J. Funct. Biomater., Volume 15, Issue 6 (June 2024) – 16 articles

Silver nanoparticles (AgNPs) are known for their antibacterial properties and their ability to promote wound healing. By incorporating silver nanoparticles into medical gauze, the resulting composite material shows promise as an advanced wound dressing. However, clinical applications are hindered by challenges related to the stability of silver nanoparticle loading on the gauze as nanoparticle leaching can compromise antibacterial efficacy. In this study, silver nanoparticles were immobilized onto polydopamine (PDA) submicron particles, which were then used to modify medical gauze. Energy dispersive spectroscopy (EDS) was employed to analyze the elemental distribution on the modified gauze, confirming successful surface modification. The antibacterial properties of the modified gauze were assessed using a laser scanning confocal microscope (CLSM). The results demonstrated a significant reduction in the adhesion rates of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) by 99.1% and 63%, respectively, on the PDA–Ag-modified gauze. Optical density (OD) measurements at 590 nm indicated that the modified gauze effectively inhibited biofilm formation, underscoring its potent antimicrobial capabilities. Further antibacterial efficacy was evaluated by diluting and plating co-cultured bacterial solutions with the modified dressing, followed by 24 h incubation and colony counting. The gauze exhibited an antibacterial efficiency of 99.99% against E. coli and 99.8% against S. aureus. Additionally, cell compatibility tests, involving the co-culture of PDA–Ag composites with human cells, demonstrated excellent biocompatibility. These findings suggest that PDA–Ag-modified medical gauze holds significant potential for the treatment of infected wounds, offering a promising solution to improve wound care through enhanced antimicrobial activity and biocompatibility. Full article

Salivary gland biofabrication represents a promising avenue in regenerative medicine, aiming to address the challenges of salivary gland dysfunction caused by various factors such as autoimmune diseases and radiotherapy. This review examines the current state of bioprinting technology, biomaterials, and tissue engineering strategies in the context of creating functional, implantable salivary gland constructs. Key considerations include achieving vascularization for proper nutrient supply, maintaining cell viability and functionality during printing, and promoting tissue maturation and integration with surrounding tissues. Despite the existing challenges, recent advancements offer significant potential for the development of personalized therapeutic options to treat salivary gland disorders. Continued research and innovation in this field hold the potential to revolutionize the management of salivary gland conditions, improving patient outcomes and quality of life. This systematic review covers publications from 2018 to April 2024 and was conducted on four databases: Google Scholar, PubMed, EBSCOhost, and Web of Science. The key features necessary for the successful creation, implantation and functioning of bioprinted salivary glands are addressed. Full article

Bone defects resulting from trauma, surgery, and congenital, infectious, or oncological diseases are a functional and aesthetic burden for patients. Bone regeneration is a demanding procedure, involving a spectrum of molecular processes and requiring the use of various scaffolds and substances, often yielding an unsatisfactory result. Recently, the new collagen sponge and its structural derivatives manufactured from European carp (Cyprinus carpio) were introduced and patented. Due to its fish origin, the novel scaffold poses no risk of allergic reactions or transfer of zoonoses and additionally shows superior biocompatibility, mechanical stability, adjustable degradation rate, and porosity. In this review, we focus on the basic principles of bone regeneration and describe the characteristics of an “ideal” bone scaffold focusing on guided bone regeneration. Moreover, we suggest several possible applications of this novel material in bone regeneration processes, thus opening new horizons for further research. Full article

The progress of digital technologies in dental prosthodontics is fast and increasingly accurate, allowing practitioners to simplify their daily work. These technologies aim to substitute conventional techniques progressively, but their real efficiency and predictability are still under debate. Many systematic reviews emphasize the lack of clinical RCTs that compare digital and traditional workflow. To address this evidence, we conducted a three-arm designed clinical RCT, which compares fully digital, combined digital, and analogic and fully analog workflows. We aimed to compare the clinical properties of each workflow regarding interproximal (IC) and occlusal contact (OC), marginal fit, impression time (IT), and patient satisfaction through a VAS scale. In total, 72 patients were included in the study. The IC and OC of the digital workflow were better than the others (p < 0.001), which obtained similar results. No difference between implant–abutment fit was observed (p = 0.5966). The IT was shorter in the digital workflow than the others (p < 0.001), which were similar. Patient satisfaction was higher in the digital workflow than in the conventional one. Despite the limitations, this study’s results support better accuracy and patient tolerance of digital workflow than of conventional techniques, suggesting it as a viable alternative to the latter when performed by clinicians experienced in digital dentistry. Full article

