Search Results (385)

Doxorubicin (DOX) is widely used for the treatment of several tumors, but considerable dose-dependent side effects on many normal tissues, including bones, have been reported. The aim of the present study was to follow for the first time the kinetics of DOX accumulation/clearance in the non-vascularized intervertebral disc (IVD), as well as to assess the drug’s biological action in the annulus fibrosus (AF) and nucleus pulposus (NP) IVD cells and tissues. DOX was administered intravenously to rabbits before the isolation of IVDs, in which DOX quantification was performed using a highly sensitive LC-HRMS/MS analytical method. The effect of the drug on IVD cells’ physiology was assessed in vitro, while IVD tissue quality post-DOX administration was studied in vivo through histological analysis. DOX delivery was found significantly lower in the IVD compared to the highly vascularized skin, declining from the outer AF to the inner NP. The low DOX concentrations reaching the IVDs had marginal effects on cells’ viability, intracellular redox status, and p38 MAPK activation, while they did not evoke cellular senescence. Most importantly, the drug did not negatively affect ECM integrity, as collagen and proteoglycan content remained stable in vitro and in vivo. Full article

Gene therapy is a groundbreaking strategy in regenerative medicine, enabling precise cellular behavior modulation for tissue repair. In situ nucleic acid delivery systems aim to directly deliver nucleic acids to target cells or tissues to realize localized genetic reprogramming and avoid issues like donor cell dependency and immune rejection. The key to success relies on biomaterial-engineered delivery platforms that ensure tissue-specific targeting and efficient intracellular transport. Viral vectors and non-viral carriers are strategically modified to enhance nucleic acid stability and cellular uptake, and integrate them into injectable or 3D-printed scaffolds. These scaffolds not only control nucleic acid release but also mimic native extracellular microenvironments to support stem cell recruitment and tissue regeneration. This review explores three key aspects: the mechanisms of gene editing in tissue repair; advancements in viral and non-viral vector engineering; and innovations in biomaterial scaffolds, including stimuli-responsive hydrogels and 3D-printed matrices. We evaluate scaffold fabrication methodologies, nucleic acid loading–release kinetics, and their biological impacts. Despite progress in spatiotemporal gene delivery control, challenges remain in balancing vector biocompatibility, manufacturing scalability, and long-term safety. Future research should focus on multifunctional “smart” scaffolds with CRISPR-based editing tools, multi-stimuli responsiveness, and patient-specific designs. This work systematically integrates the latest methodological advances, outlines actionable strategies for future investigations and advances clinical translation perspectives beyond the existing literature. Full article

Developing precision models to describe agricultural growth is a necessary step to promote sustainable agriculture and increase resource circulation. In this study, the researchers hydroponically cultivated Bibb lettuce (Lactuca sativa) across a variety of nitrogen, phosphorus, and potassium (NPK)-limited treatments and developed robust data-driven kinetic models observing nutrient uptake, biomass growth, and tissue composition based on all three primary macronutrients. The resulting Dynamic μ model is the first to integrate plant maturity’s impact on growth rate, significantly improving model accuracy across limiting nutrients, treatments, and developmental stages. This reduced error supports this simple expansion as a practical and necessary inclusion for agricultural kinetic modeling. Furthermore, analysis of nutrient uptake refines the ideal hydroponic nutrient balance for Bibb lettuce to 132, 35, and 174 mg L⁻¹ (N, P, and K, respectively), while qualitative cell yield analysis identifies minimum nutrient thresholds at approximately 26.2–41.7 mg-N L⁻¹, 3.7–5.6 mg-P L⁻¹, and 17.4–31.5 mg-K L⁻¹ to produce compositionally healthy lettuce. These findings evaluate reclaimed wastewater’s ability to offset the fertilizer burden for lettuce by 23–45%, 14–57%, and 3–23% for N, P, and K and guide the required minimum amount of wastewater pre-processing or nutrient supplements needed to completely fulfill hydroponic nutrient demands. Full article

