ABSTRACT Liver fibrosis is a common consequence of chronic liver injury and a major contributor to liver‐related mortality. Persistent hepatocellular injury promotes fibrosis initiation and progression through excessive extracellular matrix deposition. Hepatic stellate cells (HSCs), the principal source of extracellular matrix in the fibrotic liver, transition from a quiescent state to an activated myofibroblast‐like phenotype in response to profibrotic stimuli such as transforming growth factor‐beta. This transition is accompanied by transcriptional and epigenetic reprogramming involving DNA methylation, histone modifications, and regulation by non‐coding RNAs. Treating the underlying cause of liver disease, such as promoting weight loss in metabolic dysfunction‐associated steatohepatitis or eradicating viral hepatitis, remains the principal strategy for slowing or potentially reversing fibrosis. Despite substantial advances in understanding the cellular and molecular basis of liver fibrosis and HSC activation, most mechanism‐based therapeutic approaches have not yet demonstrated clinical efficacy. Further translational and clinical studies are therefore required. Recent advances in molecular biology have highlighted the potential relevance of epigenetic modifications to the diagnosis, treatment, and prognosis of chronic liver disease. In this review, we summarize the principal epigenetic changes involved in HSC activation, and initiation/progression of liver fibrosis. We also discuss recent interventions designed to modulate these epigenetic changes and evaluate their therapeutic potential in experimental models of liver fibrosis.

Multiple sclerosis (MS) is a complex, progressive neurodegenerative autoimmune disease and a major cause of neurological disability worldwide. MS affects approximately 2.8-3 million people, predominantly presenting as relapsing–remitting MS (RRMS) that frequently converts to secondary progressive disease, while effective options for progressive phenotypes remain limited. This review reframes MS immunotherapy through a tolerance-centric paradigm, distinguishing continuous maintenance disease-modifying therapies (DMTs) from immune reconstitution therapies (IRTs) and emerging antigen-specific tolerance strategies. Classical immune reconstitution therapies, including alemtuzumab, cladribine, and autologous hematopoietic stem cell transplantation (aHSCT), have demonstrated durable disease control and prolonged periods of no evidence of disease activity (NEDA) in appropriately selected patients. This is accomplished through a finite course of lymphocyte depletion followed by qualitative immune repopulation that favors tolerogenic regulatory T (Treg) and regulatory B (Breg) cells over pathogenic Th1/Th17 clones. In contrast, maintenance DMTs (interferons, sphingosine-1-phosphate modulators, anti-CD20 monoclonals, natalizumab) suppress inflammation activity during continuous administration but lack durable immune reset. Building on IRT principles, next-generation cell-based therapies (tolerogenic dendritic cells (tolDCs), autologous Tregs, and mesenchymal stromal cells/extracellular vesicles (MSCs/EVs)), aim to induce precision, antigen-specific tolerance while minimizing systemic immunosuppression. These approaches hold promise for overcoming the limitations of chronic immunosuppression, providing durable disease control, and improving long-term patient outcomes. Collectively, immune reconstitution and tolerance-inducing therapies represent an emerging shift from lifelong disease control toward durable immune resetting and the possibility of sustained drug-free remission in MS.

