Search Results (443)

Background: Obesity is associated with aggressive prostate cancer, but the links between metabolic dysregulation, inflammation, adipocyte-associated signaling, and tumor growth remain incompletely defined. This study examined whether high-fat diet (HFD)-associated systemic changes and adipocyte-derived paracrine interactions are linked to prostate cancer growth in preclinical models. Methods: An HFD xenograft model and adipocyte co-culture systems were used to evaluate systemic and local tumor-supportive features. Pharmacologic/nutritional modulation was tested using green tea or EGCG, arctigenin, and the CCR2 antagonist RS 504393, alone or in combination. Tumor growth, cell proliferation, angiogenesis-related features, circulating metabolic and cytokine levels, and selected tumor-associated signaling proteins were analyzed. Results: HFD feeding was associated with increased circulating free fatty acids, IGF-1, MCP-1, IL-6, and VEGF, together with increased tumor growth, Ki67 staining, and CD31-positive microvessel density. Adipocyte co-culture systems were used to evaluate treatment-associated changes in prostate cancer cell proliferation under adipocyte-associated conditions. Combined modulation with green tea/EGCG, arctigenin, and RS 504393 was associated with greater reductions in adipocyte-associated proliferation, tumor growth, Ki67 staining, and CD31-positive microvessel density than single or dual interventions. Antibody array analysis showed treatment-associated changes in selected stress- and apoptosis-related proteins, including cleaved caspase-7 and phosphorylated Chk1. Conclusions: HFD-associated metabolic and inflammatory alterations, adipocyte-associated interactions, proliferative activity, angiogenesis-related features, and stress/apoptosis-related signaling changes were linked within a tumor-supportive framework in preclinical prostate cancer models. Combined pharmacologic/nutritional modulation was associated with reduced tumor-supportive features under HFD conditions. Further mechanistic and translational validation is needed. Full article

Cerebral amyloid angiopathy (CAA) is a prevalent cerebral small vessel disease (CSVD) and prominent cause of vascular contributions to cognitive impairments and dementia (VCID). CAA is characterized by progressive accumulation of fibrillar amyloid β in cerebral vessel walls, leading to vascular wall degeneration, thrombotic occlusions, microbleeds, and perivascular inflammation causing cognitive deficits. Underlying mechanisms of CAA progression are poorly understood, and there exist no validated diagnostic biomarkers or therapeutic targets for CAA. Here, we performed proteomic mass spectrometry analysis of whole brain tissue and cerebral microvessel enriched fractions from the rTg-DI rat, a validated and well-characterized preclinical rat model of CAA, to identify potential targets and diagnostic markers related to vasculopathy progression in CAA. There were 92 increased and 104 decreased proteins identified in the rTg-DI whole brain samples, while 29 proteins were identified as increased in the enriched cerebral microvessel fractions. We identified several significant differentially expressed proteins in both sample types with commonly elevated proteins including HTRA1, APOE, APOD, CLU, MDK, and LAP3. This study also confirmed the differential expression of several proteins previously reported in isolated brain regions of rTg-DI rats, including ANXA3, APOD, APOE, CLU, CST3, CTSS, CTSD, CTSZ, GFAP, GSTA3, HTRA1, and ITGB2, with APOE, CLU, HTRA1, and GPC1 all reported in human CAA cases. We thus provide a “high-confidence” list of potential marker candidates from proteomic analysis in the rTg-DI rat model of capillary CAA type-1. Full article

