Hypoxia-induced factor 1α promotes osteotomy-induced regional acceleratory phenomenon via DC-STAMP mediated membrane fusion.

OBJECTIVES: The mechanisms of regional acceleratory phenomenon (RAP) induced by orthognathic osteotomy are unclear. It was not known if locally hypoxic microenvironment changes were involved in this phenomenon. METHODS: Hypoxia-induced factor-1α knockout mice harboring Cathepsin K (CTSK) Cre were used to investigate the effect of hypoxia-driven osteoclasts on alveolar bone remodeling. RAW264.7 cells were induced by CoCl2 to observe the effects of dendritic cell-specific transmembrane protein (DC-STAMP) on the fusion and differentiation of osteoclasts. RESULTS: We found mandibular osteotomy of C57 mice induced active alveolar osteoclasts and increased hypoxia-induced factor-1α (HIF-1α) positive staining areas. Alveolar bone density of the 10-week-old HIF-1α conditional knockout (CKO) mouse was increased at 10 and 14 days after bilateral mandibular osteotomy. Moreover, decreased numbers of osteoclasts and matrix metalloproteinase 9 (MMP-9)-positive cells were observed on the surface of bone resorption lacunae in the CKO group. HIF-1α could increase the expression level of DC-STAMP to enhance osteoclastogenesis and cell fusion in active RAW264.7 cells. CONCLUSION: Our data considered hypoxia-driven osteoclasts resorption to be an adaptive mechanism to permit alveolar bone loss after bilateral mandibular osteotomy of mice.

Soil nutrient levels regulate the effect of soil microplastic contamination on microbial element metabolism and carbon use efficiency.

Microplastics (MPs) are emerging environmental contaminants in soil ecosystems that disrupt the soil carbon (C) pool. Therefore, the response of microbial metabolism to MP-contaminated soil is crucial for soil-C stabilization. We undertook factorial experiments in a greenhouse with three types of soil microplastics with three levels of soil nutrients and undertook soil physiochemical analyses after 60 days. The present study revealed how the presence of degradable polylactic acid (PLA) and non-degradable polyethylene (PE) MPs affects soil microbial nutrient limitation and C use efficiency (CUE) at varying nutrient concentrations. The presence of PLA in soil with low nutrient levels led to a significant increase (29%) in the activities of nitrogen (N)-acquiring enzymes. In contrast, the presence of MPs had no effect on C- and N-acquiring enzymes. The occurrence of PE caused a 41% reduction in microbial C limitation in high-nutrient soils, and microbial nutrient metabolism was limited by the occurrence of MPs in soils amended with nutrients. A strong positive correlation between microbial C and nutrient limitation in the soil indicates that addressing C limitation followed by amendment of soil with MPs could potentially intensify microbial N limitation in soils with varying nutrients. In comparison, the microbial CUE increased by 10% with the application of degradable MPs (PLA) to soils with a low nutrient status. These findings highlight the significant influence of both degradable PLA and non-degradable PE MPs on soil microbial processes and C dynamics. In conclusion, PLA enhances metabolic efficiency in nutrient-rich soils, potentially aiding C utilization, whereas PE reduces microbial C limitation, offering promise for soil C sequestration strategies. Our findings underscore the importance of considering MPs in soil ecosystem studies and in broader sustainability efforts.

The toxicological effect on pak choi of co-exposure to degradable and non-degradable microplastics with oxytetracycline in the soil.

Microplastics and antibiotics are emerging as ubiquitous contaminants in farmland soil, harming crop quality and yield, and thus threatening global food security and human health. However, few studies have examined the individual and joint effects of degradable and/or non-degradable microplastics and antibiotics on crop plants. This study examined the individual and joint effects of polyethylene (PE) and polylactic acid (PLA) microplastics and the antibiotic oxytetracycline (OTC) on pak choi by measuring its growth, photosynthesis, antioxidant enzyme activity, and metabolite levels. Microplastics and/or oxytetracycline adversely affected root weight, photosynthesis, and antioxidant enzyme (superoxide dismutase, catalase, and ascorbate peroxidase) activities. The levels of leaf metabolites were significantly altered, causing physiological changes. Biosynthesis of plant secondary metabolites and amino acids was altered, and plant hormones pathways were disrupted. Separately and together, OTC, PE, and PLA exerted phytotoxic and antagonistic effects on pak choi. Separately and together with OTC, degradable microplastics altered the soil properties, thus causing more severe impacts on plant performance than non-degradable microplastics. This study elucidates the effects on crop plants of toxicity caused by co-exposure to degradable or non-degradable microplastic and antibiotics contamination and suggests mechanisms.

