Journal of Functional Biomaterials

Osteoconductive silk fibroin (SF) binders were fabricated for the bone repair of an alveolar cleft defect. Binders were prefigureared by mixing different ratios of a mixture of random coils and SF aggregation with SF fibrils: 100:0 (SFB100), 75:25 (SFB75), 50:50 (SFB50), 25:75 (SFB25), and 0:100 (SFB0). The gelation, molecular organization, structures, topography, and morphology of the binders were characterized and observed. Their physical, mechanical, and biological properties were tested. The SF binders showed gelation via self-assembly of SF aggregation and fibrillation. SFB75, SFB50, and SFB25 had molecular formation via the amide groups and showed more structural stability than SFB100. The morphology of SFB0 demonstrated the largest pore size. SFB0 showed a lowest hydrophilicity. SFB100 showed the highest SF release. SFB25 had the highest maximum load. SFB50 exhibited the lowest elongation at break. Binders with SF fibrils showed more cell viability and higher cell proliferation, ALP activity, calcium deposition, and protein synthesis than without SF fibrils. Finally, the results were deduced: SFB25 demonstrated suitable performance that is promising for the bone repair of an alveolar cleft defect. Full article

After bleaching, enamel surfaces are damaged, contributing to erosion and tooth sensitivity. Although fluoride is used after bleaching to try and revert alterations, it is not capable of repairing tooth structure. This study compared the effect of a self-assembly peptide (P₁₁-4), with and without fluoride, and sodium fluoride (NaF 2%) on the Knoop microhardness (KHN) and surface roughness (Ra (μm)) of bleached enamel with an in-office bleaching regimen. Enamel blocks of bovine teeth (5 × 5 × 2 mm) with standardized surface hardness were bleached with 35% carbamide peroxide, following the manufacturer’s instructions. The teeth were randomly divided into the following groups (n = 7) according to post-bleaching treatment: no treatment (negative control) (C-); 2% NaF (NaF); Curodont™ Repair (Repair); and Curodont™ Protect (Protect). Specimens were stored in artificial saliva at 37 °C. To evaluate the effect of the post-bleaching treatments, KHN and Ra were measured before bleaching (baseline) and 24 h and 7 days after bleaching. Data were submitted to repeated measures ANOVA and Bonferroni tests (α = 0.05). There were significant interactions between the study factors (p = 0.001). After 7 days, Repair (572.50 ± 79.04) and Protect (583.00 ± 74.76) specimens showed increased surface KHN, with values higher than the NaF (465.50 ± 41.50) and C- (475.22 ± 58.95) baseline values. There was no significant difference in KHN at 24 h among groups (p = 0.587). At 24 h after bleaching, Repair was significantly different from all groups (p < 0.05). Repair showed the lowest Ra (μm) values (0.133 ± 0.035). After seven days, there was no significant difference in Ra values among groups when compared to the baseline. The use of P₁₁-4-based materials after bleaching resulted in the fastest recovery to baseline enamel properties. Full article

Implant-related infection (IRI) is a major problem in orthopedics. Copper (Cu) is an essential trace element with strong bactericidal activity and, thus, presents potential for reducing IRI. The present study explored a straightforward strategy for releasing Cu²⁺ from titanium (Ti) implants, and we conducted a preliminary study to assess the feasibility of this approach in clinical translation. Polylactic acid (PLA) coatings containing different concentrations of copper ions were prepared on Ti discs. The antibacterial activity and biocompatibility of the copper ion-incorporated Ti implants were evaluated using Staphylococcus aureus (S. aureus), bone marrow mesenchymal stem cells (BMSCs) and animal models. In vitro, the coatings produced burst release of Cu²⁺ in 12 h, and inhibited S. aureus growth in a dose-dependent manner. The coatings prepared from PLA solutions containing 0.5 or 1.0 mg/mL reduced the viability and osteogenic differentiation of BMSCs, but these effects were negated after the coatings were immersed in culture medium for 6 h. Four weeks after implantation, the Cu-free K-wires challenged with S. aureus had persistent infection and inferior fracture healing to the other three groups, while Cu-coated wires had no evidence of infection. Furthermore, the Cu-coated wires placed in rabbits without S. aureus challenge showed superior fracture healing to the other three groups. These results suggest that PLA coatings containing Cu²⁺ may be an effective design for reducing IRI without adversely affecting adjacent bone healing. Full article

