterized by an average nanotube inner diameter of 25 nm (NT1) and 82 nm (NT2), as well as a two-tiered honeycomb (HC) architecture. A variable glucose content was chosen as the environmental modifier due to its well-known ability to affect specific functions of MG63 cells. Alkaline phosphatase (ALP), viability/metabolic activity and proliferation were quantified to identify the suitable preconditioning window required for dictating a change in behaviour without significantly damaging cells. Successively, a combination of immunofluorescence, colorimetric assays, live cell imaging and western blots quantified viability/metabolic activity and cell proliferation, migration and differentiation as a function of the combined effects exerted by the nanostructured substrates and the glucose content. To achieve a thorough understanding of MG63 cell adaptation and response, a comparative analysis table that includes and systematically cross-analyzes all variables from this study was used for interpretation and discussion of the results. Taken together, we have demonstrated that all surfaces mitigate the negative effects of high glucose. However, nanotubular topographies, particularly NT2, elicit a more beneficial outcome in high glucose in respect to untreated titanium. In addition, while NT1 surfaces are associated with the most stable cellular response across varying glucose levels, the NT2 and HC substrates exhibit the strongest enhancement of cell migration, viability/metabolism and differentiation. Moreover, shorter-term processes such as adhesion and proliferation are favored on untreated titanium, while anodized samples support later-term events. Lastly, the role of anodized surfaces is dominant over the effects of environmental glucose, underscoring the importance of carefully considering nanoscale surface features in the design and development of cell-instructive titanium surfaces.

Impact of Bone Morphogenetic Protein 7 and Prostaglandin receptors on osteoblast healing and organization of collagen.

This study seeks to investigate the impact of co-administering either a Prostaglandin EP2 receptor agonist or an EP1 receptor antagonist alone with a low dose BMP7 on in vitro healing process, collagen content and maturation of human osteoblasts. Human osteoblast cells were used in this study. These cells were cultured and subjected to different concentrations of Prostaglandin EP2 receptor agonist, EP1 receptor antagonist, BMP7, Control (Ct) (Vehicle alone), and various combinations treatments. Cell viability at 24, 48 and 72 hours (h) was evaluated using the XTT assay. A wound healing assay was conducted to observe the migration ability of human osteoblast cells. Additionally, Sirius red staining and Fourier-Transform Infrared Spectroscopy Imaging (FT-IR) was employed to analyze various parameters, including total protein concentration, collagen production, mature collagen concentration, and mineral content. The combination of low dose BMP7 and Prostaglandin EP2 receptor agonist resulted to the lowest cell viability when compared to both the Ct and individual treatments. In contrast, the Prostaglandin EP1 receptor antagonist alone showed the highest cellular viability at 72 h. In the wound healing assay, the combined treatment of low dose BMP7 with the Prostaglandin EP2 receptor agonist and EP1 receptor antagonist showed a decrease in human osteoblast healing after 24 h. Analysis of FT-IR data indicated a reduction in total protein content, collagen maturity, collagen concentration and mineral content in combination treatment compared to the single or Ct treatments. The combination of a Prostaglandin EP2 receptor agonist or an EP1 receptor antagonist when combined with low dose BMP7 significantly hinders both human osteoblast healing and collagen maturity/concentration in comparison to low dose BMP7 treatment alone.

Recombinant human secretory leukocyte protease inhibitor (rhSLPI) coated titanium enhanced human osteoblast adhesion and differentiation

