Hypertrophic scars resulting from pathological fibrosis and excessive collagen deposition cause significant functional and aesthetic impairment. Conventional therapies often fail to address the underlying molecular mechanisms of collagen overproduction. To overcome these limitations, we developed a novel layered dissolving microneedle system (MOF-CCH-MN) capable of delivering a synergistic therapeutic payload. This system incorporates Clostridium histolyticum collagenase (CCH) loaded within a calcium-based metal-organic framework (Ca-MOF), embedded in a γ-polyglutamic acid (γ-PGA) matrix. Characterization studies demonstrate that the microneedles possess a uniform pyramidal structure, penetrate the stratum corneum effectively, and dissolve within 25 min. Mechanistically, the Ca-MOF acts as a smart reservoir, protecting enzymatic activity while releasing Ca2+ and gallic acid to scavenge reactive oxygen species(ROS) and inhibit fibroblast - to - myofibroblast transition. In vitro assays confirmed that the system significantly downregulates pro-inflammatory cytokines (TNF-α, IL-1β) and suppresses fibrogenic gene expression (α-SMA, COL1A1, TGF-β). In vivo evaluation in a rabbit ear hypertrophic scar model demonstrated that a 20-day treatment regimen with MOF-CCH-MN significantly reduced scar thickness and promoted the reorganization of collagen fibers. These findings establish MOF-CCH-MN as a safe, effective, and minimally invasive platform for clinical scar management.

Diabetic bone regeneration is hindered by factors such as hyperglycemia, accumulation of advanced glycation end products (AGEs), and inflammation, presenting a significant clinical challenge. In this study, functionalized mesoporous silica nanoparticles (MSN) loaded with aminoguanidine (AG, MSN@AG) were incorporated into skin secretion of Andrias davidianus (SSAD) and poly (ε-caprolactone) (PCL) electrospun fibers, creating a bioactive scaffold (SSAD/PCL/MSN@AG, SPMA) that provides sustained release of AG and osteogenic factors. It effectively inhibited free radical generation, AGEs toxicity and inflammation, and recruited bone marrow mesenchymal stem cells (BMSCs) with excellent biocompatibility and better BMSCs viability, osteogenic differentiation, and mineralization under high-glucose microenvironment in vitro. Furthermore, in vivo evaluation in a diabetic mice skull defect model showed that the SPMA scaffold ameliorated local inflammation and improved vascularization, significantly accelerating new bone formation and defect repair. This study provides a simple preparation of PCL pro-osteogenic scaffolds with bioactive, hydrophilic and altering functions of the high-glucose microenvironment, which offers a new repair strategy for diabetic bone defects.

Chinese hamster ovary (CHO) cells are the primary platform for therapeutic antibody production. Although histone deacetylase inhibitors such as sodium butyrate (NaBu) can enhance recombinant expression, their growth-inhibitory and cytotoxic effects often limit volumetric productivity. Here, a structure-guided docking strategy was applied to prioritize NaBu-derived small-molecule additives (SMAs) for experimental screening in CHO antibody-producing cell lines. A lead combination (D1 +D4) increased volumetric antibody titers by approximately 2-3 fold while maintaining high cell viability (>95%) under the tested conditions. Cell-based analyses indicated reduced apoptotic markers and altered cell-cycle distributions in D1 +D4-treated cultures relative to NaBu. Metabolite profiling further revealed reduced by-product accumulation, including lactate and ammonia, together with a distinct intracellular energy and redox state compared with control cultures. Importantly, N-glycan profiles and charge variants of the produced antibodies remained comparable between control and D1 +D4-treated cultures. The productivity benefit of D1 +D4 was further maintained in bench-scale stirred-tank and wave bioreactor systems, supporting process relevance. Together, these results demonstrate that structure-guided NaBu analog screening can identify practical additives that enhance CHO antibody productivity with minimal impact on product quality.

