Tissue Engineering Applications Research Articles

USP26 Combats Age-Related Declines in Self-Renewal and Multipotent Differentiation of BMSC by Maintaining Mitochondrial Homeostasis.

Age-related declines in self-renewal and multipotency of bone marrow mesenchymal stem cells (BMSCs) limit their applications in tissue engineering and clinical therapy. Thus, understanding the mechanisms behind BMSC senescence is crucial for maintaining the rejuvenation and multipotent differentiation capabilities of BMSCs. This study reveals that impaired USP26 expression in BMSCs leads to mitochondrial dysfunction, ultimately resulting in aging and age-related declines in the self-renewal and multipotency of BMSCs. Specifically, decreased USP26 expression results in decreased protein levels of Sirtuin 2 due to its ubiquitination degradation, which leads to mitochondrial dysfunction in BMSCs and ultimately resulting in aging and age-related declines in self-renewal and multilineage differentiation potentials. Additionally, decreased USP26 expression in aging BMSCs is a result of dampened hypoxia-inducible factor 1α (HIF-1α) expression. HIF-1α facilitates USP26 transcriptional expression by increasing USP26 promoter activity through binding to the -191 - -198 bp and -262 - -269 bp regions on the USP26 promoter. Therefore, the identification of USP26 as being correlated with aging and age-related declines in self-renewal and multipotency of BMSCs, along with understanding its expression and action mechanisms, suggests that USP26 represents a novel therapeutic target for combating aging and age-related declines in the self-renewal and multipotent differentiation of BMSCs.

Bacterial Cellulose: A Multifunctional Platform for Biomedical Applications

Bacterial cellulose (BC), a biopolymer synthesized by various bacterial species, has emerged as a promising material for biomedical applications due to its unique properties, including high purity, biocompatibility, mechanical strength, and structural similarity to the extracellular matrix. This review explores the advancements in BC research over the last decade, focusing on its applications in tissue engineering, wound healing, and drug delivery systems. While BC offers numerous benefits, challenges such as large-scale production, structural modification, however regulatory approval hinder its broader clinical use. Recent studies have introduced innovative solutions, such as using agro-industrial waste to lower production costs and combining BC with other materials to enhance its bioactivity. As research progresses, BC has the potential to revolutionize the field of biomedicine, offering sustainable, versatile, and effective solutions for a wide range of medical applications.

Leveraging in vivo animal models of tendon loading to inform tissue engineering approaches.

Tendon injuries disrupt successful transmission of force between muscle and bone, resulting in reduced mobility, increased pain, and significantly reduced quality of life for affected patients. There are currently no targeted treatments to improve tendon healing beyond conservative methods such as rest and physical therapy. Tissue engineering approaches hold great promise for designing instructive biomaterials that could improve tendon healing or for generating replacement graft tissue. More recently, engineered microphysiological systems to model tendon injuries have been used to identify therapeutic targets. Despite these advances, current tissue engineering efforts that aim to regenerate, replace, or model injured tendons have largely failed due in large part to a lack of understanding of how the mechanical environment of the tendon influences tissue homeostasis and how altered mechanical loading can promote or prevent disease progression. This review article draws inspiration from what is known about tendon loading from in vivo animal models and identifies key metrics that can be used to benchmark success in tissue engineering applications. Finally, we highlight important challenges and opportunities for the field of tendon tissue engineering that should be taken into consideration in designing engineered platforms to understand or improve tendon healing.

Fabrication of porous Ni-Ti shape memory alloy via binder jet additive manufacturing and solid-state sintering

