Online First Articles

Abstract: Background: A suitable biomaterial scaffold is essential for building in vitro ovarian models. This study developed a GelMA/HAMA (GH) hydrogel to support granulosa cell culture and to construct a simplified coculture system for granulosa cell–endothelial cell interaction. Methods: GH hydrogel was characterized by rheology, tensile testing, scanning electron microscopy, swelling, degradation, and extract-based cytocompatibility. KGN granulosa cells were encapsulated in GH hydrogel, and an aging-like model was induced with H₂O₂. A Transwell system and conditioned-medium experiments were used to evaluate endothelial cell responses. Results: GH hydrogel showed light-triggered gelation, predominantly elastic behavior, higher tensile performance than GelMA, lower swelling, and slower degradation, with acceptable cytocompatibility at the tested range. In KGN-laden hydrogels, H₂O₂ treatment increased ROS and p16 expression while decreasing E2 secretion and StAR expression. Conditioned medium from aging granulosa cells increased ROS and VCAM-1 and reduced HUVEC viability. Conclusion: GH hydrogel provides a controllable matrix for granulosa cell culture and a useful in vitro platform for studying aging-related granulosa cell–endothelial cell crosstalk.

Abstract: Background: The liver tumor microenvironment is characterized by elevated reactive oxygen species (ROS), high glutathione (GSH) levels, and acidic pH, which limits the selectivity and efficacy of conventional chemotherapy. Microenvironment-responsive drug delivery systems offer a promising strategy to address these challenges. Methods: A redox-responsive thioketal-crosslinked gelatin hydrogel (TK-Gel) was prepared via dynamic covalent crosslinking and used to encapsulate doxorubicin (DOX). The rheological properties, swelling and degradation behaviors, redox responsiveness, and microenvironment-dependent drug release were systematically evaluated. In vitro antitumor performance was assessed using HepG2 liver cancer cells. Results: The TK-Gel hydrogel formed a stable and highly hydrated network under physiological conditions, while exhibiting accelerated degradation and enhanced DOX release under tumor-mimicking environments with elevated ROS and GSH. Cellular studies demonstrated good biocompatibility of the blank hydrogel and significantly enhanced cytotoxicity of DOX@TK-Gel under redox-activated conditions, accompanied by partial intracellular ROS consumption. Conclusions: Thioketal-crosslinked gelatin hydrogels enable tumor-selective drug release and redox modulation, providing a promising platform for liver cancer therapy.

Abstract: Background: Reactive oxygen species (ROS)–induced oxidative stress contributes to neuronal injury during ischemic conditions, creating a need for delivery systems that can release therapeutic molecules in response to oxidative cues. Incorporating thioketal linkages into polymeric materials provides a feasible strategy to construct ROS-degradable carriers. Methods: In this study, a triblock copolymer poly(ε-caprolactone)–thioketal–poly(ε-caprolactone) (PCL–PTK–PCL) was synthesized via ring-opening polymerization using thioketal diol as the initiator. The polymer self-assembled into nanocarriers capable of encapsulating nerve growth factor (NGF). The structural characteristics were analyzed by FTIR and TEM, while the degradation and release behaviors were evaluated under various H₂O₂ concentrations. Cytocompatibility and neuronal viability were assessed using PC12 cells. Results: The PCL–PTK–PCL nanocarriers exhibited uniform spherical morphology with an average size of ~100 nm. The presence of thioketal bonds conferred clear ROS sensitivity, as evidenced by H₂O₂-triggered swelling and accelerated NGF release. The carriers remained stable under non-oxidative conditions and showed good cytocompatibility, maintaining high neuronal cell viability after incubation. Conclusion: The synthesized PCL–PTK–PCL nanocarriers achieved ROS-triggered degradation and controlled NGF release while exhibiting minimal cytotoxicity. These findings confirm their suitability as a basic oxidation-responsive platform for further exploration in oxidative stress–related neuronal studies.