Osseointegration is vital to success in orthopedic and dental reconstructions with implanted materials. The bone matrix or cells—particularly osteoblasts—are required to achieve functional contact on the implant surface. Osteoblast induction is therefore essential for osteogenesis to occur. Enhancement of osteoblast adhesion, proliferation, and differentiation, particularly by implant surface modifications, have been found challenging to develop. Secretory Leukocyte Protease Inhibitor (SLPI), a cation ionic protein with anti-inflammatory and anti-bacterial activities, showed activation in osteoblast proliferation and differentiation. However, the effects of coating recombinant human (rh) SLPI on a titanium alloy surface on human osteoblast adhesion, proliferation, and differentiation has never been investigated. In this study, titanium alloys (Ti–6Al–4V) were coated with rhSLPI, while human osteoblast adhesion, proliferation, differentiation, actin cytoskeletal organization, and gene expressions involved in cell adhesion and differentiation were investigated. The results indicate that coating titanium with 10–100 µg/ml rhSLPI enhanced the physical properties of the Ti surface and enhanced human osteoblast (hFOB 1.19) cell adhesion, activated actin dynamic, enhanced adhesive forces, upregulated integrins α1, α2, and α5, enhanced cell proliferation, mineralization, alkaline phosphatase activity, and upregulated ALP, OCN, and Runx2. This is the first study to demonstrate that coating SLPI on titanium surfaces enhances osseointegration and could be a candidate molecule for surface modification in medical implants.

Investigating the cryoprotective efficacy of fructans in mammalian cell systems via a structure-functional perspective

Fructans have long been known with their role in protecting organisms against various stress factors due to their ability to induce controlled dehydration and support membrane stability. Considering the vital importance of such features in cryo-technologies, this study aimed to explore the cryoprotective efficacy of fructans in mammalian cell systems where structurally different fructan polymers were examined on in vitro cell models derived from organs such as the liver, frequently used in transplantation, osteoblast, and cord cells, commonly employed in cell banking, as well as human seminal fluids that are of vital importance in assisted reproductive technology. To gain insights into the fructan/membrane interplay, structural differences were linked to rheological properties as well as to lipid membrane interactions where both fluorescein leakage from unilamellar liposomes and membrane integrity of osteoblast cells were monitored. High survival rates obtained with human endothelial, osteoblast and liver cells for up to two months clearly showed that fructans could be considered as effective non-permeating cryoprotectants, especially for extended periods of cryopreservation. In trials with human seminal fluid, short chained levan in combination with human serum albumin and glycerol proved very effective in preserving semen samples across multiple patients without any morphological abnormalities.

Microgravity triggers ferroptosis and accelerates senescence in the MG-63 cell model of osteoblastic cells

In space, cells sustain strong modifications of their mechanical environment. Mechanosensitive molecules at the cell membrane regulate mechanotransduction pathways that induce adaptive responses through the regulation of gene expression, post-translational modifications, protein interactions or intracellular trafficking, among others. In the current study, human osteoblastic cells were cultured on the ISS in microgravity and at 1 g in a centrifuge, as onboard controls. RNAseq analyses showed that microgravity inhibits cell proliferation and DNA repair, stimulates inflammatory pathways and induces ferroptosis and senescence, two pathways related to ageing. Morphological hallmarks of senescence, such as reduced nuclear size and changes in chromatin architecture, proliferation marker distribution, tubulin acetylation and lysosomal transport were identified by immunofluorescence microscopy, reinforcing the hypothesis of induction of cell senescence in microgravity during space flight. These processes could be attributed, at least in part, to the regulation of YAP1 and its downstream effectors NUPR1 and CKAP2L.

Bone implant substitutes from synthetic polymer and reduced graphene oxide: Current perspective.

