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The spongy moth, Lymantria dispar, is a pest that damages various tree species throughout North America and Eurasia, has recently emerged in South Korea, threatening local forests and landscapes. The establishment of effective countermeasures against this species' outbreak requires predicting its potential distribution with climate change. In this study, we used species distribution models (CLIMEX and MaxEnt) to predict the potential distribution of the spongy moth and identify areas at risk of exposure to a sustained occurrence of the pest by constructing an ensemble map that simultaneously projected the outcomes of the two models. The results showed that the spongy moth could be distributed over the entire country under the current climate, but the number of suitable areas would decrease under a climate change scenario. This study is expected to provide basic data that can predict areas requiring intensive control and monitoring in advance with methodologically improved modeling technique.
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Monitoramento Ambiental , Mariposas , Animais , Florestas , República da CoreiaRESUMO
Under endoplasmic reticulum (ER)-stress conditions, the unfolded protein response (UPR) generates a defense mechanism in mammalian cells. The regulation of UPR signaling is important in oocyte maturation, embryo development, and female reproduction of pigs. Recent studies have shown that melatonin plays an important role as an antioxidant to improve pig oocyte maturation. However, there is no report on the role of melatonin in the regulation of UPR signaling and ER-stress during in vitro maturation (IVM) of porcine oocytes. Therefore, the objective of this study was to investigate the antioxidative effects of melatonin on porcine oocyte maturation through the regulation of ER-stress and UPR signaling. We investigated the changes in the mRNA/protein expression levels of three UPR signal genes (Bip/Grp78, ATF4, P90/50ATF6, sXbp1, and CHOP) on oocytes, cumulus cells, and cumulus-oocyte complexes (COCs) during IVM (metaphase I; 22 hours and metaphase II; 44 hours) by Western blot and reverse transcription-polymerase chain reaction analysis. Treatment with the ER-stress inducer, tunicamycin (Tm), significantly increased expression of UPR markers. Additionally, cumulus cell expansion and meiotic maturation of oocytes were reduced in COCs of Tm-treated groups (1, 5, and 10 µg/mL). We confirmed the reducing effects of melatonin (0.1 µmol/L) on ER-stress after pretreatment with Tm (5 µg/mL; 22 hours) in maturing COCs. Addition of melatonin (0.1 µmol/L) to Tm-pretreated COCs recovered meiotic maturation rates and expression of most UPR markers. In conclusion, we confirmed a role for melatonin in the modulation of UPR signal pathways and reducing ER-stress during IVM of porcine oocytes.
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Antioxidantes/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Meiose/efeitos dos fármacos , Melatonina/farmacologia , Oogênese/efeitos dos fármacos , Animais , Células do Cúmulo/efeitos dos fármacos , Feminino , Oócitos/efeitos dos fármacos , Suínos , Resposta a Proteínas não Dobradas/efeitos dos fármacosRESUMO
Gangliosides are components of the mammalian plasma membrane that help regulate receptor signaling. Ganglioside GM3, for example, plays an important role in initiating apoptosis in cancer cells; however, physiological roles for GM3 in normal processes, such as during pig oocyte maturation, are not clear. The aim of this study was to investigate the functional link between GM3 and cellular apoptosis in porcine cumulus-oocyte-complexes (COCs) during in vitro maturation. Our results indicated that denuded oocytes possess less ST3GAL5, a GM3-synthesizing enzyme, than cumulus cells or COCs after 44 hr of in vitro maturation. GM3 also affected the meiotic maturation of cultured pig oocytes, as evaluated by orcein staining. In vitro treatment of COCs with exogenous GM3 also reduced cumulus cell expansion, the proportion of meiotic maturation, and increased cumulus cell transcription of PTX3, TNFAIP6, and HAS2. Interestingly, GM3 treatment reduced the expression of Epidermal growth factor receptor (EGFR)-mediated Phosphoinositide 3-kinase/AKT signaling proteins in COCs in a concentration-dependent manner, instead increasing the abundance of pro-apoptotic factors such as AIF, activated Caspase 9, cleaved PARP1, and Caspase 3 were. Thus, GM3 might affect porcine oocyte maturation via suppression of EGFR-mediated PI3K/AKT signaling and/or induction of apoptosis during in vitro maturation.
