Search Results (82)

Brucella infection is associated with an increased risk of adverse obstetric outcomes in humans and animals. Decidualization, a process involving structural and functional changes in endometrial stromal cells, is essential for proper trophoblast implantation and placental development. Trophoblasts’ migration and their ability to invade the decidua and to undergo tubulogenesis, critical for proper implantation and placental development, are normally promoted by decidual cells. We evaluated whether Brucella infection of human endometrial stromal cells (T-HESC cell line) affects their ability to decidualize and to promote trophoblast functions. Infection of T-HESC cells with either B. abortus, B. suis, or B. melitensis resulted in deficient decidualization (as revealed by reduced prolactin levels) and an increased production of proinflammatory chemokines (C-X-C motif chemokine ligand 8 -CXCL8- and C-C motif chemokine ligand 2 -CCL2-) as compared to uninfected cells subjected to decidualization stimuli. In addition, conditioned media (CM) from infected decidualized T-HESC induced an inflammatory response (CXCL8, CCL2 and interleukin-6 -IL-6) in human trophoblasts (Swan-71 cell line) but reduced their ability to produce progesterone. Trophoblasts preincubated with this CM also had reduced migration, invasion, and tubulogenesis capacities, and this impairment was mediated, at least in part, by CXCL8 and CCL2. Moreover, infection of decidual stromal cells impaired the adhesion and spreading of blastocyst-like spheroids formed by Swan-71 cells. Brucella infection also affected the chemotactic capacity of decidual stromal cells for trophoblasts. Overall, these results suggest that Brucella infection of endometrial stromal cells impairs key processes required for successful implantation and placental development. Full article

Hydrogel is widely used as a scaffolding material for tissue engineering due to its excellent cytocompatibility and potential for biofunctionalization. However, its poor mechanical property limits its further application. Fabrication of fiber-reinforced hydrogel composite scaffolds has emerged as a solution to overcome this problem. However, existing strategies usually produce nonporous composite scaffolds, where the interfiber pores are completely filled with hydrogel. This design can hinder oxygen and nutrient exchange between seeded cells and the culture medium, thereby limiting cell invasion and colonization within the scaffold. In this study, sodium alginate (SA) hydrogel was exclusively grafted onto the surface of the constituent fibers of the melt electrowritten scaffold while preserving the porous structure. The grafted hydrogel amount and pore size were precisely controlled by adjusting the SA concentration and the crosslinking ratio (SA: CaCl₂). Experimental results demonstrated that the porous composite scaffolds exhibited superior swelling capacity, degradation ratio, mechanical properties, and biocompatibility. Notably, at an SA concentration of 0.5% and a crosslinking ratio of 2:1, the porous composite scaffold achieved optimal cell adhesion and viability. This study highlights the critical importance of preserving porous structures in composite scaffolds for tissue-engineering applications. Full article

Klebsiella pneumoniae, a zoonotic pathogen of global concern, poses significant threats to both veterinary and public health. Here, a comparative study characterized 14 clinical isolates (7 avian-derived, 7 human-derived) from Jiangsu, China, through integrated genomic and phenotypic analyses. Firstly, multilocus sequence typing (MLST) revealed distinct epidemiological patterns: the same ST type in avian isolates was circulating between different species and different regions, whereas it was not found in human isolates. In addition, hypervirulent Klebsiella pneumoniae (hvKP) phenotypes confirmed by string test were exclusive to two human isolates (KP15, KP20). Secondly, biofilm detection demonstrated 78.6% (11/14) of isolates possessed biofilm-forming capacity, with cellulose but not curli as the predominant matrix component. Human-derived KP15 and KP20 had the strongest biofilm formation ability in all isolates. Antimicrobial susceptibility profiling identified serious multidrug resistance in both avian and human isolates. Virulence gene analysis revealed striking disparities, with human isolates harboring 10–20 virulence factors (median 15) versus 6–7 (median 6.5) in avian counterparts. Finally, functional pathogenesis assessments demonstrated human-derived strains exhibited stronger epithelial cell adhesion (2-fold higher) and invasion (1.97-fold higher) in Calu-3 cell models and paradoxically showed reduced macrophage phagocytosis (2.85-fold lower at 2 h) for immune escape. In vivo models confirmed dose-dependent mortality, with human isolates demonstrating higher lethality in both Galleria mellonella and mice. Virulence gene burden positively correlated with mortality outcomes. These findings delineate critical host adaptation differences in Klebsiella pneumoniae populations and provide empirical evidence for pathogen transmission dynamics at the human-animal interface. Full article