In the context of critical challenges in curcumin-modified polyurethane synthesis—including limited curcumin bioavailability and suboptimal biodegradability/biocompatibility—a novel polyurethane material (Cur-PU) with good mechanical, shape memory, pH-responsive, and biocompatibility was synthesized via a one-pot, two-step synthetic protocol in which HO-PCL-OH served as the soft segment and curcumin was employed as the chain extender. The experimental results demonstrate that with the increase in Cur units, the crystallinity of the Cur-PU material decreases from 32.6% to 5.3% and that the intensities of the diffraction peaks at 2θ = 21.36°, 21.97°, and 23.72° in the XRD pattern gradually diminish. Concomitantly, tensile strength decreased from 35.5 MPa to 19.3 MPa, and Shore A hardness declined from 88 HA to 65 HA. These observations indicate that the sterically hindered benzene ring structure of Cur imposes restrictions on HO-PCL-OH crystallization, leading to lower crystallinity and retarded crystallization kinetics in Cur-PU. As a consequence, the material’s tensile strength and hardness are diminished. Except for the Cur-PU-3 sample, all other variants exhibited exceptional shape-memory functionality, with R_f and R_r exceeding 95%, as determined by three-point bending method. Analogous to pure curcumin solutions, Cur-PU solutions demonstrated pH-responsive chromatic transitions: upon addition of hydroxide ion (OH⁻) solutions at increasing concentrations, the solutions shifted from yellow-green to dark green and finally to orange-yellow, enabling sensitive pH detection across alkaline gradients. Hydrolytic degradation studies conducted over 15 weeks in air, UPW, and pH 6.0/8.0 phosphate buffer solutions revealed mass loss <2% for Cur-PU films. Surface morphological analysis showed progressive etching with the formation of micro-to-nano-scale pores, indicative of a surface-erosion degradation mechanism consistent with pure PCL. Biocompatibility assessments via L929 mouse fibroblast co-culture experiments demonstrated ≥90% cell viability after 72 h, while relative red blood cell hemolysis rates remained below 5%. Collectively, these findings establish Cur-PU as a biocompatible material with tunable mechanical properties, and pH responsiveness, underscoring its translational potential for biomedical applications such as drug delivery systems and tissue engineering scaffolds. Full article

CSNPs synthesized via the ionic gelation method have emerged as a promising nanoplatform in diverse fields such as pharmaceuticals, nanotechnology, and polymer science due to their biocompatibility, ease of fabrication, and tunable properties. This study focuses on the development and characterization of LL37-loaded CSNPs, designed to enhance antibacterial efficacy while maintaining biocompatibility. This study pioneers a systematic loading optimization approach by evaluating the encapsulation efficiency (%EE) of antimicrobial peptide LL37 across multiple concentrations (7.5, 15, and 30 µg/mL), thereby identifying the formulation that maximizes peptide incorporation while preserving controlled release characteristics. The multi-concentration analysis establishes a new methodological benchmark for peptide delivery system development. To achieve this, CSNPs were optimized for size and stability by adjusting parameters such as the chitosan concentration, pH, and stabilizer. LL37, a potent antimicrobial peptide, was successfully encapsulated into CSNPs at concentrations of 7.5, 15, and 30 µg/mL, yielding formulations with favorable physicochemical properties. Dynamic light scattering (DLS) and Zeta sizer analyses revealed that blank CSNPs exhibited an average particle size of 180.40 ± 2.16 nm, a zeta potential (ZP) of +40.57 ± 1.82 mV, and a polydispersity index (PDI) of 0.289. In contrast, 15-LL37-CSNPs demonstrated an increased size of 210.9 ± 2.59 nm with an enhanced zeta potential of +51.21 ± 0.93 mV, indicating an improved stability and interaction potential. Field emission scanning electron microscopy (FE-SEM) analyses exhibited the round shaped morphology of nanoparticles. The release profile of LL37 exhibited a concentration-dependent rate and showed the best fit with the first-order kinetic model. Cytocompatibility assessments using the XTT assay confirmed that both blank and LL37-loaded CSNPs did not exhibit cytotoxicity on keratinocyte cells across a range of concentrations (150 µg/mL to 0.29 µg/mL). Notably, LL37-loaded CSNPs demonstrated significant antibacterial activity against E. coli and S. aureus, with the 15-LL37-CSNP formulation exhibiting superior efficacy. Overall, these findings highlight the potential of LL37-CSNPs as a versatile antibacterial delivery system with applications in drug delivery, wound healing, and tissue engineering. Full article