Objectives: This study investigated a novel strategy for improving regenerative cartilage outcomes. It combines fractional laser treatment with the implantation of neocartilage generated from autologous dynamic Self-Regenerating Cartilage (dSRC). Methods: dSRC was generated in vitro from harvested autologous swine chondrocytes. Culture was performed for 2, 4, 8, 10, and 12 weeks to study matrix maturation. Matrix formation and implant integration were also studied in vitro in swine cartilage discs using dSRC or cultured chondrocytes injected into CO₂ laser-ablated or mechanically punched holes. Cartilage discs were cultured for up to 8 weeks, harvested, and evaluated histologically and immunohistochemically. Results: The dSRC matrix was injectable by week 2, and matrices grew larger and more solid with time, generating a contiguous neocartilage matrix by week 8. Hypercellular density in dSRC at week 2 decreased over time and approached that of native cartilage by week 8. All dSRC groups exhibited high glycosaminoglycan (GAG) production, and immunohistochemical staining confirmed that the matrix was typical of normal hyaline cartilage, being rich in collagen type II. After 8 weeks in cartilage lesions in vitro, dSRC constructs generated a contiguous cartilage matrix, while isolated cultured chondrocytes exhibited only a sparse pericellular matrix. dSRC-treated lesions exhibited high GAG production compared to those treated with isolated chondrocytes. Conclusions: Isolated dSRC exhibits hyaline cartilage formation, matures over time, and generates contiguous articular cartilage matrix in fractional laser-created microenvironments in vitro, being well integrated with native cartilage. Full article

Computer-aided design and computer-aided manufacturing (CAD/CAM) techniques are based on either subtractive (milling prefabricated blocks) or additive (3D printing) methods, and both are used for obtaining dentistry materials. Our in vitro study aimed to investigate the behavior of human gingival fibroblasts exposed to methacrylate (MA)-based CAD/CAM milled samples in comparison with that of MA-based 3D-printed samples to better elucidate the mechanisms of cell adaptability and survival. The proliferation of human gingival fibroblasts was measured after 2 and 24 h of incubation in the presence of these samples using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the membrane integrity was assessed through the lactate dehydrogenase release. The level of reactive oxygen species, expression of autophagy-related protein LC3B-I, and detection of GSH and caspase 3/7 were evaluated by fluorescence staining. The MMP-2 levels were measured using a Milliplex MAP kit. The incubation with MA-based 3D-printed samples significantly reduced the viability, by 16% and 28% from control after 2 and 24 h, respectively. There was a 25% and 55% decrease in the GSH level from control after 24 h of incubation with the CAD/CAM milled and 3D-printed samples, respectively. In addition, higher levels of LC3B-I and MMP-2 were obtained after 24 h of incubation with the MA-based 3D samples compared to the CAD/CAM milled ones. Therefore, our results outline that the MA-CAD/CAM milled samples displayed good biocompatibility during 24-h exposure, while MA-3D resins are proper for short-term utilization (less than 24 h). Full article

