Search Results (948)

Chronic kidney disease (CKD) is common in older cats, but cortical transcript-level relationships among hypoxia response, iron handling, and lipid oxygenation are poorly defined. This analysis-plan-guided, hypothesis-generating secondary analysis used public feline renal RNA-seq data (GSE303653). The internal plan fixed the gene panel, composite construction, primary inferential test, and quality-control thresholds before the review of the present expression results, but was not publicly registered. After technical quality control, 21 renal cortex samples from control, CKD 1/2, and CKD 3/4 cats were analyzed. A 23-gene panel and whole-transcriptome differential expression were evaluated using likelihood ratio testing as the primary panel-level screen, with pairwise DESeq2 contrasts, Spearman summaries, enrichment, medulla, composite, and marker-set analyses as secondary or exploratory context. VEGFA, FTL, and NCOA4 decreased with ordinal disease group, whereas ALOX5 and HIF1A increased; eight panel genes were stage-associated by likelihood ratio testing. The equal-weight composite was nonmonotonic. Advanced CKD enrichment was dominated by immune and inflammatory terms, while GPX4 and ferroptosis-pathway enrichment were not stage-significant. The findings support heterogeneous transcript-level remodeling, including ALOX5-associated inflammatory/lipid-oxygenation signal and HIF1A–VEGFA divergence, rather than evidence of ferroptotic cell death, pathway activation, or cell-specific mechanism. Full article

Background: Endothelial dysfunction and renal injury are emerging as a common feature of long COVID, especially in those with hypertension. It is not yet well characterised whether SARS-CoV-2 infection exacerbates podocyte dysfunction, fibrotic signalling and renal hemodynamic remodelling, over and above the effects of hypertension alone and there are no reliable early biomarkers in this population. Methods: We conducted a comparative cross-sectional study with prospective 6-month treatment response follow-up in 120 adult patients (aged 30–60 years) with essential hypertension (Stage I, II or III; n = 40 per stage), at Bukhara Regional Multidisciplinary Hospital. Each stage subgroup was further divided into post-COVID (3–6 months after recovery; n = 20) and non-COVID (n = 20) strata. Patients with diabetes, known chronic kidney disease, previous myocardial infarction or stroke and other major comorbidities were excluded. Serum cystatin-C, creatinine, aldosterone, TGF-β1 and VEGF-A; urinary nephrin and microalbumin; cystatin-C-derived eGFR (CKD-EPI) and oral protein-loaded renal functional reserve (RFR); and renal Doppler indices (Vps, Ved, RI, PI) of the main, segmental and interlobar arteries were assessed before and after 6 months of guideline-based renin–angiotensin–aldosterone system (RAAS) blockade (enalapril 5–10 mg or azilsartan 40–80 mg, ±eplerenone). Comparisons were made by Student’s t-test—associations by Pearson correlation. Results: At baseline, post-COVID hypertensive patients exhibited consistently higher endothelial–podocyte injury markers than non-COVID counterparts. Urinary nephrin was elevated across all stages (Stage I: 126.5 ± 9.1 vs. 91.9 ± 8.3 pg/mL, p < 0.01; Stage III: 203.3 ± 11.2 vs. 164.5 ± 9.7 pg/mL, p < 0.05), as were VEGF-A (Stage III: 286.1 ± 16.4 vs. 223.2 ± 12.6 pg/mL, p < 0.01) and TGF-β1 (Stage III: 186.4 ± 10.1 pg/mL, 1.3-fold higher; p < 0.01). The detection of microalbuminuria was 100% in Stage III post-COVID patients and 85% in non-COVID controls. The post-COVID groups had selective loss of renal functional reserve (7.8 ± 1.1% in Stage III compared to 12.5 ± 1.6% in non-COVID controls, p < 0.001). Nephrinuria correlated strongly with RFR (r = −0.824, p < 0.001), eGFR (r = −0.797, p < 0.001) and aldosterone (r = 0.613, p < 0.001). Six months of RAAS blockade reduced nephrinuria, microalbuminuria and TGF-β1 in both arms but the magnitude of biomarker reduction appeared smaller in the post-COVID group, particularly in Stage III. Conclusions: Long COVID appears to be associated with persistent endothelial dysfunction and podocyte injury in hypertensive patients. These results indicate that nephrinuria, VEGF-A, TGF-β1 and renal functional reserve are potential exploratory markers of endothelial and renal abnormalities in hypertensive patients following COVID-19. Before clinical utility can be determined, larger studies with multivariable modelling, diagnostic-performance analyses and correction for multiple testing are needed. The differences in biomarker response between groups observed in this study need to be confirmed in larger prospective studies with multivariable modelling and formal interaction analyses. Full article