Background: In recent years, thanks to the improvement of adhesive techniques, patients affected by tooth wear, related to erosion and/or parafunctional habits, can undergo restoration by adding only what has been lost of their dentition (additive approach). However, since not all clinicians are convinced that dental rehabilitation should be proposed in the early stages of exposed dentin, several treatments are often postponed. It is important to emphasize that, in the early stages, the clinical approach should remain conservative, focusing on dietary counseling, the modification of harmful habits, fluoride application, and risk factor management. Only when these preventive and non-invasive strategies prove insufficient, and the condition continues to progress, should invasive restorative treatments be considered. Unfortunately, epidemiological studies are reporting an increase in the number of young patients affected by erosive tooth wear, and not intercepting these cases earlier could lead to a severe degradation of the affected dentition. In addition, parafunctional habits are also becoming more frequent among patients. The combination of erosion and attrition can be very destructive, and may progress rapidly once dentin is exposed and the risk factors remain unaddressed. The aim of this report was to present a conservative full-mouth rehabilitation approach for severe erosive lesions and to provide a 10-year follow-up assessing the biological, functional, and esthetic outcomes. Methods: In this article, the postponed restorative treatment of a patient, suffering from severe tooth wear, is illustrated. The patient had sought dental treatment in the past; however, due to the already very compromised dentition, a conventional but very aggressive treatment was proposed and refused. Four years later, when the patient finally accepted an alternative conservative therapy, the tooth degradation was very severe, especially at the level of the maxillary anterior teeth. The combination of three different approaches, Speed-Up Therapy, BOPT (Biologically-Oriented Preparation Technique), and the 3 Step Technique, however, improved the capacity to successfully complete the difficult therapeutic task. Results: The biological goals (maintenance of the pulp vitality of all of the teeth and the minimal removal of healthy tooth structure) were accomplished, relying only on adhesive techniques. Conclusions: The overall treatment was very comfortable for the patient and less complicated for the clinician. At 10-year follow-up, biological, functional, and esthetic success was still confirmed. Full article

Pleopeltis crassinervata has demonstrated antimicrobial effects, including anti-Toxoplasma activity, which has been attributed to the presence of compounds such as terpenes and fatty acid methyl esters. In this study, the effects of P. crassinervata hexane subfraction one (Hsf1) on the Toxoplasma gondii tachyzoite ultrastructure were evaluated using TEM and SEM, and lytic cycle processes such as adhesion, invasion, and proliferation were evaluated using phase-contrast microscopy. Additionally, the antioxidant capacity of the subfraction and its main compounds (phytol and hexadecenoic acid methyl ester) were determined as well as their effects on parasite viability. Hsf1 exhibited a dose-dependent inhibitory effect on the lytic process at a concentration of 47.2 µg/mL. Among the eighteen compounds identified in this subfraction, six were evaluated, of which two (phytol and hexadecanoic acid methyl ester) significantly reduced the viability of T. gondii to 0.11% and 16.6%, respectively, at a concentration of 100 µg/mL. Additionally, Hsf1 demonstrated an antioxidant capacity of 30% as assessed using the ORAC method. The two active compounds also exhibited antioxidant properties, with antioxidant capacities of 13.33% and 33% for hexadecanoic acid methyl ester and phytol, respectively, at concentrations up to 15.4 mg/mL. Hsf1 showed membrane damage and conoid extrusion in T. gondii tachyzoites, suggesting direct interference with the lytic cycle of the parasite. These findings underscore the therapeutic potential of Hsf1 as a promising tool for controlling infections caused by T. gondii, thereby providing an alternative in the search for new antiparasitic agents. However, further research is required to determine the in vivo pharmacological effects and properties of these compounds with potential anti-Toxoplasma activity. Full article

Mastitis, an inflammatory condition of the udder, can be caused by the entry of Staphylococcus aureus, whose adhesion to the mammary epithelial cells is influenced by virulence factors such as microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) and the accessory gene regulator (agr) system. Our goal was to determine the adhesion and invasion rates of S. aureus isolates from clinical (mild and moderate) and subclinical mastitis and to assess the impact of MSCRAMM genes and agr types on disease severity. Clinical isolates predominantly carried agrII (p < 0.0083) and multiple MSCRAMM genes, correlating with high adhesion capacity but reduced invasion capacity regardless of clinical severity. Remarkably, subclinical isolates, mainly agr-negative (85.7%), showed increased cellular invasion (p < 0.0001), possibly due to reduced expression of agr-mediated virulence factors. These findings contribute to the understanding of the pathogen–host dynamics in bovine mastitis and highlight the importance of both MSCRAMMs and the agr system in modulating disease severity. These insights can inform targeted interventions for mastitis prevention and treatment. Full article