Toll-like receptor-4 activation by subgingival biofilm and periodontal treatment response.

OBJECTIVES: This study aims to investigate longitudinally the activation of Toll-like receptor-4 (TLR-4) by subgingival biofilm samples before and after nonsurgical periodontal therapy (NSPT). MATERIALS AND METHODS: Forty periodontitis patients received NSPT and were reviewed 3 and 6 months post-treatment. Subgingival biofilm was sampled from 4 teeth per patient, at baseline and each follow-up time point. TLR-4 activation was determined using the HEK-BLUE™/hTLR4 system. Changes in TLR-4 activation and probing pocket depths (PPDs) were evaluated using generalised linear models, and the association between TLR-4 activation and pocket reduction (defined as 6-month PPDs ≤ 3mm) was determined using generalised estimating equations. RESULTS: At 6 months, the mean TLR-4 activation by subgingival biofilm samples was significantly reduced from 11.2AU (95%CI 7.1AU, 15.4AU) to 3.6AU (95%CI 2.3AU, 4.8AU, p < 0.001), paralleling significant reductions in mean PPDs at sampled sites. The response to NSPT was associated with longitudinal TLR-4 activation profiles, with significantly higher TLR-4 activation by subgingival biofilm obtained from sites that did not achieve pocket reduction, compared to sites at which pocket reduction was achieved. CONCLUSIONS: The activation of TLR-4 by subgingival biofilm samples was reduced after NSPT, and this reduction was significantly associated with the clinical improvements (PPD reductions) at sampled sites. CLINICAL RELEVANCE: This study demonstrated an association between the longitudinal profile of TLR-4 activation by subgingival biofilm and periodontal treatment response. Longitudinal monitoring of TLR-4 activation by subgingival biofilm may potentially identify non-responsive sites, enabling targeted additional treatment.

Polyethyleneimine/hydrated titanium oxide-functionalized fibrous adsorbent for removing cobalt: Adsorption performance and irradiation stability.

Radionuclides of 60Co often encountered in the fields of radiation therapy, medical preparation, and equipment sterilization, which have been considered fatal. Therefore, developing efficient and irradiation-stable adsorbents for the removal of 60Co in wastewater is urgently needed. An irradiation-stable fibrous adsorbent was fabricated through the surface functionalization of collagen fibers (CFs) by polyethyleneimine (PEI) and hydrated titanium oxide (TiO) (PEI-TiO-CFs). PEI-TiO-CFs, including their adsorption performance and irradiation stability, were systematically investigated. Results showed that PEI-TiO-CFs exhibit a maximum adsorption capacity of 0.5575 mmol g-1. In addition, the adsorption capacity of PEI-TiO-CFs only demonstrated a slight decrease in the selectivity investigation of Co2+ mixed with another coexisting ion, such as Na+, K+, and NO3-, Cl-. Furthermore, breakthrough point of PEI-TiO-CFs in column is high at 80 BV (bed volume) and the PEI-TiO-CF column can be mostly regenerated using 12 BV of Na2EDTA solution. Excellent irradiation stability of PEI-TiO-CFs was confirmed by the maintained morphology and adsorption capacity after irradiation at 350 kGy of 60Co γ-ray. Results indicated that PEI-TiO-CFs are an effective adsorbent for radioactive cobalt removal from aqueous solutions.

HIF-1α regulates osteoclast activation and mediates osteogenesis during mandibular bone repair via CT-1.