Background and Objectives: Blood group antigens are well known for their importance in transfusion medicine and transplant compatibility; however, their biological role extends beyond these functions and includes associations with the risk of several diseases. In this study, we investigated the relationship between ABO blood groups and the circulating levels of 73 different molecules. Patients and Methods: Fifty-six healthy donors were enrolled, including 24 individuals with blood group O, 19 with blood group A, and 13 with blood group B. Blood samples were collected and analyzed in a single laboratory using Luminex fluorescent bead-based assay panels to determine the concentrations of 73 circulating molecules. Depending on data distribution, ANOVA or Kruskal–Wallis tests and Student’s t-test or Kolmogorov–Smirnov tests were applied to identify significant differences among groups. Associations were further assessed by binary logistic regression analysis. Results: Subjects with blood group A showed significantly higher circulating levels of IL-1R1, IL-13, IL-23, PDGF-BB, VEGF-A, VEGF-D, soluble VEGF-R2 (KDR), soluble VEGF-R3 (FLT-4), VLA-4, CD141, MMP-1, syndecan-1 (SDC-1), and mannose-binding lectin (MBL) compared with the other blood groups. In contrast, individuals with blood group B exhibited significantly higher levels of IL-22, IL-23, PDGF-BB, CD62P (P-selectin), and amyloid β1–42. Several significant associations were identified by logistic regression analysis. Conclusions: Our findings indicate that ABO blood groups are associated with distinct circulating molecular profiles, supporting the existence of biological differences that may contribute to variations in disease susceptibility among individuals with different blood types. Nevertheless, given the exploratory’s nature and limited sample size of this study, further investigations are required to validate these findings, confirm the observed associations, and clarify their potential clinical implications. Full article

Leucine, an essential branched-chain amino acid, serves not only as a substrate for protein synthesis but also as a key regulator of placental function and fetal development. This study investigated the effects of dietary supplementation with RP-Leu during late gestation on placental development and offspring performance in Hu sheep. Sixty twin-pregnant ewes at day 80 of pregnancy were randomly assigned to either a control group (fed a basal diet) or an RP-Leu group (fed a basal diet supplemented with 19 g/day RP-Leu). The feeding trial lasted for 60 d. The ewes were slaughtered at day 140 of gestation. Maternal slaughter traits and fetal organ weights were recorded. Blood and milk samples were collected for milk composition analysis and targeted metabolomic profiling. Leucine supplementation significantly increased the percentage of milk fat content, total solid content, and the birth weight of lambs (p < 0.05). Improvements in placental morphology and antioxidant capacity were observed, including a significant increase in cotyledon density and a significant enhancement of catalase (CAT) activity (p < 0.05). Gene expression analysis indicated that the NOS3, SLC38A1 and FABP4 genes in the placental cotyledons (p < 0.05), and the VEGFA, NOS3, SLC27A1 and FABP4 genes were significantly upregulated in the maternal caruncles (p < 0.05). Plasma metabolomic profiling revealed increased L-glutamic acid levels and alterations in several amino acids, with pathway enrichment indicating involvement in amino acid metabolism and membrane transport processes. Transcriptomic analysis identified 739 differentially expressed genes, which were mainly enriched in the PI3K/Akt signaling pathway, ECM–receptor interaction pathway, and cytokine–cytokine receptor interaction pathway. Collectively, these findings suggest that RP-Leu supplementation during late gestation may enhance offspring growth by modulating amino acid metabolism, promoting placental development, and improving placental nutrient transport capacity, thereby supporting fetal growth and development. Full article

