Search Results (80)

The design of biomaterial scaffolds for bone tissue engineering requires a balance between bioactivity, porosity, mechanical stability, and osteoinductivity. Kappa- (KC) and iota-carrageenan (IC) have been explored for scaffold fabrication due to their biocompatibility and structural similarity to glycosaminoglycans. However, there are limited reports on how their distinct sulfation degree affects the osteogenic differentiation of cells cultured on them. While laponite has been reported as an osteoinductive nanoclay, its combined effect with different carrageenan types and its concentration-dependent effect on scaffold functionality remain unexplored. Therefore, we developed composite scaffolds comprising poly(vinyl alcohol) (PVA) and gelatin (GEL), reinforced with kappa- or iota-carrageenan (KC, IC) and functionalized with two different concentrations of laponite (LAP), 0.5 and 1% w/v, to monitor composition-structure-function relationships. The scaffolds were fabricated via lyophilization and dual crosslinking, and characterized for their physicochemical, structural, mechanical, and biological properties. The incorporation of both carrageenans into scaffolds, maintained high swelling ratios of 600% after 24 h, and increased porosity without altering their apparent density (0.09–0.11 g/cm³), whereas LAP preserved interconnectivity, densified pore walls, raised their compressive modulus at >220 kPa, and improved stability (>60% mass retained after 40 days). In vitro validation using MC3T3-E1 pre-osteoblastic cells demonstrated robust cytocompatibility, with the LAP-containing scaffolds significantly promoting cell adhesion, proliferation, and osteogenic differentiation, evidenced by elevated alkaline phosphatase activity, calcium production and collagen secretion. Direct comparison between KC and IC scaffolds confirmed that differences in sulfate substitution modulated scaffold stiffness, swelling, and degradation, while variation in LAP concentration affected the biological response, with the 0.5 wt% concentration favoring early cell proliferation, whereas the 1 wt% significantly promoted the osteogenic differentiation. This compositional strategy demonstrates how tuning the interplay between carrageenan and laponite can balance scaffold hydration, mechanical and biological properties, thereby guiding the design of scaffolds for bone repair. Full article

Bone regeneration relies on the coordinated interplay between mechanical and biological cues. Piezoelectric composites, capable of converting mechanical strain into electrical signals, offer a promising approach to stimulate osteogenesis. This study aimed to develop and characterize polycaprolactone (PCL) and barium titanate (BaTiO₃) composite scaffolds fabricated through thermally induced phase separation (TIPS), and to systematically evaluate the effects of polymer concentration and ceramic incorporation on scaffold morphology, porosity, mechanical properties, and cytocompatibility were systematically evaluated. The resulting scaffolds exhibited a highly porous, interconnected architecture, with 9% PCL formulation showing the most uniform morphology and consistent mechanical and biological behavior. Incorporation of BaTiO₃ did not alter pore structure or compromise cytocompatibility but slightly enhanced stiffness and surface uniformity. SEM-based image analysis confirmed homogeneous BaTiO₃ dispersion across all formulations. MTT assays and confocal microscopy demonstrated robust pre-osteoblast adhesion and spreading, particularly on denser composite scaffolds, confirming that the inclusion of BaTiO₃ supports a favorable environment for cell proliferation. Overall, optimizing polymer concentration and ceramic dispersion enables fabrication of structurally coherent, cytocompatible scaffolds. The findings establish structure–property–biology relationships that serve as a baseline for future investigations into the electromechanical behavior of PCL/BaTiO₃ scaffolds and their potential to promote osteogenic differentiation under physiological loading. Full article

Nanostructured TiO₂/Cu coatings were deposited on Ti6Al4V alloy by a two-step glow-discharge sputtering process and evaluated for their structural, electrochemical, and biological properties. Dual-acid etching produced microroughened substrates before TiO₂ layer deposition, followed by surface Cu sputtering with varied deposition times. Characterisation by AFM, OM, SEM/EDS, and XRD confirmed the formation of TiO₂ with Cu/Cu₂O-containing hybrid coatings with good adhesion to the substrate. Increasing Cu deposition enhanced surface hydrophobicity and copper ion release. EIS measurements proved that the coatings retained stable protective behaviour in simulated body fluid (SBF). Antibacterial tests against Escherichia coli showed up to 98% improved efficacy compared to bare Ti6Al4V, confirming the strong antimicrobial role of copper. However, MG63 osteoblast-like cells exhibited reduced viability even after pre-immersion in PBS, suggesting that cytotoxicity was associated not only with excess Cu ion release but also with direct interaction between cells and surface Cu nanostructures. Overall, the results indicate that TiO₂/Cu coatings provide excellent antimicrobial activity, good protection and strong adhesion, but their limited biocompatibility highlights the need for fine-tuned copper incorporation in future biomedical implant applications. Full article