OBJECTIVES: Hypoxia is one of the characteristics of microenvironmental changes after orthognathic surgery for fractures. HIF-1α is a main regulator of the hypoxic response and plays a crucial role in bone formation, remodelling, and homeostasis. Osteoclasts participate in bone absorption and affect osteogenesis, and osteoclasts differentiate in a path from the oxygen-rich bone marrow to oxygen-deficient bone lesions. Thus, we aimed to study the key functions of HIF-1α in osteoclasts during mandibular healing after osteotomy. MATERIALS AND METHODS: The function of HIF-1α in osteoclasts during fracture healing in osteoclast-specific HIF-1α-conditional-knockout mice was investigated in mandibular osteotomy. Primary osteoclasts were used to explore the expression of HIF-1α and cardiotrophin-1 (CT-1) at both the mRNA and protein levels. The ability of BMSCs co-cultured with conditioned media from osteoclast-specific HIF-1α-knockout primary osteoclasts was detected using osteoclast-mediated osteogenesis experiments. RESULTS: Hypoxia-inducible factor-1α increased osteoclastogenesis and bone resorption, and a delay in bone healing was found in osteoclast-specific HIF-1α-conditional-knockout mice compared with normal mice. HIF-1α-knockout primary osteoclasts inhibited bone resorption and CT-1 expression, and HIF-1α enhanced the osteoclast-mediated stimulation of BMSC differentiation by secreting CT-1. CONCLUSIONS: Hypoxia-inducible factor-1α can play a key role in the physiology and pathogenesis of bone resorption by promoting osteoclastogenesis during fracture and influencing osteogenesis through CT-1 during bone healing.

Bone-targeted erythrocyte-cancer hybrid membrane-camouflaged nanoparticles for enhancing photothermal and hypoxia-activated chemotherapy of bone invasion by OSCC.

BACKGROUND: Jaw bones are the most common organs to be invaded by oral malignancies, such as oral squamous cell carcinoma (OSCC), because of their special anatomical relationship. Various serious complications, such as pathological fractures and bone pain can significantly decrease the quality of life or even survival outcomes for a patient. Although chemotherapy is a promising strategy for bone invasion treatment, its clinical applications are limited by the lack of tumor-specific targeting and poor permeability in bone tissue. Therefore, it is necessary to develop a smart bone and cancer dual targeting drug delivery platform. RESULTS: We designed a dual targeting nano-biomimetic drug delivery vehicle Asp8[H40-TPZ/IR780@(RBC-H)] that has excellent bone and cancer targeting as well as immune escape abilities to treat malignancies in jaw bones. These nanoparticles were camouflaged with a head and neck squamous cell carcinoma WSU-HN6 cell (H) and red blood cell (RBC) hybrid membrane, which were modified by an oligopeptide of eight aspartate acid (Asp8). The spherical morphology and typical core-shell structure of biomimetic nanoparticles were observed by transmission electron microscopy. These nanoparticles exhibited the same surface proteins as those of WSU-HN6 and RBC. Flow cytometry and confocal microscopy showed a greater uptake of the biomimetic nanoparticles when compared to bare H40-PEG nanoparticles. Biodistribution of the nanoparticles in vivo revealed that they were mainly localized in the area of bone invasion by WSU-HN6 cells. Moreover, the Asp8[H40-TPZ/IR780@(RBC-H)] nanoparticles exhibited effective cancer growth inhibition properties when compared to other TPZ or IR780 formulations. CONCLUSIONS: Asp8[H40-TPZ/IR780@(RBC-H)] has bone targeting, tumor-homing and immune escape abilities, therefore, it is an efficient multi-targeting drug delivery platform for achieving precise anti-cancer therapy during bone invasion.

The effects of mandibular osteotomy on maxillary orthodontic tooth movement and bone remodelling in a rat model.

