Synthetic biomaterials with osteoconductive or osteoinductive capabilities continue to evolve in functionality and expand their potential as viable substitutes for bone autografts. Recent developments with nanomaterial lattices have seen scaffold systems, especially brittle calcium phosphates, optimized for biological and mechanical performance. The addition of 2D atomic layers of carbon (graphene) to ceramic scaffolds is a well-studied avenue that potentially improves strength, functionality, and osteogenic potency; however, concerns of poor bioresorption and concentration-dependent toxicity have limited their translational value. The recent discovery of hematene, an ultrathin 2D nanosheet of iron oxide (Fe2O3) with its unique physiochemical properties and potential to be bioresorbable, could present a promising opportunity. In this work, exfoliated ultrathin hematene nanosheets were incorporated into porous monetite implants. In vitro testing with MC3T3-E1 preosteoblasts showed that hematene-loaded scaffolds supported cell infiltration and adhesion with a 24% increase in proliferative activity. The activity of the bioscaffold on cell proliferation and differentiation was evaluated. Findings showed that hematene-loading significantly improved the mechanical compression performance of monetite cements by 20%, without impacting degradation profiles. Furthermore, hematene loading significantly enhanced the osteogenic potency of monetite scaffolds with heightened bone biomarker expression levels of ALP, RUNX2, and SPARC. This preliminary report uncovers the therapeutic potential of hematene derivatives for the first time, particularly as a promising scaffold for bone repair.

Introduction The increase in e-cigarette use in the population has led to substantial interest in the health impacts associated with e-cigarette smoking. E-cigarette smoking represents a key external environmental cell stressor. Whilst there have been several studies to investigate the effect of nicotine-containing e-cigarette fluid, there is still a significant lack of understanding of how nicotine-free e-cigarette smoking can impact individuals. However, preliminary studies indicate that nicotine-free e-cigarette smoking can cause impaired endothelial function in humans. Materials and Methods In the present study, we therefore used a common brand of nicotine-free e-cigarette and human umbilical vein endothelial cells to assess angiogenic processes in vitro . Results We observed a significant upregulation in endothelial cell adhesion, migration and new tube formation with exposure to nicotine-free e-cigarette condensate (eVape) which was abrogated with exposure to the antioxidant, N-acetyl cysteine. Proteome analysis demonstrated that eVape exposure increased expression of the pro-angiogenic factors, angiogpoeitin-2, endoglin (CD105), PIGF and VEGF, as well as the ADP ribosylation factor, ARF6, and ARF6-GEF, ARNO. Chemical inhibition of ARNO reduced eVape-induced oxidative stress, angiogenic processes, and release of angiogpoeitin-2, endoglin (CD105) and VEGF. Discussion These findings demonstrate that nicotine-free eVape causes aberrant upregulated angiogenesis in an in vitro model of the human endothelium through ARNO-dependent signalling. This study is the first to demonstrate the molecular mechanisms in response to the cellular stressor, nicotine-free eVape which underlie impaired vascular function.

Tissue-resident macrophages (RTMs) form during embryogenesis, self-renew locally, and regulate tissue homeostasis by clearing dead cells and debris1-6. During tissue damage, however, bone-marrow-derived monocytes enter tissues and differentiate into RTMs, repairing the tissue and replenishing macrophages in the niche1. The universal cell-intrinsic mechanisms that control the monocyte-to-RTM transition and the maintenance of mature RTMs across tissues remain elusive3. Here we show that deoxyhypusine synthase (DHPS), an enzyme that mediates spermidine-dependent hypusine modification of translation factor eIF5A5,7, is required for RTM differentiation and maintenance. Mice with myeloid cell lack of DHPS (Dhps-ΔM mice) had a global defect in RTMs across tissues, resulting in persistent but ultimately futile monocyte influx. Transcriptional analyses of DHPS-deficient macrophages indicated a block in their ability to differentiate into mature RTMs, whereas proteomics revealed defects in cell adhesion and signalling pathways. Sequencing of ribosome-engaged transcripts identified a subset of mRNAs involved in cell adhesion and signalling that rely on DHPS for efficient translation. Imaging of DHPS-deficient macrophages in tissues showed differences in morphology and tissue interactions, which were correlated with their failed RTM differentiation. DHPS-deficient macrophages were also defective in critical homeostatic RTM functions including efferocytosis and tissue maintenance. Together, our results demonstrate a cell-intrinsic, tissue-agnostic pathway that drives differentiation of monocyte-derived macrophages into RTMs.