Fucoidan, a sulfated polysaccharide, is known for its beneficial bioactive effects, for example antioxidant, anti-inflammatory, and vascular modulatory effects. Such a bioactive compound may also be useful for treating neurodegenerative diseases like age-related macular degeneration (AMD). Our research focuses on AMD-related pathomechanisms using primary porcine retinal pigment epithelium (RPE) cells in vitro and zebrafish (Danio rerio) models in vivo. We tested the bioactivity of a commercially available fucoidan (FVs) from bladderwrack with regard to pathomechanisms of AMD. We performed multiplex assays, RT-qPCR and fluorescence-based assays for the formation of nitric oxide (DAF-FM assay) and reactive oxygen species (DCF-DA assay) to analyze angiogenesis-related chemokines and pro-inflammatory cytokines as well as protection against oxidative stress and inflammatory insult. Our results showed that FVs significantly reduced the secretion of pro-angiogenic vascular endothelial growth factor A (VEGF-A) and follistatin as well as the pro-inflammatory cytokines interleukin 8 (IL-8) after lipopolysaccharide (LPS) and polyinosinic/polycytidylic acid (PIC) induction. Interleukin 6 (IL-6) was also reduced in the supernatant of the RPE cells. Additionally, in zebrafish, fucoidan decreased the production of NO and ROS. Gene expression of zebrafish embryos revealed anti-inflammatory effects by suppressing pro-inflammatory genes and significantly downregulating, e.g., interleukin 1 beta (IL-1β). These findings indicate modulation of oxidative stress, inflammatory responses, and VEGF secretion of the used FVs. This study demonstrates that fucoidan possesses AMD-relevant bioactivities in vitro and in vivo, suggesting fucoidan warrants further investigation in AMD-related research and related pathological mechanisms. Full article

Anti-vascular endothelial growth factor agents (anti-VEGFs) are the cornerstone of treatment for neovascular age-related macular degeneration (AMD). Resveratrol is a natural polyphenol with well-established antioxidant and anti-apoptotic properties. This study investigated whether resveratrol exerts cytoprotective effects when combined with anti-VEGFs on ARPE-19 cells in vitro. Cells were treated with ranibizumab (RNZ), aflibercept (AFL), or ziv-aflibercept (ZFL), either alone or in combination with resveratrol. Mitochondrial and cytosolic reactive oxygen species (MitROS and CytROS), mitochondrial membrane depolarization (MitDep), caspase-3, -8 and -9 activities, cell viability, apoptosis, and VEGF-A levels were evaluated using confocal microscopy, plate reader-based assays, and ELISA techniques. Anti-VEGFs induced tolerable oxidative or apoptotic stress in ARPE-19 cells but did not exhibit intrinsic antioxidant and cytoprotective effects. The addition of resveratrol significantly enhanced beneficial effects by reducing oxidative stress, preserving mitochondrial integrity, and suppressing intrinsic apoptotic signalling, while increasing cell viability. VEGF-A levels were effectively reduced by anti-VEGF treatment, and this suppression was further augmented by resveratrol without compromising cellular survival. These findings indicate that resveratrol acts as an additive modulator that strengthens the cellular effects of anti-VEGFs on ARPE-19 cells. The combination strategy may represent a supportive approach to optimize long-term anti-VEGF therapy in AMD. Full article

Ovarian tissue cryopreservation (OTC) has emerged as the only viable fertility preservation strategy for prepubertal girls and adolescent cancer patients facing gonadotoxic treatments. While OTC has transitioned from an experimental procedure to an established clinical practice, the functional longevity of transplanted grafts remains limited by massive follicle depletion. This review synthesizes recent technological advances in OTC for female children, with a particular focus on the underlying molecular mechanisms and innovative protective strategies. We systematically evaluate pre-cryopreservation assessments, surgical harvesting techniques such as medulla-sparing biopsies, and the comparative efficacy of slow freezing versus vitrification in preserving stromal and follicular integrity. Central to this discussion are the molecular drivers of post-transplantation injury, including ischemia–reperfusion-induced oxidative stress and the iatrogenic over-activation of the PI3K/Akt/mTOR signaling pathway, which leads to follicular “burnout.” Furthermore, we explore targeted pharmacological interventions, such as the dual-drug application of VEGFA and rapamycin, alongside emerging bioengineering frontiers including decellularized extracellular matrix scaffolds and 3D-printed bioprosthetic ovaries. Clinical outcomes are also summarized, highlighting high rates of endocrine recovery (~95%) and promising live birth rates (~28%), predominantly through natural conception. By integrating deep molecular insights with advanced tissue engineering, this review provides a comprehensive framework for optimizing long-term fertility restoration and improving the quality of survivorship for young female cancer survivors. Full article