BACKGROUND:Hypercholesterolemia is a common metabolic disorder characterized by elevated low-density lipoprotein cholesterol (LDL-C) levels, a major risk factor for cardiovascular disease. MicroRNAs (miRNAs) are key post-transcriptional regulators of gene expression and lipid metabolism. METHODS AND RESULTS:In this study, in silico analysis revealed that miR-30b-3p could potentially target Apolipoprotein B (APOB), Proprotein convertase subtilisin/kexin type 9 (PCSK9), and Cholesteryl ester transfer protein (CETP). miR-30b-3p binding sites in the 3'-untranslated regions (UTRs) of PCSK9, APOB, and CETP were predicted and validated by dual luciferase reporter assays. Luciferase assays indicated significant reducing reporter activity (PCSK9: 49%, APOB: 39%, CETP: 86%). miR-30b-3p overexpression decreased mRNA levels of PCSK9, APOB, and CETP, with reduced PCSK9/ApoB protein but unchanged CETP protein. miR-30b-3p could play a regulatory role in lipid metabolism by targeting PCSK9 and APOB, reducing LDL levels in hepatocytes. Huh-7 cells underwent miR-30b-3p overexpression via lentiviral transduction and the expression of genes/proteins were assessed by quantitative PCR and western blot. LDL and high-density lipoprotein (HDL)-associated proteins in the culture media supernatant were measured to evaluate functional lipid changes. LDL-associated protein decreased in the conditioned medium, whereas HDL-associated readouts remained unchanged, leading to a lower LDL/HDL ratio, indicating impact on lipid homeostasis. CONCLUSIONS:This study aimed to investigate the effects of miR-30b-3p on the expression and function of the mentioned genes and the outcome of this intervention on lipid metabolism. These findings propose miR-30b-3p as a promising therapeutic candidate for hypercholesterolemia, warranting further in vivo validation.

ABSTRACT Liver fibrosis is the common consequence of liver injury caused by a variety of chronic liver disorders. This condition leads to the development of more severe complications, particularly cirrhosis and hepatocellular carcinoma. Despite abundant studies, the fundamental cell and molecular mechanisms of liver fibrosis are still unknown. There are many key players involved in the initiation and progression of liver fibrosis. Thus, the specific type of underlying disease and the study's objectives should be considered while choosing suitable models for liver fibrosis. Numerous in vitro and in vivo models have been generated to investigate liver fibrosis and proposed for drug screening and toxicology; however, there are no ideal in vitro models for drug discovery yet. In this review, we introduced the available in vitro models and highlighted certain platforms such as organoids and liver‐on‐a‐chip for investigating liver fibrosis. Furthermore, we discussed the current challenges and potential application of each model.

In recent years, cell-based medicinal products (CMPs) have emerged as novel therapeutics with specific potential to treat a wide range of diseases. These products should be produced in accordance with good manufacturing practices (GMPs) and follow specific guidelines to ensure their safety and meet standard quality control criteria. This manuscript reviews current standard methods for quality control and validation specific to CMPs intended for clinical applications. We summarize critical quality attributes, including safety assessments such as sterility, endotoxin, mycoplasma, viral testing, tumorigenicity, and genetic stability; quantitative parameters, including cell counts and dose determination; and quality characteristics encompassing cell viability, morphology, growth kinetics, and immunophenotyping. We also address purity evaluation, potency assays, and the importance of validating analytical methods to guarantee reproducible and reliable test results. Furthermore, this article discusses necessary considerations for donor screening, raw material sourcing, manufacturing environment monitoring, and stability testing to maintain product integrity throughout production and storage. Emphasis is placed on adherence to GMP and relevant regulatory guidelines as defined by international pharmacopeias and authorities, such as the Food and Drug Administration and the European Medicines Agency. Finally, we highlight challenges faced in standardizing quality control for CMPs and underscore the need for continued development of rapid and robust testing methods tailored to their unique characteristics. This comprehensive overview aims to support academic and industrial stakeholders in implementing effective quality control strategies for advanced cell-based therapies.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a major chronic liver disorder and a growing global health concern driven in part by oxidative stress. Activation of antioxidant pathways, particularly the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway, by vitamin E represents a promising therapeutic strategy. In this study, we developed a biomimetic co-culture platform to evaluate the impact of vitamin E on oxidative stress and steatosis progression in a MASLD model. The model consisted of a co-culture of Huh-7 and LX-2 cells (4:1) seeded on plates coated with 100 μ g/mL liver extracellular matrix-derived hydrogel (LEMgel) from decellularized sheep liver. MASLD was induced by the co-culture treatment with Oleic acid (330 µM) and Palmitic acid (165 µM) for 2 days. Subsequently, cells were treated with 100 μM vitamin E for 4 days. Oil Red O staining and gene expression analysis of CD36, SREBP-1c, and CPT-1 confirmed successful steatosis induction and transcriptome alterations consistent with a MASLD phenotype. Vitamin E treatment significantly improved cell viability, reduced intracellular lipid accumulation, and downregulated CPT-1, NOX4, SREBP-1c, CD36, and PPAR γ , while upregulating HO-1, NQO1, SOD, and GSH, indicating enhanced antioxidant capacity. It also modulated hepatic markers (albumin, alpha-fetoprotein), suppressed LX-2 activation via TGF-β inhibition, activated Nrf2 signaling, and induced expression of downstream target genes, including the antioxidant/carboxylesterase gene CES1. Overall, vitamin E attenuated oxidative stress and steatosis, and the biomimetic platform provides a physiologically relevant model for MASLD pathogenesis and drug screening.