Oxytetracycline (OTC) has served as an antibiotic to treat various bacterial infections in fish raised in aquaculture. Nonetheless, administering OTC in overly high doses can lead to adverse side effects in fish and also negatively impact on their surroundings. The objective of this work was to evaluate the expression levels of immune markers such as TLR-1, TLR-2, IκB-α, MyD88, NF-κB, IFN-γ, and IL-6 in intestinal cell primary culture (foregut, midgut, and hindgut) using qRT-PCR, and in addition, to assess the in vivo response to different doses of OTC in coho salmon at different times. The expression levels of all genes increased significantly after 1 h on day 1 with high doses of OTC compared with control conditions in all tissues under both approaches (in vivo and in vitro). However, the transcriptional responses decreased to 3, 6, and 12 h in vitro and day 3 in vivo. In conclusion, the transcriptional responses were differentially modulated by OTC in the three intestinal portions under both experimental conditions. These results demonstrate for the first time in primary cell culture fish that the expression of immune biomarkers in all tissues induces a differential response of these genes, depending on the concentration of OTC and the kinetics of time. This study offers valuable insights that can be applied to enhance aquaculture, determine optimal drug doses, and improve fish health. Full article

This study aimed to design dual-responsive chitosan–polylactic acid nanosystems (PLA@CS NPs) for controlled and targeted ledipasvir (LED) delivery to HepG2 liver cancer cells, thereby reducing the systemic toxicity and improving the therapeutic selectivity. Two formulations were developed utilizing ionotropic gelation and w/o/w emulsion techniques: LED@CS NPs with a size of 143 nm, a zeta potential of +43.5 mV, and a loading capacity of 44.1%, and LED-PLA@CS NPs measuring 394 nm, with a zeta potential of +33.3 mV and a loading capacity of 89.3%, with the latter demonstrating significant drug payload capacity. Since most drugs work through interaction with DNA, the in vitro affinity of DNA to LED and its encapsulated forms was assessed using stopped-flow and other approaches. They bind through multi-modal electrostatic and intercalative modes via two reversible processes: a fast complexation followed by a slow isomerization. The overall binding activation parameters for LED (cordination affinity, K_a = 128.4 M⁻¹, K_d = 7.8 × 10⁻³ M, ΔG = −12.02 kJ mol⁻¹), LED@CS NPs (K_a = 2131 M⁻¹, K_d = 0.47 × 10⁻³ M, ΔG = −18.98 kJ mol⁻¹) and LED-PLA@CS NPs (K_a = 22026 M⁻¹, K_d = 0.045 × 10⁻³ M, ΔG = −24.79 kJ mol⁻¹) were obtained with a reactivity ratio of 1/16/170 (LED/LED@CS NPs/LED-PLA@CS NPs). This indicates that encapsulation enhanced the interaction between the DNA and the LED-loaded nanoparticle systems, without changing the mechanism, and formed thermodynamically stable complexes. The drug release kinetics were assessed under tumor-mimetic conditions (pH 5.5, 10 mM GSH) and physiological settings (pH 7.4, 2 μM GSH). The LED@CS NPs and LED-PLA@CS NPs exhibited drug release rates of 88.0% and 73%, respectively, under dual stimuli over 50 h, exceeding the release rates observed under physiological conditions, which were 58% and 54%, thereby indicating that the LED@CS NPs and LED-PLA@CS NPs systems specifically target malignant tissue. Release regulated by Fickian diffusion facilitates tumor-specific payload delivery. Although encapsulation did not enhance the immediate cytotoxicity compared to free LED, as demonstrated by an in vitro cytotoxicity in HepG2 cancer cell lines, it significantly enhanced the therapeutic index (2.1-fold for LED-PLA@CS NPs) by protecting non-cancerous cells. Additionally, the nanoparticles demonstrated broad-spectrum antibacterial effects, suggesting efficacy in the prevention of chemotherapy-related infections. The dual-responsive LED-PLA@CS NPs allowed controlled tumor-targeted LED delivery with better selectivity and lower off-target toxicity, making LED-PLA@CS NPs interesting candidates for repurposing HCV treatments into safer cancer nanomedicines. Furthermore, this thorough analysis offers useful reference information for comprehending the interaction between drugs and DNA. Full article

