Neuroblastoma (NB) is a highly malignant pediatric extracranial tumor with genetic and epigenetic factors influencing its pathogenesis. Key genetic contributors include germline mutations in ALK and PHOX2B, as well as somatic amplification of MYCN. The epigenetic regulator EZH2, a histone methyltransferase of the PRC2 complex, is overexpressed in NB and linked to poor prognosis in advanced cases. Targeting EZH2 offers a promising therapeutic approach due to its role in tumor proliferation, survival, and maintenance of undifferentiated states. This study explores the use of acetylated human serum albumin nanoparticles (HSANPs) loaded with 4O4HPR for epigenetic therapy in NB, tested in vitro and in vivo using nude mice xenograft models. The nanoparticles enter cells via clathrin-mediated endocytosis and induce G2-M cell cycle arrest, mitochondrial depolarization, and the production of reactive oxygen species, which activate Caspase 3 and trigger apoptosis. Mechanistic studies show increased p53 acetylation, stabilizing p53 and inhibiting EZH2's epigenetic silencing. Chromatin Immunoprecipitation reveals the disruption of EZH2 binding to the E-Cadherin promoter, which suppresses cell migration and EMT-like transition. Western blotting confirms upregulation of epithelial and downregulation of mesenchymal markers, with inhibited wound closure in SH-SY5Y cells. This nanotherapy disrupts EZH2-E-Cadherin interaction, offering novel translational potential for NB treatment.

Neuroblastoma (NB) is a neuroendocrine tumor derived from neural crest progenitor cells, commonly arising along the sympathetic nervous system, especially in the adrenal medulla. Despite therapeutic advances, the prognosis for advanced-stage NB remains poor, necessitating improved treatment options. 4HPR has demonstrated cytotoxicity in various tumors, including NB, with low systemic toxicity; however, its clinical use is restricted by poor solubility and bioavailability. To address this, we developed a human serum albumin-based nanoformulation of 4HPR using a simple desolvation method. This formulation effectively induced apoptosis in NB cells, marked by increased ROS generation, elevated Bax/Bcl-2 ratio, and enhanced cell detachment. Notably, we identified for the first time that MAPKAPK3 downregulation leads to reduced Bax phosphorylation and increased mitochondrial translocation. Co-immunoprecipitation confirmed a direct MAPKAPK3-Bax interaction, indicating MAPKAPK3 regulates Bax via phosphorylation. Our nanoformulation modulates this cross-talk, demonstrating promising translational potential as a novel therapeutic strategy for neuroblastoma.

Preclinical models are widely utilized to gain a deeper understanding of disease processes as well as develop novel therapies for inflammatory bowel disease (IBD). However, routinely used histological scoring of these specimens lack standardized guidelines, are subject to inter-reader variability, as well as being labor-intensive for pathologists. We present initial results of developing a comprehensive computational framework to automatically quantify histological components of inflammatory impact via digitized murine pathology samples. Our framework includes pre-analytical processing including quality evaluation, stain normalization, and background extraction. Next, a U-net segmentation model is utilized to segment out mucosa vs muscularis propria, with morphological operations to yield refined and accurate annotations. Finally, intuitive computerized scores were developed to quantify inflammatory impact in terms of tissue properties. When evaluated on a cohort of 20 murine pathology samples, the U-net model yielded an average dice similarity coefficient of 0.853 +/- 0.219 for segmenting mucosa tissue regions. Computational scores demonstrated statistically significant differences between treated vs disease control samples, following intuitive trends for inflammatory impact in the mucosa. Our framework represents a critical first step toward more quantitative, robust evaluation of IBD therapies via preclinical models.