Graphene-based materials have attracted much attention due to their fascinating properties such as hydrophilicity, high dispersion in aqueous media, robust size, high biocompatibility, and surface functionalization ability due to the presence of functional groups and interactions with biomolecules such as proteins and nucleic acid. Modified methods were developed for safe, direct, inexpensive, and eco-friendly synthesis. However, toxicity to the environment and animal health has been reported, raising concerns about their utilization. This review focuses primarily on the synthesis methods of graphene-based materials already developed and the unique properties that make them so interesting for different applications. Different applications are presented and discussed with particular emphasis on biological fields. Furthermore, antimicrobial potential and the factors that affect this activity are reviewed. Finally, questions related to toxicity to the environment and living organisms are revised by highlighting factors that may interfere with it. Full article

Minimally invasive exodontia is among the long-sought-for development aims of safe dental medicine. In this paper, we aim, for the first time, to examine whether the enzymatic disruption of the periodontal ligament fibers reduces the force required for tooth extraction. To this end, recombinantly expressed clostridial collagenase G variant purified from Escherichia coli was injected into the periodontal ligament of mesial and distal roots of the first and second split porcine mandibular premolars. The vehicle solution was injected into the corresponding roots on the contralateral side. Following sixteen hours, the treated mandibles were mounted on a loading machine to measure the extraction force. In addition, the effect of the enzyme on the viability of different cell types was evaluated. An average reduction of 20% in the applied force (albeit with a large variability of 50 to 370 newton) was observed for the enzymatically treated roots, reaching up to 50% reduction in some cases. Importantly, the enzyme showed only a minor and transient effect on cellular viability, without any signs of toxicity. Using an innovative model enabling the analytical measurement of extraction forces, we show, for the first time, that the enzymatic disruption of periodontal ligament fibers substantially reduces the force required for tooth extraction. This novel technique brings us closer to atraumatic exodontia, potentially reducing intra- and post-operative complications and facilitating subsequent implant placement. The development of novel enzymes with enhanced activity may further simplify the tooth extraction process and present additional clinical relevance for the broad range of implications in the oral cavity. Full article

The fabrication of patient-specific scaffolds for bone substitutes is possible through extrusion-based 3D printing of calcium phosphate cements (CPC) which allows the generation of structures with a high degree of customization and interconnected porosity. Given the brittleness of this clinically approved material, the stability of open-porous scaffolds cannot always be secured. Herein, a multi-technological approach allowed the simultaneous combination of CPC printing with melt electrowriting (MEW) of polycaprolactone (PCL) microfibers in an alternating, tunable design in one automated fabrication process. The hybrid CPC+PCL scaffolds with varying CPC strand distance (800–2000 µm) and integrated PCL fibers featured a strong CPC to PCL interface. While no adverse effect on mechanical stiffness was detected by the PCL-supported scaffold design; the microfiber integration led to an improved integrity. The pore distance between CPC strands was gradually increased to identify at which critical CPC porosity the microfibers would have a significant impact on pore bridging behavior and growth of seeded cells. At a CPC strand distance of 1600 µm, after 2 weeks of cultivation, the incorporation of PCL fibers led to pore coverage by a human mesenchymal stem cell line and an elevated proliferation level of murine pre-osteoblasts. The integrated fabrication approach allows versatile design adjustments on different levels. Full article