Pseudomonas aeruginosa is an opportunistic pathogenic bacterium, responsible for several life-threatening infections due to its multiple virulence factors and problematic multi-drug resistance, hence the necessity to find alternatives such as competitive probiotics. Pediococcus pentosaceus MZF16 is an LAB strain, isolated from traditional dried meat “Ossban”, with high probiotic potential. Our study investigated the capacity of P. pentosaceus MZF16 to counteract P. aeruginosa H103 using several tests on intestinal cells (analysis of cytotoxicity, inflammation, adhesion/invasion) and on the in vivo Caenorhabditis elegans model. The effect of MZF16 on the quorum sensing of the pathogen was also examined. We found that P. pentosaceus MZF16 was able to reduce H103 cytotoxicity and inflammatory activity and prevented pathogen colonization and translocation across Caco-2/TC7 cells. MZF16 also exerted an anti-virulence effect by attenuating quorum-sensing (QS) molecules and pyoverdine production and extended C. elegans lifespan. The obtained results highlight the potential of P. pentosaceus MZF16 probiotic strain as an anti-Pseudomonas aeruginosa alternative and establish a basis for elucidating the mechanisms of P. pentosaceus MZF16 involved in countering P. aeruginosa virulence. Full article

Objectives: To develop and evaluate graphene oxide/gelatin/alginate scaffolds for advanced wound therapy capable of mimicking the native extracellular matrix (ECM) and bio-stimulating all specific phases of the wound healing process, from inflammation and proliferation to the remodeling of damaged skin tissue in three dimensions. Methods: The scaffolds were engineered as interpenetrating polymeric networks by the crosslinking reaction of gelatin in the presence of alginate and characterized by structural, morphological, mechanical, swelling properties, porosity, adhesion to the skin tissue, wettability, and in vitro simultaneous release of the active agents. Biocompatibility of the scaffolds were evaluated in vitro by MTT test on fibroblasts (MRC5 cells) and in vivo using Caenorhabditis elegans assay. Results: The scaffolds exhibited a highly porous interconnected morphology with adjustable porosity (93–96%) and mechanical strength (1.10–2.90 MPa), hydrophilic nature with high capacity to absorb physiological fluids, and stable adhesion to the skin tissue. The obtained results of MRC5 cell viability indicate that the scaffolds are safe for biomedical applications. No mortality was detected among the Caenorhabditis elegans throughout the incubation period, indicating that the scaffolds are not toxic. The results of in vitro release study of allantoin, quercetin, and caffeic acid confirm the scaffolds’ significant potential for simultaneous release. Conclusion: The graphene oxide/gelatin/alginate scaffolds are promising candidates for non-invasive, dual ECM-mimetic, and multi-target wound therapy, offering an innovative strategy to address the complexities of wound healing process. Full article

Background and Objectives: Autologous bone grafting is the first choice for reconstructive surgery in bone defects due to trauma or malignant tumors. However, there is an increasing demand for minimally invasive alternatives involving bone regeneration using artificial materials. Biomimetic materials that replicate the body’s microscopic structure, such as Cellnest^®, are gaining attention. Cellnest is a xeno-free recombinant peptide based on human type I collagen, containing a rich Arg-Gly-Asp (RGD) motif related to cell adhesion. The aim of this study was to compare the effects of Cellnest with existing collagen materials (Pelnac^®, Integra^®, Terudermis^®) on bone regeneration and elucidate the underlying mechanisms. Materials and Methods: In vivo experiments involved a rat model of calvarial bone defects, in which Cellnest and other collagen materials were implanted into the defect area. Bone formation was assessed after 4 weeks using micro-computed tomography (micro-CT) and histological analysis. In vitro experiments included the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), adhesion, and migration assays, and a real-time polymerase chain reaction using rapidly expanding cells (RECs) to explore the mechanisms of Cellnest’s bone regenerative capacity. Results: The micro-CT analysis showed that the regenerated bone area was significantly greater in the Cellnest group (72.3%) than in the Pelnac^® (25.5%), Integra^® (31.6%), and Terudermis^® (38.3%) groups. The histological analysis confirmed similar trends, with Cellnest showing 42.2% bone regeneration, outperforming the other materials. The in vitro assays revealed that Cellnest promoted cell proliferation, adhesion, and migration. Gene expression analysis demonstrated that Cellnest significantly increased the levels of the bone formation markers ALP and COL1. Conclusions: Cellnest, a human type I collagen-like peptide rich in RGD motifs, enhances bone regeneration by promoting MSC adhesion and migration, and bone formation-related gene expression. The findings suggest its potential as an effective material for bone defect reconstruction. Full article