Liver fibrosis, characterized by the excessive deposition of extracellular matrix (ECM) driven by hepatic stellate cells (HSCs) activation, remains a critical challenge due to its progression to cirrhosis and hepatocellular carcinoma (HCC). This review clarifies the complex crosstalk between the immune system and HSCs, highlighting key cellular players including macrophages, natural killer (NK) cells, regulatory T cells (Tregs), and their cytokine-mediated signaling pathways that regulate fibrogenesis and fibrosis resolution. We describe pivotal molecular mechanisms such as transforming growth factor (TGF)-β, platelet-derived growth factor (PDGF), Wnt/β-catenin, and NF-κB signaling in HSCs modulation, emphasizing their interplay with immune responses. Novel therapeutic strategies targeting this complex immune–HSCs interaction, ranging from immunomodulatory agents, macrophage polarization, and NK cell-based therapies, to stem cell-derived exosomes, offer promising opportunities for preventing and reversing fibrosis. We further discuss innovative combination therapies integrating immunotherapies with antifibrotic agents, personalized strategies based on immune profiling, and the challenges of immune heterogeneity in fibrosis management. This review discusses recent advances in molecular interplay of immune system and HSCs, highlighting novel therapeutic targets, and future perspectives for managing chronic liver diseases.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a common chronic liver disease, with an increasing incidence globally. Characterized by excessive lipid deposition, MASLD triggers chronic inflammation that can progress to metabolic dysfunction-associated steatohepatitis (MASH). Despite its growing burden, no approved treatment protocol exists, highlighting the need for novel therapeutic modalities. Due to lack of appropriate experimental models, research and development of innovative medications remains challenging. In this study, multicellular liver microtissues were generated by encapsulating human hepatoma (Huh-7), hepatic stellate (LX-2) and monocyte (THP-1) cell lines and umbilical vein endothelial (HUVEC) cells in a liver extracellular-derived hydrogel, mimicking the native hepatic microenvironment. MASLD pathogenesis was induced by exposing microtissues to defined concentrations of Oleic and Palmitic acids for 3 days. We evaluated the therapeutic potential of human placenta extract (hPE) on this biomimetic model. On day 4, post-MASLD-model formation, microtissues received 5 mg/ml hPE for 48 h. Assessments on day 6 showed significant improved cell viability, reduced lipid droplets, and restored hepatic synthetic functions, evidenced by enhanced albumin, alpha-fetoprotein, and urea levels. Additionally, hPE treatment declines lipid uptake and synthesis related genes (CD36 and SREBP) while upregulating fatty acid oxidation related gene (CPT1), decreased pro-inflammatory cytokines (IL-6 and TNF-α), and increased cytochrome P450 3A4 (CYP3A4) expression. Overall, hPE could alleviate MASLD complications in this bioengineered liver microtissue model and may be proposed as a potential candidate for further preclinical/clinical studies in MASLD.