Research on diabetic retinopathy (DR) usually emphasizes hyperglycemia and other causes like dyslipidemia, which are still not well understood. This study examined the effects of palmitic acid (PA) exposure, alone and combined with high glucose (G25), on Müller Glial Cell (MGC) dysfunction and angiogenic signaling. Primary MGC cultures were treated with G25 (25 mM), PA (250 µM), or PA + G25 for 24 and 48 h, followed by assessments of cell viability and analysis of the Vascular Endothelial Growth Factor (VEGFA)/VEGFA receptor 2 (VEGFR2) pathway through immunofluorescence, Western blot, and ELISA. Additionally, Gaussian mixture models (GMMs) were used to identify phenotypic subpopulations based on fluorescence intensity. The results showed that while hyperglycemia did not cause significant changes, PA and PA + G25 induced apoptosis-related cell death and significantly increased the expression of VEGFA, VEGFR2, HIF-α, and SP1. Although broad phenotypic diversity was observed at 24 h, by 48 h, a distinct shift towards an angiogenic phenotype was noted, with significantly elevated VEGFA/VEGFR2 levels. In summary, this research demonstrates that PA acts as a critical inducer of an angiogenic secretory phenotype in MGCs, indicating that lipid-mediated signaling plays a vital role in neovascularization in DR, possibly independent of glucose levels. Full article

Single-domain antibodies (sdAbs) are the smallest antigen-binding antibody (Ab) fragments (12–15 kDa) and have emerged as a versatile therapeutic platform. Their compact size, high solubility, stability, and ability to access cryptic epitopes distinguish them from conventional monoclonal Abs (mAbs) and larger Ab fragments. These properties are particularly attractive in ophthalmology, where molecular size, tissue penetration, and formulation constraints critically influence therapeutic performance. This narrative review summarizes the structural features, engineering strategies, immunogenicity considerations, and production platforms of sdAbs, with a focus on ocular applications. Preclinical studies demonstrate promising efficacy in retinal vascular diseases through targeting of VEGFA, ANG2, TNFα, and complement components, as well as in inflammatory and anterior segment disorders. SdAbs can be formatted as multimeric or Fc-fused constructs to extend intraocular half-life or delivered via gene therapy vectors as a sustained intraocular “biofactory” approach. Notably, recent work demonstrates the feasibility of vector-encoded sdAbs targeting complement C3 in vivo. While challenges remain regarding immunogenicity, pharmacokinetics, and regulatory pathways, the approval of several sdAb-based drugs in other fields underscores their clinical potential. SdAbs represent a promising next-generation modality for ocular therapeutics, enabling innovative strategies beyond conventional antibody formats. Full article

Background. This rat study reveals a new point: the considerable impact of unilateral adrenalectomy, severe vascular and multiorgan failure, occlusion/occlusion-like syndrome, and the stable gastric pentadecapeptide BPC 157 therapy. Based on the recent Fourier transform infrared (FTIR) spectroscopy vascular disturbance studies, particularly those after unilateral adrenalectomy in rats, the noted cytoprotective vascular recovery effect of the BPC 157 therapy may be useful. Methods. In rats, unilateral adrenalectomy (at 15 min, 5 h, 24 h) leads to integrated gross and morphological changes, vascular alterations, oxidative stress parameters, molecular markers and occlusion/occlusion-like syndrome and BPC 157 as useful therapy (/kg ig) (10 µg, 10 ng). Results. Peripherally and centrally, counteraction includes the lesions (adrenal, brain, heart, lung, liver, kidney, gastrointestinal tract), organ hemorrhage, and thrombosis. Attenuated/eliminated were arrhythmias, intracranial (superior sagittal sinus), portal, caval hypertension, and aortic hypotension. Significant resolution occurred via activation of collateral pathways, the azygos vein (direct blood flow delivery), and the recovered peduncle of the inferior suprarenal artery and superior suprarenal vein. Virchow’s triad circumstances were reversed. Occlusion/occlusion-like syndrome was counteracted as a whole. Also, BPC 157 counteracted adrenal lesions (lipid depletion, congestion). There were higher cortisol values, but still very low, and a shift toward the left of the adrenal compensatory weight increase. For the indicative conclusion along with previous studies, mechanistically, BPC 157 therapy exhibits the NO-system modulation/oxidative stress balance, increases NO-level, counteracts oxidative stress (malondialdehyde (MDA)), upregulates NOS1–3, and VEGF-A expression. Conclusions. These effects of BPC 157 therapy and its easy applicability deserve further consideration. Full article