This paper explores the additive manufacturing of nickel-titanium (Ni-Ti) shape memory alloys (SMAs) using binder jetting and subsequent solid-state sintering under an Ar atmosphere. The consolidated 3D-printed Ni-Ti parts exhibit a correlation between pore morphology, pore fraction, and phase formation at different solid-state sintering temperatures. At a sintering temperature of 1175 °C, NiTi grains with TiC precipitates along grain boundaries are observed, accompanied by irregular, interconnected pores constituting ∼15 % of the volume. Increasing the sintering temperature to 1185 °C leads to the formation of a minor phase of Ni4Ti3 within NiTi grains, along with grain boundary TiC precipitates, and isolated pores with a volume fraction of ∼5 %. The higher sintering temperature corresponds to a higher average nanohardness value (8.1 GPa or 763 Hv compared to 6.9 GPa or 654 Hv), indicating the presence of a greater abundance of secondary phases and higher densification at the elevated sintering temperature. While the transformation temperatures (TTs) were undetectable in the bulk-printed samples, a different structure featuring designed channels and thin struts, sintered under similar conditions, exhibited detectable TTs, with a martensite start temperature of 34 °C. Biocompatibility tests demonstrated cell spreading and attachment on both sintered Ni-Ti samples. These findings offer valuable insights into the potential use of binder jetted Ni-Ti for medical implants and tissue engineering applications.

Electrospun "Hard-Soft" Interpenetrating Nanofibrous Tissue Scaffolds Facilitating Enhanced Mechanical Strength and Cell Proliferation.

"Soft" hydrogel-based macroporous scaffolds have been widely used in tissue engineering and drug delivery applications due to their hydrated interfaces and macroporous structures, but have drawbacks related to their weak mechanics and often weak adhesion to cells. In contrast, "hard" poly(caprolactone) (PCL) electrospun fibrous networks have desirable mechanical strength and ductility but offer minimal interfacial hydration and thus limited capacity for cell proliferation. Herein, we demonstrate the fabrication of interpenetrating nanofibrous networks based on coelectrospun PCL and poly(oligoethylene glycol methacrylate) (POEGMA) nanofibers that exhibit the mechanical benefits of PCL but the interfacial hydration benefits of hydrogels. The electrospinning process results in partially aligned but interpenetrating fiber network with minimal internal phase separation, leading to anisotropic but strong mechanical properties even in the hydrated state; apparent ultimate tensile strengths of the swollen scaffolds ranged from 429 ± 39 kPa in the direction of fiber alignment (longitudinal) to 86 ± 25 kPa perpendicular to fiber alignment (cross-longitudinal), typical of PCL-based scaffolds and enabling efficient suture retention in different directions. However, contact angle measurements indicate hydrogel-like interfacial properties due to the presence of the interpenetrating POEGMA network. C2C12 myoblast proliferation in the PCL-POEGMA scaffolds was 50% higher than that observed on PCL-only scaffolds, a result attributed to the presence of the more hydrophilic POEGMA interpenetrating nanofiber network. Overall, this method is demonstrated to represent a facile single-step strategy to fabricate strong macroporous but still interfacially hydrophilic scaffolds for tissue engineering applications.

Fabrication and characterization of nanohydroxyapatite/chitosan/decellularized placenta scaffold for bone tissue engineering applications

Novel biomaterials are necessary to fabricate biomimetic scaffolds for bone tissue engineering. In the present experiment, we aimed to fabricate and evaluate the osteogenic properties of nanohydroxyapatite/chitosan/decellularized placenta (nHA.Cs.dPL) composite scaffolds. The human placenta was decellularized (dPL), characterized, and digested in pepsin to form the hydrogel. nHA.Cs.dPL scaffolds were fabricated using salt leaching/freeze drying and evaluated for their morphology, chemical composition, swelling, porosity, degradation, mechanical strength, and biocompatibility. Saos-2 cells were seeded on scaffolds, and their osteogenic properties were investigated by evaluating alkaline phosphatase (ALP), osteocalcin (OCN), collagen type 1 (COL I) expression, and calcium deposition under osteogenic differentiation. The dPL was prepared with minimized DNA content and a well-preserved porous structure. Scaffolds were highly porous with interconnected pores and exhibited appropriate swelling and degradation rates supporting saos-2 cell attachment and proliferation. dPL improved scaffold physicochemical features and increased cell proliferation, ALP, OCN, COL I expression, and calcium deposition under osteogenic differentiation induction. nHA.Cs.dPL composite scaffolds provide a 3D microenvironment with superior physicochemical features that support saos-2 cell adhesion, proliferation, and osteogenic differentiation.

Biomimetic bilayer scaffold from Bombyx mori silk materials for small diameter vascular applications in tissue engineering.