Feather follicles are specialized skin appendages that are essential for thermoregulation, protection, and down production in birds, forming through complex genetic and epigenetic interactions during embryogenesis. In this study, we examined skin and follicle development in Hungarian white goose embryos, focusing on dynamic epigenetic-transcriptomic changes. Histology showed smooth epidermis at E10, feather buds at E13, and columnar follicles with medullary tissue and secondary follicles at E18. Transcriptomics revealed 1327 and 1847 DEGs enriched in epidermal development, differentiation, and adhesion. Primordial initiation at E10-E13 featured Wnt, TGF-β, and melanogenesis, whereas follicle formation at E13-E18 involved lipid metabolism and VEGF signaling. Keratinization genes were continuously upregulated, and Wnt, Shh, and muscle pathways were activated late. Key regulators included LEF1, MSX2, and FOXN1. ATAC-seq showed dynamic chromatin accessibility, stage-specific promoter openness, and motifs for YY1, KLF5, and KLF4. Differentially accessible regions enriched genes in Wnt and TGF-β signaling, cell adhesion, and mitophagy, shifting from proliferation and basic metabolism at E10-E13 to differentiation, lipid metabolism, and homeostasis at E13-E18. Integrated analyses linked fatty acid metabolism, MAPK, and FoxO signaling to differentiation, while downregulation of redox and migration pathways preserved homeostasis. Some fatty acid metabolism and cell polarity genes increased expression despite reduced accessibility at E13-E18, indicating epigenetic pre-programming and post-transcriptional interplay. This work delineates coordinated epigenetic-transcriptional regulation of goose embryonic skin and feather follicle morphogenesis, offering insights for avian and vertebrate skin appendage studies.

Mural cells, such as pericytes, integrate with endothelial cells (ECs) lining the capillaries, which are pivotal in vascular development and stabilization as well as in supporting EC function. Dental pulp stem cells (DPSCs) were recently revealed to have intimacy with pericytes in the dental pulp microenvironment and are regulated by EphB4/ephrinB2 signaling. However, how EphB4/ephrinB2 signaling regulates DPSC pericyte biology and their interactions with ECs remains unknown. In this study, single-cell RNA sequencing data analysis and immunofluorescence staining of healthy human dental pulp were used to demonstrate the roles of mesenchymal stem cells (MSCs) as pericytes and ephrinB phosphorylation between MSCs and ECs interaction. Bulk RNA-seq further showed transcriptomic similarity between DPSCs and human brain vascular pericytes. In vitro coculture of DPSCs and ECs further confirmed ephrinB2 activation at cell-cell contact sites. To investigate how ephrinB2 influenced DPSCs' pericyte function, EFNB2 was either knocked down using small hairpin RNA or overexpressed via open reading frame (ORF) lentiviral transduction. Focal adhesion protein assessment was conducted, and a 3-dimensional (3D) fibrin beads assay was established to visualize the interaction between DPSCs and ECs in vitro. EFNB2 knockdown in DPSCs significantly reduced cell proliferation, adhesion, and transwell migration but increased contractility. Conversely, EFNB2 overexpression enhanced proliferation and adhesion but reduced migration and contractility. Interestingly, EFNB2 overexpression significantly increased ECs' sprouting capability, improving pericyte coverage in 3D fibrin beads assays. These effects were mediated through the enhanced focal adhesion pathway involving Src, paxillin and FAK phosphorylation, and vinculin. Our findings demonstrate that ephrinB2 signaling regulates critical pericyte functions of DPSCs through the Src/FAK/paxillin signaling pathway, thus modulating their adhesion toward ECs. Targeting ephrinB2 signaling may therefore represent a promising strategy to enhance vascular formation and functional recovery in dental pulp tissue engineering.