Bioactive glasses are often designed as porous implantable templates in which newly-formed bone can grow in three dimensions (3D). This research work aims to investigate the bone regenerative capability of silicate bioactive glass scaffolds produced by robocasting in comparison with powder and granule-like materials (oxide system: 47.5SiO₂-10Na₂O-10K₂O-10MgO-20CaO-2.5P₂O₅, mol.%). Morphological and compositional analyses performed by scanning electron microscopy (SEM), combined with energy dispersive spectroscopy (EDS) after the bioactivity studies in a simulated body fluid (SBF) confirmed the apatite-forming ability of the scaffolds, which is key to allowing bone-bonding in vivo. The scaffolds exhibited a clear osteogenic effect upon implantation in rabbit femur and underwent gradual resorption followed by ossification. Full resorption in favor of new bone growth was achieved within 6 months. Osseous defect healing was accompanied by the formation of mature bone with abundant osteocytes and bone marrow cells. These in vivo results support the scaffold’s suitability for application in bone tissue engineering and show promise for potential translation to clinical assessment. Full article

The development of antimicrobial devices and surfaces requires the setup of suitable materials, able to store and release active principles. In this context, zeolites, which are microporous aluminosilicate minerals, hold great promise, since they are able to serve as a reservoir for metal-ions with antimicrobial properties. Here, we report on the preparation of Linde Type A zeolites, partially exchanged with combinations of metal-ions (Ag⁺, Cu²⁺, Zn²⁺) at different loadings (0.1–11.9 wt.%). We combine X-ray fluorescence, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray diffraction to monitor the metal-ion contents, distribution, and conservation of the zeolite structure after exchange. Then, we evaluate their antimicrobial activity, using agar dilution and optical-density monitoring of Escherichia coli cultures. The results indicate that silver-loaded materials are at least 70-fold more active than the copper-, zinc-, and non-exchanged ones. Moreover, zeolites loaded with lower Ag⁺ concentrations remain active down to 0.1 wt.%, and their activities are directly proportional to the total Ag content. Sequential exchanges with two metal ions (Ag⁺ and either Cu²⁺, Zn²⁺) display synergetic or antagonist effects, depending on the quantity of the second metal. Altogether, this work shows that, by combining analytical and quantitative methods, it is possible to fine-tune the composition of bi-metal-exchanged zeolites, in order to maximise their antimicrobial potential, opening new ways for the development of next-generation composite zeolite-containing antimicrobial materials, with potential applications for the design of dental or bone implants, as well as biomedical devices and pharmaceutical products. Full article

Biodegradable metals have been extensively studied due to their potential use as temporary biomedical devices, on non-load bearing applications. These types of implants are requested to function for the healing period, and should degrade after the tissue heals. A balance between mechanical properties requested at the initial stage of implantation and the degradation rate is required. The use of temporary biodegradable implants avoids a second surgery for the removal of the device, which brings high benefits to the patients and avoids high societal costs. Among the biodegradable metals, iron as a biodegradable metal has increased attention over the last few years, especially with the incorporation of additive manufacturing processes to obtain tailored geometries of porous structures, which give rise to higher corrosion rates. Withal by mimic natural bone hierarchical porosity, the mechanical properties of obtained structures tend to equalize that of human bone. This review article presents some of the most important works in the field of iron and porous iron. Fabrication techniques for porous iron are tackled, including conventional and new methods highlighting the unparalleled opportunities given by additive manufacturing. A comparison among the several methods is taken. The effects of the design and the alloying elements on the mechanical properties are also revised. Iron alloys with antibacterial properties are analyzed, as well as the biodegradation behavior and biocompatibility of iron. Although is necessary for further in vivo research, iron is presenting satisfactory results for upcoming biomedical applications, as orthopaedic temporary scaffolds and coronary stents. Full article