The myocardium possesses one of the highest vascular densities in the body. The outermost wall layer of large and medium-sized vessels, the adventitia, forms a critical interface between the vasculature and the myocardium and serves as a reservoir for stem and progenitor cells capable of differentiating into all vascular wall lineages as well as innate immune cells, including macrophages. Current cardiac organoid models intrinsically develop networks of endothelial cords and small capillary-like structures that resemble cardiac microvessels. However, these microvessels mostly lack an adventitial compartment in vivo. Here, we present a potential alternative assembloid strategy that combines vascular segments from mouse and human origin with either cardiomyocytes or cardiac spheroids derived from human induced pluripotent stem cells, thereby incorporating large diameter vessels and the vascular adventitia into a cardiac tissue model. Within the assembloids, the myocardial component remained contractile and connected to the vascular adventitia, which displayed cellular sprouting toward the hiPSC-derived cardiac tissue. Immunostaining for vascular and immune markers revealed that the adventitia gave rise to endothelial sprouts and macrophage-like cells which integrated into the myocardial tissue. In summary, we present proof of concept for complex assembloids composed of vessel segments and human iPSC-derived cardiomyocytes which contain and maintain an in vivo-like adventitial compartment. We suggest this model may serve as a platform for investigating myocardial–stromal interactions, cardiac tissue repair, and functional remodeling under both physiological and pathological conditions. Furthermore, the incorporation of large-lumen vessel segments may enable future experimental perfusion, rendering the model particularly suitable for drug testing via intravascular delivery. Full article

Objectives: To investigate the clinicopathologic characteristics and molecular biomarkers of atypical vasoproliferative retinal tumor (VPRT) with hemangioblastoma-like histopathologic features and concomitant von Willebrand factor (VWF) abnormalities. Methods: A 48-year-old woman undergoing phacoemulsification and 25-gauge pars plana vitrectomy with tumor resection was evaluated. Histopathological findings and immunohistochemical study of the resected tumor were performed using CD34, α-smooth muscle actin (αSMA), and glial fibrillary acidic protein (GFAP) markers. Preoperative plasma and intraoperative vitreous fluid VWF antigen levels, as well as ristocetin cofactor activity, were quantified using latex immunoturbidimetry. Results: Ultra-widefield imaging and angiography demonstrated a peripheral retinal tumor with intense vascular leakage and surrounding capillary nonperfusion. Histopathology showed hyalinized vascular components supportive of VPRT, along with abundant CD34/α-SMA-positive microvessels and scant GFAP-positive glial cells. Notably, numerous foamy vacuolated poorly differentiated cells suggested mixed hemangioblastoma-like features. Preoperative plasma VWF antigen (182.6%) and ristocetin cofactor activity (147.7%) were elevated, and vitreous VWF antigen was successfully detected at a low but distinct level (7.7%).and suggests that VWF abnormalities in the plasma and vitreous may reflect endothelial activation and/or blood–retinal barrier disruption in a subset of vascularized retinal tumors. Conclusions: Our findings demonstrate that VPRT may exhibit mixed clinicopathologic features, including hemangioblastoma-like components, which underscores the necessity of immunohistochemical assessment for definitive diagnosis. Furthermore, the quantification of VWF abnormalities in the plasma and vitreous suggests that VWF serves as a potential biomarker reflecting endothelial activation and/or blood–retinal barrier disruption in vascularized retinal tumors. Full article

Intraoral soft-tissue defects are traditionally managed with autogenous connective tissue grafts, though donor site morbidity has driven interest in xenogeneic collagen matrices as alternatives. However, the impact of matrix architecture on soft-tissue integration remains poorly understood. This study compared soft-tissue responses to a sheet-form collagen matrix (ShCM) and a spongy collagen matrix (SpCM) placed beneath full-thickness flaps in a beagle mandibular defect model. Standardized bilateral defects were created in 23 skeletally mature female beagles. Defects were then assigned to serve as the negative control (sham) or were treated with porcine collagen matrix in (i) sheet form (ShCM) (Regenity Biosciences, Oakland, NJ, USA), or (ii) porous/spongy form (SpCM) (Fibro-Gide^®, Geistlich Pharma North America, West Windsor Township, NJ, USA). The mandibular sites that received no surgical intervention served as positive controls. Experimental conditions were randomized and interpolated within each animal to minimize anatomical site bias and evaluated histologically at 4- (n = 7), 8- (n = 7), and 12-weeks (n = 9) postoperatively. Histologic sections were evaluated for matrix presence, inflammation, subepithelial healing, and matrix thickness. At 4 weeks, both matrices were present, though SpCM showed significantly higher inflammation scores (p = 0.013). By 8 weeks, ShCM demonstrated greater resorption (p = 0.003) alongside an organized collagen layer with fibroblasts and new microvessels, while SpCM remained thick and porous, with a persistent fibrous capsule and elevated inflammation versus both ShCM (p = 0.002) and sham (p = 0.009). At 12 weeks, inflammation declined and subepithelial healing improved similarly across matrix groups. These findings suggest matrix architecture influences soft-tissue healing outcomes in the oral cavity. Sheet-form matrices may be preferable where biocompatibility and predictable integration are priorities, while spongy matrices may better support long-term space maintenance and tissue ingrowth. However, clinical studies are needed to confirm these translational implications. Full article