Enhancing the biocompatibility and mechanical stability of small diameter vascular scaffolds remain significant challenges. To address this challenge, small-diameter tubular structures were electrospun from silk fibroin (SF) from silk textile industry discarded materials to generate bilayer scaffolds that mimic native blood vessels, but derived from a sustainable natural material resource. The inner layer was obtained by directly dissolving SF in formic acid, while the middle layer (SF-M) was achieved through aqueous concentration of the protein. Structural and biological properties of each layer as well as the bilayer were evaluated. The inner layer exhibited nano-scale fiber diameters and 57.9% crystallinity, and degradation rates comparable with the SF-M layer. The middle layer displayed micrometer-scale fibers diameters with an ultimate extension of about 274%. Both layers presented contact angles suitable for cell growth and cytocompatibility, while the bilayer material displayed an intermediate mechanical response and a reduced enzymatic degradation rate when compared to each individual layer. The bilayer material emulates many of the characteristics of native small-diameter vessels, thereby suggesting further studies towards in vivo opportunities.

In Vitro Evaluation of Electrospun PCL Bioscaffold with Zinc-Doped Bioactive Glass Powder Addition.

Preparing electrospun fibers by applying a potential difference between a polymeric solution and a contacting substrate is increasingly attracting attention in tissue engineering applications. Among the numerous polymers, polycaprolactone (PCL) bioscaffold has been widely investigated due to its biocompatibility and biodegradability. Bioactive powder can be added to further improve its performance. In the present study, bioactive glass powder modified by adding 0-6 wt.% antibacterial zinc element (coded as ZBG) was prepared through the sol-gel process. Furthermore, PCL bioscaffolds with various ZBG additions were prepared using the electrospinning technique. The zinc-doped bioactive glass powder and electrospun PCL/ZBG bioscaffolds were evaluated using scanning electron microscopy, X-ray diffraction and Fourier-transform infrared spectroscopy to determine their structural properties. Additionally, in vitro bioactivity, biocompatibility and antibacterial performance were investigated. Experimental results showed that sol-gelled ZBG powder possessed superior bioactivity and 0.8 g ZBG was the optimal addition to prepare PCL/ZBG bioscaffolds with. All the electrospun PCL/ZBG bioscaffolds were biocompatible and their antibacterial performance against two S. aureus strains (SA133 and Newman) improved with increasing zinc concentration. Electrospun PCL/ZBG bioscaffolds exhibited excellent bioactivity and have great potential for biomedical application.

A hybrid construct with tailored 3D structure for directing pre-vascularization in engineered tissues

Hybrid 3D constructs combining different structural components afford unique opportunities to engineer functional tissues. Creating functional microvascular networks within these constructs is crucial for promoting integration with host vessels and ensuring successful engraftment. Here, we present a hybrid 3D system in which poly (ethylene oxide terephthalate)/poly (butylene terephthalate) fibrous scaffolds are combined with pectin hydrogels to provide internal topography and guide the formation of microvascular beds. The sequence/method of seeding human endothelial cells (EC) and mesenchymal stromal cells (MSC) into the system had a significant impact on microvessel formation. Optimal results were obtained when EC were directly seeded onto the fibrous scaffold, followed by the addition of hydrogel-embedded MSC. This approach facilitated the development of highly oriented microvascular networks along the fibers. These networks were lumenized, supported by a basement membrane, and stabilized by pericyte-like cells, persisting for at least 28 days in vitro. Furthermore, culture under pro-angiogenic and osteoinductive conditions induced MSC osteogenic differentiation without impairing microvessel formation. Upon subcutaneous implantation in mice, the pre-vascularized constructs were infiltrated by host vessels, and human microvessels were still present after 2 weeks. Overall, the proposed hybrid 3D system, combined with an optimized cell-seeding protocol, offers an effective approach for directing the formation of robust and geometrically oriented microvessels, making it promising for tissue engineering applications.