Lung cancer remains the cancer with the highest mortality worldwide, largely due to a limited understanding of the precise molecular mechanisms that drive its progression. microRNAs (miRNAs) have emerged as crucial regulators of lung cancer progression by influencing key cellular processes, notably the epithelial–mesenchymal transition (EMT). EMT is a complex and potentially reversible process where epithelial cells lose their polarity and adhesion, reorganize their cytoskeleton, and transition to a mesenchymal phenotype, enhancing their migratory and invasive capacities. While EMT plays an essential role in normal physiological contexts such as tissue development and wound healing, it is also a critical mechanism underlying the progression and metastasis of lung cancer. This review aims to summarize the latest research findings on the role of endogenous and exosome-derived microRNAs in regulating EMT in lung cancer, focusing on studies conducted over the past five years. It also provides an overview of EMT’s essential molecular mechanisms to better understand how miRNAs regulate EMT in lung cancer. Full article

Kinin receptors B1 and B2 are involved in migration and invasion in gastric, glioma, and cervical cancer cells, among others. However, the role of kinin receptors in breast cancer cells has been poorly studied. We aimed to reveal the impact of B1 and B2 receptors on migration and invasion in breast cancer cells and demonstrate their capacity to modulate in vivo tumor growth. MDA-MB-231, MCF-7, and T47D cells treated with Lys-des[Arg⁹]bradykinin (LDBK) or bradykinin (BK) were used to evaluate migration and invasion. Des-[Arg⁹]-Leu⁸-BK and HOE-140 were used as antagonists for the B1 and B2 receptors. MDA-MB-231 cells incubated or not with antagonists were subcutaneously inoculated in BALBc NOD/SCID mice to evaluate tumor growth. LDBK and BK treatment significantly increased migration and invasion in breast cancer cells, effects that were negated when antagonists were used. The use of antagonists in vivo inhibited tumor growth. Moreover, the migration and invasion induced by kinins in breast cancer cells were inhibited when focal adhesion kinase (FAK) and Src inhibitors were used. The novelty revealed in our work is that B1 and B2 receptors activated by LDBK and BK induce migration and invasion in breast cancer cells via a mechanism that involves the FAK–Src signaling pathway, and the antagonism of both receptors in vivo impairs breast tumor growth. Full article

The impressive adhesive capacity of marine mussels has inspired various fascinating designs in biomedical fields. Mussel-inspired injectable adhesive hydrogels, as a type of promising mussel-inspired material, have attracted much attention due to their minimally invasive property and desirable functions provided by mussel-inspired components. In recent decades, various mussel-inspired injectable adhesive hydrogels have been designed and widely applied in numerous biomedical fields. The rational incorporation of mussel-inspired catechol groups endows the injectable hydrogels with the potential to exhibit many properties, including tissue adhesiveness and self-healing, antimicrobial, and antioxidant capabilities, broadening the applications of injectable hydrogels in biomedical fields. In this review, we first give a brief introduction to the adhesion mechanism of mussels and the characteristics of injectable hydrogels. Further, the typical design strategies of mussel-inspired injectable adhesive hydrogels are summarized. The methodologies for integrating catechol groups into polymers and the crosslinking methods of mussel-inspired hydrogels are discussed in this section. In addition, we systematically overview recent mussel-inspired injectable adhesive hydrogels for biomedical applications, with a focus on how the unique properties of these hydrogels benefit their applications in these fields. The challenges and perspectives of mussel-inspired injectable hydrogels are discussed in the last section. This review may provide new inspiration for the design of novel bioinspired injectable hydrogels and facilitate their application in various biomedical fields. Full article