The invasive tumor front (ITF) of colorectal carcinoma (CRC) is a biologically active interface where epithelial dissociation, stromal activation, immune remodeling, and angiogenesis converge. This retrospective histopathological association study examined whether tumor budding combined with selected ITF microenvironmental markers could better characterize aggressive clinicopathological features of non-metastatic CRC. Fifty-two surgically resected primary CRCs were analyzed after exclusion of patients with distant metastatic disease at diagnosis or surgery. Tumor budding was scored according to ITBCC 2016 criteria. Immunohistochemistry evaluated stromal podoplanin (D2-40) expression, CD34-based microvessel density, the CD163/CD68 ratio, and CD117-positive mast cell density. An unweighted integrated ITF microenvironment score (0–4) was generated by summing high values for these four parameters. High histological grade, lymphovascular invasion, high tumor budding, and node-positive disease were significantly associated with multiple microenvironment-related alterations. The integrated ITF score was significantly higher in high-grade, lymphovascular invasion-positive, node-positive, and high-budding CRCs, suggesting cumulative stromal, immune, and vascular remodeling at the invasive front. These findings support the ITF as an integrated histopathological compartment in CRC progression. However, the score remains hypothesis-generating and requires validation in larger, independent, molecularly and outcome-annotated cohorts before prognostic or clinical use. Full article

COVID-19 remains a challenge to the global healthcare despite the end of the pandemic, including due to the significant involvement of children in the epidemic process. During the pandemic period, an increase in the incidence of Sudden Unexpected Infant Death (SUID) and Sudden Infant Death Syndrome (SIDS) was observed. Currently, their rates remain elevated compared to the prepandemic period. The pathogenetic mechanisms underlying the fulminant course of infection in infants leading to fatal outcomes remain insufficiently understood. In this study, we report for the first time the results of histological and immunohistochemical examination of the lungs in a case of COVID-19-associated SUID in a 2-month-old infant. The absence of similar studies in the available literature limits opportunities for analyzing the pathogenesis of SUID. Our data allow a detailed characterization of the histological changes in the lungs, the localization and range of SARS-CoV-2 nucleocapsid protein expression, the identification of molecular mechanisms underlying apoptosis in the pulmonary microvascular endothelium, and the elucidation of the role of endothelial dysfunction. Particular attention in this article is devoted to the role of cytokines (IL-6, TNF-α, and IFN-γ) in the pathogenesis of hyperacute viral infection. The obtained data demonstrate substantial differences between the observed changes and the classic presentation of COVID-19 in older children. These findings offer prospects for improving prevention strategies and developing targeted therapy for fulminant forms of COVID-19, while also contributing to the understanding of SIDS pathogenesis. Full article

Natural bioactive compounds present promising avenues for the prevention and therapeutic intervention of cancer. Octacosanol has garnered significant attention for its distinctive biological properties, yet its specific antitumor effects and underlying mechanisms remain unclear. This study systematically evaluated its antitumor effects and elucidated the associated molecular mechanisms. We confirmed that it dose-dependently inhibited A549 cell proliferation in vitro. It also remarkably suppressed cell invasion and migration by downregulating MMP2 and MMP9 expression, an effect that was associated with reduced phosphorylation of JAK3/STAT3 and PI3K/AKT, suggesting a potential regulatory role of these signalling cascades. Meanwhile, it significantly inhibited tumor cell VEGF secretion and VEGF-mediated neoangiogenesis by modulating the PI3K/AKT signaling axis. Mouse experiments demonstrated that octacosanol significantly reduced tumor p-AKT, MMP2, and MMP9 levels, indicating its in vivo anti-metastatic effect. It also remarkably decreased tumor microvessel density, alongside reduced VEGF and vascular endothelial marker CD31 expression, further verifying its potent anti-angiogenic activity. This work provides evidence of octacosanol’s dual anti-metastatic and anti-angiogenic effects in lung cancer and offers novel mechanistic insights into its activity against this highly prevalent malignancy. These findings establish a solid foundation for further exploration and development of octacosanol as a promising adjuvant for clinical antitumor therapy. Full article