Background/Objectives: Largemouth bass rhabdovirus (Micropterus salmoides rhabdovirus, MSRV) disease causes high mortality in largemouth bass farming. Therefore, vaccine development is critical for largemouth bass prevention against MSRV. Methods: An attenuated strain, denoted as MSRV-0509, was selected through intraperitoneal injection and immersion challenge assays, followed by plaque purification. The biological characteristics of MSRV-0509, including optimal inoculation dose, replication kinetics, thermostability, pH resistance, chloroform tolerance, and storage viability, were determined via viral titration. Spatiotemporal distribution patterns in largemouth bass post-intraperitoneal injection or immersion infection were quantified by qPCR. Immunoprotective efficacy was evaluated through intraperitoneal and immersion vaccination. Mechanistic insights were explored via relative qPCR and serum neutralization assays. Safety was assessed by single-dose overdose immunization and virulence reversion experiments. Results: An attenuated strain MSRV-0509 was screened through a challenge assay, exhibiting complete avirulence in largemouth bass compared to the virulent strain SCRV-T6. MSRV-0509 demonstrated optimal replication at low MOI (0.0001) in CPB cells, with peak titers (10^8.3 TCID₅₀/mL) at 96 h post-infection. The virus showed susceptibility to high temperatures, lipid solvents and acidic conditions, with prolonged stable storage viability at −80 °C. Tissue distribution revealed the spleen as the primary target after intraperitoneal injection, while immersion restricted infection to gills, with rapid clearance by 3–6 dpi. Vaccination trials identified 5 × 10² TCID₅₀/fish via intraperitoneal injection and 10^6.0 TCID₅₀/mL via immersion as effective immunizing doses, providing 100% relative survival post-challenge. Immune gene expression and serum neutralization showed Th1 and Th2 activation via intraperitoneal injection (elevated IL-12, IFN-γ, IL-10, IgM), whereas only the Th1 response was activated after vaccine immersion. No abnormality and mortality were observed in single overdose vaccination and virulence reversion experiments, confirming that MSRV-0509 was safe. Conclusions: These results proved that MSRV-0509 could be a promising vaccine candidate to protect largemouth bass from MSRV disease. Full article

Stem-cell-based therapies rely on the transplantation of stem cells or stem-cell-derived organotypic cells into injured tissues in order to improve or restore tissue function that has been impaired by various diseases. The potential of induced pluripotent stem cells has created many applications in the field of cell therapy, for example. Some applications, for example, those in cardiac cell therapy, suffer from low or very low efficiencies of cell engraftment. Therefore, magnetic cell targeting can be discussed as a method for capturing superparamagnetic nanoparticle-labelled cells in the tissue. Here, we employ superparamagnetic iron oxide nanoparticles (SPIONs) for the intracellular magnetic loading of mesenchymal stem cells (MSCs). In addition, we test a novel strategy of labelling MSCs with ferromagnetic particles. The adhesion assays demonstrate a faster adhesion kinetic of SPIONs-loaded MSC spheroids when a magnetic field was applied, resulting in >50% spheroid adhesion after 30 min. Clustering of cells inside the magnetic field is a second potential mechanism of magnetic cell retention and >80% of cells were found to be aggregated in clusters when placed in a magnetic field for 10 min. SPIONs-loaded and ferromagnetic-particle-loaded cells performed equally in the cell clustering assay. In conclusion, the clustering of SPION-labelled cells explains the observation that magnetic targeting reaches maximal efficiency in vivo after only 10 min of magnetic field application. This has significant implications for magnetic-targeting-assisted stem cell and cell replacement therapies. Full article

Ortho-R (ChitogenX Inc., Kirkland, QC, Canada) is a lyophilized chitosan formulation that also contains calcium chloride and trehalose. Ortho-R was designed to be solubilized in autologous platelet-rich plasma (PRP), a blood-derived component, in order to become an injectable implant that augments the surgical repair of soft tissues. The Ortho-R/PRP formulation coagulates post-application, similarly to blood. Having the ability to predict the speed of coagulation of an Ortho-R/PRP mixture prepared with PRP isolated from a specific patient would be an advantage in the operating room. The purpose of this study was to investigate whether human donor characteristics (age, sex, body mass index, habits) and autologous PRP properties would have an impact on Ortho-R/PRP mixture coagulation. Clot maximal amplitude and shear elastic modulus were significantly positively correlated with body mass index and platelet concentration in the isolated PRPs. Clot formation time, maximal amplitude and shear elastic modulus were all negatively correlated with PRP red blood cell concentration (and associated hemoglobin and hematocrit content). Donor characteristics were not good predictors of coagulation kinetics in Ortho-R/PRP mixtures. Some of the isolated PRP properties were better predictors of Ortho-R/PRP coagulation kinetics. However, predicting how an Ortho-R/PRP mixture from a particular patient will coagulate is very difficult since all PRP isolation devices yield heterogeneous PRPs and analysis of the isolated PRPs occurs post-administration. Full article