The use of endosseous dental implants may become unfeasible in the presence of significant maxillary bone atrophy; thus, surgical techniques have been proposed to promote bone regeneration in such cases. However, such techniques are complex and may expose the patient to complications. Subperiosteal implants, being placed between the periosteum and the residual alveolar bone, are largely independent of bone thickness. Such devices had been abandoned due to the complexity of positioning and adaptation to the recipient bone site, but are nowadays witnessing an era of revival following the introduction of new acquisition procedures, new materials, and innovative manufacturing methods. We have analyzed the changes induced in gene and protein expression in C-12720 human osteoblasts by differently surface-modified TiO₂ materials to verify their ability to promote bone formation. The TiO₂ materials tested were (i) raw machined, (ii) electropolished with acid mixture, (iii) sand-blasted + acid-etched, (iv) AlTiColorTM surface, and (v) anodized. All five surfaces efficiently stimulated the expression of markers of osteoblastic differentiation, adhesion, and osteogenesis, such as RUNX2, osteocalcin, osterix, N-cadherin, β-catenin, and osteoprotegerin, while cell viability/proliferation was unaffected. Collectively, our observations document that presently available TiO₂ materials are well suited for the manufacturing of modern subperiosteal implants. Full article

The management and reconstruction of critical-sized segmental bone defects remain a major clinical challenge for orthopaedic clinicians and surgeons. In particular, regenerative medicine approaches that involve incorporating stem cells within tissue engineering scaffolds have great promise for fracture management. This narrative review focuses on the primary components of bone tissue engineering—stem cells, scaffolds, the microenvironment, and vascularisation—addressing current advances and translational and regulatory challenges in the current landscape of stem cell therapy for critical-sized bone defects. To comprehensively explore this research area and offer insights for future treatment options in orthopaedic surgery, we have examined the latest developments and advancements in bone tissue engineering, focusing on those of clinical relevance in recent years. Finally, we present a forward-looking perspective on using stem cells in bone tissue engineering for critical-sized segmental bone defects. Full article

The objective was to evaluate the change in color, hardness, and roughness produced by carbamide peroxide (CP) at two different concentrations on two resins. The 16% or 45% CP was applied to 66 resin discs with and without Bis-GMA. The color was measured with a spectrophotometer, and ΔE_ab and ΔE₀₀ were calculated. Microhardness tester and SEM were used. In both composites, the a* and b* coordinates tended to be red and yellow, respectively, and were significant in the Bis-GMA group (p < 0.05). The ΔE_ab and ΔE₀₀ were higher in the composite with Bis-GMA, regardless of the treatment received (p < 0.05). The microhardness was reduced in both composites regardless of the PC concentration compared to the control (p < 0.05). The 45% CP reduced the microhardness in the resin group with Bis-GMA compared to 16% CP (p < 0.001) but was not significant in the resin without Bis-GMA (p = 1). An increase in roughness was directly proportional to the concentration of CP, and it was more notable in the composite without Bis-GMA. The composite with Bis-GMA showed a greater tendency to darken than the one without Bis-GMA. The surface hardness of the composite was reduced in both composites and was not influenced by CP concentration in the composite without Bis-GMA. Bleaching is a common procedure nowadays. It is important to know how CP affects composites to establish a prognosis of the treatments in terms of color change, roughness, and hardness. Full article

This contribution gives basic information about the mechanical behavior of the facial part of the human skull cranium, i.e., the splanchnocranium, associated with external loads and injuries caused mainly by brachial violence. The main areas suffering from such violence include the orbit, frontal, and zygomatic bones. In this paper, as a first approach, brachial violence was simulated via quasi-static compression laboratory tests, in which cadaveric skulls were subjected to a load in a testing machine, increasing till fractures occurred. The test skulls were also used for research into the dynamic behavior, in which experimental and numerical analyses were performed. A relatively high variability in forces inducing the fractures has been observed (143–1403 N). The results lay the basis for applications mainly in forensic science, surgery, and ophthalmology. Full article