This study investigates the role of extracellular vesicles (EVs) in predicting melanoma patients' responses to anti-PD1 immunotherapy. Nine patients with advanced melanoma provided blood samples at three stages: before treatment, before the second dose, and either at disease progression or nine months later. EVs were isolated from serum and analyzed using mass-spectrometry proteomics, followed by network and enrichment analyses. Six out of nine patients progressed despite treatment. Before therapy, responders exhibited higher levels of adaptive immune and cell adhesion proteins, while proteins related to UV radiation response were deplected. An eight-protein signature and cellular adhesion markers correlated with longer progression-free survival. After treatment, non-responders had EV proteins enriched in proteasome activity and metabolic pathways, especially glycolysis. Finally, dynamic changes in EV protein over time showed decreased coagulation proteins, along with an increase in MHC proteins in patients with progressive disease. Overall, EV protein profiles differed between responders and non-responders both before and during therapy. These findings suggest that EVs could provide predictive biomarkers and insights into resistance mechanisms, potentially guiding more effective melanoma treatment strategies.

Breast cancer is one of the most common malignant tumors among women worldwide, and its high heterogeneity poses significant challenges for early diagnosis and precise treatment. Despite the continuous development of molecular typing and targeted therapy, how to precisely distinguish the benign and malignant regions of tumors at the spatial level and analyze their molecular characteristics remains an urgent problem to be solved. This study is based on spatial transcriptomics technology on the 10x Genomics Visium platform. Systematic transcriptional analyses were conducted on human breast cancer tissue sections. Through differential expression analysis of DCIS/LCIS and IDC regions, a total of 1,143 significantly differentially expressed genes were identified, among which genes such as S100A6, STC2, and TFF3 were significantly upregulated in breast cancer progression, suggesting a key role in malignant transformation of tumors. Gene ontology enrichment analysis revealed that the differentially expressed genes were significantly aggregated in key functional modules such as metabolic pathways, MAPK signaling pathways, cell cycle and cancer-related pathways. in particular, the significant enrichment of the Rap1 signaling pathway and Pathways in cancer revealed the synergistic activation mechanism of multi-level biological processes such as cell adhesion, migration, and matrix degradation during the transformation of breast cancer from benign to malignant. In summary, this study revealed the molecular heterogeneity in the benign and malignant regions of breast cancer and the spatial distribution characteristics of key signaling pathways at the tissue level through spatial transcriptomics, providing new theoretical basis and spatial omics evidence for spatial molecular typing of breast cancer, targeted drug development, and optimization of individualized treatment strategies.

Precision medicine aims to develop 3D tumor models to validate new therapies and investigate disease mechanisms by isolating the extracellular matrix as a foundation for recreating tumors in vitro . The use of decellularized tumor matrices offers a promising and versatile platform for in vitro cancer research and therapeutic testing. The tumor microenvironment (TME) is the surrounding milieu of cancerous tissues and contains an intricate network of extracellular matrix (ECM) components and signaling proteins that regulate tumorigenesis, invasion, and metastasis. However, the decellularization techniques process can be disruptive and often damage essential macromolecules and proteins, potentially compromising the restoration of a biologically relevant microenvironment during recellularization. This review explores the most relevant macromolecules and proteins within the TME, emphasizing their roles in tumors and metastasis. Here, the potential of reinstating these components into decellularized tumor scaffolds to enhance their biological relevance and functionality is highlighted. Key macromolecules, including collagen, fibronectin, hyaluronic acid (HA), and laminin, are discussed alongside the contributions of proteins such as integrins, matrix metalloproteinases (MMPs), and growth factors to ECM remodeling, cell adhesion, migration, and proliferation. The strategic reintroduction of these elements will improve the recellularization process and create more realistic TME models. These improved models hold promise for cancer research, medication discovery, and therapeutic testing, providing a deeper understanding of tumor biology and enabling the development of more effective treatment strategies.