The 5-year overall survival rate for hepatocellular carcinoma (HCC) patients remains below 20%. Alterations in the extracellular matrix (ECM) are increasingly recognized as central drivers of HCC initiation and progression. This study applied a system biology framework integrating omics data and machine learning to analyze gene expression and regulatory networks in HCC using The Cancer Genome Atlas. Eight ECM-associated genes (CSPG4, CD34, C1orf35, ESM1, MAPT, PLXDC1, STC2, and THBS4) were identified as upregulated diagnostic biomarkers with strong discriminatory power. Among them, MAPT, PLXDC1, and STC2 showed significant associations with poor overall survival, defining a prognostic subset. Validation in the GSE104310 and GSE144269 datasets confirmed consistent expression patterns across cohorts. Functional enrichment linked these genes to tissue remodeling and angiogenesis. Single-cell RNA sequencing revealed MAPT upregulation in T cells, PLXDC1 enrichment in cancer-associated fibroblasts, and mild STC2 elevation in tumor-associated macrophages and endothelial cells. These findings identify key ECM-based biomarkers with potential for early detection, prognosis, and therapeutic targeting in HCC.

Vector-borne diseases such as malaria are a threat to global public health and the economy. These diseases were proposed to be managed and controlled by new preventive strategies such as paratransgenesis. This is an innovative technique that makes use of symbiotic microorganisms to influence vector or targeted pathogens. The performed studies on Anopheles stephensi and Anopheles gambiae demonstrated that the carboxypeptidase-B1 enzyme plays a vital role in the sexual development of the Plasmodium parasite in the mosquito midgut by its enzymatic activity. Therefore, inhibiting its enzymatic activity could be a target for preventing approaches. Potato Carboxypeptidase Inhibitor (PCI) has desirable characteristics that make it a promising effector molecule for paratransgenesis. In this study, the inhibitory effect of PCI on Carboxypeptidase-1 from An. stephensi (CPBAs1) was evaluated. The coding sequence of the cpbas1 and pci genes were cloned into the pET-23a expression vector, expressed, and purified. Finally, the inhibitory effect of the PCI on CPBAs1 was evaluated in parallel with the 1,10-phenanthroline as the commercial-specific inhibitor. Our findings revealed that PCI could inhibit the enzymatic activity of the CPBAs1 efficiently in low concentrations. Given PCI’s remarkable inhibition activity against the CPBAs1 and its suitable structural features, PCI could be considered as a potential effector molecule for use in the paratransgenesis approach in future related studies.

Parkinson's disease (PD) ranks as the second most prevalent neurodegenerative disorder, primarily characterized by motor dysfunction resulting from the degeneration of dopaminergic neurons. Early and accurate diagnosis is crucial for effective treatment; however, the overlap of symptoms with other disorders frequently results in misdiagnosis. This study aims to identify reliable biomarkers for the early PD diagnosis through a comprehensive literature review and bioinformatics analysis. We initially identified 32 genes strongly associated with PD, from published studies and database annotations. Further bioinformatics validation using protein-protein interaction networks and external gene expression datasets revealed additional candidate genes, including GBA1 and LRRK2, which are relevant to both familial and sporadic forms of PD. Enrichment analyses of these genes, emphasizing pathways related to mitochondrial function, autophagy and neurodegeneration-related pathways. Our findings highlight the promise of genetic biomarkers in improving diagnostic precision and guiding therapeutic approaches, thereby enhancing clinical outcomes for patients with PD. Ongoing validation of these results is essential for integrating biomarkers into standard clinical practice, with the ultimate goal of revolutionizing the diagnosis and management of PD.