Yeasts resistant to antifungals have become an increasing risk to human health. One of the best antimicrobial properties is reported to be present in silver nanoparticles (AgNPs); however, little is known about the antimicrobial potential of AgNPs produced using thermophilic bacteria. How AgNPs cause cell death is different depending on the type of the cell, and the mode of death induced is cell-type specific. Apoptosis, one of the types of regulated cell death, can be extremely useful in the fight against infection because surrounding cells that have phagocytic activity can efficiently absorb the apoptotic bodies formed during apoptosis. In the course of this work, for the first time, comprehensive antifungal studies of AgNPs were performed using thermophilic Geobacillus spp. bacteria against Candida guilliermondii, also with the addition of the model yeast Saccharomyces cerevisiae. The determined minimal inhibitory concentrations (MICs) were 10 μg/mL against C. guilliermondii and 50 μg/mL against S. cerevisiae for Geobacillus sp. strain 25 AgNPs, and for Geobacillus sp. 612 the MICs were 5 μg/mL and 25 μg/mL, respectively. It was shown for the first time that the exposure of the yeast cells leads to caspase activation in both S. cerevisiae and C. guilliermondii after exposure to Geobacillus spp. AgNPs. Also, a statistically significant change in the number of cells with permeable membranes was detected. Moreover, it was shown that the antimicrobial effect of the AgNPs is related to ROS generation and lipid peroxidation in C. guilliermondii yeast. Full article

The recombinant structural protein described in this study was designed based on sequences derived from elastin and silk. Silk–elastin hybrid copolymers are characterized by high solubility while maintaining high product flexibility. The phase transition temperature from aqueous solution to hydrogel, as well as other physicochemical and mechanical properties of such particles, can differ significantly depending on the number of sequence repeats. We present a preliminary characterization of the EJ17zipR protein obtained in high yield in a prokaryotic expression system and efficiently purified via a multistep process. Its addition significantly improves biomaterial’s rheological and mechanical properties, especially elasticity. As a result, EJ17zipR appears to be a promising component for bioinks designed to print spatially complex structures that positively influence both shape retention and the internal transport of body fluids. The results of biological studies indicate that the addition of the studied protein creates a favorable microenvironment for cell adhesion, growth, and migration. Full article

Tissue engineering aims to develop bionic scaffolds as alternatives to autologous vascular grafts due to their limited availability. This study introduces a novel wet-electrospinning fabrication technique to create small-diameter, uniformly aligned tubular scaffolds. By combining this innovative method with conventional electrospinning, a bionic tri-layer scaffold that mimics the zonal structure of vascular tissues is produced. The inner and outer layers consist of PCL/Gelatin and PCL/PLGA fibers, respectively, while the middle layer is crafted using PCL through Wet Vertical Magnetic Rod Electrospinning (WVMRE). The scaffold’s morphology is analyzed using Scanning Electron Microscopy (SEM) to confirm its bionic structure. The mechanical properties, degradation profile, wettability, and biocompatibility of the scaffold are also characterized. To enhance hemocompatibility, the scaffold is crosslinked with heparin. The results demonstrate sufficient mechanical properties, good wettability of the inner layer, proper degradability of the inner and middle layers, and overall good biocompatibility. In conclusion, this study successfully develops a small-diameter tri-layer tubular scaffold that meets the required specifications. Full article

Effective root canal disinfection and the subsequent release of natural growth factors from dentin are crucial to the success of regenerative endodontic procedures. This study evaluated the effect of newly introduced calcium silicate-based temporary intracanal medicament Bio-C Temp and calcium hydroxide-based material UltraCal XS on the release of transforming growth factor β1 (TGF-β1) from root canal dentin. Twenty-two intact and fully developed human premolars from patients aged 15–18 were shaped and irrigated according to the current clinical recommendations. The teeth were then gently split in half, and the root canal dentin of paired samples was covered with Bio-C Temp or UltraCal XS. After 3 weeks of incubation, the specimens were conditioned with 17% EDTA and the collected solution was subjected to the quantification of the released TGF-β1 by performing an ELISA. One-way analysis of variance (ANOVA), followed by Tukey’s test, was selected to determine the statistically significant differences between the groups at the 0.95 confidence level. The highest mean value of released TGF-β1 (1993.1 pg/mL) was detected in the control group, where the root canal dentin was conditioned with 17% EDTA alone. Regarding the experimental groups, Bio-C Temp released a statistically significantly higher amount of TGF-β1 (282.14 pg/mL) compared to UltraCal XS (114.28 pg/mL; p = 0.0158). Bio-C Temp affected the release of growth factors from root canal dentin less than UltraCal XS and may therefore serve as an intracanal medicament for regenerative endodontic procedures. Full article