OBJECTIVES: The accelerated tooth movement phenomenon after orthognathic surgery has been observed. However, the underlying mechanism remains unclear. There is no experimental study showing the effect of orthognathic surgery on orthodontic tooth movement of the opposing jaw. Therefore, the present study aimed at investigating if mandibular osteotomy enhances maxillary tooth movement and bone remodelling. MATERIALS AND METHODS: Fifty-four male Sprague-Dawley rats were randomly divided into two groups: maxillary tooth movement (TM) and maxillary tooth movement + mandibular surgery (TM + MS). The orthodontic force was delivered to move the left maxillary first molar mesially. The surgical intervention was performed on the left mandible. Microcomputed tomography, histological analysis, enzyme-linked immunosorbent assay, and quantitative real-time polymerase chain reaction were used to assess changes at 3, 7, and 21 days after surgery. RESULTS: The mandibular osteotomy accelerates the rate of maxillary tooth movement with decreased bone volume fraction on the seventh day. Bone resorption was observed on the third and seventh day after mandibular osteotomy. It was found that serum interleukin-1ß level increased significantly in the TM + MS group compared with the TM group, as well as the high expression level of cathepsin K and tumour necrosis factor receptor-associated factor 5 of the orthodontic tooth on the third and seventh day after mandibular osteotomy. CONCLUSION: Data from the present study suggested that mandibular osteotomy accelerates maxillary osteoclast activity and post-operative tooth movement, providing evidence for accelerated tooth movement phenomenon after orthognathic surgery.

Two-year follow-up of a randomized multicenter study comparing a drug-coated balloon with a drug-eluting stent in native small coronary vessels: The RESTORE Small Vessel Disease China trial.

OBJECTIVES: To report the clinical outcomes of the RESTORE drug-coated balloon (DCB; Cardionovum, Bonn, Germany) for treatment of de novo small vessel disease (SVD) beyond 1 year. BACKGROUND: Previous reports have demonstrated the noninferiority of the RESTORE DCB to the RESOLUTE Integrity drug-eluting stent (DES; Medtronic, Minneapolis, Minnesota) in terms of 9-month in-segment percent diameter stenosis. METHODS: In the prospective, multicenter, noninferiority RESTORE SVD China trial, 230 patients with visually-estimated reference vessel diameter (RVD) ≥2.25 and ≤2.75 mm were randomized to DCB or DES in a 1:1 ratio stratified by diabetes and number of lesions treated. Furthermore, 32 patients with RVD ≥2.00 and <2.25 mm were enrolled in a nested very small vessel (VSV) registry. Clinical follow-up were performed at 2 years to evaluate target lesion failure (TLF) in both groups and the VSV cohort. RESULTS: Overall, 256 (97.7%) patients (115 and 109 in the DCB and DES groups, respectively, and 32 in the VSV cohort) completed 2 years of follow-up. There was no significant difference in TLF between the DCB and DES groups (5.2 vs. 3.7%, p = .75). Target lesion revascularization was acceptable at 1 month, 1 year, and 2 years, and did not differ significantly with DCB from that in the DES group (0.9 vs. 0%, p = 1.0, 4.4 vs. 2.6%, p = .72, 5.2 vs. 2.8%, p = .50, respectively). CONCLUSIONS: Compared to the second-generation DES, the RESTORE DCB did not increase the risk of clinical outcomes. Late catch-up phenomen requiring revascularization was not significant in this study.

HIF-1α facilitates osteocyte-mediated osteoclastogenesis by activating JAK2/STAT3 pathway in vitro.

Osteocytes, entrapped within the mineralized bone matrix, has been found to have numerous functions such as acting as an orchestrator of bone remodeling through regulation of both osteoclast and osteoblast activity and also functioning as an endocrine cell. Due to a specialized morphology and surrounding structure, osteocytes are more tolerant to hypoxia during osteoporosis, fracture, osteoarthritis, and orthodontic-orthognathic combination therapy. Hypoxia-inducible factor-1α (HIF-1α) is one of the master regulators of hypoxia reactions, playing an important role in bone modeling, remodeling, and homeostasis. This study aimed to investigate the pivotal functional role of HIF-1α in osteocytes initiating of bone remodeling under hypoxia. In the present study, the osteoclasts formation induced by RAW264.7 was significantly promoted in conditioned media (CM) from osteocytic MLO-Y4 exposed to hypoxia in vitro. Therefore, hypoxic MLO-Y4 cells simulated by 100 µmol/L CoCl2 or 2% O2 stably expressed HIF-1α proteins and upregulated the expression of receptor activator of nuclear factor-κB ligand (RANKL) at both the messenger RNA (mRNA) and protein level. Furthermore, with the Knockdown of HIF-1α, the expression of RANKL mRNA and protein decreased after transient transfection. In addition, the phosphorylation of Janus kinase 2 (JAK2) and signal transducer and activator of transcription (STAT3) was also correlated with HIF-1α and RANKL levels under hypoxia. Then AG490, a JAK2 inhibitor, inhibited p-JAK2, p-STAT3 and RANKL expression. It was possible that AG490 disturbed the contact of HIF-1α and RANKL by JAK2/STAT3 pathway, influencing osteoclastogenesis. Our findings suggested that HIF-1α promoted the expression of RANKL by activating JAK2/STAT3 pathway in MLO-Y4 cells, and enhanced osteocyte-mediated osteoclastic differentiation in vitro.