Abstract The repair of mandibular bone defects remains a significant clinical hurdle, and tissue engineering offers an alternative strategy for reconstructive therapy. Nevertheless, conventional direct ossification is heavily dependent on a permissive microenvironment, which severely limits the regenerative outcome in large-scale bone defects. Endochondral ossification (EO)-based approaches, together with bone organoid microcarrier technology, have emerged as potential resolutions to this limitation. In the present work, cartilage regeneration units (CRUs) were fabricated by seeding decellularized bone matrix microparticles (DBM/COL-MPs) with bone marrow-derived mesenchymal stem cells (BMSCs), followed by chondrogenic priming in vitro. Upon implantation into mandibular defects in rabbits, these CRUs yielded a bone volume fraction (BV/TV) of 51.6% at 4 weeks—a value 1.5-fold higher than that of the control group. Moreover, the CRUs promoted vascular infiltration and accomplished robust and durable bone regeneration alongside satisfactory osseointegration through the EO pathway, both in ectopic and orthotopic mandibular defect settings. Collectively, this strategy furnishes a viable solution for the regenerative restoration of mandibular bone defects.

Nonunion fractures present a significant clinical challenge because of their complex microenvironment, which includes poor vascularization, insufficient osteogenesis, infection, and separation of fracture ends. The current clinical treatments have certain limitations. Inspired by this phenomenon, sandcastle worms secrete adhesive proteins that bind sand grains, shell fragments, and mineral particles, thereby constructing their “castles.” In this study, we developed an injectable bone cement using methacryloyl chitosan (CSMA) combined with a specific concentration of oyster shell nanoparticles (OS-np) to treat nonunion fractures. Oyster shells are composed primarily of calcium carbonate, which releases ions that promote angiogenesis and osteogenesis. The in vivo results at 8 weeks showed that the expression of BMP2, RUNX2, and VEGF in the OS-np/CSMA group was increased by 5.47, 4.38, and 3.54 times, respectively, compared to the control group, significantly enhancing vascularization and bone repair in the bone nonunion model. The injectability of the OS-np/CSMA bone cement ensures that it can adapt well to the complex structures of nonunion sites, providing a supportive matrix for new bone formation. Both in vivo and in vitro osteogenesis experiments demonstrated that the OS-np/CSMA bone cement significantly enhanced vascularization and bone repair in nonunion models, which was because of the synergistic effects of ion release and the bioactive properties of the oyster shell nanoparticles. This study highlights the potential of OS-np/CSMA injectable bone cement as a promising treatment strategy for complex nonunion fractures that effectively promotes angiogenesis and osteogenesis.

Diabetic nonunion, a significant clinical challenge with notably increasing incidence, arises primarily from cellular metabolic dysregulation and chronic inflammation induced by persistent hyperglycemia, leading to compromised self-stabilization at the nonunion site, diminished osteogenic capacity, and impaired angiogenesis. Currently, there are no effective clinical interventions established for managing diabetic nonunion. To address these issues, this work proposes an injectable, rapidly photocurable bone cement composed of methacrylated gelatin (GelMA) and methacrylated hyaluronic acid (HAMA), which is unconstrained by local geometry. This system is integrated with glucose oxidase (GOx)-loaded mesoporous nano-flower stacked particles derived from oyster shell powder (nHAP), referred to as GOx@nHAP-GelMA&HAMA. The incorporation of nHAP enhances the mechanical properties of GelMA&HAMA, while its slow degradation characteristics provide essential elements for bone growth and pro-angiogenic metal ions. GOx loaded on nHAP is gradually released to metabolize local glucose, achieving a moderate reduction in regional blood glucose levels and synergistically promoting angiogenesis. In summary, this study establishes a multifunctional platform for diabetic nonunion therapy, combining biomechanical support with microenvironmental modulation to offer a promising strategy for diabetic bone regeneration through localized intervention.