Background: Crohn's disease (CD) is a chronic inflammatory disorder of the gastrointestinal tract with an elusive etiology. There are multiple studies which reported dysregulated metabolites in IBD patients. However, there lies a great variability in the patient population, disease location, disease state, biological samples, and metabolite detection techniques across the studies. All these variables add to the great heterogeneity in the detected metabolites, making it difficult to achieve a disease specific comprehensive signature of the metabolites. Objective: We aimed to analyze CD specific metabolomic studies and available datasets to provide a most accurate and comprehensive signature of dysregulated metabolites and metabolic pathways implicated in human CD. Design: A comprehensive, systematic review search was carried out using Medline and Embase databases to search the studies (inception to May 2024) which used analytical chemistry techniques to quantify the metabolites in different biological samples of Crohn's patients and non-IBD controls. Metabolites significantly altered in Crohn's patients and reported in at least 2 studies were included and considered for further analysis. Results: The systematic search yielded 3,632 studies, with 88 selected for data extraction. Across these studies, 79 metabolites were found to be significantly altered in CD patients in two or more studies. These metabolites differentiate between Crohn's patients and non-IBD controls, showing a distinct signature within the biological samples of CD patients and their importance in pathophysiology of CD. Conclusion: This systematic review provides a comprehensive and categorical signature of dysregulated metabolites across biological samples and provide detailed insight into the perturbed metabolic pathways involved in CD. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement This work was supported by UC Davis School of Medicine TriP program (MD and APA). The funding agencies had no role in the study analysis or writing of the manuscript. Its contents are solely the responsibility of the authors. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The study used only publicly available human data. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present study are available upon reasonable request to the authors

Schematic represents 4O4HPR loaded HSANPs directing the proteasomal degradation of the polycomb group of the protein, EZH2, by triggering oxidative stress and PKC-δ activation, leading to the killing of neuroblastoma cancer cells.

Mesenchymal stem cells (MSCs) are novel therapeutics for the treatment of Crohn’s disease. However, their mechanism of action is unclear, especially in disease-relevant chronic models of inflammation. Thus, we used SAMP-1/YitFc (SAMP), a chronic and spontaneous murine model of small intestinal inflammation, to study the therapeutic effects and mechanism of action of human bone marrow-derived MSCs (hMSC). hMSC dose-dependently inhibited naïve T lymphocyte proliferation via prostaglandin E2 (PGE2) secretion and reprogrammed macrophages to an anti-inflammatory phenotype. We found that the hMSCs promoted mucosal healing and immunologic response early after administration in SAMP when live hMSCs are present (until day 9) and resulted in a complete response characterized by mucosal, histological, immunologic, and radiological healing by day 28 when no live hMSCs are present. hMSCs mediate their effect via modulation of T cells and macrophages in the mesentery and mesenteric lymph nodes (mLN). Sc-RNAseq confirmed the anti-inflammatory phenotype of macrophages and identified macrophage efferocytosis of apoptotic hMSCs as a mechanism that explains their long-term efficacy. Taken together, our findings show that hMSCs result in healing and tissue regeneration in a chronic model of small intestinal inflammation and despite being short-lived, exert long-term effects via sustained anti-inflammatory programming of macrophages via efferocytosis.

Hydrogels have emerged as a versatile platform for a numerous biomedical application due to their ability to absorb a huge quantity of biofluids. In order to design hydrogels, natural polymers are an attractive option owing to their biocompatibility and biodegradability. Due to abundance in occurrence, cost effectiveness, and facile crosslinking approaches, alginate has been extensively investigated to fabricate hydrogel matrix. Management of cancer and chronic wounds have always been a challenge for pharmaceutical and healthcare sector. In both cases, curcumin have been shown significant improvement and effectiveness. However, the innate restraints like poor bioavailability, hydrophobicity, and rapid systemic clearance associated with curcumin have restricted its clinical translations. The current review explores the cascade of research around curcumin encapsulated alginate hydrogel matrix for wound healing and cancer therapy. The focus of the review is to emphasize the mechanistic effects of curcumin with its fate inside the cells. Further, the review discusses different approaches to designed curcumin loaded alginate hydrogels along with the parameters that regulates their release behavior. Finally, the review is concluded with emphasize on some key aspect on increasing the efficacy of these hydrogels along with novel strategies to further develop curcumin loaded alginate hydrogel matrix with multifacet applications.