Surgical sutures are widely used biomaterials in clinical practice. Like all other biomaterials, they induce a foreign body response after implantation that involves inflammation and angiogenesis. Although it is well known that these processes differ in males and females, sex-specific differences in the tissue response to sutures have not been investigated so far. To do this in the present study, polypropylene sutures were implanted into the dorsal skinfold chamber and subcutaneous flank tissue of male and female mice to assess their acute and chronic effects on the local tissue microenvironment using intravital fluorescence microscopy and immunohistochemistry over 14 and 28 days, respectively. Microhemodynamic parameters and the numbers of rolling and adherent leukocytes in venules next to the implants were comparable in male and female mice. Immunohistochemical analyses on day 14 revealed a stronger neutrophilic (myeloperoxidase (MPO)⁺ cells: 526 ± 29 mm⁻²) and macrophage (CD86⁺ cells: 188 ± 21 mm⁻²; CD163⁺ cells: 269 ± 25 mm⁻²) response, as well as reduced T-cell activation (CD3⁺ cells: 31 ± 4 mm⁻²) in females when compared to males (MPO⁺ cells: 221 ± 25 mm⁻²; CD86⁺ cells: 120 ± 15 mm⁻²; CD163⁺ cells: 101 ± 19 mm⁻²; CD3⁺ cells: 62 ± 13 mm⁻²), while microvessel density and collagen deposition in the forming granulation tissue around the implants did not differ between sexes. In the flank model, there were no detectable sex-specific differences in the chronic foreign body response. These findings demonstrate that polypropylene sutures provoke a stronger early activation of the innate immune system in females, whereas the chronic foreign body response to the implants is comparable in both sexes. Full article

Background: Angiogenesis is critical for tumor progression. Microvessel density (MVD) is commonly assessed using CD31 and CD34, which detect both mature and newly formed vessels and therefore cannot distinguish active neoangiogenesis from stable, quiescent vasculature. Nestin, an intermediate filament protein expressed preferentially in proliferating endothelial cells, has been proposed as a complementary marker of active angiogenesis and has been investigated in several solid tumor types, including pancreatic, colorectal, and breast carcinomas. However, no studies have quantitatively compared nestin-positive MVD across AK, BCC, and SCC using standardized methods. Methods: Immunohistochemistry for nestin, CD31, and CD34 was performed on 118 patient samples collected in 2015–2019 and diagnosed with AK, BCC, or SCC. MVD was quantified by averaging vessel counts in three representative “hot spot” areas. Results: Nestin-positive MVD was significantly lower in patients with AK compared to patients with BCC and SCC (p < 0.001). The mean MVD of nestin-positive vessels was significantly lower in AK than in BCC and SCC (p < 0.0001). In all three groups, nestin-positive MVD demonstrated a strong, positive correlation with both CD34 and CD31. Conclusions: Nestin-positive MVD was significantly elevated in BCC and SCC compared to AK lesions and demonstrated strong correlations with standard angiogenic markers. These findings suggest that nestin may warrant further investigation as a complementary marker of angiogenesis in non-melanoma skin cancer. Whether nestin-positive MVD offers independent diagnostic or prognostic value in this context remains to be determined in larger, prospective, multicentre studies. Full article