Hepatocellular carcinoma (HCC) is a prevalent and aggressive liver malignancy with limited therapeutic options. Circular RNAs (circRNAs) have emerged as critical regulators in various cancers, including HCC, but their roles in HCC progression remain largely unexplored. Here, the role of circZNF79(5) in HCC progression and its underlying mechanisms is investigated. CircZNF79(5) expression in HCC tissues, cell lines and the serum exosomes is analyzed using qRT-PCR and FISH, and evaluated its effects on cell proliferation, migration, invasion and apoptosis using CCK8, colony formation, EdU, Transwell and Flow cytometry. CircZNF79(5)'s is verified to be upregulated in HCC, and found that it can promote HCC cells proliferation, migration and invasion, while inhibiting the apoptosis. Mechanistically, circZNF79(5) is found to stabilizes the oncogenic protein YBX1 by recruiting BRCC36, a K63 chain deubiquitinating enzyme, thereby preventing YBX1 from p62-mediated selective autophagic degradation via the AMPK/mTOR signaling pathway. In vivo studies using subcutaneous and orthotopic tumor models confirmed that circZNF79(5) knockdown reduced tumor growth and YBX1 expression. The findings reveal a novel mechanism by which circZNF79(5) promotes HCC progression through YBX1 stabilization and selective autophagy regulation, highlighting the circZNF79(5)-YBX1-BRCC36 axis as a potential therapeutic target for HCC.

Epilepsy is one of the most prevalent central nervous system disorders, with antiepileptic drugs (AEDs) as the mainstay of treatment. While effective in reducing seizure frequency in approximate to 70% of patients, traditional AEDs are limited by blood concentration fluctuations, drug resistance, and cognitive side effects. Neuromodulation, particularly electrical stimulation, has emerged as a promising alternative by reversibly regulating abnormal neural circuits. However, conventional systems require implanted electrodes and external power sources, increasing the risk of trauma and infection. Piezoelectric nanomaterials offer a non-invasive strategy by converting endogenous biomechanical forces or ultrasound stimulation into localized electric currents, inducing neuronal hyperpolarization to suppress excitation. Based on this mechanism, a biomimetic piezoelectric nanoplatform is developed capable of ultrasound-triggered electrical stimulation for targeted neuromodulation without surgical implantation. Additionally, these nanoplatforms can co-deliver AEDs, enabling a dual therapeutic approach that combines localized stimulation with sustained drug release, enhancing efficacy while minimizing systemic exposure. This synergistic integration of ultrasound-responsive piezoelectric nanoplatforms and pharmacotherapy represents a transformative paradigm for safe, effective, and non-invasive epilepsy treatment.

Osteoarthritis (OA) is the most prevalent joint disease, yet effective disease-modifying OA drugs (DMOADs) remain elusive. Targeting macrophage polarization has emerged as a promising avenue for OA treatment. This study identified skatole through high-throughput screening as an efficient modulator of macrophage polarization. In vivo experiments demonstrated that skatole administration markedly reduced synovitis and cartilage damage in both destabilization of medial meniscus (DMM)-induced OA mice and monosodium iodoacetate (MIA)-induced OA rats. Mechanistically, skatole activated signal transducer and activator of transcription 6 (Stat6) signaling, promoting M2 macrophage polarization, while inhibiting nuclear factor-κB (NFκB) and mitogen-activated protein kinase (MAPK) signaling pathways to suppress M1 polarization. RNA-sequencing analysis, targeted metabolomics, and mitochondrial stress tests further revealed that skatole treatment shifted macrophages toward oxidative phosphorylation for energy production. Additionally, it up-regulated genes associated with glutathione metabolism and reactive oxygen species (ROS) pathways, reducing intracellular ROS production. The CUT&Tag assay results indicated that the downstream transcription factor p65 of NFκB can directly bind to gene loci related to inflammation, oxidative phosphorylation, and glutathione metabolism, thereby modulating gene expression. This regulatory process is inhibited by skatole. At the chondrocyte level, conditional medium from skatole-treated M1 macrophages balanced anabolism and catabolism in mouse chondrocytes and inhibited apoptosis. In IL1β-treated chondrocytes, skatole suppressed inflammation and catabolism without affecting apoptosis or anabolism. Overall, skatole maintains immune microenvironment homeostasis by modulating macrophage polarization in joints and preserves cartilage function by balancing chondrocyte anabolism and catabolism, effectively alleviating OA. These findings suggest skatole’s potential as a DMOAD.