Background: Pancreatic ductal adenocarcinoma (PDAC) has dismal survival, and vascular invasion is strongly associated with dissemination and poor outcomes; however, its molecular basis remains unclear. Methods: Transcriptomic and clinical data from The Cancer Genome Atlas-Pancreatic Adenocarcinoma (TCGA-PAAD) cohort were integrated with Genotype-Tissue Expression (GTEx) normal pancreas data. Vascular invasion-associated candidate genes were identified using subgroup-specific differential expression filtering. A three-gene prognostic signature was constructed using Cox and least absolute shrinkage and selection operator (LASSO) regression and validated in an independent PACA-AU cohort. Nucleoporin 35 (NUP35) was functionally evaluated by shRNA knockdown, phenotypic assays, endothelial assays using conditioned medium, and Western blotting of extracellular signal-regulated kinase (ERK)-vascular endothelial growth factor A (VEGFA) signaling. Results: We identified 172 vascular invasion-associated candidate genes and developed a three-gene model comprising NUP35, GMNN, and KLK11. The model stratified TCGA patients into risk groups with significantly different overall survival (OS; log-rank p = 0.014) and good predictive performance, with areas under the receiver operating characteristic curve (AUC) of 0.659, 0.722, and 0.796 for 1-, 3-, and 5-year OS, respectively. Consistent trends were observed in PACA-AU. Risk group transcriptomes were enriched for proliferative and tumor progression programs. Among the signature genes, NUP35 was prioritized because higher NUP35 expression was associated with poorer survival and positively correlated with VEGFA expression. NUP35 knockdown suppressed malignant phenotypes, reduced endothelial migration and tube formation, and decreased phosphorylated ERK and VEGFA without altering total ERK levels. Conclusions: A vascular invasion-related three-gene signature enables prognostic stratification in PDAC. NUP35 is associated with malignant and pro-angiogenic phenotypes and may regulate angiogenic activity partly through ERK-VEGFA signaling, supporting its potential as a prognostic biomarker and candidate therapeutic vulnerability. Full article

Background: Low-dose aspirin (LDA) reduces preeclampsia (PE) risk by up to 40%, yet its molecular effects on chorion trophoblast cells (CTCs), a fetal membrane lineage at the feto-maternal interface, remain obscure. CTCs form a structural and immunoregulatory barrier whose dysfunction drives inflammation-associated membrane pathology in PE. Extracellular vesicles (EVs) released by CTCs may encode cellular stress and adaptation states, offering a molecular window into aspirin’s timing-dependent effects on PE risk modification. Methods: Human CTCs were challenged with cigarette smoke extract (CSE) to model oxidative stress-driven PE pathology. Two paradigms were tested: (1) prophylactic aspirin (4 and 40 µg/mL) before and/or flanking the CSE, and (2) therapeutic aspirin after the CSE challenge. The EVs were isolated via ultracentrifugation and size-exclusion chromatography, characterized by nanoparticle tracking and immunoblotting, and profiled by quantitative mass spectrometry. A network pathway analysis and machine learning biomarker selection defined the EV-encoded molecular states. Results: The CTC-derived EVs from the CSE-exposed cells carried a PE-like proteomic signature marked by suppressed VEGF/ECM remodeling, activated TNF-p53 apoptotic signaling, and heightened inflammation. Prophylactic low-dose aspirin shifted the EV cargo toward an EV-encoded signature consistent with preserved angiogenic potential (enrichment of VEGFA, COL1A1, and MMP14) and predicted attenuation of apoptotic and NF-κB pathway activity by an Ingenuity Pathway Analysis. High-dose aspirin produced broad transcriptional suppression without an accompanying pro-angiogenic EV signature. Therapeutic (post-injury) aspirin partially attenuated the injury-associated EV cargo but did not restore the angiogenic EV signature. An exploratory machine learning analysis of EV proteomes identified a candidate prophylactic biomarker panel anchored by HSPA8, SERPINF2, COL4A1, and PLOD1, mapped to the predicted angiogenic recovery and redox-balance pathways. These EV cargo readouts represent the predicted molecular states and require functional validation before clinical interpretation. Conclusions: The CTC-derived EV proteomic signatures capture the dose- and timing-dependent aspirin effects in this in vitro CTC model, positioning the chorion as a candidate pharmacological “secondary responder” favoring cellular resilience over classical anti-inflammatory suppression. As an exploratory hypothesis-generating study, EV-based molecular profiling could provide a foundation for future investigations aimed at stratifying aspirin responders from non-responders, although clinical validation in maternal plasma cohorts will be required before any translational application. Full article