Mandibular osteotomy-induced hypoxia enhances osteoclast activation and acid secretion by increasing glycolysis.

The rapid bone remodeling after osteotomy has been reported for a long time. However, the underlying mechanism promoting the active bone reconstruction was still to be elucidated. Since not only the bone, blood vessels, and supportive tissues, but also the local microenvironment were destroyed, if the changes on the cell metabolism was contributed to the accelerated bone remodeling came into sight. In present study, we found that the mandibular osteotomy in rabbit activated osteoclasts, as well as the expression of hypoxia-inducible factor 1α (HIF-1α) in alveolar bone. Hypoxia or HIF-1α could enhanced osteoclastogenesis, bone absorption, and lactic acid concentration in receptor activator of nuclear factor κΒ ligand-induced RAW264.7 cells. Coincided with the upregulated HIF-1α expression, HIF-driven glycolytic enzymes, such as lactate dehydrogenase A (LDHA), glucokinase (GCK), pyruvate kinase M2 (PKM2), and phosphofructokinase1 (PFK1), were found massively increased in both hypoxic RAW264.7 cells and the alveolar HIF-1α-positive osteoclasts after mandibular osteotomy. Knockdown of HIF-1α suppressed not only the hypoxia-mediated glycolysis, but also the hypoxia-induced acid secretion and bone resorption in RAW264.7 cells. Application of inhibitor on glycolysis gave rise to the similar results as HIF-1α knockdown. Our findings suggested that hypoxia-driven glycolysis in osteoclasts was an adaptive mechanism to permit alveolar bone remodeling after mandibular osteotomy.

Transcriptomic insights into citrus segment membrane's cell wall components relating to fruit sensory texture.

BACKGROUND: During fresh fruit consumption, sensory texture is one factor that affects the organoleptic qualities. Chemical components of plant cell walls, including pectin, cellulose, hemicellulose and lignin, play central roles in determining the textural qualities. To explore the genes and regulatory pathways involved in fresh citrus' perceived sensory texture, we performed mRNA-seq analyses of the segment membranes of two citrus cultivars, Shiranui and Kiyomi, with different organoleptic textures. RESULTS: Segment membranes were sampled at two developmental stages of citrus fruit, the beginning and end of the expansion period. More than 3000 differentially expressed genes were identified. The gene ontology analysis revealed that more categories were significantly enriched in 'Shiranui' than in 'Kiyomi' at both developmental stages. In total, 108 significantly enriched pathways were obtained, with most belonging to metabolism. A detailed transcriptomic analysis revealed potential critical genes involved in the metabolism of cell wall structures, for example, GAUT4 in pectin synthesis, CESA1, 3 and 6, and SUS4 in cellulose synthesis, CSLC5, XXT1 and XXT2 in hemicellulose synthesis, and CSE in lignin synthesis. Low levels, or no expression, of genes involved in cellulose and hemicellulose, such as CESA4, CESA7, CESA8, IRX9 and IRX14, confirmed that secondary cell walls were negligible or absent in citrus segment membranes. A chemical component analysis of the segment membranes from mature fruit revealed that the pectin, cellulose and lignin contents, and the segment membrane's weight (% of segment) were greater in 'Kiyomi'. CONCLUSION: Organoleptic quality of citrus is easily overlooked. It is mainly determined by sensory texture perceived in citrus segment membrane properties. We performed mRNA-seq analyses of citrus segment membranes to explore the genes and regulatory pathways involved in fresh citrus' perceived sensory texture. Transcriptomic data showed high repeatability between two independent biological replicates. The expression levels of genes involved in cell wall structure metabolism, including pectin, cellulose, hemicellulose and lignin, were investigated. Meanwhile, chemical component contents of the segment membranes from mature fruit were analyzed. This study provided detailed transcriptional regulatory profiles of different organoleptic citrus qualities and integrated insights into the mechanisms affecting citrus' sensory texture.