Abstract: Background: Oxidative stress is a major contributor to granulosa cell dysfunction and follicular atresia, with excessive reactive oxygen species (ROS) impairing mitochondrial activity and cell survival. Hydrogels based on gelatin methacryloyl (GelMA) are biocompatible and easily photocrosslinked, but they lack intrinsic antioxidant function. Methods: A GelMA–tannic acid (TA) hydrogel was synthesized by visible light curing in the presence of TA. Structural features were confirmed by FT-IR and ¹H NMR. Photorheology was used to determine gelation kinetics, and scanning electron microscopy assessed morphology. Antioxidant performance was evaluated by DPPH, ABTS, FRAP, and H₂O₂ assays. The protective effect on human granulosa-like KGN cells under H₂O₂ stress was examined by CCK-8 viability, DCFH-DA ROS detection, JC-1 mitochondrial membrane potential, and Live/Dead staining. Results: GelMA–TA exhibited a rapid gelation time of 8.5 s and a plateau modulus of 1.8 kPa, higher than the 1.5 kPa modulus of GelMA alone. In antioxidant assays, GelMA–TA showed significant radical scavenging activity with DPPH (86.1%), ABTS (91.4%), and FRAP (1.15 mmol Fe²⁺ equivalent), as well as nearly complete H₂O₂ removal (94.8%). In KGN cells, GelMA–TA reduced intracellular ROS by 60%, restored mitochondrial membrane potential (Δψm) to 0.9 (compared to 0.5 for H₂O₂ treatment), and improved cell viability by 30%. Conclusion: These findings demonstrate that GelMA–TA forms a fast-curing, antioxidant hydrogel capable of maintaining a low-ROS microenvironment and protecting granulosa cells, offering a promising platform for ovarian tissue engineering and related regenerative applications.

Abstract: Background: The fabrication of vascular-mimetic hydrogel scaffolds with precise luminal geometry, suitable elasticity, and good cytocompatibility remains a major challenge in tissue engineering. Digital light processing (DLP) printing offers high resolution and rapid fabrication, but over-curing and limited structural fidelity in hollow constructs still restrict its application in vascular-like scaffold fabrication. Methods: In this study, GelMA-based tubular scaffolds with Y-shaped and curved vascular geometries were fabricated by DLP printing. To improve printing precision, 0.02% (w/v) tartrazine was introduced as a light-absorbing agent to regulate light penetration and curing depth during the photopolymerization process. The printed scaffolds were characterized by optical imaging and scanning electron microscopy, while their mechanical properties were evaluated through tensile and compression tests. Cytocompatibility was assessed using CCK-8 assay, Live/Dead staining, and quantitative cell survival analysis. Results: The incorporation of tartrazine effectively reduced excessive light penetration during printing, enabling the formation of continuous tubular structures with improved lumen definition and structural integrity. The DLP-printed GelMA scaffolds showed high geometric fidelity, with smooth and stable inner channels of approximately 1 mm in diameter. Mechanical testing demonstrated a nonlinear J-shaped tensile response and strain-stiffening behavior under compression, indicating favorable elasticity and resistance to deformation. In vitro biological evaluation further showed high cell viability, with CCK-8 results remaining above the control level and Live/Dead staining confirming a predominance of viable cells on the scaffold surface. Conclusion: The incorporation of a small amount of photoabsorber enabled high-resolution DLP printing of vascular-like GelMA scaffolds with excellent mechanical flexibility and biocompatibility. These constructs hold strong potential as perfusable, elastic hydrogel platforms for microvascular regeneration, endothelialization studies, and organ-on-chip applications.

Abstract: Background: Thermosensitive hydrogels have been widely investigated for localized drug delivery; however, their in vitro physicochemical stability, degradation behavior, and cell-level anti-inflammatory performance require systematic evaluation before further translational studies. In particular, prostate epithelial inflammation remains an underexplored application scenario for such delivery platforms. Methods: In this study, a PLGA–PEG–PLGA thermosensitive hydrogel was evaluated as a sustained delivery system for dexamethasone. The copolymer was characterized by GPC, FTIR, and ¹H NMR. Sol–gel transition behavior and viscoelastic properties were assessed using micro-DSC and rheological analysis. The microstructure of the drug-loaded hydrogel was examined by SEM. Degradation behavior was investigated under physiological (PBS, 37°C) and accelerated alkaline conditions. Cytocompatibility and anti-inflammatory effects were evaluated in RWPE-1 prostate epithelial cells. Results: SEM revealed an interconnected porous network structure in the drug-loaded hydrogel. Degradation studies showed high structural stability in PBS with minimal mass loss over 14 days, while rapid degradation occurred under alkaline conditions, confirming hydrolytic degradability. The hydrogel enabled sustained dexamethasone release and maintained good cytocompatibility. Notably, dexamethasone-loaded hydrogels significantly reduced IL-6 and IL-8 secretion, whereas blank hydrogels showed no intrinsic anti-inflammatory effect. Conclusion: This work provides a comprehensive in vitro evaluation of a thermosensitive PLGA–PEG–PLGA hydrogel for sustained dexamethasone delivery at the cellular level. The results clarify the relationship between hydrogel microstructure, degradation behavior, and diffusion-dominated drug release, establishing a solid foundation for future in vivo investigations.