Objectives: Diminished ovarian reserve (DOR) significantly compromises in vitro fertilization (IVF) success. Although systemic markers such as anti-Müllerian hormone (AMH) serve as valuable clinical indicators of the ovarian reserve, they lack the sensitivity to reflect the qualitative deterioration of the follicular microenvironment. Therefore, in this study, we aimed to characterize the inflammatory proteome of follicular fluid (FF) to establish a high-performance auxiliary diagnostic model for DOR. Methods: Utilizing the ultra-sensitive Olink proximity extension assay, we quantified 92 inflammation-related proteins in the FF of 88 participants (67 with DOR and 21 normal controls). Differentially expressed proteins (DEPs) were identified, and their relationships with key clinical indices were evaluated. A robust predictive signature was refined through integrated Least Absolute Shrinkage and Selection Operator (LASSO) regression and Random Forest algorithms, with diagnostic performance assessed via 10-fold cross-validation. Results: Thirty-five DEPs were significantly dysregulated in the FF of patients with DOR, demonstrating strong associations with serum AMH and basal estradiol concentrations. A minimized diagnostic panel comprising four core proteins, adenosine deaminase (ADA), vascular endothelial growth factor A (VEGFA), eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), and matrix metalloproteinase-1 (MMP-1), was established. This multivariable model achieved an excellent area under the receiver operating characteristic curve (AUC) of 0.953. Conclusions: The identified four-protein signature reflects localized chronic inflammation and early pathophysiological shifts in the DOR follicular microenvironment. As a high-performance molecular index, this panel could complement conventional systemic assessments, provide a reliable means of evaluating follicular viability, and optimize individualized therapeutic strategies. Full article

Astragalus root, a traditional Chinese herbal remedy, has shown potential benefits against diabetic nephropathy (DN). However, the mechanisms driving its effects remain poorly understood. This study explored the molecular pathways through which Astragalus root improves DN. To identify possible targets and mechanisms of Astragalus root in DN treatment, we applied network pharmacology, molecular docking, molecular dynamics simulation, and in vitro assays. Network pharmacology screening uncovered 46 overlapping targets between Astragalus root and DN. Protein–protein interaction (PPI) network analysis identified five core candidate targets: CASP3, VEGFA, CTNNB1, MYC, and PRKCB (PKCβ). KEGG pathway analysis indicated that the AGE-RAGE signaling pathway was the most significantly enriched. Molecular docking revealed that quercetin, β-carotene, daidzein, capsaicin, and kaempferol—potential bioactive components of Astragalus root—bound strongly to each of the five core targets. Molecular dynamics simulations further confirmed the conformational stability of kaempferol when complexed with these target proteins. In vitro experiments showed that kaempferol markedly reduced protein levels of α-SMA, Col I, and Col IV; lowered secretion of TNF-α, IL-6, and IL-1β; and decreased ROS and MDA content. Additionally, kaempferol’s therapeutic effects were mediated through suppression of the AGE-RAGE-PKCβ-TGF-β1 signaling axis. This work identified kaempferol, a bioactive ingredient of Astragalus root, as a potential therapeutic agent against DN, along with its target pathways. These findings provide a scientific foundation for its clinical translation. Full article

Introduction: platelet-rich fibrin (PRF) is a second-generation platelet concentrate which is known for promoting cell migration, tissue repair, angiogenesis and bone formation. In contrast, the specific effects of trauma-activated PRF on mesenchymal stromal cells (MSC) are not yet fully understood. The present study investigates systemic effects of surgical trauma-activated PRF on MSCs in vitro, analyzing their metabolic activity, inflammatory responses, and regenerative capacity to optimize advanced treatment concepts for severe fractures and injuries. Material & Methods: PRF membranes (T-PRF from trauma patients, C-PRF from healthy controls) were generated. After co-incubation with MSC cells for 24, 72, and 120 h, further investigations of metabolic activity (MTT assay) and gene expression analyses were performed. Results: for MTT assay, results especially showed a significantly higher metabolic activity of T-PRF after 120 h. ELISA-results measuring cytokine levels (CXCL10, IL-6, VEGF, and IDO) exposed a frequent peak in T-PRF group at 72 h, declining slightly at 120 h. In the gene expression analyses, T-PRF exerted a comparatively stronger stimulating effect on MAPK14 and VEGFA after 24 h, while a decrease in gene expression for MAPK8, MAPK14, and RUNX2 was observed over time. Conclusion: surgical trauma-activated PRF seems to be a powerful inducer of early inflammatory and stress responses in MSCs with preserved angiogenic but limited osteogenic signaling. Therefore, a targeted balance between inflammatory activation and sustainable regeneration, as well as optimized preparation and possible combination with immunomodulatory approaches, appear to be crucial for the therapeutic success of PRF-based strategies. Full article