Periostin promotes migration, proliferation, and differentiation of human periodontal ligament mesenchymal stem cells.

OVERVIEW: Periostin (POSTN) is critical to bone and dental tissue morphogenesis, postnatal development, and maintenance; however, its roles in tissue repair and regeneration mediated by human periodontal ligament mesenchymal stem cells (PDLSCs) remain unclear. The present study was designed to evaluate the effects of POSTN on hPDLSCs in vitro. MATERIALS AND METHODS: hPDLSCs were isolated and characterized by their expression of the cell surface markers CD44, CD90, CD105, CD34, and CD45. Next, 100 ng/mL recombinant human POSTN protein (rhPOSTN) was used to stimulate the hPDLSCs. Lentiviral POSTN shRNA was used to knockdown POSTN. The cell counting kit-8 (CCK8) and scratch assay were used to analyze cell proliferation and migration, respectively. Osteogenic differentiation was investigated using an alkaline phosphatase (ALP) activity assay, alizarin staining, and quantitative calcium analysis and related genes/protein expression assays. RESULTS: Isolated hPDLSCs were positive for CD44, CD90, and CD105 and negative for CD34 and CD45. In addition, 100 ng/mL rhPOSTN significantly accelerated scratch closure, and POSTN-knockdown cells presented slower closure at 24 h and 48 h. Furthermore, the integrin inhibitor Cilengitide depressed the scratch closure that was enhanced by POSTN at 24 h. The CCK8 assay showed that 100 ng/mL rhPOSTN promoted hPDLSC proliferation. Moreover, 100 ng/mL rhPOSTN increased the expression of RUNX2, OSX, OPN, OCN, and VEGF and enhanced ALP activity and mineralization. POSTN silencing decreased the expression of RUNX2, OSX, OPN, OCN, and VEGF and inhibited ALP activity and mineralization. CONCLUSIONS: POSTN accelerated the migration, proliferation, and osteogenic differentiation of hPDLSCs.

Immediate implant placement and complete mouth rehabilitation with CAD-CAM titanium frameworks and cemented crowns for a patient with severe periodontal disease: A clinical report.

The technique of immediate implantation has been widely used to reduce treatment time and bone loss after extraction. However, immediate implant placement in infected extraction sockets is generally contraindicated. This clinical report describes a treatment protocol for immediate implantation after the extraction of teeth with generalized chronic periodontitis. The technique used for the oral rehabilitation used computer-assisted design and computer-assisted manufacturing (CAD-CAM) titanium frameworks and cemented zirconia crowns. The titanium frameworks overcame suboptimal implant position and the cemented crowns provided excellent function and esthetics despite the locations of screw-access openings. No clinical complications occurred during a 13-month follow-up.

Shear-Induced Breakup of Cellulose Nanocrystal Aggregates.

The flow properties of two kinds of cellulose nanocrystal (CNC) rods with different aspect ratios and similar zeta potentials in aqueous suspensions have been investigated. The aqueous CNC suspensions undergo a direct transition from dilute solution to colloidal glass instead of phase separation with the increasing CNC concentration. The viscosity profile shows a single shear-thinning behavior over the whole range of shear rates investigated. The shear-thinning behavior becomes stronger with the increasing CNC concentration. The viscosity is much higher for the unsonicated suspension when compared with the sonicated suspensions. The CNC rods appear arrested without alignment with an increasing shear rate from the small-angle light scattering patterns. The arrested glass state results from electric double layers surrounding the CNC rods, which give rise to long-ranged repulsive interactions. For the first time, we demonstrate that, within a narrow range of CNC concentrations, a shear-induced breakup process of the CNC aggregates exists when the shear rate is over a critical value and that the process is reversible in the sense that the aggregates can be reformed. We discuss the competition between the shear-induced breakup and the concentration-driven aggregation based on the experimental observations. The generated aggregate structure during the breakup process is characterized by a fractal dimension of 2.41. Furthermore, we determine two important variables-the breakup rate and the characteristic aggregate size-and derive analytical expressions for their evolution during the breakup process. The model predictions are in quantitative agreement with the experimental results.