Background: Three-dimensional (3D) cell models may bridge the gap between two-dimensional (2D) cell cultures and animal models. Technical advances have led to the development of 3D-bioprinted cell models, characterized by greater reproducibility and the ability to mimic in vivo conditions. Glioblastoma multiforme (GBM) is a highly aggressive brain tumor with poor clinical outcomes due to its heterogeneity, angiogenic activity, and invasiveness. Src family kinases (SFKs) play a crucial role in GBM progression, making them attractive targets for drug development. Here, we show results about the pharmacological profile of a new prodrug synthesized from a Src inhibitor, SI306. Methods: Three-dimensional-bioprinted GBM cell models were used in predicting the antitumor activity of the prodrug SI306-PD2 with respect to its precursor, SI306. Results: Since the prodrug releases the active inhibitor through the cleavage by specific enzymes, SI306-PD2 was analyzed for stability and release kinetics in various media, including fetal bovine serum (FBS), which is normally used in cell culture. In comparison to SI306, SI306-PD2 demonstrated higher solubility in water, higher permeability across gastrointestinal and blood–brain barrier membranes, and the ability to release the drug in the presence of FBS progressively. In the 2D GBM cell model, using U87 and U251 cell lines, both compounds similarly reduced tumor cell viability. In 3D-bioprinted cell models, in the presence of an FBS-free medium, SI306-PD2 exhibited a more effective antitumor activity compared to SI306, reducing the proliferation and diameter of U251 spheroids grown within the bioprinted scaffold in a statistically significant manner. The analysis of proteins extracted from 3D scaffolds confirmed that SI306-PD2 inhibited Src activation more efficiently than SI306. Conclusions: Our study suggests that, when tissue permeability represents a discriminating characteristic, bioprinted cell models can provide a valid alternative for studying the cytotoxicity of new antitumor compounds. This approach has permitted us to ascertain the potential of the prodrug SI306-PD2 as a therapeutic agent for GBM, demonstrating better tissue penetration and antiproliferative efficacy compared to the precursor compound SI306. Full article

Background: Birnaviruses infect a wide range of aquatic and terrestrial hosts, including several economically important fish species. This study aimed to isolate and characterize a novel birnavirus strain from mandarin fish (Siniperca chuatsi), a high-value freshwater species in Chinese aquaculture. Methods: A novel strain, designated mandarin fish birnavirus (MFBV), was isolated from diseased fish and propagated in SCK cells. The complete genome was determined using high-throughput sequencing and RACE. Viral replication kinetics, tissue distribution, and pathogenicity were assessed through in vitro infection, RT-qPCR, histopathology, and experimental challenges. In addition, disinfectant sensitivity and environmental stability were evaluated. Results: The MFBV genome comprises two segments (A: 3539 bp; B: 2719 bp), and phylogenetic analysis revealed close relatedness to largemouth bass birnavirus (LBBV) and Lates calcarifer birnavirus (LCBV). MFBV displayed rapid replication in SCK cells, completing a replication cycle in 8–10 h. In juvenile and fry fish, an experimental infection caused acute disease with cumulative mortality ranging from 41.8% to 83.6%, with fry showing higher susceptibility. Viral RNA was detected in multiple tissues (7.9 × 10⁶–7.9 × 10⁷ copies/μg RNA), and histopathological lesions were observed in the intestine, spleen, and kidney. MFBV was highly sensitive to glutaraldehyde (20 ppm), while other disinfectants showed reduced efficacy. Viral half-life ranged from 36.5 to 144.5 h at room temperature. Conclusions: These findings demonstrate that MFBV can induce acute systemic infection in mandarin fish. The results offer new insights into the genomic and biological features of birnaviruses, contributing to improved disease management and viral taxonomy. Full article