Objective:Mesenchymal stem cells (MSCs) are novel therapeutics for treatment of Crohn's disease. However, their mechanism of action is unclear, especially in disease-relevant chronic models of inflammation. Thus, we used SAMP-1/YitFc, a chronic and spontaneous murine model of small intestinal inflammation, to study the therapeutic effect and mechanism of human bone marrow-derived MSCs (hMSC).Design:hMSC immunosuppressive potential was evaluated through in vitro mixed lymphocyte reaction, ELISA, macrophage co-culture, and RT-qPCR. Therapeutic efficacy and mechanism in SAMP were studied by stereomicroscopy, histopathology, MRI radiomics, flow cytometry, RT-qPCR, small animal imaging, and single-cell RNA sequencing (Sc-RNAseq).Results:hMSC dose-dependently inhibited naïve T lymphocyte proliferation in MLR via PGE 2 secretion and reprogrammed macrophages to an anti-inflammatory phenotype. hMSC promoted mucosal healing and immunologic response early after administration in SAMP model of chronic small intestinal inflammation when live hMSCs are present (until day 9) and resulted in complete response characterized by mucosal, histological, immunologic, and radiological healing by day 28 when no live hMSCs are present. hMSC mediate their effect via modulation of T cells and macrophages in the mesentery and mesenteric lymph nodes (mLN). Sc-RNAseq confirmed the anti-inflammatory phenotype of macrophages and identified macrophage efferocytosis of apoptotic hMSCs as a mechanism of action that explains their long-term efficacy.Conclusion:hMSCs result in healing and tissue regeneration in a chronic model of small intestinal inflammation. Despite being short-lived, exert long-term effects via macrophage reprogramming to an anti-inflammatory phenotype.Data Transparency Statement:Single-cell RNA transcriptome datasets are deposited in an online open access repository 'Figshare' (DOI: https://doi.org/10.6084/m9.figshare.21453936.v1 ).

With the persistent escalating exigence for new avenues in drug delivery strategies and imaging applications, different novel and efficient nanomaterials have surfaced. The league of these nanomaterials demonstrates many unique properties of which exceptional optical signals are exceedingly investigated. The confinement within the nano range endows exclusive optical attributes that can be explored in the domain of bioimaging and fluorescence-based detections. Despite the innovation in engineering fluorescent nanoparticles (FNPs) with exceptional properties, still, the appetite to develop highly efficient FNPs with multimodal utility can be perceived. Up recently, hydrogels have been projected as an excellent entrant for biomedical applications, due to their biocompatible and biodegradable nature with excellent absorption capacity along with tunable mechanical features. These hydrogels are capable to provide a favourable microenvironment for cellular growth and proliferation. In this critical review article, FNPs conjugated hydrogels (FH) have been critically evaluated for their role in drug delivery and imaging applications. The article will describe the rationale for the origin of fluorescence in FNPs and will also highlight different fabrication methodologies for their synthesis. Further, the utility of these hybrid FH in terms of drug delivery and imaging will be critically discussed and evaluated in detail. Finally, attention will be given to the limitations of these fascinating hybrid hydrogels that limit their clinical translations followed by their prospects that will be decisive for their clinical implications.

Stimulation of cells with electrical cues is an imperative approach to interact with biological systems and has been exploited in clinical practices over a wide range of pathological ailments. This bioelectric interface has been extensively explored with the help of piezoelectric materials, leading to remarkable advancement in the past two decades. Among other members of this fraternity, colloidal perovskite barium titanate (BaTiO 3 ) has gained substantial interest due to its noteworthy properties which includes high dielectric constant and excellent ferroelectric properties along with acceptable biocompatibility. Significant progression is witnessed for BaTiO 3 nanoparticles (BaTiO 3 NPs) as potent candidates for biomedical applications and in wearable bioelectronics, making them a promising personal healthcare platform. The current review highlights the nanostructured piezoelectric bio interface of BaTiO 3 NPs in applications comprising drug delivery, tissue engineering, bioimaging, bioelectronics, and wearable devices. Particular attention has been dedicated toward the fabrication routes of BaTiO 3 NPs along with different approaches for its surface modifications. This review offers a comprehensive discussion on the utility of BaTiO 3 NPs as active devices rather than passive structural unit behaving as carriers for biomolecules. The employment of BaTiO 3 NPs presents new scenarios and opportunity in the vast field of nanomedicines for biomedical applications.

The intrinsic architecture and complexity of the brain restricts the capacity of therapeutic molecules to reach their potential targets, thereby limiting therapeutic possibilities concerning neurological ailments and brain malignancy. As conventional models fail to recapitulate the complexity of the brain, progress in the field of microfluidics has facilitated the development of advanced in vitro platforms that could imitate the in vivo microenvironments and pathological features of the blood–brain barrier (BBB). It is highly desirous that developed in vitro BBB-on-chip models serve as a platform to investigate cancer metastasis of the brain along with the possibility of efficiently screening chemotherapeutic agents against brain malignancies. In order to improve the proficiency of BBB-on-chip models, hydrogels have been widely explored due to their unique physical and chemical properties, which mimic the three-dimensional (3D) micro architecture of tissues. Hydrogel-based BBB-on-chip models serves as a stage which is conducive for cell growth and allows the exchange of gases and nutrients and the removal of metabolic wastes between cells and the cell/extra cellular matrix (ECM) interface. Here, we present recent advancements in BBB-on-chip models targeting brain malignancies and examine the utility of hydrogel-based BBB models that could further strengthen the future application of microfluidic devices in oncology research.