Blastocyst vitrification has significantly improved embryo transfer methods, leading to higher implantation success rates and better pregnancy outcomes in subsequent frozen embryo transfer cycles. This study aimed to simulate the transcriptional changes caused by vitrifying human blastocysts using mouse blastocysts as a model and to further investigate these changes’ effects. Utilizing a human vitrification protocol, we implanted both vitrified and fresh embryos into mice. We observed the implantation success rates and performed transcriptomic analysis on the blastocysts. To validate the results from messenger RNA sequencing, we conducted reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) to measure the expression levels of specific genes. Based on mRNA profiling, we predicted the microRNAs responsible for the regulation and used qPCR basic microRNA assays for validation. Our observations revealed a higher implantation success rate for vitrified embryos than fresh embryos. Transcriptomic analysis showed that vitrified–warmed blastocysts exhibited differentially expressed genes (DEGs) primarily associated with thermogenesis, chemical carcinogenesis-reactive oxygen species, oxidative phosphorylation, immune response, and MAPK-related signaling pathways. RT-qPCR confirmed increased expression of genes such as Cdk6 and Nfat2, and decreased expression of genes such as Dkk3 and Mapk10. Additionally, gene-microRNA interaction predictions and microRNA expression analysis identified twelve microRNAs with expression patterns consistent with the predicted results, suggesting potential roles in uterine epithelial cell adhesion, trophectoderm development, invasive capacity, and immune responses. Our findings suggest that vitrification induces transcriptomic changes in mouse blastocysts, and even small changes in gene expression can enhance implantation success. These results highlight the importance of understanding the molecular mechanisms underlying vitrification to optimize embryo transfer techniques and improve pregnancy outcomes. Full article

Listeria monocytogenes (L. monocytogenes) is a foodborne pathogen that causes listeriosis in humans and other animals. Surface proteins with the LPXTG motif have important roles in the virulence of L. monocytogenes. Lmo0159 is one such protein, but little is known about its role in L. monocytogenes virulence, motility, and biofilm formation. Here, we constructed and characterized a deletion mutant of lmo0159 (∆lmo0159). We analyzed not only the capacity of biofilm formation, motility, attachment, and intracellular growth in different cell types but also LD₅₀; bacterial load in mice’s liver, spleen, and brain; expression of virulence genes; and survival time of mice after challenge. The results showed that the cross-linking density of the biofilm of ∆lmo0159 strain was lower than that of WT by microscopic examination. The expression of biofilm-formation and virulence genes also decreased in the biofilm state. Subsequently, the growth and motility of ∆lmo0159 in the culture medium were enhanced. Conversely, the growth and motility of L. monocytogenes were attenuated by ∆lmo0159 at both the cellular and mouse levels. At the cellular level, ∆lmo0159 reduced plaque size; accelerated scratch healing; and attenuated the efficiency of adhesion, invasion, and intracellular proliferation in swine intestinal epithelial cells (SIEC), RAW264.7, mouse-brain microvascular endothelial cells (mBMEC), and human-brain microvascular endothelial cells (hCMEC/D3). The expression of virulence genes was also inhibited. At the mouse level, the LD₅₀ of the ∆lmo0159 strain was 10^0.97 times higher than that of the WT strain. The bacterial load of the ∆lmo0159 strain in the liver and spleen was lower than that of the WT strain. In a mouse model of intraperitoneal infection, the deletion of the lmo0159 gene significantly prolonged the survival time of the mice, suggesting that the lmo0159 deletion mutant also exhibited reduced virulence. Thus, our study identified lmo0159 as a novel virulence factor among L. monocytogenes LPXTG proteins. Full article

Cadherins are cell–cell adhesion proteins which have been strongly implicated in cancer invasion, dissemination and metastasis capacity; thus, they are key players in the epithelial-to-mesenchymal transition (EMT) program. However, their role in glioblastoma (GBM), a primary central nervous system aggressive tumor, remains to be clarified. N-, E- and P-cadherin expression was analyzed on a large series of GBMs, characterized with clinical, imaging and neuropathological parameters, as well as with patients’ survival data. In addition, cadherins’ expression was studied in match-recurrent cases. Using TCGA data, cadherin expression profiles were also evaluated according to GBM transcription subtypes. N-cadherin expression was observed in 81.5% of GBM, followed by E-cadherin in 31% and P-cadherin in 20.8%. Upon tumor recurrence, P-cadherin was the only significantly upregulated cadherin compared with the primary tumor, being positive in 65.8% of the cases. Actually, P-cadherin gain was observed in 51.4% of matched primary-recurrent cases. Cadherins’ co-expression was also explored. Interestingly, E- and N-cadherin co-expression identified a GBM subgroup with frequent epithelial differentiation and a significant survival benefit. On the other hand, subgroups with P-cadherin expression carried the worse prognosis. P- and N-cadherin co-expression correlated with the presence of a mesenchymal phenotype. Expressions of isolated P-cadherin or E- and P-cadherin co-expression were associated with imaging characteristics of aggressiveness, to highly heterogeneous tumors, an d to worse patient survival. Classical cadherins co-expression subgroups present consistent clinical, imaging, neuropathological and survival differences, which probably reflect different states of an EMT-like program in GBM. Full article