Nuclear-targeting TAT-PEG-Asp8-doxorubicin polymeric nanoassembly to overcome drug-resistant colon cancer.

AIM: Drug efflux-associated multidrug resistance (MDR) is a main obstacle to effective cancer chemotherapy. Large molecule drugs are not the substrates of P-glycoprotein, and can circumvent drug efflux and be retained inside cells. In this article we report a polymer-drug conjugate nanoparticulate system that can overcome MDR based on size-related exclusion effect. METHODS: Doxorubicin was coupled with the triblock polymeric material cell-penetrating TAT-PEG-poly(aspartic acid). The amphiphilic macromolecules (termed TAT-PEG-Asp8-Dox) could self-assemble into nanoparticles (NPs) in water. The antitumor activity was evaluated in drug-resistant human colon cancer HCT8/ADR cells in vitro and in nude mice bearing HCT8/ADR tumor. RESULTS: The self-assembling TAT-PEG-Asp8-Dox NPs were approximately 150 nm with a narrow particle size distribution, which not only increased the cellular uptake efficiency, but also bypassed P-glycoprotein-mediated drug efflux and improved the intracellular drug retention, thus yielding an enhanced efficacy for killing drug-resistant HCT8/ADR colon cancer cells in vitro. Importantly, the TAT-PEG-Asp8-Dox NPs enhanced the intranuclear disposition of drugs for grater inhibition of DNA/RNA biosynthesis. In nude mice bearing xenografted HCT8/ADR colon cancers, intravenous or peritumoral injection of TAT-PEG-Asp8-Dox NPs for 22 d effectively inhibited tumor growth. CONCLUSION: TAT-PEG-Asp8-Dox NPs can increase cellular drug uptake and intranuclear drug delivery and retain effective drug accumulation inside the cells, thus exhibiting enhanced anticancer activity toward the drug-resistant human colon cancer HCT8/ADR cells.

The tetramerization domain potentiates Kv4 channel function by suppressing closed-state inactivation.

A-type Kv4 potassium channels undergo a conformational change toward a nonconductive state at negative membrane potentials, a dynamic process known as pre-open closed states or closed-state inactivation (CSI). CSI causes inhibition of channel activity without the prerequisite of channel opening, thus providing a dynamic regulation of neuronal excitability, dendritic signal integration, and synaptic plasticity at resting. However, the structural determinants underlying Kv4 CSI remain largely unknown. We recently showed that the auxiliary KChIP4a subunit contains an N-terminal Kv4 inhibitory domain (KID) that directly interacts with Kv4.3 channels to enhance CSI. In this study, we utilized the KChIP4a KID to probe key structural elements underlying Kv4 CSI. Using fluorescence resonance energy transfer two-hybrid mapping and bimolecular fluorescence complementation-based screening combined with electrophysiology, we identified the intracellular tetramerization (T1) domain that functions to suppress CSI and serves as a receptor for the binding of KID. Disrupting the Kv4.3 T1-T1 interaction interface by mutating C110A within the C3H1 motif of T1 domain facilitated CSI and ablated the KID-mediated enhancement of CSI. Furthermore, replacing the Kv4.3 T1 domain with the T1 domain from Kv1.4 (without the C3H1 motif) or Kv2.1 (with the C3H1 motif) resulted in channels functioning with enhanced or suppressed CSI, respectively. Taken together, our findings reveal a novel (to our knowledge) role of the T1 domain in suppressing Kv4 CSI, and that KChIP4a KID directly interacts with the T1 domain to facilitate Kv4.3 CSI, thus leading to inhibition of channel function.

Clickable protein nanocapsules for targeted delivery of recombinant p53 protein.