Background and Objective : Myeloid-derived suppressor cells (MDSCs) are a crucial and diverse group of cells found in the tumor microenvironment (TME) that facilitate progression, invasion, and metastasis within solid tumors. CD84, a homophilic adhesion molecule expressed on MDSCs, plays a critical role in their accumulation and function within the TME. This study aims to investigate the protein-protein interactions of CD84 using molecular dynamics simulations and to explore potential therapeutic strategies targeting these interactions. Methods : Through computational techniques, we generated highly potent mutated CD84 mini-proteins and peptides as antagonists with significantly higher affinity for CD84 to mimic the key features of the IgV-like domain of the protein. Additionally, we engineered an antibody capable of blocking CD84. Binding affinities were assessed using dissociation constant (Kd) calculations. Results : Data analysis shows that the Kd values for the designed peptides ranged from 10 to 100 times stronger than those of the natural CD84 interactions, indicating efficient inhibition of CD84 interactions. Additionally, mutagenesis of the Ig-like V domain of CD84 resulted in variants with improved binding stability, with a Gibbs free energy change (ΔΔG) indicating enhanced interaction potential. Conclusions : This study provides insights into CD84 interactions and their implications for immunotherapy targeting MDSCs in solid tumors. However, experimental validation is necessary to confirm the findings of this study and evaluate peptide selectivity as potential molecular therapeutics.

Liver fibrosis (LF) is a pathological condition resulting from a chronic inflammatory response to multiple etiological factors, including viral infections, excessive alcohol consumption, and metabolic disorders. The important role of macrophages in this process, especially the M2 subtype, has attracted attention as a potential target for macrophage-based immunotherapy. M2 macrophages have anti-inflammatory and reparative properties that enable them to modulate the immune response and facilitate repairing damaged tissues. They participate in reducing fibrogenic features in term of gene expression and histological markers associated with LF. These cells phagocytose apoptotic cells and matrix components. M2 macrophage-based immunotherapy has shown great potential in ameliorating LF through mechanisms involving the IL-10/STAT3 and TGF-β/SMAD signaling pathways, which are essential in suppressing the pro-inflammatory response and supporting tissue regeneration. However, significant challenges such as individual resistance to therapy and the potential for promoting fibrosis suggest that further development and research are needed to optimize the safety and efficacy of this therapy in clinical applications. This study provides comprehensive insights into the role of M2 macrophages in LF and explores their potential as an innovative therapeutic approach in treating LF.

Globally, liver cancer is reported to be the third leading cause of cancer-related mortality. The most common type of these cancers is hepatocellular carcinoma (HCC). Current preventive strategies, including lifestyle modifications, antiviral therapies, and surveillance, are limited in their effectiveness. Mitochondria play critical roles in regulating cellular metabolism, oxidative stress, and apoptosis. Mitochondrial dysfunction can accelerate HCC progression, particularly in patients with liver diseases such as metabolic-associated fatty liver disease (MAFLD) and metabolic dysfunction-associated steatohepatitis (MASH). In this review, we discuss the mechanisms of mitochondrial dysfunction in HCC from a molecular point of view, including oxidative stress, mitophagy dysregulation, mitochondrial dynamics dysregulation, and mitochondrial DNA (mtDNA)-mediated dysregulation of innate immune responses. Additionally, we explore molecular-targeted therapies aimed at restoring mitochondrial function. Critical approaches include targeting reactive oxygen species pathways through agents such as iridium (III) complexes and Mito Rh S, which induce cancer cell death through apoptosis and ferroptosis. Other compounds, including dehydrocrenatidine, enhance oxidative phosphorylation and promote apoptosis. Inhibitors of dynamin-related protein 1 (Drp1) target mitochondrial fission to reduce tumor growth. Furthermore, mitophagy modulators, such as SIRT1 activators, improve mitochondrial quality control, minimize the negative effects of oxidative stress, and reduce cancer development. Clinical trials are ongoing for the mitochondrial enzyme-targeting agents CPI-613 and Gamitrinib, a heat shock protein-targeting agent, which have hence shown great promise for these therapies. With further investigation, mitochondrial-targeted interventions could be promising for preventing or reducing HCC incidence and recurrence, increasing long-term survival, and improving the quality of life of patients with advanced-stage disease.