In the present work, bone implant materials (BIM) were produced, in sheet form which comprises epoxy resin (synthetic polymer) (ER), calcium carbonate (CaCO3), and reduced graphene oxide (R-GO), by open mold method, for the possibility uses in bone tissue engineering. The developed BIM was analyzed for its physico-chemical, mechanical, bioactivity test, antimicrobial study, and biocompatibility. The BIM had excellent mechanical properties such as tensile strength (194.44 + 0.21 MPa), flexural strength (278.76 + 0.41 MPa), and water absorption (02.61 + 0.24%). A pore size distribution study using the HR-SEM has proved the 180 and 255 μm average pore was observed in the BIM structure. The Bioactivity test of BIM was examined after being immersed in a simulated body fluids (SBF) solution. The result of BIM formed an excellent deposition of bone tube apatite crystals. High-resolution scanning electron microscopy (HR-SEM) morphology of the bone tube apatite crystals revealed the diameter size in the range from 100 ± 159 to 210 ± 188 nm. BIM has excellent antimicrobial characteristics against E. coli (8.75 + 0.06 mm) and S. aureus (9.82 + 0.08 mm). The biocompatibility of the study MTT (3-(4, 5-dimethyl) thiazol-2-yl-2, 5-dimethyl tetrazolium bromide) assay using the MG-63 (human osteoblast cell line) has proven to be the 78% viable cell presence in BIM. After receiving the necessary approval, the scaffold with the required strength and biocompatibility could be tested as a bone implant material in large animals.

Enhanced Proliferation and Differentiation of Human Osteoblasts by Remotely Controlled Magnetic-Field-Induced Electric Stimulation Using Flexible Substrates.

With the progressive aging of the population, bone fractures are an increasing major health concern. Diverse strategies are being studied to reduce the recovery times using nonaggressive treatments. Electrical stimulation (either endogenous or externally applied electric pulses) has been found to be effective in accelerating bone cell proliferation and differentiation. However, the direct insertion of electrodes into tissues can cause undesirable inflammation or infection reactions. As an alternative, magnetoelectric heterostructures (wherein magnetic fields are applied to induce electric polarization) could be used to achieve electric stimulation without the need for implanted electrodes. Here, we develop a magnetoelectric platform based on flexible kapton/FeGa/P(VDF-TrFE) (flexible substrate/magnetostrictive layer/ferroelectric layer) heterostructures for remote magnetic-field-induced electric field stimulation of human osteoblast cells. We show that the use of flexible supports overcomes the clamping effects that typically occur when analogous magnetoelectric structures are grown onto rigid substrates (which preclude strain transfer from the magnetostrictive to the ferroelectric layers). The study of the diverse proliferation and differentiation markers evidence that in all the stages of bone formation (cell proliferation, extracellular matrix maturation, and mineralization), the electrical stimulation of the cells results in a remarkably better performance. The results pave the way for novel strategies for remote cell stimulation based on flexible platforms not only in bone regeneration but also in many other applications where electrical cell stimulation may be beneficial (e.g., neurological diseases or skin regeneration).

Twin-screw extrusion of vitamin D3/iron-blend granules in corn and lentil composite flours: Stability, microstructure, and interaction of vitamin D3 with human osteoblast cells.

Vitamin D3 (VD3) and iron-blend granules were blended with corn and lentil composite flour (75/25, w/w) and fed into a pilot-scale twin-screw extruder to produce ready-to-eat snacks. The morphology and microstructure of extruded snacks were examined using scanning electron microscopy with energy-dispersive X-ray (SEM-EDX), X-ray powder diffraction, and FT-IR. Differential scanning calorimetry and thermogravimetric analysis measured the melting temperature and thermal stability of the extrudates. SEM and FT-IR analysis demonstrate that micronutrients are mixed well in formulations used in extrudates at high shear and high temperatures. The SEM-EDX exhibited the presence of iron, whereas high performance liquid chromatography measurements confirmed the significant retention of VD3 in the extruded snacks. The interaction between VD3 and human osteoblast cells was studied using live imaging and the MMT assay. Overall, for the first time, VD3 and Fe2+ blend granules have been used in an extrusion platform, which has significant potential for the intervention of VD3 and iron deficiencies. PRACTICAL APPLICATION: For the first time, we reported the use of VD3/iron-blend granules in extruded products. The findings of this work demonstrated the thermal stability and capability of providing adequate quantities of VD3 and iron in corn flour/lentil flour/VD3-iron blend extruded snacks. Furthermore, the interaction of VD3 with osteoblast cells highlights the potential health benefits of the extrudates.