Endometriosis is a chronic inflammatory disorder affecting approximately 10% of reproductive-age women, yet non-hormonal therapeutic options remain limited. This study investigates the role of the TLR4/NF-κB/NLRP3 inflammasome axis in endometriosis pathogenesis and evaluates the therapeutic potential of pharmacologic TLR4 inhibition. Ectopic endometriotic tissues, eutopic endometrium, and peritoneal fluid were collected from 15 patients with ovarian endometriosis and 15 control subjects. The endometriotic epithelial cell line 11Z was stimulated with LPS and ATP with or without the TLR4 inhibitor TAK-242. A murine endometriosis model was established in wild-type C57BL/6 and TLR4⁻/⁻ mice treated with TAK-242. Expression of TLR4, p-p65, NLRP3, caspase-1, cleaved caspase-1 (p20), GSDMD-N, IL-1β, PCNA, and CD31 was assessed by qPCR, Western blot, IHC, and ELISA. Ectopic lesions showed significantly elevated TLR4/NF-κB/NLRP3/IL-1β signaling compared with eutopic and control endometrium (all p < 0.05). Peritoneal fluid IL-1β was increased in patients, indicating a localized pelvic inflammatory response. In vitro, TAK-242 suppressed LPS/ATP-induced NF-κB/NLRP3 activation, pyroptosis, and IL-1β secretion (p < 0.05). Furthermore, the NLRP3-specific inhibitor MCC950 confirmed the essential role of NLRP3 inflammasome activation in IL-1β maturation. In vivo, TLR4 deletion or TAK-242 treatment reduced lesion weight, PCNA proliferation, and CD31 microvessel density (all p < 0.05). TLR4 inhibition blocks NF-κB nuclear translocation and subsequent inflammasome activation, suggesting a potential role in attenuating inflammation and angiogenesis. The TLR4/NF-κB/NLRP3 axis may drive endometriosis progression by linking innate immunity, inflammasome activation, pyroptosis, with possible involvement in angiogenesis warranting further investigation. Pharmacological inhibition of TLR4 attenuates lesion growth, supporting TLR4 as a promising non-hormonal therapeutic target for endometriosis. Full article

The aorta is the largest vascular conduit in humans, comprising three layers and a multitude of varying cell types collectively maintaining homeostasis and normal aortic wall function. Amongst these layers, the tunica adventitia is the external-most layer, where microvessels, termed vasa vasorum, can be found. These comprise pericytes and endothelial cells (ECs) and provide nourishment to the tunica adventitia and the outer media layers in the thoracic aorta. Adjacent to these microvessels, stem/progenitor group populations can be found, together forming a perivascular niche. Eventually, however, many of these cells and components can become dysregulated and contribute to development of thoracic aortic aneurysm (TAA). The purpose of this narrative review is to evaluate the recent literature related to marker gene expression in tunica adventitia pericytes, as well as the contribution of these populations to the development of aneurysm in the thoracic aorta. Pericytes in TAA generally exhibit phenotypic changes, which could be driven, in part, by loss of fibroblast growth factor (FGF) signaling. These changes eventually lead to vasa vasorum remodeling in the thoracic aorta, in turn contributing to the development of TAA. Full article

Diabetic wounds are a common and severe complication of diabetes mellitus, characterized by delayed healing due to persistent inflammation, impaired angiogenesis, and cellular dysfunction. Conventional therapeutic approaches remain limited in efficacy. In recent years, exosomes have attracted considerable attention in wound healing and regenerative medicine because of their crucial role in intercellular communication and tissue repair. However, rapid clearance of exosomes in vivo greatly limits their therapeutic efficacy. To address this critical limitation, we engineered a decellularized extracellular matrix (dECM)-based hydrogel system functionalized with exosomes derived from skin-derived precursor cells (SKPs). This biomimetic scaffold was designed to serve as a local exosome-delivery platform at the wound site, with the aim of improving exosome utilization and augmenting their regenerative effects. Comprehensive in vitro characterization demonstrated that the exosome-loaded composite hydrogels exhibited robust pro-angiogenic activity, as evidenced by enhanced endothelial cell proliferation, migration, and tube formation. Moreover, the hydrogels displayed significant antibacterial effects against wound-relevant pathogens and potent reactive oxygen species (ROS)-scavenging capacity, thereby mitigating oxidative damage. Notably, the composite hydrogels also promoted the phenotypic polarization of macrophages toward the pro-regenerative M2 phenotype. In parallel, in vivo studies using a streptozotocin-induced diabetic rat wound model confirmed that treatment with the composite hydrogels significantly accelerated wound closure rates compared to control groups. Histological and immunohistochemical analyses revealed enhanced angiogenesis, as evidenced by increased CD31-positive microvessel density, as well as improved collagen deposition, re-epithelialization, and an attenuated local inflammatory microenvironment characterized by reduced pro-inflammatory cytokine expression and elevated M2 macrophage infiltration. Collectively, the SKPs exosome-loaded dECM based composite hydrogels developed in this study represent a potential therapeutic strategy for the treatment of diabetic wounds. Full article