Liver tissue function relies on cells' spatial organization and interactions within a 3D microenvironment. While previously reported liver-on-a-chip models effectively provide basic structural organization and multilayered arrangements using artificial barriers, they fall short in replicating the higher-order organization of diverse cell types. Herein, we fabricated a membrane-free liver-on-a-chip (MF-LOC) usingCGRGDS peptide-modified PEGDA hydrogel that not only provides a native-like 3D microenvironment for encapsulated HepG2 (Hepatocytes) cells and NIH-3T3 (fibroblasts) but also offers a platform for HUVEC monolayer formation. The immobilization of the CGRGDS peptide on the hydrogel surface, which operates at a nanoscale level to enhance cell adhesion and signaling via integrin binding, strengthens HUVEC adhesion and prevents cell detachment caused by the shear stress of direct tangential flow. In MF-LOC, nutrients easily diffuse through the HUVEC monolayer and hydrogel pores to sustain cell functions. Live/Dead imaging and cell tracking showed HepG2 clusters associated with neighboring NIH-3T3 fibroblasts, with HUVECs forming a surface monolayer which replicate native liver structure. Functional validation confirmed prolonged albumin and urea secretion, with MF-LOC exhibiting sustained CYP1A1 enzyme activity compared to gold standard microsomes, highlighting its ability to replicate liver-like metabolism. MF-LOC has the potential for predicting drug-induced liver injury (DILI) and may provide a powerful platform for disease modeling.

Large-scale proteomics provides an opportunity to understand chronic kidney disease (CKD) and cardiovascular disease, yet research in this field is limited. This study utilized proteomics to inform biology and risk stratification for these diseases. This cohort study included 44,779 participants free of prevalent CKD, and 3,749-4,272 participants with prevalent CKD from the UK Biobank. The Olink Explore 3072 platform quantified 2,923 plasma proteins. Cox proportional hazards models were used to assess associations of proteins with kidney diseases including CKD and end stage kidney disease, and cardiovascular diseases including coronary heart disease (CHD), stroke, and heart failure (HF). Mendelian randomization examined genetic associations, pathway analyses identified biological pathways, and predictive models were developed for incident diseases. Median follow-up periods were 12.2-12.6years. We identified 598 (20.5%) proteins shared across ≥ 2 diseases, with 595 (20.4%) showing consistent directions of associations, and 471 (16.1%) unique to a single disease. CKD and HF specifically shared the largest number of 279 (9.6%) proteins. POLR2F, TNFRSF10B, and IGFBP2 were positively associated with all five diseases, with Mendelian randomization supporting genetic associations of POLR2F with CHD and IGFBP2 with hypertensive renal disease. Pathway analyses highlighted cell adhesion, signal transduction, and cytokine-cytokine receptor interaction for disease-associated proteins. Incorporating predictive proteins into clinical models improved risk prediction for CKD, CHD, stroke, and HF, yielding Harrell's C indices of 0.750-0.818 (corresponding increases of 0.027-0.090). This study deepens insights into disease biology and provides a foundation for early detection and integrated risk stratification in CKD and cardiovascular disease.

Titanium (Ti) and its alloys are widely used in biomedical applications due to their biocompatibility and corrosion resistance; however, surface modifications are required to enhance biological functionality. Surface functionalization using natural biomolecules has emerged as a promising strategy to improve early cell–surface interactions and biocompatibility of implant materials. In this study, Ti6Al4V alloy surfaces were biofunctionalized using Spirulina platensis (S. platensis) biomass and protein extract to evaluate morphological, chemical, and biological effects. The functionalization process involved activation with piranha solution, silanization with 3-aminopropyltriethoxysilane (APTES), and subsequent biomolecule immobilization. Surface characterization by scanning electron microscopy (SEM), inductively coupled plasma mass spectrometry (ICP-MS), energy-dispersive X-ray spectroscopy (EDS), and Fourier transform infrared spectroscopy (FTIR) confirmed the successful incorporation of microalgal components, including nitrogen-, phosphorus-, and oxygen-rich organic groups. Biomass-functionalized surfaces exhibited higher phosphorus and oxygen content, while protein-coated surfaces showed nitrogen-enrich chemical signatures, reflecting the distinct molecular compositions of the immobilized biomolecules. Cell adhesion assays demonstrated enhanced early cell attachment on biofunctionalized surfaces, particularly in samples functionalized with 5 g/L biomass for three hours, which showed significantly greater cell attachment than both the control and protein-treated samples. These findings highlight the complementary yet distinct roles of S. platensis biomass and protein extract in modulating surface chemistry and cell–material interactions, emphasizing the importance of tailoring biofunctionalization strategies to optimize early biological responses on titanium-based implants.