Injectable hydrogels, as carriers, offer great potential to incorporate cells or growth factors for dental tissue regeneration. Notably, the development of injectable hydrogels with appropriate structures and properties has been a challenging task, leaving much to be desired in terms of cytocompatibility, antibacterial and self-healing properties, as well as the ability to support dental stem cell functions. This paper presents our study on the development of a novel self-cross-linkable hydrogel composed of oxidized alginate and carboxymethyl chitosan and its characterization as a cell carrier for dental enamel regeneration in vitro. Oxidized alginate was synthesized with 60% theoretical oxidation degree using periodate oxidation and characterized by Fourier Transform Infrared spectroscopy, proton nuclear magnetic resonance spectroscopy, and Ultraviolet-visible absorption spectroscopy. Then, hydrogels were prepared at three varying weight ratios of oxidized alginate to carboxymethyl chitosan (4:1, 3:1, and 2:1) through Schiff base reactions, which was confirmed by Fourier Transform Infrared spectroscopy. The hydrogels were characterized in terms of gelation time, swelling ratio, structure, injectability, self-healing, antibacterial properties, and in vitro characterization for enamel regeneration. The results demonstrated that, among the three hydrogels examined, the one with the highest ratio of oxidized alginate (i.e., 4:1) had the fastest gelation time and the lowest swelling ability, and that all hydrogels were formed with highly porous structures and were able to be injected through a 20-gauge needle without clogging. The injected hydrogels could be rapidly reformed with the self-healing property. The hydrogels also showed antibacterial properties against two cariogenic bacteria: Streptococcus mutans and Streptococcus sobrinus. For in vitro enamel regeneration, a dental epithelial cell line, HAT-7, was examined, demonstrating a high cell viability in the hydrogels during injection. Furthermore, HAT-7 cells encapsulated in the hydrogels showed alkaline phosphatase production and mineral deposition, as well as maintaining their round morphology, after 14 days of in vitro culture. Taken together, this study has provided evidence that the oxidized alginate-carboxymethyl chitosan hydrogels could be used as an injectable cell carrier for dental enamel tissue engineering applications. Full article

Tissue engineering concepts, which are concerned with the attachment and growth of specific cell types, frequently employ immobilized ligands that interact preferentially with cell types of interest. Creating multicellular grafts such as heart valves calls for scaffolds with spatial control over the different cells involved. Cardiac heart valves are mainly constituted out of two cell types, endothelial cells and valvular interstitial cells. To have control over where which cell type can be attracted would enable targeted cell settlement and growth contributing to the first step of an engineered construct. For endothelial cells, constituting the outer lining of the valve tissue, several specific peptide ligands have been described. Valvular interstitial cells, representing the bulk of the leaflet, have not been investigated in this regard. Two receptors, the integrin α9β1 and CD44, are known to be highly expressed on valvular interstitial cells. Here, we demonstrate that by covalently grafting the corresponding peptide and polysaccharide ligand onto an erodible, polycaprolactone (PCL), and a non-degradable, polytetrafluoroethylene (PTFE), polymer, surfaces were generated that strongly support valvular interstitial cell colonization with minimal endothelial cell and reduced platelet adhesion. The technology for covalent binding of corresponding ligands is a key element towards tissue engineered cardiac valves for in vitro applications, but also towards future in vivo application, especially in combination with degradable scaffold material. Full article

During an angioscopy operation, a transparent liquid called dextran is sprayed out from a catheter to flush the blood away from the space between the camera and target. Medical doctors usually inject dextran at a constant flow rate. However, they often cannot obtain clear angioscopy visibility because the flushing out of the blood is insufficient. Good flushing conditions producing clear angioscopy visibility will increase the rate of success of angioscopy operations. This study aimed to determine a way to improve the clarity for angioscopy under different values for the parameters of the injection waveform, endoscope position, and catheter angle. We also determined the effect of a stepwise waveform for injecting the dextran only during systole while synchronizing the waveform to the cardiac cycle. To evaluate the visibility of the blood-vessel walls, we performed a computational fluid dynamics (CFD) simulation and calculated the visible area ratio (VAR), representing the ratio of the visible wall area to the total area of the wall at each point in time. Additionally, the normalized integration of the VAR called the area ratio (AR_VAR) represents the ratio of the visible wall area as a function of the dextran injection period. The results demonstrate that the AR_VAR with a stepped waveform, bottom endoscope, and three-degree-angle catheter results in the highest visibility, around 25 times larger than that under the control conditions: a constant waveform, a center endoscope, and 0 degrees. This set of conditions can improve angioscopy visibility. Full article