Investigating the impact of curcumin extract incorporation on PVA/Collagen/Chitosan/HAp nanofiber scaffold: morphological, mechanical, wettability, degradation rate, and antibacterial activity

This study investigates the impact of curcumin extract incorporation on the properties of PVA/Collagen/Chitosan/HAp nanofiber scaffolds for bone tissue engineering applications. Nanofibers were fabricated via electrospinning using PVA, collagen, chitosan, hydroxyapatite (HAp), and curcumin. Their morphology, mechanical properties, wettability, degradation rate, and antibacterial activity were comprehensively evaluated. Fourier transform infrared (FTIR) spectroscopy confirmed the successful incorporation of all components into the nanofibers and the potential for interactions between their functional groups. Scanning Electron Microscopy (SEM) revealed bead-free, fine nano-fibers with a decrease in diameter ranging from 139 nm to 108 nm. Simultaneously, enhanced mechanical properties were observed, with ultimate strength increasing from 2.8 MPa to 8.8 MPa as the diameter decreased. Notably, 15% curcumin extract maintained favorable hydrophilicity and degradation rates, crucial factors for tissue regeneration. Antibacterial activity was also enhanced, with an inhibition zone of 6.71 mm against E.coli observed at 15% curcumin treatment. Staphylococcus aureus exhibited the largest zone of inhibition (8.74 mm) at 15% curcumin concentration. This research demonstrates the feasibility of incorporating curcumin into nanofiber scaffolds for bone tissue engineering applications.

Piezoelectric Scaffolds as Smart Materials for Bone Tissue Engineering.

Bone repair and regeneration require physiological cues, including mechanical, electrical, and biochemical activity. Many biomaterials have been investigated as bioactive scaffolds with excellent electrical properties. Amongst biomaterials, piezoelectric materials (PMs) are gaining attention in biomedicine, power harvesting, biomedical devices, and structural health monitoring. PMs have unique properties, such as the ability to affect physiological movements and deliver electrical stimuli to damaged bone or cells without an external power source. The crucial bone property is its piezoelectricity. Bones can generate electrical charges and potential in response to mechanical stimuli, as they influence bone growth and regeneration. Piezoelectric materials respond to human microenvironment stimuli and are an important factor in bone regeneration and repair. This manuscript is an overview of the fundamentals of the materials generating the piezoelectric effect and their influence on bone repair and regeneration. This paper focuses on the state of the art of piezoelectric materials, such as polymers, ceramics, and composites, and their application in bone tissue engineering. We present important information from the point of view of bone tissue engineering. We highlight promising upcoming approaches and new generations of piezoelectric materials.

Optimization of Polyvinylidene Fluoride/Polyvinyl Alcohol Electrospun using Taguchi Method for Tissue Engineering Application

Composite nanofibers for tissue engineering applications were prepared with polyvinylidene fluoride and polyvinyl alcohol via a horizontal electrospinning process. The effect of the processing parameters (polymer concentration, voltage, feed rate and tip-to-collector distance) on the fibre diameter was studied using L9 orthogonal arrays of the Taguchi Method, S/N ratio, and ANOVA to produce uniform and finest fibre. The optimum parameters were at 90 wt% PVDF ratios for polymer concentration, applied voltage at 10 kV, feed rate at 0.5 mL/h and tip-to-collector distance at 15 cm. The PVDF/PVA electrospun compromises 171.60 nm nanofiber diameter with homogenous morphology and no bead formed. MTT and live/dead kit assay analysis toward human (HSF) cells confirmed the potential of the PVDF/PVA electrospun to be used as an alternative tissue template to allow new cell or tissue formation in the future.

How to select agroforestry waste biomass for electrospinning and its potential application in bone tissue engineering

The high value-added utilization of agroforestry waste biomass is urgent. Herein, a feasible approach was provided to obtain biodegradable cellulose fibrous films from agroforestry waste biomass. The cellulose used was extracted from agroforestry waste biomass and then the cellulose fibrous film was obtained by direct electrospinning. Lignin-carbohydrate complex (LCC) structure was considered as the key factor for the dissolution of lignocellulose while cellulose molecular weight >335,664 was suitable for electrospinning. Bamboo cellulose was chosen as an example to verify the potential application of the electrospun cellulose films from agroforestry waste biomass. The as-prepared electrospun bamboo cellulose fibrous film exhibited a tensile strength of 24.12 MPa, which outperformed most of the reported electrospun nanofibrous films. Moreover, the film possessed a super-wetting surface and outstanding cytocompatibility. These excellent properties offer the film with immense potential for application in bone tissue engineering. In addition, this work provides a new route for transforming agroforestry waste into high value-added products.