In order to address the fundamental drawbacks of conventional surgical sutures, such as infection susceptibility and scarring potential, an enzyme‐enhanced hyaluronic acid composite hydrogel (HANB‐MOF‐TG) is developed through a dual‐crosslinking strategy integrating tissue transglutaminase (TG) catalysis and calcium‐metal–organic frameworks (Ca‐MOFs) coordination. The bioadhesive system is engineered by encapsulating TGase within Ca‐MOFs, providing a sustained reductive environment to maintain enzymatic activity, followed by dynamic covalent crosslinking with nitrophenylboronic acid‐functionalized hyaluronic acid. This hierarchical design conferred exceptional tissue adhesion strength, broad‐spectrum antibacterial efficacy, and pro‐regenerative functionality. In a full‐thickness skin incision model in mice, the hydrogel enabled immediate wound closure compared to the blank group and achieved more precise wound edge alignment and improved healing outcomes relative to the suture and medical glue groups, owing to TG‐mediated collagen fiber alignment and extracellular matrix remodeling. Meanwhile, the hydrogel effectively inhibited scar formation, as evidenced by ≈30% reduction in α‐SMA expression at day 14 compared to day 7. Mechanistic studies revealed that the sustained release of Ca 2 ⁺ from MOF structures synergized with enzymatic crosslinking to activate fibroblast migration and angiogenesis. This work establishes a paradigm for next‐generation bioadhesives that unify mechanical robustness, biological functionality, and scarless healing in suture‐free wound management.

Osteochondral defects are still facing a significant challenge in clinical surgery, making post-trauma repair difficult. Tissue engineering has provided a promising approach to solving these defects. However, existing scaffolds cannot replicate the complex biphasic cartilage-bone microenvironment with accuracy. We aimed to develop a biphasic biomimetic scaffold with regionally regulated vascularization that promoted chondrogenesis and osteogenesis through bidirectional regulation of endochondral ossification. This scaffold consisted of pre-chondrogenic microspheres (PCMs) and a decalcified bone frame prepared by decalcifying the cartilage layer and bone layer of the scaffold to varying degrees. Incorporation of PCMs into the cartilage layer created a microenvironment that promoted cartilage regeneration while axitinib was modified to inhibit vascularization and enhance cartilage regeneration. The bone layer provided a microenvironment that promoted endochondral ossification and facilitated bone repair. In vitro studies have shown that axitinib-modified cartilage layers significantly inhibit the VEGF expression of pre-chondrogenic cells, while decalcified bone powder from the bone layer significantly promotes the ossification of PCMs. In vivo experiments indicated that this decalcified bone frame controls the endochondral ossification of PCMs through regionalized angiogenesis, promoting the integrated regeneration and reconstruction of osteochondral defects in rabbit knee joints. These results suggest that our designed demineralized bone frame can precisely engineer the osteochondral regeneration microenvironment, providing theoretical guidance for the integrated regeneration and repair of anisotropic tissue injuries.

Diabetic fracture frequently results in delayed healing or nonunion due to hyperglycemia-induced vascular impairment and compromised osteogenic capacity. Current clinical interventions face dual challenges in regulating pathological microenvironments and enhancing bone regeneration efficacy. This study innovatively integrated three key components: (i) The gelatin methacryloyl/methacrylated hyaluronic acid (GelMA/HAMA) double-network (DN) hydrogel provides enhanced mechanical robustness; (ii) Ti3C2Tx MXene nanosheets (MXene) with superior photothermal conversion efficiency for mild hyperthermia induction and mechanical reinforcement of the hydrogel; (iii) Oyster shell-derived bioinspired hydroxyapatite nanoflowers (nHAP) enabling sustained release of osteogenic ions (Mg2+, Sr2+, Ca2+, and Si4+) while further augmenting compressive strength. Collectively, these components formed an injectable near-infrared (NIR) photothermal responsive bone cement, designated as nHAP-MXene-GelMA/HAMA. Synergistic therapeutic benefits were validated through both cellular and animal models. The functional components collectively enhanced mechanical performance of the material. Additionally, the combination of nHAP-mediated ion release and NIR-triggered photothermal therapy (PTT, 40-43 degrees C) significantly upregulated angiogenesis markers and osteogenic differentiation indicators in diabetic fracture models. Radiographic and histological analyses confirmed accelerated callus formation and complete cortical bone remodeling within 8 weeks. This bone cement presents a novel strategy for diabetic fracture treatment through microenvironment regulation and osteogenesis promotion, providing potential solutions for clinical applications in internal fixation and bone regeneration.