A Lactococcus (L.) lactis strain producing antimicrobial and anti-inflammatory biomolecules (mainly 1,4-Diaza-2,5-dioxobicyclo[4.3.0]nonanes and pyrazine-derivatives) was tested for its capacity to cure clinical endometritis in buffaloes compared to conventional antibiotic-based treatment. Clinical endometritis-diagnosed buffaloes (n = 16/group) were infused intrauterine with four doses of 10⁹ CFU-free (FLC group) or nanoencapsulated L. lactis (NLC group) and compared to those that received three doses of saline + a single dose of 500 mg cephapirin benzathin (AB group) or four doses of saline (control, C group) every other day. Endometrium samples were analyzed for cytological (polymorphonuclear cells, PMN), bacteriological, and proinflammatory mRNA expression. Uterine wash and blood samples were collected to determine proinflammatory cytokine concentrations and metabolites in the blood samples. The reproductive performance of buffaloes was assessed. Compared to the C group, the AB and NLC groups had the lowest percentage of PMN, followed by those in the FLC group (p < 0.05). All treated buffaloes had significantly lower numbers of pathogens than the control buffaloes. Compared to control, all treatments significantly down-regulated endometrial proinflammatory encoding mRNA expression. The concentrations of IL1B, TNFAIP7, and leukocyte esterase activity in the uterine washings were significantly decreased in the AB and NLC groups compared to the C and FLC groups. All treatments significantly decreased concentrations of serum proinflammatory cytokines compared to control. Both the AB and NLC groups had significantly lower concentrations of serum NEFA than the C and FLC groups. The percentage of control buffaloes having an echogenic uterus and PVD score > 2 was significantly higher than those in the treated buffaloes with higher numbers of corpora lutea, higher conception rates, and shorter days open than control buffaloes (p < 0.05). In conclusion, L. lactis-producing antimicrobial and anti-inflammatory metabolites reduce uterine inflammatory responses and improve fertility in buffaloes. Full article

Introduction: With the popularity of clear aligners, the sequential distalization protocol has been more commonly used for molar distalization. However, the amount of molar distalization that can be achieved, as well as the accompanying side effects on the sagittal dimension, are unclear. Methods: Registered with PROSPERO (CRD42023447211), relevant original studies were screened from seven databases (MEDLINE [PubMed], EBSCOhost, Web of Science, Elsevier [SCOPUS], Cochrane, LILACS [Latin American and Caribbean Health Sciences Literature], and Google Scholar) supplemented by a manual search of the references of the full-reading manuscripts by two investigators independently. A risk of bias assessment was conducted, relevant data were extracted, and meta-analysis was performed using RStudio. Results: After the screening, 13 articles (11 involving maxillary distalization, two involving mandibular distalization) met the inclusion criteria. All studies had a high or medium risk of bias. The meta-analysis revealed that the maxillary first molar (U6) mesiobuccal cusp was distalized 2.07 mm [1.38 mm, 2.77 mm] based on the post-distalization dental model superimposition, and the U6 crown was distalized 2.00 mm [0.77 mm, 3.24 mm] based on the post-treatment lateral cephalometric evaluation. However, the U6 mesiobuccal root showed less distalization of 1.13 mm [−1.34 mm, 3.60 mm], indicating crown distal tipping, which was validated by meta-analysis (U6-PP angle: 2.19° [1.06°, 3.33°]). In addition, intra-arch anchorage loss was observed at the post-distalization time point (U1 protrusion: 0.39 mm [0.27 mm, 0.51 mm]), which was corrected at the post-treatment time point (incisal edge-PTV distance: −1.50 mm [−2.61 mm, −0.39 mm]). Conclusion: About 2 mm maxillary molar distalization can be achieved with the sequential distalization protocol, accompanied by slight molar crown distal tipping. Additional studies on this topic are needed due to the high risk of bias in currently available studies. Full article