Characterization and physiochemical of PCL/extracted collagen blend coated nanostructure Sodium-Alginate substrate for skin tissue engineering application

Abstract This study involves fabrication a nano-membrane of collagen and polycarbolactone by electrospinning and depositing into alginate films prepared by casting method to serve as a scaffold for tissue engineering. Collagen extracted from bovine skin showed poor ability to electrospun, so polycaprolactone (PCL), a synthetic polymer commonly used in tissue engineering scaffolds was chosen to improve the electrospinning process and obtain continuous fibers without beads suitable for application in tissue engineering. The scaffolds were analyzed using Field emission scanning electron microscopy, Fourier transformtion infrared spectroscopy, swelling degree testing, and wettability measurements. FESEM results showed that blending PCL with collagen led to improving the electrospinning process and obtaining uniform, continuous fibers (with average fiber diameter 44.97 ± 1.61 nm) without beads and more crosslinking compared to the polycarbolactone scaffold. The results of the wettability and degree of swelling also showed the effect of collagen on increasing the hydrophilicity of the scaffold, and reducedthe water contact angle to (66.66°) with degree of swelling (1256%), that making it suitable for tissue engineering applications.

SonoPrint: Acoustically Assisted Volumetric 3D Printing for Composites.

Advances in additive manufacturing in composites have transformed aerospace, medical devices, tissue engineering, and electronics. A key aspect of enhancing properties of 3D-printed objects involves fine-tuning the material by embedding and orienting reinforcement within the structure. Existing methods for orienting these reinforcements are limited by pattern types, alignment, and particle characteristics. Acoustics offers a versatile method to control the particles independent of their size, geometry, and charge, enabling intricate pattern formations. However, integrating acoustics into 3D printing has been challenging due to the scattering of the acoustic field between polymerized layers and unpolymerized resin, resulting in unwanted patterns. To address this challenge, SonoPrint, an innovative acoustically assisted volumetric 3D printer is developed that enables simultaneous reinforcement patterning and printing of the entire structure. SonoPrint generates mechanically tunable composite geometries by embedding reinforcement particles, such as microscopic glass, metal, and polystyrene, within the fabricated structure. This printer employs a standing wave field to create targeted particle motifs-including parallel lines, radial lines, circles, rhombuses, hexagons, and polygons-directly in the photosensitive resin, completing the print in just a few minutes. SonoPrint enhances structural properties and promises to advance volumetric printing, unlocking applications in tissue engineering, biohybrid robots, and composite fabrication.

Impact of Polydeoxyribonucleotides on the Morphology, Viability, and Osteogenic Differentiation of Gingiva-Derived Stem Cell Spheroids

Background and Objectives: Polydeoxyribonucleotides (PDRN), composed of DNA fragments derived from salmon DNA, is widely recognized for its regenerative properties. It has been extensively used in medical applications, such as dermatology and wound healing, due to its ability to enhance cellular metabolic activity, stimulate angiogenesis, and promote tissue regeneration. In the field of dentistry, PDRN has shown potential in promoting periodontal healing and bone regeneration. This study aims to investigate the effects of PDRN on the morphology, survival, and osteogenic differentiation of gingiva-derived stem cell spheroids, with a focus on its potential applications in tissue engineering and regenerative dentistry. Materials and Methods: Gingiva-derived mesenchymal stem cells were cultured and formed into spheroids using microwells. The cells were treated with varying concentrations of PDRN (0, 25, 50, 75, and 100 μg/mL) and cultivated in osteogenic media. Cell morphology was observed over seven days using an inverted microscope, and viability was assessed with Live/Dead Kit assays and Cell Counting Kit-8. Osteogenic differentiation was evaluated by measuring alkaline phosphatase activity and calcium deposition. The expression levels of osteogenic markers RUNX2 and COL1A1 were quantified using real-time polymerase chain reaction. RNA sequencing was performed to assess the gene expression profiles related to osteogenesis. Results: The results demonstrated that PDRN treatment had no significant effect on spheroid diameter or cellular viability during the observation period. However, a PDRN concentration of 75 μg/mL significantly enhanced calcium deposition by Day 14, suggesting increased mineralization. RUNX2 and COL1A1 mRNA expression levels varied with PDRN concentration, with the highest RUNX2 expression observed at 25 μg/mL and the highest COL1A1 expression at 75 μg/mL. RNA sequencing further confirmed the upregulation of genes involved in osteogenic differentiation, with enhanced expression of RUNX2 and COL1A1 in PDRN-treated gingiva-derived stem cell spheroids. Conclusions: In summary, PDRN did not significantly affect the viability or morphology of gingiva-derived stem cell spheroids but influenced their osteogenic differentiation and mineralization in a concentration-dependent manner. These findings suggest that PDRN may play a role in promoting osteogenic processes in tissue engineering and regenerative dentistry applications, with specific effects observed at different concentrations.