Chinese hamster ovary (CHO) cells are major expression platforms for the transient production of recombinant therapeutic proteins (RTPs). Most improvement strategies have focused on promoting transcriptional expression in CHO cells. However, methods for promoting the yield of RTPs through translational regulation remain unclear. In this study, we investigated characteristics of the 5'-untranslated region (UTR) that influence recombinant protein expression in CHO cells and identified sequences that have positive effects on protein expression using ribosome sequencing. Some elements and characteristics of 5'-UTR differentially affected the translation of the main open reading frame and increased recombinant protein expression by 1.5-fold in CHO cells. The findings may help relieve the bottleneck of the yield of RTPs on translation enhancement.

The pathogenesis of cartilage injury and degeneration is exceptionally complex. In addition to being associated with osteoarthritis and trauma, factors such as age, gender, obesity, inflammation, and apoptosis of chondrocytes are also considered significant influencing factors. Due to the lack of direct blood supply, lymphatic circulation, and neural innervation, coupled with low metabolic activity, the self-repair capability of cartilage after injury is extremely limited, making its treatment quite challenging. Recent research indicated that ncRNA, a class of RNA transcribed from the genome that does not encode proteins, played a crucial regulatory role in various disease processes. Particularly noteworthy is its positive regulatory role in cartilage regeneration, achieved through the modulation of the inflammatory microenvironment, promotion of chondrocyte proliferation, inhibition of chondrocyte degradation, and facilitation of the recruitment and differentiation of bone marrow mesenchymal stem cells into chondrocytes. In the earlier phase, we conducted a review and outlook on therapeutic strategies for the regeneration of articular cartilage injuries. This article specifically focuses on summarizing the regulatory roles and research advancements of ncRNA in cartilage regeneration, as well as its contributions to the clinical application of gene therapy for cartilage defects.

Meniscus is a crescent-shaped fibrocartilage tissue for providing structural congruence and absorbing mechanical forces. Currently, the development of material-guided regeneration medicine strategy has emerged as a promising alternative for meniscus treatment. However, it often presents more complex pathological conditions of immune-inflammatory responses, and thus inevitably causes a harsh microenvironment that extremely hinders fibrocartilage regeneration. Therefore, there is an urgent need to develop bioactive materials to achieve cartilaginous immunomodulatory throughout the whole regenerative periods. In this study, we develop a novel dynamic-covalent hybrid (DCH) hydrogel with cartilaginous immune microenvironment (CIME) to temporally regulate meniscus regeneration. By combining dynamic boronic ester crosslinking and covalent photopolymerization reactions, DCH hydrogels exhibit favorable injectability, self-healing, and tissue adhesion properties for practical operation. Furthermore, CIME is successfully created by the introduction of a temporally on-demand regulatory system: naproxen anti-inflammatory drugs are preferentially released to regulate M1/M2 macrophage polarization through PI3K/Akt/mTOR signaling pathway at early stage, while TGFβ3/CTGF growth factors are on-demand released to promote fibrochondrogenic differentiation of stem cells in the post-regulatory microenvironment at later stage. Finally, in vivo experiments demonstrate the satisfactory repair of meniscus cartilage defects in rabbits by activating the endogenous repair of stem cells homing based on our established cartilaginous immunomodulatory strategy.