Background/Objectives: Immune checkpoint inhibitors (ICIs) improve outcomes in metastatic non-squamous non-small cell lung cancer (NSCLC), yet predictive biomarkers remain limited. This study investigated whether CD34-stained microvessel density (MVD) is associated with ICI efficacy. Methods: Patients with metastatic non-squamous NSCLC and tumor specimen suitable for anti-CD34 immunohistochemistry were included. In the derivation cohort, ROC analysis for objective response rate (ORR; CR + PR) identified a Youden-optimized threshold of 14.5 vessels/HPF (×400) (sensitivity 67%, specificity 93%; AUC 0.744, 95% CI 0.53–0.95; p = 0.021), which was applied without re-optimization to an internal validation cohort and a chemotherapy-only comparator. Results: In the derivation cohort, (n = 25), ORR was higher with MVD ≥ 14.5 than <14.5 (88.9% vs. 25.0%; p = 0.004) and OS/TFST were longer (OS: 53.0 vs. 17.1 months, p = 0.037; TFST: 50.0 vs. 6.0 months, p = 0.014). In the validation cohort (n = 13), MVD ≥ 14.5 was associated with longer TFST (9.0 vs. 4.0 months; p = 0.035) and numerically longer OS (12.0 vs. 7.0 months; p = 0.059). As a cut-off-independent sensitivity analysis, continuous MVD (per five vessels/HPF) was associated with longer TFST (HR 0.663, 95% CI 0.454–0.969; p = 0.034). No association was observed in the chemotherapy-only cohort. Conclusions: Higher CD34-MVD may be a feasible vascular metric associated with ICI outcomes that warrants prospective validation. Full article

Tumor-associated Tie2-expressing monocytes/macrophages (TEMs) have been implicated in promoting angiogenesis and metastasis in colorectal cancer (CRC), yet the molecular mechanisms linking TEMs infiltration to tumor metastasis and progression remain incompletely defined. This study investigated the distribution of TEMs in CRC and their association with gene expression profiles, microvessel density (MVD), and clinical outcomes. Immunohistochemistry on 30 formalin-fixed paraffin-embedded (FFPE) primary CRC samples revealed that TEMs, which characteristically express tyrosine kinase with immunoglobulin and epidermal growth factor homology domains 2 (Tie2) receptor and CD14, preferentially localize to perivascular regions and are associated with higher histological grade, tumor size, lymph node metastasis, and increased MVD. However, Tie2⁻/CD14⁺ macrophages and CD68⁺ tumor-associated macrophages (TAMs) showed uniform stromal distribution. Gene set enrichment analysis (GSEA) of in silico transcriptomic datasets of metastatic CRC (mCRC) identified enrichment of pathways related to cell–cell recognition, calcium signaling, transcription regulation, and metalloexopeptidase activity in Tie2⁺/CD14⁺ tumors. Subsequent qRT-PCR validation on FFPE primary CRC samples confirmed significant upregulation of C-C chemokine receptor 7 (CCR7), platelet-derived growth factor A (PDGFRA), CBP/p300-interacting transactivator with glutamic acid/aspartic acid-rich carboxyl-terminal domain 2 (CITED2), and carboxypeptidase E (CPE) in TEMs⁺ regions. Notably, angiopoietin1 (Ang1), but not angiopoietin2 (Ang2), was significantly elevated in TEMs⁺ primary tumors. Kaplan–Meier analysis on 1336 CRC patients indicated that high expression of CITED2, CPE, and Ang2 is associated with reduced overall survival. Collectively, these findings suggest that TEM infiltration is linked to transcriptional regulation, biological processes, and enzymatic programs in CRC, potentially contributing to tumor progression and poor prognosis, and highlight CCR7, PDGFRA, CITED2, CPE, and Ang1 as candidate biomarkers for further mechanistic exploration. Full article