Exploring the role of lactylation-related genes in osteosarcoma: A deep dive into prognostic significance and therapeutic potential.

Osteosarcoma (OS), notorious for its complex pathogenesis and formidable prognosis, represents a significant medical quandary. This research embarked on a quest to unravel the implications of lactylation-related genes (LRGs) in OS, offering a novel lens through which to interpret its intricacies. A meticulous evaluation of 329 LRGs within the TARGET dataset spotlighted 27 paramount genes, intricately intertwined with survival. These genes highlighted metabolic processes-particularly amino acid metabolism-as key players, as evidenced in both GO and KEGG analyses. Utilizing consensus clustering and principal component analysis, the 93 OS samples were segmented into two distinct groups, differing notably in overall and event-free survival. Cluster 2 demonstrated a heightened immune response, contrasting the other cluster. Machine learning techniques, like generalized boosted model, CoxBoost, and RSF, spotlighted MYC and GOT2 as critical genes. Using multivariate Cox regression, a risk model was developed, categorizing patients into high and low-risk groups, each displaying varied survival patterns. Additionally, a contrast was observed between MYC and GOT2's associations with HLA molecules, emphasizing their distinct roles in antigen presentation. Potential therapeutic avenues were identified for each risk group, with special attention to mutations in MYC, particularly amplifications, hinting at its role in tumor progression. Finally, delving deeper into the role of MYC, Western blot analyses exhibited amplified myc protein levels in OS cells U-2 and MG-63 when juxtaposed against human osteoblastic cells Hfob1.19. A focused knockdown of myc in OS cells subsequently confirmed its influence on cell proliferation and migration, with reduced myc expression resulting in inhibited cell activities. Furthermore, immunofluorescence assays corroborated myc's heightened expression in OS cells relative to normal osteoblastic cells. In summary, this study accentuates the vital role of LRGs and specifically MYC in OS, ushering in a horizon of tailored therapeutic strategies.

Freeze casting of hydroxyapatite-titania composites for bone substitutes.

Hydroxyapatite (HA) is commonly used as a bone substitute material, but it lacks mechanical strength when compared to native bone tissues. To improve the efficacy of HA as a bone substitute by improving the mechanical strength and cell growth attributes, porous composite scaffolds of HA and titania (HA-TiO2 ) were fabricated through a freeze-casting process. Three different compositions by weight percent, 25-75 HA-TiO2 , 50-50 HA-TiO2 , and 75-25 HA-TiO2 , were custom-made for testing. After sintering at 1250°C, these composite scaffolds exhibited improved mechanical properties compared to porous HA scaffolds. Substrate mixing was observed, which helped reduce crystal size and introduced new phases such as β-TCP and CaTiO3 , which also led to improved mechanical properties. The composition of 50-50 HA-TiO2 had the highest ultimate compressive strength of 3.12 ± 0.36 MPa and elastic modulus 63.29 ± 28.75 MPa. Human osteoblast cell proliferation assay also increased on all three different compositions when compared to porous HA at 14 days. These results highlight the potential of freeze casting composites for the fabrication of bone substitutes, which provide enhanced mechanical strength and biocompatibility while maintaining porosity.

Chitin scaffolds derived from the marine demosponge Aplysina fistularis stimulate the differentiation of dental pulp stem cells.