Encapsulating anticancer protein therapeutics in nanocarriers is an attractive option to minimize active drug destruction, increase local accumulation at the disease site, and decrease side effects to other tissues. Tumor-specific ligands can further facilitate targeting the nanocarriers to tumor cells and reduce nonspecific cellular internalization. Rationally designed non-covalent protein nanocapsules incorporating copper-free "click chemistry" moieties, polyethylene glycol (PEG) units, redox-sensitive cross-linker, and tumor-specific targeting ligands were synthesized to selectively deliver intracellular protein therapeutics into tumor cells via receptor-mediated endocytosis. These nanocapsules can be conjugated to different targeting ligands of choice, such as anti-Her2 antibody single-chain variable fragment (scFv) and luteinizing hormone releasing hormone (LHRH) peptide, resulting in specific and efficient accumulation within tumor cells overexpressing corresponding receptors. LHRH-conjugated nanocapsules selectively delivered recombinant human tumor suppressor protein p53 and its tumor-selective supervariant into targeted tumor cells, which led to reactivation of p53-mediated apoptosis. Our results validate a general approach for targeted protein delivery into tumor cells using cellular-responsive nanocarriers, opening up new opportunities for the development of intracellular protein-based anticancer treatment.

Suture cartilage formation pattern varies with different expansive forces.

INTRODUCTION: Midpalatal suture expansion could induce osteogenesis to correct maxillary insufficiency; cartilage formation could also be induced, and lower-magnitude forces might generate a preferable response pattern. In this study, we aimed for an enhanced understanding of the cartilage formatting effects of expansion. METHODS: Thirty 6-week-old male C57BL/6 mice were randomly and evenly assigned to 3 groups; the animals in each group received a sustained suture expansion at 0, 10, and 20 g, respectively. Ten additional mice were fed the same as the baseline controls and received no expansion. After 7 days, the animals were killed; coronal paraffin sections were stained using toluidine blue and safranin-O. The proliferating cell nucleus antigen, the nuclear antigen Ki-67, alkaline phosphatase, and matrix metalloproteinase 13 expressions were visualized with immunohistochemistry. All data were analyzed statistically, and the differences were considered significant at P <0.05. RESULTS: Compared with the control, the cartilage matrix volume was significantly increased by the 20-g expansion, showing increased cartilage matrix and hypertrophic chondrocytes with the highest matrix metalloproteinase 13 expression. The 10-g expansion formed condensed proliferating chondrocyte masses, within which the highest percentages of proliferating cell nucleus antigen and Ki-67 positive cells were present. The 10-g and the 20-g expansions equally intensified the alkaline phosphatase expression. CONCLUSIONS: The lower expansion (10 g) promoted chondrocyte proliferation and induced a more preferable suture cartilage response pattern compared with the higher expansion (20 g), which just increased the cartilage matrix production.

Desired properties of polymeric hydrogel vitreous substitute.

Vitreous replacement is a commonly employed method for treating a range of ocular diseases, including posterior vitreous detachment, complex retinal detachment, diabetic retinopathy, macular hole, and ocular trauma. Various clinical substitutes for vitreous include air, expandable gas, silicone oil, heavy silicone oil, and balanced salt solution. However, these substitutes have drawbacks such as short retention time, cytotoxicity, high intraocular pressure, and the formation of cataracts, rendering them unsuitable for long-term treatment. Polymeric hydrogels possess the potential to serve as ideal vitreous substitutes due to their structure-mimicking to natural vitreous and adjustable mechanical properties. Replacement with hydrogels as the tamponade can help maintain the shape of the eyeball, apply pressure to the detached retina, and ensure the metabolic transport of substances without impairing vision. This literature review examines the required properties of artificial vitreous, including the optical properties, rheological properties, expansive force action, and physiological and biochemical functions of chemically and physically crosslinked hydrogels. The strategies for enhancing the biocompatibility and injectability of hydrogels are also summarized and discussed. From a clinical ophthalmology perspective, this paper presents the latest developments in vitreous replacement, providing clinicians with a comprehensive understanding of hydrogel clinical applications, which offers guidance for future design directions and methodologies for hydrogel development.