Atrophic nonunion, a particularly challenging complication of diaphyseal bone fractures, arises from a deteriorated regenerative microenvironment characterized by insufficient vascularization and pathological accumulation of reactive oxygen species (ROS). To address this clinical challenge, a dual-function bioactive scaffolds is developed that simultaneously blocks disease progression and promotes tissue regeneration. The scaffolds design incorporates three key components: First, MnO2-Cu2+ (MC) nanoparticles are synthesized to combat the pathological microenvironment through dual mechanisms of ROS scavenging and angiogenesis promotion. Second, circular bone morphogenetic protein 2 (circBMP2) is engineered for sustained BMP2 expression to enhance osteogenic differentiation, with polyethylenimine-mediated surface conjugation onto MC nanoparticles. Finally, these MC-circBMP2 (MCB) complexes are integrated into a dopamine-modified demineralized bone matrix scaffolds to create a hierarchically structured regenerative platform. In vivo experiments showed that the bioactive scaffolds promoted the healing of atrophic nonunion of radial bone defects in rabbits by improving angiogenesis, scavenging ROS, and accelerating the osteogenic differentiation of endogenous cells. The above results indicate that the designed bioactive scaffolds can precisely block the pathology and rapidly initiate the regeneration of damaged tissue, providing theoretical guidance for the regeneration of pathological refractory tissue injuries.

Chinese hamster ovary cells are the main expression system for recombinant therapeutic proteins. During the production of these proteins, certain host cell proteins are secreted, broken down, and released by host cells in the culture along with the proteins of interest. These host cell proteins are often difficult to remove during the downstream purification process, and thus affect the quality, safety, and effectiveness of recombinant protein biopharmaceutical products and increase the production cost of recombinant therapeutic proteins. Therefore, host cell protein production must be reduced as much as possible during the production process and eliminated during purification. This article reviews the harm caused by host cell proteins in the production of recombinant protein drugs using Chinese hamster ovary cell, factors affecting host cell proteins, the monitoring and identification of these proteins, and methods to reduce their type and quantity in the final product.

Osteoarthritis (OA) is the most prevalent cartilage degenerative and low-grade inflammatory disease of the whole joint. However, there are currently no FDA-approved drugs or global regulatory agency-approved treatments OA disease modification. Therefore, it’s essential to explore novel effective therapeutic strategies for OA. In our study, we investigated the effects of AFK-PD, a novel pyridone agent, on the development of OA induced by destabilization of the medial meniscus (DMM) in vivo, and its impact on the function of chondrocytes treated with IL-1β in vitro. Our results demonstrated AFK-PD alleviated OA progression through inhibiting cartilage degeneration, articular inflammation and osteophyte formation. Notably, AFK-PD inhibited chondrocyte inflammation and synovial macrophage M1 polarization, leading to the attenuation of articular inflammation. Additionally, AFK-PD promoted chondrocyte anabolism while mitigating catabolism and apoptosis, effectively inhibiting cartilage degeneration. Mechanistically, AFK-PD suppressed the expression of key signaling molecules involved in the MAPK pathway, such as p-ERK1/2 and p-JNK, as well as the NF-κB signaling molecule p-p65, in IL-1β-induced chondrocytes. These findings suggest AFK-PD ameliorates the development of OA by protecting chondrocyte functions and inhibiting articular inflammation in chondrocytes and synovial macrophages. Overall, our study highlights AFK-PD as a promising therapeutic candidate for the treatment of OA.

Tissue damage often causes considerable suffering to patients due to slow recovery and poor prognosis. The use of electroactive materials to deliver biophysical signals plays a key role in regulating tissue regeneration processes. Among these materials, piezoelectric materials have unique electromechanical conversion capabilities, making them suitable for use as cell scaffolds. They can deform and emit electrical signals in response to external stimuli, thereby regulating cell proliferation and differentiation. In this review, recent advances are presented in piezoelectric materials as physical signaling mediators that regulate cell differentiation. The basic mechanisms, classification of these materials, and their different applications in tissue regeneration are described. Finally, a comprehensive discussion of current challenges and prospects in the field is provided. Together, existing experimental results basically show that piezoelectric materials can improve the process and effect of tissue repair, providing new technical options for the development of tissue engineering in the future.