This study presents the development of titanium-based implants coated with zeolite layers for controlled delivery of epigallocatechin gallate (EGCG), a polyphenolic compound with osteogenic, antiresorptive, and antibacterial properties. Zeolite coatings were modified with divalent ions (Zn2+, Mg2+, Ca2+) to investigate their influence on EGCG adsorption and release under neutral (pH 7.4, SBF) and acidic (pH 5.0, acetate buffer) conditions. Comprehensive characterization using SEM, EDS, FT-IR, UV-vis spectroscopy, and surface profilometry confirmed uniform zeolite formation, effective EGCG loading, and tunable release profiles. Zinc-containing zeolite exhibited the highest EGCG adsorption but demonstrated cytotoxicity toward hFOB 1.19 osteoblasts. Magnesium-zeolite-coated implants provided controlled EGCG release, were nontoxic, and did not support cell adhesion, making them suitable for temporary internal fixation in the management of orthopedic trauma. Release studies revealed pH-dependent kinetics, with accelerated EGCG release under acidic conditions simulating osteoclast activity. These findings demonstrate the potential of Mg-zeolite-coated titanium implants as functional devices that provide mechanical support, enable localized drug delivery, and promote bone regeneration while minimizing tissue damage during removal.

Core fucosylation of N-glycoproteins plays pivotal roles in regulating many cellular events such as receptor-ligand binding and cell adhesion. Here, we developed a chemoenzymatic method combining selective enrichment, enzymatic reactions, and multiplexed proteomics to systematically quantify the core fucosylation stoichiometries of glycoproteins in human cells. The results demonstrated that the core fucosylation stoichiometries vary dramatically in different subcellular compartments with the lowest in the lysosome and the highest in the extracellular matrix. Different core fucosylation stoichiometries were observed among glycosylation sites in various protein domains, and more aromatic and hydrophobic residues neighboring glycosylation sites are associated with lower core fucosylation stoichiometry. The method was applied to quantify the core fucosylation stoichiometry changes in the epithelial-to-mesenchymal transition (EMT), and some glycoproteins involved in extracellular matrix organization and ligand recognition displayed marked stoichiometry changes. Furthermore, the core fucosylation stoichiometries in embryonic human kidney cells (HEK293T) were compared with those in kidney cancer cells (A498). The average stoichiometry in HEK293T cells was much higher than that of A498 cells, indicating that core fucosylation may be a critical regulator in embryonic development. Without any sample restriction, this method can be extensively applied to investigate core fucosylation changes in various biological samples.