INTRODUCTION:Tumor cell's resistance, high recurrence rate, and low overall survival rate have made hepatocellular carcinoma (HCC) a major health concern. The combination of advanced targeted therapies such as immunotherapy, with conventional treatments has gained traction for application on HCC. Immunotoxins (ITs) represent a category of biomolecules that combine the targeted affinity of antibodies with the cytotoxic properties of toxins. AREAS COVERED:This study highlights Glypican3 (GPC3) as a potential candidate for targeted therapeutic interventions using ITs. It presents a comprehensive overview of the advantages and challenges associated with these modalities, and their promising outcomes in HCC treatment. A systematic literature review was conducted using PubMed, Web of Science and Scopus from 2015 to 2024. EXPERT OPINION:Despite potential applicability, many concerns should be addressed before the employment of GPC3-based ITs. These include improving efficient penetration of ITs into the solid tumors, considering neutralizing antibodies against the drugs, and enhancing serum half-life of ITs. Furthermore, the ITs potential in eliminating cancer stem cells (CSCs) and residual tumor cells is discussed. The ability to target CSCs can significantly reduce the likelihood of recurrence and improve overall survival rate. This could make ITs a pivotal component in the future of HCC treatment.

Helicobacter pylori, a significant factor in the development of gastric cancer and peptic ulcers, poses challenges for drug development due to its resilience. Computational approaches offer potential solutions for effective vaccine development targeting its antigens while ensuring stability and safety. The four critical antigenic proteins included in this study's innovative vaccine design are neuraminyllactose-binding hemagglutinin (HpaA), catalase (KatA), urease (UreB), and vacuolating toxin (VacA). Advanced immunoinformatics methods identified the possibility of triggering an immunological reaction. An adjuvant (50S ribosomal protein L7/L12) was fused to the vaccine sequence's N-terminus to improve immunogenicity. GROMACS molecular dynamics simulations with the OPLS-AA force field further improved the structure. The vaccine design and human Toll-like receptor 5 (TLR5) demonstrated a strong binding in docking tests. A model of simulating immune response confirmed the vaccine's efficacy and predicted how it would affect the immune system. Using the optimal restriction sites of the pET28b (+) expression vector, the vaccine candidate was cloned in silico. To validate the findings, this vaccine design will be synthesized in a bacterial system, and in experimental studies will be conducted in the following phase.

Objective:Despite the remarkable advances in approved therapeutic approaches, the recurrence rate of hepatocellular carcinoma (HCC) is very high after treatment. Therefore, introducing innovative therapeutic modalities such as targeted molecular therapies is inevitable. Lysine demethylase 6A (KDM6A) is a member of the KDM6 family with histone demethylase activity. This gene frequently mutates in different cancers, and its mutations are associated with the increased likelihood of carcinogenesis. This study is aimed at evaluating if inducing KDM6A expression could attenuate cancerous features of HCC cells. Method:A lentiviral-based vector was used to induce KDM6A expression in Huh-7 cells. The impact of KDM6A overexpression on the cancerous phenotype of HCC cells was assessed by measuring proliferation rate, migration and colony formation capacity, and differentiation induction toward hepatocytes. Results:KDM6A overexpression significantly altered cellular morphology, proliferation rate, cell cycle pattern, colony formation, and migration capacity of HCC cells. In addition, induction of differentiation toward hepatocytic fate resulted in down/upregulation of epithelial-mesenchymal transition (EMT) markers associated with the cadherin switch. Furthermore, the expressions of ALB and HNF4α, key hepatocytic hallmarks, were increased. Conclusion:Overexpression of KDM6A could be used as a potential noninvasive molecular therapeutic strategy to prevent metastasis and recurrence rate in HCC.