Abstract: Background: Recurrent pregnancy loss (RPL) associated with endometrial dysfunction remains clinically challenging due to the lack of localized, multifunctional therapeutic strategies. Restoring endometrial receptivity, vascularization, and immune balance is key to successful intervention. Methods: An injectable bioactive hydrogel was developed by crosslinking aldehyde-modified hyaluronic acid (HA-CHO) with chitosan. The hydrogel was loaded with LIF (Leukemia Inhibitory Factor), VEGF (Vascular Endothelial Growth Factor), IL-11 (Interleukin-11), and valproic acid to enhance regenerative activity. Its effects were assessed in vitro via cell proliferation (CCK-8), tube formation (HUVEC assay), cytokine expression (THP-1 qPCR), and endometrial gene profiling (hEMSC qPCR). Results: The hydrogel exhibited rapid gelation, good biocompatibility, and factor-loading capacity. It significantly enhanced hEMSC proliferation and HUVEC tube formation. Pro-inflammatory cytokines (TNF-α, IL-6) were downregulated, while IL-10 (Interleukin-10) was upregulated in macrophages. The hydrogel also increased expression of LIF and IGFBP1 (Insulin-like Growth Factor Binding Protein 1), but not PRL, indicating enhanced receptivity without full decidualization. Conclusion: This HA-CHO/chitosan hydrogel supports endometrial regeneration through coordinated promotion of proliferation, angiogenesis, immune modulation, and receptivity. It holds strong potential for localized treatment of RPL with endometrial insufficiency.

Abstract: This in vitro study performed a comparative mechanical evaluation of three classes of direct restorative materials: resin composites, polyacid-modified composite resins (components), and Glass Ionomer Cements (GICs). The objective was to clarify the mechanical position of components relative to composites and glass ionomer cements (GICs). Specimens from two materials per class including the components Dyract AP and Dyract Extra were fabricated using standardized molds. Compressive, diametral tensile, and three-point flexural strength tests were conducted according to ADA and ISO specifications, followed by statistical analysis using ANOVA and Tukey’s HSD test. Results established a consistent mechanical hierarchy: composite resins exhibited the highest strength values in all tests, followed by components, with GICs demonstrating the lowest. Within the component group, the thirdgeneration Dyract Extra showed superior properties compared to Dyract AP. The findings confirm that components occupy an intermediate mechanical position, offering significantly greater strength than GICs but not matching composites. Thus, components suit low-to-moderate stress applications where fluoride release and handling ease are beneficial, while composites remain indicated for high-stress areas. This study provides clear evidence for evidence-based clinical material selection.

Abstract: Background: Gastric cancer remains one of the leading causes of cancer-related deaths worldwide, and the development of effective, targeted drug delivery systems is crucial to improve therapeutic outcomes. Graphene oxide (GO)-based nanocarriers have shown promise for controlled drug release, yet their biological evaluation remains limited. Methods: We synthesized a composite nanoparticle system by electrostatic self-assembly of chitosan (CS) onto graphene oxide (GO), followed by doxorubicin (DOX) loading. The resulting GO–CS–DOX nanoparticles were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential, and pHresponsive release profiles. Preliminary biological performance was evaluated in gastric cancer cells (AGS), including dose–response cytotoxicity and fluorescence-based uptake studies. Results: GO–CS– DOX nanoparticles showed a clear pH-dependent DOX release behavior, with accelerated release under mildly acidic conditions. DLS and zeta potential measurements confirmed successful drug loading and changes in surface charge. In vitro, GO–CS–DOX demonstrated comparable or slightly enhanced cytotoxicity relative to free DOX at specific concentrations. Cellular uptake of DOX was observed under acidic conditions, consistent with lysosomal trafficking. However, only preliminary in vitro data were collected and no mechanistic apoptosis studies were performed. Conclusion: This study presents the design and initial evaluation of a pH-responsive GO–CS–DOX nanocarrier. While the in vitro results indicate potential for controlled drug release and tumor-targeted delivery, the biological findings are still limited and should be interpreted as preliminary. Further in-depth studies, including apoptosis assays and in vivo validation, are necessary to fully establish therapeutic efficacy.