Calcium phosphate coatings are known for their osteoconductive prowess. In this work, calcium phosphate coatings were studied on a model biodegradable magnesium alloy of AZ31, primarily to provide improved corrosion protection and, more importantly, to confer in vitro cytocompatibility to the AZ31 alloy. Correspondingly, an aqueous-based approach was developed to deposit Sr²⁺-substituted calcium phosphates on micro-arc oxidized AZ31. Micro-arc oxidation was used mainly as a pretreatment technique due to improved homogeneity and adhesion strength in comparison to the coatings formed by the traditionally used alkaline and acidic pretreatment. Calcium phosphate coatings with up to 11.5 mol. % Sr were formed on micro-arc oxidized AZ31 substrates. Despite observation of greater than the intended 10 mol. % Sr to the calcium phosphate coatings as measured within the measurement error, biphasic mixtures of dicalcium phosphate dihydrate and poorly crystalline hydroxyapatite were formed. Micro-arc oxidation treatment, nevertheless, provided a slight improvement in corrosion protection compared to uncoated AZ31. However, much-improved corrosion protection was provided by the calcium phosphate coatings prepared either with or without Sr²⁺. The calcium phosphate coatings prepared with Sr²⁺ were also observed to support improved MC3T3-E1 murine pre-osteoblast cell proliferation compared to the calcium phosphate coated substrates prepared without Sr²⁺. Full article

The integration of nanomaterials within hydrogel scaffolds offers significant promise in bone tissue engineering by improving mechanical performance and modulating cellular responses through mechanotransductive and biochemical signaling. Previous studies have demonstrated that nanodiamonds (NDs) incorporated in electrospun microfibrillar meshes enhance cellular adhesion, spreading, and cytoskeletal organization through localized mechanical reinforcement. However, the effects of ND loading into soft, bioinert three-dimensional hydrogel matrices remain underexplored. Here, we developed nanostructured 3D printing inks composed of gellan gum (GG) supplemented with a low content of ND nanoadditive (0–3% w/v). ND integration improved the shear-thinning properties of the formulation, enabling consistent filament formation and reliable extrusion-based 3D printing. Structural and mechanical assessments confirmed enhanced scaffold morphology, reduced deformation, and improved morphostructural integrity under compression and increased local stiffness at 2% ND loading (GG_ND2%). Biological assessments revealed that increasing ND content enhanced murine preosteoblast viability, proliferation, and attachment, particularly in GG_ND2%. Furthermore, bioactivation of the GG_ND2% formulation with icariin (ICA), a bioflavonoid known for its osteogenic and angiogenic activity, amplified the beneficial cellular responses of MG-63 cells to ND loading, promoting enhanced surface mineralization and improved cell–matrix interactions. Collectively, these findings highlight the potential of ND-reinforced GG scaffolds bioactivated with ICA, integrating structural reinforcement and biological functionalities that may support osteogenic responses. Full article

Objectives: To prepare porous scaffolds combining hydrogel and hydroxycarbonateapatite, enriched with a promising therapeutic agent, gold nanoparticles, to improve bone regeneration. The fabrication procedure is conducted under mild conditions, without toxic or aggressive chemicals, at physiological pH, and low temperatures; Methods: Gold nanoparticles (15–20 nm), were obtained by the Turkevith method. The scaffolds were fabricated by the GELPOR3D method, which has demonstrated its ability to integrate thermal labile molecules, during the scaffold fabrication process. The role of these nanoparticles in promoting cell adhesion, proliferation, and mineralization processes in vitro has been studied using osteoprogenitor MC3T3-E1 cells; Results: The scaffold fabrication conditions, combined with the surface functionalization of the gold nanoparticles with poly(ethylene glycol), ensure their uniform distribution throughout the scaffold and facilitate their gradual release over 48 h in a physiological medium. A significant increase in the mean cell area and a significant decrease in the circularity index during the early stages of osteoblast differentiation are observed. These pieces of evidence suggest that adequate cell spreading could lead to enhanced proliferation and matrix deposition activity; Conclusions: Scaffolds containing these gold nanoparticles exhibited a marked improvement in adhesion, proliferation, and mineralization of preosteoblasts (MC3T3 cells) at the concentrations studied. The functionalization of the nanoparticles, along with the shaping procedure employed, is critical for their homogeneous dispersion throughout the scaffold and their progressive release. The findings confirm the crucial role of gold nanoparticles in the early stages of osteoblast differentiation, which is essential for the transition from premature osteoblasts to mature osteoblasts. Full article