Nanocoatings for the biomineralization membrane were developed on a gelatin (Gel) and carbonated hydroxyapatite (CHA) composite, namely, Gel/CHA membrane. The membrane, which contains bioactive polymer and a mineral phase, was further modified using plasma to deposit thin functional polymeric films. These surface-tailored nanocoatings improved the membrane's performance by increasing its swelling capacity to better mimic a tissue-like environment, strengthening its mechanical stability during application, and modulating interactions with biological components such as proteins to support tissue engineering and regeneration. Four types of thin films were explored, namely, 2-methyl-2-oxazoline (OX), 1,7-octadiene, acrylic acid (AC), and allylamine (AA). The physicochemical, mechanical, and biological properties of these plasma-functionalized membranes were systematically investigated. Fourier-transform infrared spectroscopy, energy-dispersive X-ray spectroscopy (EDX), and X-ray photoelectron spectroscopy (XPS) confirmed the successful deposition of functional coatings, while scanning electron microscopy (SEM) and atomic force microscopy demonstrated variations in the surface morphology and roughness. Water contact angle and swelling ratio measurements revealed tunable hydrophilicity and water uptake properties, with AC- and OX-coated membranes exhibiting enhanced hydration. Mechanical testing in dry state indicated that the OX-coated membranes achieved an optimal balance between tensile strength and elongation, thereby enhancing their mechanical resilience. Protein adsorption studies highlighted increased biomolecular interactions with hydrophobic surfaces demonstrating higher protein affinity. Cytocompatibility assessments, following ISO 10993-5:2009 standards, confirmed that all plasma-coated membranes were noncytotoxic to osteoblast MC-3T3-E1 cell lines, with cell viability exceeding 80%. The in vitro evaluation of MC-3T3-E1 cell adhesion was confirmed using DAPI staining and SEM observation and revealed that the MC3T3-E1 cells attached on control, AC, and AA membranes with a flatter shape. Additionally, the mineralization potential was assessed via simulated body fluid immersion for up to 360 min. EDX analysis showed time-dependent deposition of calcium and phosphate, with AA-coated membranes exhibiting the highest mineral accumulation. These findings demonstrate that plasma-assisted functionalization can significantly improve the membrane properties, making it a promising candidate for advanced bone tissue engineering applications.

BackgroundCardiovascular diseases are the leading cause of mortality worldwide, with coronary artery bypass grafting being the most effective treatment for severe cases. While autografts are preferred, donor veins are often limited. Human umbilical arteries (hUAs) show promise as an alternative. However, to make vascular graft by decellularization, a traditional chemical method can damage tissue structure and function.ObjectiveThis study aims to evaluate the shortening of treatment time and the hUA decellularization efficiency of the perfusion bioreactor systems.MethodshUAs were perfused with 1% Triton X-100 for 6 h, followed by two different concentrations of (0.5% and 1%) SDS for 18 h, and subsequently subjected to a washing procedure. The decellularization process was evaluated using histological staining and DNA quantification, along with tests for cytotoxicity, cell adhesion and proliferation.ResultsThe 0.5% SDS protocol proved most effective, reducing residual DNA to below 50 ng/mg of dry weight while preserving collagen structure. It showed no cytotoxicity to L929 cells, SEM analysis confirmed human umbilical vein endothelial cell (HUVEC) attachment and CCK-8 testing showed promoted HUVECs proliferation.ConclusionThe decellularization protocol of perfusing through 1% TX for 6 h and 0.5% SDS for 18 h through the perfusion bioreactor system is efficient in the intact hUAs tissue. This sets the stage for future in vivo studies and potential clinical applications.

Cell-cell adhesion is crucial for maintaining cell functions and the integrity of tissue structure in organisms. However, cell-cell adhesion cues have not been effectively replicated in biomaterials and the associated mechanisms that enhance neural regeneration remained largely unexplored. Here, we present a diffusive N-cadherin functionalized hydrogel system, which provided cell-cell adhesion cues to modulate intercellular communications to significantly promote the formation of active neural network via thrombospondin-1 mediated neural communication and activation of TGF-β/Smad pathway. The dynamic assembly of N-cadherin at cell-hydrogel interface driven by adhered neurons effectively facilitated the reshaping of membrane protrusions to initiate intercellular adherens junctions. Further, this hydrogel system promisingly promoted neurological function recovery in rats following traumatic brain injury. Our study provides the principle of replicating diffusive cell adhesion molecules to mediate cell-cell adhesion in hydrogels, which may have broad applications in developing engineered biomaterials aimed at modulating cell fates in regeneration of various tissues.