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Apoptose/efeitos dos fármacos , Células do Cúmulo/efeitos dos fármacos , Receptores ErbB/metabolismo , Gangliosídeo G(M3)/farmacologia , Oogênese/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Animais , Células do Cúmulo/citologia , Feminino , Modelos Biológicos , Oócitos/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , SuínosRESUMO
A new Schiff-base colorimetric chemosensor 1 was developed for the detection of Cu2+, Co2+ and S2-. Sensor 1 could simply monitor Cu2+ and Co2+ by a color change from colorless to yellow. The binding modes of 1 to Cu2+ and Co2+ were determined to be a 2 : 1 complexation stoichiometry through Job's plot and ESI-mass spectrometry analysis. The detection limits (0.02 µM and 0.63 µM) for Cu2+ and Co2+ were lower than the recommended values (31.5 µM and 1.7 µM) by the World Health Organization (WHO) for Cu2+ and the Environmental Protection Agency (EPA) for Co2+, respectively. Importantly, 1 could detect and quantify Cu2+ in real water samples. In addition, the Cu2+-2·1 complex could be used as a highly selective colorimetric sensor for S2- in the presence of other anions without any interference. Moreover, the sensing mechanisms of Cu2+ and Co2+ by 1 were explained by theoretical calculations.
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[Purpose] The present study aimed to determine the changes in the muscle activities of the gluteus medius, latissimus dorsi, and gluteus maximus at different gait speeds, to collect basic data for the study of the gluteus medius. [Subjects and Methods] The subjects were 18 young and healthy male adults whose mean age, height, and weight were 26.4â years, 173.37â cm, and 72.5â kg, respectively. Electromyograpy was used to measure the maximum voluntary isometric contraction of each muscle three times and the values averaged. Then, the subjects walked on a treadmill at gait speeds of 1.5 m/s, 2.5 m/s, and 3.5 m/s and the muscle activity of each muscle was measured. [Results] The gluteus medius showed no significant difference in muscle activity among the different gait speeds. [Conclusion] For selectively strengthening the gluteus medius, to establish the external stability of the pelvis during walking, weight loading or sloped treadmills are effective interventions. However, different gait speeds exert no significant effect on the selective strengthening of the gluteus medius.
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Despite the remarkable advances of dermal fillers that reduce wrinkles caused by dermis thickness reduction, they still lack effective hydrogel systems that stimulate collagen generation along with injection convenience. Here, we develop a stem cell-derived extracellular vesicle (EV)-bearing thermosensitive hydrogel (EVTS-Gel) for effective in vivo collagen generation. The TS-Gel undergoes sol-gel transition at 32.6 °C, as demonstrated by the storage and loss moduli crossover. Moreover, the TS-Gel and the EVTS-Gel have comparable rheological properties. Both hydrogels are injected in a sol state; hence, they require lower injection forces than conventional hydrogel-based dermal fillers. When locally administered to mouse skin, the TS-Gel extends the retention time of EVs by 2.23 times. Based on the nature of the controlled EV release, the EVTS-Gel significantly inhibits the dermis thickness reduction caused by aging compared to the bare EV treatment for 24 weeks. After a single treatment, the collagen layer thickness of the EVTS-Gel-treated dermis becomes 2.64-fold thicker than that of the bare EV-treated dermis. Notably, the collagen generation efficacy of the bare EV is poorer than that of the EVTS-Gel of a 10× lesser dose. Overall, the EVTS-Gel shows potential as an antiaging dermal filler for in vivo collagen generation.
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Colágeno , Derme , Vesículas Extracelulares , Hidrogéis , Animais , Camundongos , Derme/metabolismo , Derme/efeitos dos fármacos , Colágeno/química , Hidrogéis/química , Hidrogéis/farmacologia , Vesículas Extracelulares/química , Vesículas Extracelulares/metabolismo , Humanos , Células-Tronco/citologia , Células-Tronco/metabolismo , Células-Tronco/efeitos dos fármacos , Preenchedores Dérmicos/química , Preenchedores Dérmicos/farmacologiaRESUMO
"Smart" biomaterials that are responsive to pathological abnormalities are an appealing class of therapeutic platforms for the development of personalized medications. The development of such therapeutic platforms requires novel techniques that could precisely deliver therapeutic agents to the diseased tissues, resulting in enhanced therapeutic effects without harming normal tissues. Among various therapeutic platforms, injectable pH-responsive biomaterials are promising biomaterials that respond to the change in environmental pH. Aqueous solutions of injectable pH-responsive biomaterials exhibit a phase transition from sol-to-gel in response to environmental pH changes. The injectable pH-responsive hydrogel depot can provide spatially and temporally controlled release of various bioactive agents including chemotherapeutic drugs, peptides, and proteins. Therapeutic agents are imbibed into hydrogels by simple mixing without the use of toxic solvents and used for long-term storage or in situ injection using a syringe or catheter that could form a stable gel and acts as a controlled release depot in a minimally invasive manner. Tunable physicochemical properties of the hydrogels, such as biodegradability, ability to interact with drugs and mechanical properties, can control the release of the therapeutic agent. This review highlights the advances in the design and development of biodegradable and in situ forming injectable pH-responsive biomaterials that respond to the physiological conditions. Special attention has been paid to the development of amphoteric pH-responsive biomaterials and their utilization in biomedical applications. We also highlight key challenges and future directions of pH-responsive biomaterials in clinical translation.