Titanium and its alloy scaffolds are widely utilized in clinical settings; however, their biologically inert surfaces and inherent mechanical characteristics impede osteogenesis and soft tissue integration, thereby limiting their application. Selective laser melting (SLM) was employed to fabricate scaffolds with matched cortical bone mechanical properties, achieving a composite coating of hydroxyapatite complexed with trace elements of silicon, strontium, and fluoride (mHA), along with type I collagen (Col I) and fibrinogen (Fg), thus activating the scaffold surface. Initially, we utilized the excellent adhesive properties of dopamine to co-deposit mHA and polydopamine (PDA) onto porous Ti-6Al-4V scaffolds, which was followed by immobilization of type I collagen and fibrinogen onto PDA. This bioinorganic/bioprotein composite coating, formed via PDA bonding, exhibits excellent stability. Moreover, in vitro cell experiments demonstrate excellent biocompatibility of the porous Ti-6Al-4V scaffold with composite bioactive coatings on its surface. Preosteoblasts (MC3T3-E1) and human keratinocytes (HaCaT) exhibit enhanced adhesion and proliferation activity, and the osteogenic performance of the scaffold is significantly improved. The PDA-mHA-Col I-Fg composite-coated porous titanium alloy scaffold holds significant promise in enhancing the efficacy of percutaneous bone transplantation and requires further investigation. Full article

Bioactive glasses in the SiO₂-CaO-P₂O₅ system represent emerging materials for hard-tissue-regeneration applications. This article focuses on the synthesis, characterization, and biological interaction of glasses doped with Mg²⁺ and/or Zn²⁺, with an emphasis on their effects on biomineralization, antibacterial behavior, and interactions with preosteoblasts from the MC3T3-E1 cell line. The bioglasses were synthesized using the sol-gel method, and the vitreous nature remained predominant even after thermal treatment at 600 °C for 2 h. From an in vitro perspective, the synthesized bioglasses demonstrated strong cell adhesion and proliferation (notably in the case of Mg²⁺ doping), low cytotoxicity, and antibacterial properties (especially in Zn²⁺-doped samples). Additionally, the simultaneous doping with Mg²⁺ and Zn²⁺ of the bioactive glass matrix is a prospective strategy for developing biomaterials with a “dual” biological characteristics–both osteoinductive and antibacterial. Full article

The development of biomaterials that enhance bone healing and integrate with native bone tissue has gained significant interest. Metal-organic frameworks (MOFs) have emerged as promising candidates due to their unique surface properties and biocompatibility. While various bioactive element-incorporated MOFs have been studied, the osteogenic potential of lithium (Li)-modified MOFs remains largely unexplored. This study presents the synthesis and characterization of a nanosized calcium-based MOF incorporating Li⁺ ions to enhance osteoinductive properties. The MOFs were evaluated in vitro for apatite mineralization, degradation, ion release, protein adsorption, cell adhesion, viability, and osteogenic differentiation using pre-osteoblast cells. The synthesized MOFs promoted apatite formation under simulated physiological conditions, facilitated by their surface nucleation properties, controlled degradation, and sustained Li⁺ and Ca²⁺ ion release. Cytocompatibility assays confirmed excellent pre-osteoblast adhesion and viability. Furthermore, CaMOF nanoparticles stimulated osteogenic differentiation by enhancing alkaline phosphatase (ALP) activity, even in the absence of osteogenic supplements. Among tested MOFs, Li/CaMOF exhibited the highest osteoinductive potential. These findings highlight lithium-modified MOFs as promising biomaterials for bone regeneration. However, further in vivo studies are necessary to assess their long-term stability, bone integration, and clinical applicability. Full article