Abstract: Background: Postoperative wound sealing and localized drug delivery are critical needs in nasopharyngeal carcinoma (NPC) management. Natural polysaccharide-based films offer a biocompatible platform for addressing these challenges. Methods: A composite film was prepared by Schiff base crosslinking of chitosan and oxidized pullulan. The film was characterized by FTIR, and its adhesion, drug release, cytocompatibility, and effects on cell migration were evaluated using in vitro assays. Results: The composite film exhibited a distinct C=N peak in FTIR spectra and significantly enhanced wet adhesion (55 kPa) compared to individual components. Cisplatin-loaded films showed sustained release over 72 h and reduced the viability of CNE-2 cells to 28%. The drug-free film was non-cytotoxic. Extracts from the composite film promoted nasopharyngeal epithelial cell migration, as shown by RTCA assay. Conclusion: This study explored the in vitro characteristics of a chitosan/oxidized pullulan film and evaluated its basic biological performance at the cellular level capable of localized drug release and supporting cell-level healing responses. Further validation in more complex models is warranted.

Abstract: Background: Antiphospholipid syndrome (APS) is a major cause of pregnancy morbidity, characterized by antiphospholipid antibody–induced platelet activation and placental thrombosis. Integrin αIIbβ3 is a key mediator of platelet aggregation in this pathological process. Methods: We developed PEGDA-based hydrogels functionalized with acrylated RGD peptides to competitively bind αIIbβ3 and inhibit platelet aggregation. The effects of hydrogel and αIIbβ3 antibody treatments were evaluated in vitro using integrin-binding assays, turbidity-based platelet aggregation and clotting assays, ELISA for cytokine release, and transwell migration of trophoblasts. Results: RGD-modified hydrogels showed dose-dependent binding to αIIbβ3. Both hydrogel and antibody treatments significantly reduced antiphospholipid antibodies (aPL)-induced platelet aggregation, delayed fibrin clot formation, and suppressed IL-6 and TNF-α release from trophoblasts. Cell migration assays confirmed that these interventions preserved trophoblast motility impaired by aPL exposure. Conclusion: Targeting integrin αIIbβ3 with RGD-functionalized biomaterials or neutralizing antibodies effectively attenuated aPL-mediated thromboinflammation. This approach represents a promising localized intervention strategy to prevent APS-related pregnancy complications.

Abstract: Background: Alzheimer’s disease (AD) progresses silently, with pathological changes such as amyloid-beta (Aβ) accumulation occurring years before cognitive symptoms emerge. Early intervention is considered critical to modifying disease progression. However, conventional therapeutics often lack pathological specificity and cause systemic side effects, highlighting the need for disease-responsive strategies. Methods: We designed a BACE1-cleavable peptide precursor that self-assembles into nanofibers in Aβ-rich environments. Enzyme-triggered assembly was characterized using dynamic light scattering (DLS) and Thioflavin T (ThT) fluorescence. Neuroprotective effects were assessed via MTT assay and qPCR of inflammatory markers. Results: The responsive peptide formed stable nanostructures upon BACE1 activation, significantly suppressed Aβ aggregation, improved neuronal viability, and reduced IL-1β expression. Conclusion: This study validates the use of enzyme-responsive peptide nanofibers as a selective and multifunctional therapeutic strategy for early intervention in Alzheimer’s disease, combining disease-environment activation with anti-aggregation, anti-inflammatory, and cognitive protective effects.

Abstract: Background: Parkinson’s disease involves progressive dopaminergic neuron degeneration and elevated oxidative stress. Targeted delivery of neurotrophic factors like glial cell line-derived neurotrophic factor (GDNF) remains a therapeutic challenge due to the need for site-specific, stimulusresponsive release. Methods: We developed a reactive oxygen species (ROS)-responsive hydrogel designed to release GDNF upon exposure to oxidative signals. The hydrogel’s degradation behavior and release kinetics were assessed under physiological and oxidative conditions. Bioactivity of the released GDNF was evaluated in vitro using ROS-damaged neuronal cells. Results: The hydrogel showed high stability under normal conditions but degraded rapidly in the presence of H₂O₂, enabling dose-dependent GDNF release. Released GDNF retained biological activity, promoting neuronal survival and neurite extension. Conclusion: This ROS-responsive hydrogel represents a promising platform for oxidative stress-triggered neurorepair and site-specific neuromodulation in Parkinson’s disease therapy.