Background/objective: Mucinous borderline tumors of the ovary (MBOTs) are characterized by their unique histological features and intermediate malignant potential; however, the factors underlying their molecular carcinogenesis and tumor biology remain largely unknown. Developing cell lines from these tumors presents an ongoing challenge. The purpose of this study is to establish MBOT cell lines and characterize their biological features. Methods: Epithelial cells were collected and purified from surgically removed MBOT samples and then stably maintained with an extended life span by overexpressing CyclinD1/CDK4 in combination with human telomerase reverse transcriptase. The characterization of resulting cell lines was defined by morphology, growth kinetics, functional analysis, whole-exome sequencing, and tumorigenicity in mice. Results: Two independent cell lines, HMucBOT-1 and HMucBOT-2, were successfully established from the tissues of a patient with an MBOT, with the latter showing more aggressive growth capacity. In the patient-derived xenograft model, HMucBOT-1 cells retained the original morphological characteristics of the MBOT, whereas HMucBOT-2 cells displayed a transition to mucinous carcinoma accompanying undifferentiated carcinoma, suggestive of dedifferentiated carcinoma. Genetic analysis of the original tumor sample and HMucBOT-2 cells revealed shared oncogenic mutations. However, KRAS amplification and certain copy number alterations were uniquely observed in the HMucBOT-2 cells. Conclusions: The above results indicate that HMucBOT-1 can serve as a preclinical model for investigating the biological behavior of and potential targeted therapies for human MBOTs, with HMucBOT-2 serving as a valuable tool for studying the heterogeneity and genetic diversity of this tumor and explaining the potential causes of treatment failure or relapse. Full article

Effective molecular imaging and targeted cancer therapy rely on receptor-specific targeted delivery systems that are both metabolically stable and kinetically inert for optimal in vivo performance. Until now, no single metal complexing agent has demonstrated the versatility to coordinate metals across the periodic table while maintaining the kinetic inertness required for clinical theranostic applications. Therefore, enhancing the in vivo kinetic stability of radiolabeled, cell-targeting, biologically active compounds remains a critical goal to minimize unintended accumulation of radioactivity in collateral tissues. This review describes the usage of NOTA [NOTA = 1,4,7-triazacyclononane-1,4,7-triacetic acid] and derivatives of NOTA, a metal complexing agent that has been found to have the ability to effectively coordinate with a wide range of radiometals, including metal-radiohalogens, to form stable complexes. This enables the development of new cell-targeting small molecule and peptide conjugates with the potential to resist demetallation in vivo, thereby reducing radionuclide uptake in non-target tissues. Herein, we discuss the design and development of NOTA-based, cell-targeting, small molecules having very high affinity and selectivity for the GRPR (Gastrin-Releasing Peptide Receptor), the SSTR2 (Somatostatin Receptor Subtype 2), and the MC1R (Melanocortin-1) receptors that are present on the surfaces of numerous solid primary human tumors and their metastatic counterparts. Full article

With the concurrent circulations of SARS-CoV-2 omicron and influenza A viruses in the community, there is evidence showing co-infection with both viruses. However, disease severity may vary due to the complex immunity landscape of the patients and the neutralizing antibody waning status. The intrinsic dynamic relationship and pathological significance for such co-infections remain largely unknown. The replication kinetics and innate immune responses from the co-infections of SARS-CoV-2 (Omicron BA.1 and D614G variant) and influenza A viruses (pandemic H1N1, seasonal H3N2 and highly pathogenic avian H5N1) were characterized in human respiratory tissue explants, human airway, and alveolar epithelial cells. SARS-CoV-2 reduced the replication of influenza A viruses, but not vice versa, during co-infections in human bronchial tissues and airway epithelial cells. In lung tissues, the co-infections showed minimal effects on each other, but the viral replications of the two viruses were mutually reduced except for H1N1pdm in the alveolar epithelial cells irrespective of the enhancement of the ACE2 receptor. Notably, the co-infections showed a significant upregulation of the innate immune responses of SARS-CoV-2 in comparison to single infections in both respiratory epithelial cells, suggesting that co-infections of influenza A viruses potentially lead to more severe damage to the host than SARS-CoV-2 single infections. Full article