Despite the advent of promising technologies in tissue engineering, finding a biomimetic 3D bio-construct capable of enhancing cell attachment, maintenance, and function is still a challenge in producing tailorable scaffolds for bone regeneration. Here, osteostimulatory effects of the butterfly wings as a naturally porous and non-toxic chitinous scaffold on mesenchymal stromal cells are assessed. The topographical characterization of the butterfly wings implied their ability to mimic bone tissue microenvironment, whereas their regenerative potential was validated after a 14-day cell culture. In vivo analysis showed that the scaffold induced no major inflammatory response in Wistar rats. Topographical features of the bioconstruct upregulated the osteogenic genes, including COL1A1, ALP, BGLAP, SPP1, SP7, and AML3 in differentiated cells compared to the cells cultured in the culture plate. However, butterfly wings were shown to provide a biomimetic microstructure and proper bone regenerative capacity through a unique combination of various structural and material properties. Therefore, this novel platform can be confidently recommended for bone tissue engineering applications. Full article

In the current study, cornstarch (CS) and eggshell powder (ESP) were combined using a casting technique to develop a biodegradable film that was further morphologically and physicochemically characterized using standard methods. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to characterize the morphology of the ESP/CS film, and the surface of the film was found to have a smooth structure with no cracks, a spherical and porous irregular shape, and visible phase separation, which explains their large surface area. In addition, the energy dispersive X-ray (EDX) analysis indicated that the ESP particles were made of calcium carbonate and the ESP contained carbon in the graphite form. Fourier Transform Infrared Spectroscopy indicated the presence of carbonated minerals in the ESP/CS film which shows that ESP/CS film might serve as a promising adsorbent. Due to the inductive effect of the O–C–O bond on calcium carbonate in the eggshell, it was discovered that the ESP/CS film significantly improves physical properties, moisture content, swelling power, water solubility, and water absorption compared to the control CS film. The enhancement of the physicochemical properties of the ESP/CS film was principally due to the intra and intermolecular interactions between ESP and CS molecules. As a result, this film can potentially be used as a synergistic adsorbent for various target analytes. Full article

The aim of this clinical case study was to observe biopsy tissues at 5 months after an autograft of a partially demineralized dentin/cementum matrix (pDDM) into a tooth-extracted socket exhibiting healing failure. A 66-year-old female presented with healing failure in the cavity for 2 months after the extraction (#36). Initial X-ray photos showed a clear remainder of lamina dura (#36), a residual root (#37), and a horizontal impaction (#38). The vital tooth (#38) was selected for pDDM. The third molar crushed by electric mill was decalcified in 1.0 L of 2.0% HNO₃ for 20 min and rinsed in cold distilled water. The pDDM granules (size: 0.5–2.0 mm) were grafted immediately into the treated socket. X-ray views just after pDDM graft showed radio-opaque granules. At 5 months after pDDM graft, the surface of regenerated bone was harmonized with the mandibular line, and bone-like radio-opacity was found in the graft region. The biopsy tissue (diameter: 3.0 mm) at 5 months after pDDM graft showed that mature bone was interconnected with the remaining pDDM. The novel histological evidence highlighted that newly formed bone was connected directly with both dentin-area and cementum-area matrix of pDDM. We concluded that pDDM contributed to the regeneration of bone in the unhealed socket, and this regeneration prepared the socket for implant placement. Autogenous pDDM could be immediately recycled as an innovative biomaterial for local bone regeneration. Full article

Monetite and brushite are regarded with increasing interest for the preparation of biomaterials for applications in the musculoskeletal system. Herein, we investigated the influence of strontium substitution in the structures of these two phosphates on bone cell response. To achieve this aim, co-cultures of human primary osteoclasts and human osteoblast-like MG63 cells were tested on strontium-substituted monetite and strontium-substituted brushite, as well as on monetite and brushite, as controls. In both structures, strontium substitution for calcium amounted to about 6 at% and provoked enlargement of the cell parameters and morphologic variations. Cumulative release in physiological solution increased linearly over time and was greater from brushite (up to about 160 and 560 mg/L at 14 days for Sr and Ca, respectively) than from monetite (up to about 90 and 250 mg/L at 14 days for Sr and Ca, respectively). The increasing viability of osteoblast-like cells over time, with the different expression level of some typical bone markers, indicates a more pronounced trigger toward osteoblast differentiation and osteoclast inhibition by brushite materials. In particular, the inhibition of cathepsin K and tartrate-resistant acid phosphatase at the gene and morphological levels suggests strontium-substituted brushite can be applied in diseases characterized by excessive bone resorption. Full article