PurposeAniridia-associated keratopathy (AAK) leads to loss of corneal transparency because of epithelial, inflammatory, and pathological vascular changes. Here, we sought to understand this process at the transcriptomic level while evaluating an experimental pharmacotherapy for potential modulatory effects.Method17 Pax6+/− Small-eye (Sey) heterozygous mice with p.Gly208* Pax6 mutation and 10 wild-type 129S1/SvImJ mice at four months of age were examined to identify dysregulated genes and pathways in established AAK. We next evaluated the potential efficacy of 10 µM duloxetine administered as eye drops twice daily for 90 days, assessing outcomes at the transcriptomic level via microarray and protein level with Western blot and immunostaining.ResultsTranscriptomic analysis of the cornea revealed enrichment of Ccl21 gene family members associated with lymphangiogenesis, along with upregulation of genes involved in inflammation, cell adhesion, differentiation, motility, and keratinization, and downregulation of drug metabolism with significantly dysregulated genes emerging as potential therapeutic targets, including Gpha2, Chrnb3, Epgn, Cnfn, kallikreins and inflammation mediators Il18r1 and classical complement factors. Duloxetine therapy failed to regress AAK in adult corneas; however, transcriptomic profiling indicated duloxetine suppressed inflammatory genes and promoted anti-inflammatory and protective activity while modulating drug metabolism, suggesting potential beneficial effects in the cornea.ConclusionsTranscriptomics reveals multiple unexplored pathways and genes altered in the AAK mouse model. Although clinical results with duloxetine are promising, our current regimen and delivery method did not improve established disease. Duloxetine's therapeutic potential requires further study.

DNA-PAINT resolves E-cadherin-independent cross-junctional F-actin organization in Drosophila embryonic tissue.

Cell adhesion tightly controls cell proliferation and homeostasis in stratified epithelia by mechanisms that remain unclear. Focal adhesion kinase (FAK) transduces cell adhesion signals, is frequently deregulated in epithelial cancer, and it has been associated with proliferation and resistance to treatments. The mechanisms by which FAK controls the epithelial cell cycle are still intriguing. We previously unraveled a mitosis-differentiation checkpoint that is the limiting factor in the keratinocyte cell cycle. To investigate whether FAK plays a role in this checkpoint, we inactivated the protein in normal human oral keratinocytes by specific shRNAs or by the specific inhibitor defactinib. Inactivation of FAK very rapidly and strikingly blocked entry into mitosis and triggered a differentiation response. This response was independent of DNA damage. Tumor suppressor P53 was induced shortly after inhibition of FAK, while mitotic Cyclin B was not translocated into the nucleus. Human epidermal N-TERT cells that were synchronized in prometaphase failed to execute mitosis. Concomitant inhibition of FAK-downstream Rho-associated kinase (Rock) rescued mitotic progression. The results unveil a rapid Rock-dependent mitosis switch upon inactivation of FAK, inducing terminal differentiation, pointing at a mitotic automatic mechanism of epithelia to suppress suprabasal proliferation of precancerous cells.

Purpose This study aims to fabricate a polylactic acid (PLA)/polypropylene carbonate (PPC) composite tracheal scaffold using embedded 3D printing and demonstrate its triple-shape memory functionality. Design/methodology/approach Three tracheal scaffolds of different geometry (cylinder, bellow and spiral-shaped) were investigated using mechanical (radial and longitudinal compression tests) and compared with that of the native goat trachea. As a case study, the bellow-shaped scaffold was implanted inside the lumen of the native goat trachea and the triple-shape memory behavior was demonstrated. To further assess the biocompatibility of the samples, MTT, live/dead and cell adhesion assays were conducted using fibroblast cells. Findings The thermal characterization revealed that the effect of Laponite nano-silicate on the PLA/PPC composite surface had negligible effect on the thermal transitions and shape-memory ratios. Comparative studies between the three scaffolds highlighted that the bellow-shaped scaffold was found to closely mimic the mechanical properties (radial and longitudinal compression) of the native goat trachea. The PLA/PPC composite samples fabricated using embedded 3D printing were found to have a shape-fixity ratio of 96.07 ± 3.06% and a shape-recovery ratio of 95.88 ± 2.61%. The in vitro cytotoxicity of the PLA/PPC composite demonstrated excellent biocompatibility. Originality/value The proposed methodology of embedded 3D printing allows for fabrication of complex geometry structures using a wide variety of polymers by leveraging the solvent–water interaction capability.