The metabolic poise, or balance, between glycolysis and fatty acid oxidation (FAO) has recently been found to play a critical role in osteogenic differentiation and homeostasis. While simulated microgravity (SMG) is known to impede osteoblast differentiation (OBD) by inhibiting the Wnt/β-catenin pathway, how it affects osteoblast metabolism in this context remains unclear. We previously analyzed the effect of SMG on the differentiation of pre-osteoblast MC3T3-E1 cells and found that it reduced focal adhesion kinase (FAK) activity. This, in turn, downregulated Wnt/β-catenin and two of its downstream targets critical for OBD bone morphogenic protein-2 (BMP2) and type-1 collagen (COL1) formation, leading to a reduction in alkaline phosphatase (ALP) activity and cell matrix mineralization. In this study, we further analyzed how SMG-induced alterations in energy metabolism contribute to the inhibition of OBD in MC3T3-E1 cells. Consistent with our earlier findings, we demonstrated that SMG inhibits OBD by downregulating the collective activity of FAK and the Wnt/β-catenin-BMP2-COL1 transcriptional pathway. Interestingly, we observed that SMG also reduces the abundance of sirtuin-1 (SIRT1), peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) and carnitine palmitoyl transferase-1α (CPT1A), which are all key metabolic factors regulating mitochondrial number and FAO capacity. Accordingly, we found that the mitochondrial content and FAO potential of MC3T3-E1 cells were lower upon exposure to SMG but were both rescued upon administration of the FAK activator cytotoxic necrotizing factor-1 (CNF1), thereby allowing cells to overcome SMG-induced inhibition of OBD. Taken together, our study indicates that the metabolic regulator SIRT1 may be a new target for reversing SMG-induced bone loss. Full article

A key principle of guided bone regeneration (GBR) is the use of a barrier membrane to prevent cells from non-osteogenic tissues from interfering with bone regeneration in patients with hard tissue deficiencies. The aim of the study was to investigate whether the osteoinductive properties of bone-conditioned medium (BCM) obtained from cortical bone chips harvested at the surgical site can be transferred to a native bilayer collagen membrane (nbCM). BCM extracted within 20 or 40 min, which corresponds to a typical implant surgical procedure, and BCM extracted within 24 h, which corresponds to BCM released from the autologous bone chips in situ, contained significant and comparable amounts of TGF-β1, IGF-1, FGF-2, VEGF-A, and IL-11. Significant (p < 0.001) quantities of BMP-2 were only detected in the 24-h BCM preparation. The bioactive substances contained in the BCM were adsorbed to the nbCMs with almost 100% efficiency. A fast but sequential release of all investigated proteins occurred within 6–72 h, reflecting their stepwise involvement in the natural regeneration process. BCM-coated nbCM significantly (p < 0.05) increased the migratory, adhesive, and proliferative capacity of primary human bone-derived cells (hBC), primary human periodontal ligament cells (hPDLC), and an osteosarcoma-derived osteoblastic cell line (MG-63) compared to cells cultured on BCM-free nbCM. The high proliferative rates of MG-63 cells cultured on BCM-free nbCM were not further potentiated by BCM, indicating that BCM-coated nbCM has no detrimental effects on cancer cell growth. BCM-coated nbCM caused significant (p < 0.05) induction of early osteogenic marker gene expression and alkaline phosphatase activity, suggesting an important role of BCM-functionalized nbCM in the initiation of osteogenesis. The 24-h BCM loaded on the nbCM was the only BCM preparation that caused significant induction of late osteogenic marker gene expression. Altogether, our data define the pre-activation of collagen membranes with short-term-extracted BCM as a potential superior modality for treating hard tissue deficiencies via GBR. Full article

The demand for dental implants has increased, establishing them as the standard of care for replacing missing teeth. Several factors contribute to the success or failure of an implant post-placement. Modifications to implant surfaces can enhance the biological interactions between bone cells and the implant, promoting better outcomes. Surface coatings have been developed to electrochemically alter implant surfaces, aiming to reduce healing time, enhance bone growth, and prevent bacterial adhesion. Quaternized silicon carbon nitride (QSiCN) is a novel material with unique electrochemical and biological properties. This study aimed to assess the influence of QSiCN, silicon carbide nitride (SiCN), and silicon carbide (SiC) coatings on the viability of osteoblast cells on nanostructured titanium surfaces. The experiment utilized thirty-two titanium sheets with anodized TiO₂ nanotubes featuring nanotube diameters of 50 nm and 150 nm. These sheets were divided into eight groups (n = 4): QSiCN-coated 50 nm, QSiCN-coated 150 nm, SiCN-coated 50 nm, SiCN-coated 150 nm, SiC-coated 50 nm, SiC-coated 150 nm, non-coated 50 nm, and non-coated 150 nm. Preosteoblast MC3T3-E1 Subclone 4 cells (ATCC, USA) were used to evaluate osteoblast viability. After three days of cell growth, samples were assessed using scanning electron microscopy (SEM). The results indicated that QSiCN coatings significantly increased osteoblast proliferation (p < 0.005) compared to other groups. The enhanced cell adhesion observed with QSiCN coatings is likely due to the positive surface charge imparted by N⁺. Full article