Cryogenic 3D Printing of GelMA/Graphene Bioinks: Improved Mechanical Strength and Structural Properties for Tissue Engineering.

Tissue engineering aims to recreate natural cellular environments to facilitate tissue regeneration. Gelatin methacrylate (GelMA) is widely utilized for its biocompatibility, ability to support cell adhesion and proliferation, and adjustable mechanical characteristics. This study developed a GelMA and graphene bioink platform at concentrations of 1, 1.5, and 2 mg/mL to enhance scaffold properties for tissue engineering applications. Graphene was incorporated into GelMA matrices to improve mechanical strength and electrical conductivity of the bioinks. The compressive strength and thermal stability of the resulting GelMA/graphene scaffolds were assessed through DSC analysis and mechanical testing. Cytotoxicity assays were conducted to determine cell survival rates. Cryoprinting at -30°C was employed to preserve scaffold structure and function. The chorioallantoic membrane (CAM) assay was used to evaluate biocompatibility and angiogenic potential. The integration of graphene significantly amplified the compressive strength and thermal stability of GelMA scaffolds. Cytotoxicity assays indicated robust cell survival rates of 90%, confirming the biocompatibility of the developed materials. Cryoprinting effectively preserved scaffold integrity and functionality. The CAM assay validated the biocompatibility and angiogenic potential, demonstrating substantial vascularization upon scaffold implantation onto chick embryo CAM. Integrating graphene into GelMA hydrogels, coupled with low-temperature 3D printing, represents a potent strategy for enhancing scaffold fabrication. The resultant GelMA/graphene scaffolds exhibit superior mechanical properties, biocompatibility, and pro-vascularization capabilities, making them highly suitable for diverse tissue engineering and regenerative medicine applications.

3D melt blowing of Elastollan thermoplastic polyurethane for tissue engineering applications: A pilot study

Scaffolds, in addition to being biocompatible, should possess structural and mechanical properties similar to the natural tissues they intend to replace. Many tissue engineering applications require porous 3D scaffolds characterized by unique microfibrous organization and mechanical anisotropy. Manufacturing process principles and process parameter-biomaterial interactions ultimately govern the properties that can be achieved in the scaffold. In this study, we investigate a recently developed nonwoven scaffold fabrication process, 3D melt blowing (3DMB), for processing Elastollan®, a thermoplastic polyurethane with basic mechanical properties suitable for musculoskeletal tissue engineering. The range of feasible processing parameters was screened and the effects of two sets of critical process parameters (fiber deposition offset and surface velocity of the collector) that produced contrasting scaffold morphologies were assessed. Results showed that scaffolds of Group B that were fabricated at the higher fiber deposition offset (90 %) and higher surface velocity of the collector (6 × 105 mm/min) possessed significantly smaller fiber diameter and higher porosity and degree of fiber alignment along the principal direction of collector rotation during 3DMB (all p < 0.05) compared to Group A scaffolds (fabricated at 50 % offset and 1 × 105 mm/min surface velocity). Although both groups possessed similar tensile stiffness, the elongation at failure was significantly different (p < 0.0001). The higher elongation at failure of Group B correlated with the higher degree of fiber alignment in these scaffolds. In contrast, the more isotropic fibrous organization of Group A contributed to their higher compressive stiffness (p = 0.004). The introduction of NaOH treatment to improve hydrophilicity of the scaffolds resulted in a significant reduction of tensile stiffness of Group A (p < 0.05) but not Group B. This treatment did not significantly affect the elongation at failure or compressive stiffness of both groups. With NaOH-treatment, both groups demonstrated good biocompatibility when seeded with fibroblast cells over 14 days. This study confirms the ability to fabricate via 3DMB, biocompatible, micro-fibrous, Elastollan scaffolds relevant for musculoskeletal tissue engineering.