Neuroblastoma (NB) is an extracranial pediatric tumor with highly invasive growth of cancer biomass and frequent metastases. During the differentiation process in embryonic development, altered epigenetic modifications lead to dysregulated expression of pluripotency markers, resulting in epithelial-mesenchymal transition (EMT) progression. Currently, available chemotherapies have provided a limited solution to this problem due to systemic toxicities and drug resistance. Epigenetic therapeutic molecules like histone deacetylase inhibitors are still in the initial stages of development. We have developed a retinoid (N-(4-hydroxyphenyl) retinamide, 4HPR) loaded acetylated human serum albumin (HSA) nanoformulation to address the epigenetic imbalance and chemoresistance in NB. The idea was conceived to deliver an acetyl pool along with a chemotherapeutic drug, 4HPR, to restrict the invasiveness of NB by maintaining the balance between histone acetylation and trimethylation. The therapeutic efficacy of the formulation was successfully evaluated in the in vitro and in vivo xenograft mouse model system of neuroblastoma. The synthesized nanoparticles show high biocompatibility and therapeutic efficacy in treating neuroblastoma subcutaneous xenografts in nude mice.

Aims: Inflammatory Bowel Disease is characterised by abdominal pain, diarrhoea, rectal bleeding and weight loss. Sometimes it may leads to severe health complications resulting in death of an individual. Current research efforts to highlight the role of melatonin in regulating EZH2, a master epigenetic regulator and its beneficiary effect in case of IBD management.Material methods: Murine macrophages (RAW 264.7) were treated with lipopolysaccharides (LPS) to activate them for generating inflammatory response to investigate efficacy of melatonin in-vitro models. Similarly, for developing in vivo models, Dextran sodium sulphate (36-50 kDa) was used. Evaluations of anti-inflammatory activities were carried out by nitrite assay, western blotting, q-PCR, immunofluorescence, and histological studies.Key findings: Reduction of epigenetic target, EZH2 by melatonin significantly improves the clinical symptoms of dextran sodium sulphate induced colitis and may be implicated as a potential therapeutic target in IBD management. The present study evaluates the efficacy of melatonin by epigenetic regulation in IBD models. Down regulation of EZH2 by melatonin reduced the chemical induced inflammatory insults in in vitro and in vivo models. Exploration of molecular pathways has revealed interlink of EZH2 and NOS2, a hallmark of inflammation. Molecular mechanistic action of melatonin is attributed to inhibition of the expression and physical interaction of EZH2 and NOS2.Significance: Our study highlights melatonin therapeutic effect via attenuating interaction between EZH2 and NOS2 which is beneficial in managing IBD treatment.

Neuroblastoma (NB) is a highly invasive and clinically challenging form of tumor to treat; despite multiple treatment strategies, a concrete treatment plan has not been developed so far. Here, for the first time, we have fabricated 4oxo-fenretinide-loaded human serum albumin nanoparticles with acetyl group modification on their surface (4OFnANP) using acetic anhydride as an acetylation agent. 4OFnANP are evaluated for their therapeutic potential for inducing apoptosis and modulating the histone epigenetic modifiers in NB. 4OFnANP show multi-phase cell cycle arrest with >75% inhibition in cancer cell proliferation after 72 h of treatment, which is 25-30% higher than the bare 4OFn drug response for a similar treatment time. 4OFnANP show an impressive reduction in the volume and size of NB xenografts with significant decrease in the expression of mesenchymal marker fibronectin and increased expression of epithelial marker Ecadherin, showing inhibition of epithelial mesenchymal transition (EMT). The nanoformulation marked the efficacy in NB by targeting the slug and E-cadherin axis and downregulating the expression of histone methyltransferase EZH2 and histone demethylase JMJD3 [histone H3 lysine 27 (H3K27) demethylase]; aberrant expression of JMJD3 and EZH2 is linked to carcinogenesis and EMT. Histopathological evaluation of major body organs of mice treated with the nanoformulation shows no systemic toxicities of the treatment. This study confers a novel, biocompatible, and effective way for targeting the EMT in NB.