Due to various concerns about the use of metal-on-metal that is detrimental to users, the use of metal as acetabular cup material was later changed to ultra high molecular weight polyethylene (UHMWPE). However, the wear on UHMWPE releases polyethylene wear particles, which can trigger a negative body response and contribute to osteolysis. For reducing the wear of polyethylene, one of the efforts is to investigate the selection of metal materials. Cobalt chromium molybdenum (CoCrMo), stainless steel 316L (SS 316L), and titanium alloy (Ti6Al4V) are the frequently employed materials. The computational evaluation of contact pressure was carried out using a two-dimensional axisymmetric model for UHMWPE acetabular cup paired with metal femoral head under gait cycle in this study. The results show Ti6Al4V-on-UHMWPE is able to reduce cumulative contact pressure compared to SS 316L-on-UHMWPE and CoCrMo-on-UHMWPE. Compared to Ti6Al4V-on-UHMWPE at peak loading, the difference in cumulative contact pressure to respective maximum contact pressure is 9.740% for SS 316L-on-UHMWPE and 11.038% for CoCrMo-on-UHMWPE. Full article

Tissue engineering using scaffolds is a promising strategy to repair damaged articular cartilage, whose self-repair is inefficient. Cellulose properties have been recognized for their application in the biomedical field. The aim of this study was to fabricate and characterize novel scaffolds based on poly(ɛ-caprolactone) (PCL) and sustainable cellulose. Thus, the performance of corncob-derived cellulose (CC) in scaffolds as an alternative to wood cellulose (WC) was also investigated to reduce the environmental footprint. Two concentrations of CC in scaffolds were tested, 1% and 2% (w/w), and commercial WC using the same concentrations, as a control. Morphologically, all the developed scaffolds presented pore sizes of ~300 µm, 10 layers, a circular shape and well-dispersed cellulose. Thus, all of these characteristics and properties provide the manufactured scaffolds suitable for use in cartilage-replacement strategies. The use of 2% CC results in higher porosity (54.24%), which promotes cell infiltration/migration and nutrient exchange, and has similar mechanical properties to WC. As for the effects of enzymatic degradation of the scaffolds, no significant changes (p > 0.05) were observed in resistance over time. However, the obtained compressive modulus of the scaffold with 2% CC was similar to that of WC. Overall, our results suggest that the integration of 2% corncob cellulose in PCL scaffolds could be a novel way to replace wood-cellulose-containing scaffolds, highlighting its potential for cartilage-replacement strategies. Full article

The combination of titania nanofilms and silver nanoparticles (NPs) is a very promising material, with antibacterial and osseointegration-induced properties for titanium implant coatings. In this work, we successfully prepared TiO₂ nanolayer/Ag NP structures on titanium disks using atomic layer deposition (ALD). The samples were studied by scanning electron microscopy (SEM), X-ray diffraction, X-ray photoelectron spectroscopy (XPS), contact angle measurements, and SEM-EDS. Antibacterial activity was tested against Staphylococcus aureus. The in vitro cytological response of MG-63 osteosarcoma and human fetal mesenchymal stem cells (FetMSCs) was examined using SEM study of their morphology, MTT test of viability and differentiation using alkaline phosphatase and osteopontin with and without medium-induced differentiation in the osteogenic direction. The samples with TiO₂ nanolayers, Ag NPs, and a TiO₂/Ag combination showed high antibacterial activity, differentiation in the osteogenic direction, and non-cytotoxicity. The medium for differentiation significantly improved osteogenic differentiation, but the ALD coatings also stimulated differentiation in the absence of the medium. The TiO₂/Ag samples showed the best antibacterial ability and differentiation in the osteogenic direction, indicating the success of the combining of TiO₂ and Ag to produce a multifunctional biocompatible and bactericidal material. Full article