Biomolecule-functionalized exfoliated-graphene and graphene oxide as heteronucleants of nanocrystalline apatites to make hybrid nanocomposites with tailored mechanical, luminescent, and biological properties

Nanocrystalline apatite (Ap), known for its exceptional biological properties, faces limitations in hard tissue engineering due to its poor mechanical properties. To overcome this limitation, we investigated the preparation of nanocomposites through heterogeneous nucleation of calcium phosphate on exfoliated graphene (G) and graphene oxide (GO) flakes, selected for their outstanding mechanical properties. The flakes were treated (functionalized) with amino acids of varying isoelectric points—namely L-Arginine (Arg), L-Alanine (Aln) and L-Aspartic acid (Asp)— as well as citrate (Cit) molecules. Furthermore, Tb3+ was incorporated into the formulations to introduce luminescence and further enrich the functionality of the composite. The synthesis was conducted using the sitting drop vapor diffusion method. Functionalized GO/Ap nanocomposites significantly improved roughness, adhesion forces and elastic modulus compared to Ap and G-based particles. GO-Asp-Ap-Tb nanocomposites exhibited the highest roughness (163.8 ± 116.2 nm), while G-Cit-Ap had the lowest (6.8 ± 5.6 nm). In terms of adhesion force, GO-Cit-Ap-Tb reached the highest value (31.06 ± 13.3 nN), while G-Arg-Ap had the lowest (3.7 ± 1.8 nN) compared to Ap (13.6 ± 3.2 nN). For the elastic modulus, GO-Aln-Ap-Tb demonstrated the greatest stiffness (3489 ± 101.01 MPa) compared to Ap (30.2 ± 6.5 MPa), while G-Aln-Ap-Tb showed the lowest (17.2 ± 8.4 MPa). Concerning their luminescence, regardless of G/Ap and GO/Ap, the relative luminescence intensities depended on the biomolecule used and decreased in the order Arg > Aln >Asp and Cit. Furthermore, G/Ap and GO/Ap nanocomposites demonstrated good biocompatibility on murine mesenchymal stem cells at low concentrations, showing cell viabilities exceeding 80% at 0.1 μg/mL. This research offers a novel approach to enhancing the mechanical properties of apatites while preserving their good biocompatibility properties and introducing new functionalities (i.e. luminescence) in the composites, thereby expanding their range of applications in hard tissue engineering.

Influence of manufacturing parameters on bioactive glass 45S5: Structural analysis and applications in bone tissue engineering

Bioactive glass (BG-45S5) production through the melting process is affected by a wide variety of parameters. This study investigated the synthesis of BG-45S5 granules and the process variables to produce a bioactive and osteoinductive BG for bone grafting applications. The melting process was initially analyzed by varying parameters such as crucible type and pouring environment using P2O5 as phosphorus precursor. The obtained products were characterized by crystalline phases, characteristic chemical groups, particle size distribution, and chemical composition. Materials poured into graphite or steel molds resulted in particle sizes more suitable for applications in granular form. Using a platinum crucible yielded a chemical composition closer to the target when compared with another ceramic crucible. Subsequently, the melting process was evaluated to different phosphorus precursor (P2O5 or Na2HPO4) and melting duration (1 or 2 h) in a platinum crucible verifying their effects on the thermal behavior, chemical composition and structure of BG-45S5. Employing Na2HPO4 as a precursor led to higher glass transition and crystallization temperatures as compared to P2O5, enhancing glass homogeneity and structural stability. The product with better characteristics in terms of composition and structure was further characterized for bioactivity and cell culture behavior, showing a greater amount of mineralization nodules when compared to commercial hydroxyapatite. This is particularly due to its behavior as the solubility and interaction in biological environments.