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Materiais Biocompatíveis , Hidrogéis , Preparações de Ação Retardada/química , Hidrogéis/química , Materiais Biocompatíveis/farmacologia , Proteínas , Concentração de Íons de HidrogênioRESUMO
We herein report manufacturing of dental crowns made of 5-mol% yttria partially stabilized zirconia (5Y-PSZ) with desired mechanical properties, optical translucency and dimensional accuracy using digital light processing (DLP). To this end, all processing parameters were carefully controlled and optimized. First, 5Y-PSZ particles with a bimodal distribution were prepared via calcination of as-received granules and subsequent ball-milling and then used to formulate 5Y-PSZ suspensions with a high solid loading of 50 vol% required for high densification after sintering. Dispersant content was also optimized. To provide high dimensional accuracy, initial dimensions of dental crowns for 3D printing were precisely determined by considering increase and decrease in dimensions during photopolymerization and sintering, respectively. Photopolymerization time was also optimized for a given layer thickness of 50 µm to ensure good bonding between layers. A multi-step debinding schedule with a slow heating rate was employed to avoid formation of any defects. After sintering at 1500 °C for 2 h, 5Y-PSZ could be almost fully densified without noticeable defects within layers and at interfaces between layers. They had high relative densities (99.03 ± 0.39%) with a high cubic phase content (59.1%). These characteristics allowed for achievement of reasonably high mechanical properties (flexural strength = 625.4 ± 75.5 MPa and Weibull modulus = 7.9) and % transmittance (31.4 ± 0.7%). In addition, 5Y-PSZ dental crowns showed excellent dimensional accuracy (root mean square (RMS) for marginal discrepancy = 44.4 ± 10.8 µm and RMS for internal gap = 22.8 ± 1.6 µm) evaluated by the 3D scanning technique.
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The western conifer seed bug (WCSB) Leptoglossus occidentalis (Heidemann) (Heteroptera: Coreidae) is a pest insect that causes significant losses of coniferous trees worldwide. In this study, we sought to project the potential distribution of the WCSB based on dual CLIMEX modeling and random forest (RF) analysis to obtain basic data for WCSB monitoring strategies. The CLIMEX model, a semimechanistic niche model that responds to climate-based environmental parameters, is a species distribution model that focuses on regional climatic suitability. Given that this model can be used to select areas that are likely to reflect the climatically favorable spread of species, which we initially used CLIMEX to evaluate the potential distribution of the WCSB. The RF algorithm was used to predict the potential occurrence of WCSB and to evaluate the relative importance of environmental variables for WCSB occurrence. Using the RF model, land cover was found to be the most important variable for classifying the presence/pseudo-absence of the WCSB, with an accuracy of 77.1%. Climatic suitability for the WCSB was predicted to be 2.4-fold higher in Southern Europe than in Western Europe, and the WCSB was predicted to occur primarily near coniferous forests. Given that CLIMEX and RF analyses yielded different prediction results, using the findings of both models may compensate for the shortcomings of these models when used independently. Consequently, to ensure greater prediction reliability, we believe that it would be beneficial to base predictions on the combined potential distribution data obtained using both modeling approaches.
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OBJECTIVE: This study explored microdosing methods for evaluating the distribution and pharmacokinetics (PK) of a central nervous system (CNS) drug candidate. METHODS: We used sertraline as a model drug. In this open-label, one-arm, three-period, multiple-dosing study, 10 healthy male volunteers received 6-day administrations of sertraline at doses of 5, 25 or 50 mg/d in three different periods. Before the first dose of Period 1, and 24 h after the last dose of each period, an intravenous bolus of [11C]sertraline was injected for positron emission tomography (PET) scanning. After the sixth dose in each period, serial blood samples were collected at scheduled intervals over 48 h; then serum sertraline concentrations were determined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). RESULTS: Sertraline was distributed in the brain within 20 min, and it was highly distributed in the putamen, cingulate, and thalamus. Linearity in steady-state Cmax and AUClast were observed in the 5 - 50 mg dose range. The results suggested that microdosing with PET was a useful method for exploring the bloodbrain- barrier penetration and distribution of a candidate CNS drug. CONCLUSIONS: This study described a microdosing method that combined PET with LC-MS/MS for determining the brain distribution and PK characteristics of a CNS drug candidate.