Matrix extracellular phosphoglycoprotein (Mepe), present in bone and dentin, plays important multifunctional roles in cell signaling, bone mineralization, and phosphate homeostasis. Mepe expression in bone cells changes in response to pulsating fluid shear stress (PFSS), which is transmitted into cells through integrin-based adhesion sites, i.e., α and β subunits. Whether and to what extent PFSS influences Mepe expression through the modulation of integrin α and/or β subunit expression in pre-osteoblasts is uncertain. Therefore, we aimed to test whether low and/or high PFSS affects Mepe expression via modulation of integrin α and/or β subunit expression. MC3T3-E1 pre-osteoblasts were treated with ± 1 h PFSS (magnitude: 0.3 Pa (low PFSS) or 0.7 Pa (high PFSS); frequency: 1 Hz). Single integrin fluorescence intensity in pre-osteoblasts was increased, but single integrin area was decreased by low and high PFSS. Expression of two integrin α subunit-related genes (Itga1 and Itga5 2) was increased by low PFSS, and one (Itga5 2) by high PFSS. Expression of five integrin β subunit genes (Itgb1, Itgb3, Itgb5, Itgb5 13, and Itgb5 123) was increased by low PFSS, and three (Itgb5, Itgb5 13, and Itgb5 123) by high PFSS. Interestingly, Mepe expression in pre-osteoblasts was only modulated by low, but not high, PFSS. In conclusion, both low and high PFSS affected integrin α and β subunit expression in pre-osteoblasts, while integrin β subunit expression was more altered by low PFSS. Importantly, Mepe gene expression was only affected by low PFSS. These results might explain the different ways that Mepe-induced changes in pre-osteoblast mechanosensitivity may drive signaling pathways of bone cell function at low or high impact loading. These findings might have physiological and biomedical implications and require future research specifically addressing the precise role of integrin α or β subunits and Mepe during dynamic loading in bone health and disease. Full article

Titanium and titanium alloys are the prevailing dental implant materials owing to their favorable mechanical properties and biocompatibility, but how roughness dictates the biological response is still a matter of debate. In this study, laser texturing was used to generate eight paradigmatic roughened surfaces, with the aim of studying the early biological response elicited on MC3T3-E1 pre-osteoblasts. Prior to cell tests, the samples underwent SEM analysis, optical profilometry, protein adsorption assay, and optical contact angle measurement with water and diiodomethane to determine surface free energy. While all the specimens proved to be biocompatible, supporting similar cell viability at 1, 2, and 3 days, surface roughness could impact significantly on cell adhesion. Factorial analysis and linear regression showed, in a robust and unprecedented way, that an isotropic distribution of deep and closely spaced valleys provides the best condition for cell adhesion, to which both protein adsorption and surface free energy were highly correlated. Overall, here the authors provide, for the first time, a thorough investigation of the relationship between roughness parameters and osteoblast adhesion that may be applied to design and produce new tailored interfaces for implant materials. Full article

The strategies used to associate peptide arginylglycylaspartic acid (RGD) with calcium phosphate grafts to enhance cell–biomaterial interactions have been controversial in the literature. Several works have demonstrated that RGD-functionalized hydroxyapatite (HA) surfaces improve cell adhesion, whereas others claim that RGD-loaded HA has an inhibitory effect when serum is present in the biological medium. To investigate such contradictory results, we associated RGD with the HA surface using physical adsorption and chemical bonding methods and evaluated the cell adhesion and spreading in pre-osteoblasts culture with and without fetal bovine serum (FBS). The effect of functionalization methods on the physicochemical characteristics of both surfaces was analyzed using multiscale techniques. Adsorption assays of serum allowed us to estimate the impact of the association method on the HA surface’s reactivity. Physically adsorbed RGD did not increase the number of adhered cells due to the weak interactions between the peptide and the surface. Although chemical binding stabilizes RGD on the HA, the functionalization procedure covered the surface with molecules such as (3-aminopropyl)triethoxysilane (APTEs) and carbodiimide, changing the surface’s chemical activity. Serum protein adsorption decreased by 90%, revealing a significant reduction in the surface interactions with molecules of the biological medium. The present study’s findings showed that the RGD’s physical association with HA did not improve cell adhesion and that this phenomenon is highly dependent on the presence of serum proteins. Full article