Hypoimmunogenic pluripotent stem cells (hPSCs) represent a transformative innovation in regenerative medicine, offering solutions to the longstanding challenge of immune rejection in cell-based therapies. Through advanced gene-editing techniques, particularly CRISPR/Cas9, hPSCs are engineered to downregulate or eliminate the expression of major histocompatibility complex (MHC) molecules while upregulating immunomodulatory proteins such as HLA-G, PD-L1, and CD47. These modifications enhance immune evasion and create the foundation for universal donor cells. Compared to conventional cell therapies that rely on lifelong immunosuppression, hPSC-based strategies offer safer, more sustainable, and patient-friendly solutions by minimizing the risks of infection, malignancy, and drug toxicity. Beyond immune compatibility, critical challenges persist, including the risk of tumorigenicity, off-target genetic alterations, and ethical considerations surrounding genome editing. Recent advances, such as the integration of suicide gene systems and sensitive monitoring assays, offer promising strategies to enhance the safety and functional stability of hPSC-derived therapies. This review comprehensively discusses the molecular engineering of hPSCs, their biomedical applications, safety strategies, ethical implications, and the evolving regulatory frameworks needed for clinical translation. By addressing both the scientific and societal dimensions, hPSCs have the potential to revolutionize personalized and off-the-shelf regenerative treatments, provided that rigorous safeguards are implemented.

It has been observed that five members of Secreted Frizzled-Related proteins act as antagonists for the Wnt signaling pathway in humans. These glycoproteins have two functional domains: the cysteine-rich domain (CRD) and the netrin-related domain (NTR), with a completely conserved disulfide bond in the CRD domain. Phylogenetic analysis revealed that this protein family can be divided into two subgroups, SFRP1/SFRP2/SFRP5 versus SFRP3/SFRP4. The SFRP3/SFRP4 group was found to be more closely related to the sponge Lubomirskia baicalensis, which is believed to represent the ancient origin of SFRPs. The model evaluation demonstrated high-quality conformational homology modeling in the predicted Human SFRP models compared to the Sizzled crystal structure of Xenopus laevis. The molecular dynamic simulation illustrated that SFRP1 and SFRP2 exhibit the most stable structures during 100 ns of simulation. Multiple sequence alignment and conservation analysis of Human SFRPs showed that the CRD domain of SFRPs is more conserved than the NTR domain. The docking result indicated that SFRP3 has the highest binding affinity to Wnt3, while SFRP1 and SFRP5 have the lowest. Despite the lower affinity of SFRP1/SFRP5 for Wnt3, a higher positive charge in their NTR domains leads to an increase in their local concentration near the secreting cells and an enhancement in the antagonistic activity. In contrast, SFRP3/SFRP4 can act as an antagonist in distant cells due to less positive regions in their NTR domain and weakly binding to the heparin of the intercellular matrix.

Despite remarkable progress in the clinical management of hepatocellular carcinoma (HCC), complications such as heterogenicity of HCC cells and characteristics of cancer stem cells (CSCs) contribute to frequent relapse and treatment resistance. Lack of proper in vitro models has limited developing novel approaches to evaluate innovative therapeutic settings to overcome these challenges. To address current limitations for mimicking cancer microenvironments; various three-dimensional (3D) platforms have been developed, such as tumoroids, patient-derived xenograft (PDX) models, microfluidics-based cancer chip devices, and bio-printed microtissues. Notably, 3D bio-printing technology has enabled researchers to produce scalable complex multicellular tissue models with accurate matrix composition and cellular organization. These microtissues provide precise platforms studying liver regeneration pathways, fibrosis reversal, and cellular responses to therapeutic interventions. This paper, a systematic literature search of databases covering publications from 2000 to 2025, uniquely highlights how these advances enable precise recapitulation of tumor heterogeneity and microenvironmental complexity, thereby offering transformative platforms for personalized drug screening and elucidating mechanisms of liver tissue repair and regeneration. We discussed current challenges and future directions for translating 3D bio-printed liver models into clinically relevant tools, potentially accelerating therapeutic advances and their potential applications in regenerative medicine in terms of personalized medicine and drug screening.