The use of stem cells for tissue regeneration is a prominent trend in regenerative medicine and tissue engineering. In particular, dental pulp stem cells (DPSCs) have garnered considerable attention. When exposed to specific conditions, DPSCs have the ability to differentiate into osteoblasts and odontoblasts. Scaffolds are critical for cell differentiation because they replicate the 3D microenvironment of the niche and enhance cell adhesion, migration, and differentiation. The purpose of this study is to present the biological responses of human DPSCs to a purified 3D chitin scaffold derived from the marine demosponge Aplysina fistularis and modified with hydroxyapatite (HAp). Responses examined included proliferation, adhesion, and differentiation. The control culture consisted of the human osteoblast cell line, hFOB 1.19. Electron microscopy was used to examine the ultrastructure of the cells (transmission electron microscopy) and the surface of the scaffold (scanning electron microscopy). Cell adhesion to the scaffolds was determined by neutral red and crystal violet staining methods. An alkaline phosphatase (ALP) assay was used for assessing osteoblast/odontoblast differentiation. We evaluated the expression of osteogenic marker genes by performing ddPCR for ALP, RUNX2, and SPP1 mRNA expression levels. The results show that the chitin biomaterial provides a favorable environment for DPSC and hFOB 1.19 cell adhesion and supports both cell proliferation and differentiation. The chitin scaffold, especially with HAp modification, isolated from A. fistularis can make a significant contribution to tissue engineering and regenerative medicine.

Mechanical Properties, Degradation Behavior, and Cytocompatibility of Zn–Mg–Graphene Nanoplatelets Composite for Orthopedic‐Implant Applications

Zinc (Zn)‐based materials reveal inadequate mechanical properties as orthopedic biodegradable implantable materials, which limits their biomedical applications. Herein, Zn–xMg (magnesium) composites (x = 0.5, 1.0, 1.5, and 2.0 wt%) reinforced with 0.2 wt% graphene nanoplatelets (GNP) are obtained via powder metallurgy and hot‐pressing sintering. The addition of 0.5–2.0 wt% Mg into Zn–0.2 wt% GNP composite resulted in the formation of Mg2Zn11 and MgZn2 phases without any additional intermetallic carbide phases. The hot‐pressing sintered (HPS) Zn–0.5Mg–0.2GNP composite exhibits a compressive yield strength of 169 ± 18 MPa, an ultimate compressive strength of 270 ± 39 MPa, a compressive strain of 17 ± 6%, and a microhardness of 86 ± 2 HV. Electrochemical corrosion testing reveals that corrosion resistance of HPS Zn–xMg–0.2GNP composites decreases with increasing Mg content, while immersion tests in Hanks’ balanced salt solution for 30 d indicate that the degradation rate increases from 0.032 to 0.141 mm y−1 by increasing Mg content from 0 to 2.0 wt%. In vitro cytotoxicity assessments using SaOS2 human osteoblast cells show >90% viability following exposure over 5 d to 12.5% extract concentrations of all HPS composites. The HPS Zn–0.5Mg–0.2GNP composite exhibits the appropriate material properties for biodegradable bone‐implant applications.

Small‐Molecule Chemistry Effect on the Functionalization of Polydimethylsiloxane with Hydroxyapatite

Herein, polydimethylsiloxane (PDMS) surfaces are coated with hydroxyapatite (HA) using immobilization and biomineralization techniques with the aid of small molecules, namely, l‐aspartic acid (AA), l‐glutamic acid, and citric acid, in an effort to develop biointeractive surfaces for bone tissue‐related research. The efficacy of biomineralization and immobilization techniques and the impact of small molecules on HA deposition on PDMS surfaces are investigated by chemical and morphological analysis of surfaces, besides in vitro cell culture studies. Characteristic peaks of phosphate groups at wavelengths of 569 and 603 cm−1 in Fourier transform infrared spectroscopy analysis and HA's characteristic patterns identified at 26.1°, 31.7°, and 45.5° in X‐ray diffraction analysis demonstrate the successful deposition of HA particles on PDMS surfaces which are also observed on scanning electron microscopy micrographs. The interaction of human fetal osteoblast cells (hFOB 1.19) with HA‐deposited PDMS surfaces is evaluated with in vitro cell culture studies, which reveals increased cell metabolic activity and growth across all groups as compared to plain PDMS, with HA deposited on AA‐PDMS surfaces through biomineralization being the most stimulating group. Overall, small‐molecule effect on deposition of HA on PDMS substrates merits further investigation, particularly on substrates mimicking bone surface chemical and morphological features.

Co-precipitation-hydrothermal mediated synthesis, characterization and toxicity evaluation of 2D Zn–Al LDHs in human osteoblast cells

Co-precipitation-hydrothermal mediated synthesis, characterization and toxicity evaluation of 2D Zn–Al LDHs in human osteoblast cells

Evaluation of Polypeptide Complexes from Marine Sources with Relevance in Initiation and Propagation Mechanisms of Osteoarticular Dysfunctions

The aim of the present study was to obtain a protein fraction rich in enzymes with proteolytic activity such as serine proteases, chymotrypsins and cysteine proteases and to investigate preliminary pharmacological effects with therapeutic potential in osteoarticular diseases. The processing of marine raw material by applying established and optimized sequences of operations (selective precipitation followed by fractionation by FPLC) led to well-defined protein fractions rich in proteolytic enzymes. To evaluate the biological effect of the isolated polypeptide fractions, an in vitro model on normal human osteoblast (HOB) and osteoarthritic (HOB-OA) cell lines was carried out in which the modulation of cell proliferation under the action of the polypeptide complexes was followed.

Interplay between Cultured Human Osteoblastic and Skeletal Muscle Cells: Effects of Conditioned Media on Glucose and Fatty Acid Metabolism.

The interplay between skeletal muscle and bone is primarily mechanical; however, biochemical crosstalk by secreted mediators has recently gained increased attention. The aim of this study was to investigate metabolic effects of conditioned medium from osteoblasts (OB-CM) on myotubes and vice versa. Human skeletal muscle cells incubated with OB-CM showed increased glucose uptake and oxidation, and mRNA expression of the glucose transporter (GLUT) 1, while fatty acid uptake and oxidation, and mRNA expression of the fatty acid transporter CD36 were decreased. This was supported by proteomic analysis, where expression of proteins involved in glucose uptake, glycolytic pathways, and the TCA cycle were enhanced, and expression of several proteins involved in fatty acid metabolism were reduced. Similar effects on energy metabolism were observed in human bone marrow stromal cells differentiated to osteoblastic cells incubated with conditioned medium from myotubes (SKM-CM), with increased glucose uptake and reduced oleic acid uptake. Proteomic analyses of the two conditioned media revealed many common proteins. Thus, our data may indicate a shift in fuel preference from fatty acid to glucose metabolism in both cell types, induced by conditioned media from the opposite cell type, possibly indicating a more general pattern in communication between these tissues.

DP2, a Carbohydrate Derivative, Enhances In Vitro Osteoblast Mineralisation.

Bone fracture healing is a complex biological process involving four phases coordinated over time: hematoma formation, granulation tissue formation, bony callus formation, and bone remodelling. Bone fractures represent a significant health problem, particularly among the elderly population and patients with comorbidities. Therapeutic strategies proposed to treat such fractures include the use of autografts, allografts, and tissue engineering strategies. It has been shown that bone morphogenetic protein 2 (BMP-2) has a therapeutic potential to enhance fracture healing. Despite the clinical efficacy of BMP-2 in osteoinduction and bone repair, adverse side effects and complications have been reported. Therefore, in this in vitro study, we propose the use of a disaccharide compound (DP2) to improve the mineralisation process. We first evaluated the effect of DP2 on primary human osteoblasts (HOb), and then investigated the mechanisms involved. Our findings showed that (i) DP2 improved osteoblast differentiation by inducing alkaline phosphatase activity, osteopontin, and osteocalcin expression; (ii) DP2 induced earlier in vitro mineralisation in HOb cells compared to BMP-2 mainly by earlier activation of Runx2; and (iii) DP2 is internalized in HOb cells and activates the protein kinase C signalling pathway. Consequently, DP2 is a potential therapeutical candidate molecule for bone fracture repair.