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Radioisótopos de Carbono , Tomografia por Emissão de Pósitrons/métodos , Inibidores Seletivos de Recaptação de Serotonina/farmacocinética , Sertralina/farmacocinética , Adulto , Humanos , Masculino , Sertralina/administração & dosagemRESUMO
BACKGROUND: Changes in dietary patterns have led to a decrease in rice consumption, raising demands for the cultivation of alternative crops that meet the current requirements. Potatoes are highly productive and can be stored for a relatively long period, thereby ensuring adequate income for farmers; however, optimal cultivation is necessary to maximize yield. OBJECTIVE: This study proposes optimal cultivation regions for potato considering climate and soil conditions. MATERIALS AND METHODS: The CLIMEX model was developed to evaluate climatic suitability, while the soil suitability was scored based on five soil characteristics. The final areal suitability for potato cultivation was classified into 4 levels: very suitable, suitable, marginal, and unsuitable. RESULTS: Overall, 36.5% of South Korea had very suitable climate, but areas with the best values for soil conditions were approximately 10% of the climatically suitable areas. When considering the climate and soil conditions simultaneously, climatic suitability and soil condition were inversely related, resulting in only 1.2% of optimal areas with Jeju Island as the most suitable area. CONCLUSIONS: Because both climate and soil conditions need to be suitable for growing crops, this study can provide potential paddy-cultivation areas for potato cultivation and a method for evaluating suitable areas for crop cultivation.
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Solo , Solanum tuberosum , Mudança Climática , Produtos Agrícolas , República da CoreiaRESUMO
Background: Despite remarkable advances in sonodynamic therapy (SDT) of cancer, the low reactive oxygen species (ROS) quantum yield of the sonosensitizer remains a critical concern in glutathione (GSH)-overexpressing cancer cells. Methods: For enhanced SDT, we report hydrophilized self-immolative polymer (SIP)-decorated TiO2 nanoparticles (HSIPT-NPs) to achieve on-demand GSH depletion and ROS generation. Results: Upon intracellular delivery of HSIPT-NPs into hydrogen peroxide-rich cancer cells, SIP is degraded through electron transfer to produce GSH-depleting quinone methide, reprogramming GSH high cancer cells into GSH low phenotype. In the presence of ultrasound, compared to conventional TiO2 NPs, HSIPT-NPs induce significantly higher oxidative stress to cancer cells by incapacitating their antioxidant effects. SDT with HSIPT-NPs effectively inhibit tumor growth in mice via the synergistic effects of GSH depletion and ROS generation. Conclusion: On the basis of their ability to reprogram cancer cells, HSIPT-NPs offer considerable potential as a nanosensitizer for enhanced SDT.
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Nanopartículas , Neoplasias , Camundongos , Animais , Espécies Reativas de Oxigênio/metabolismo , Glutationa/metabolismo , Antioxidantes/farmacologiaRESUMO
Extracellular vesicles (EVs) have shown great potential in disease diagnosis and treatment; however, their clinical applications remain challenging due to their unsatisfactory long-term stability and the lack of effective delivery strategies. In this study, we prepared human adipose stem cell-derived EV (hASC-EV)-loaded hyaluronic acid dissolving microneedles (EV@MN) to investigate the feasibility of EVs for their clinical applications. The biological activities of the EVs in this formulation were maintained for more than six months under mild storage conditions, especially at temperatures lower than 4 °C. Moreover, the EV@MN enabled precise and convenient intradermal delivery for sustained release of EVs in the dermis layer. Therefore, EV@MN significantly improved the biological functions of hASC-EVs on dermal fibroblasts by promoting syntheses of proteins for the extracellular matrix such as collagen and elastin, enhancing fibroblast proliferation, and regulating the phenotype of fibroblast, compared with other administration methods. This research revealed a possible and feasible formulation for the clinical application of EVs.
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Hyaluronic acid-based hydrogels (Hyal-Gels) have the potential to reduce wrinkles by physically volumizing the skin. However, they have limited ability to stimulate collagen generation, thus warranting repeated treatments to maintain their volumizing effect. In this study, stem cell-derived extracellular vesicle (EV)-bearing Hyal-Gels (EVHyal-Gels) were prepared as a potential dermal filler, ameliorating the dermis microenvironment. No significant differences were observed in rheological properties and injection force between Hyal-Gels and EVHyal-Gels. When locally administered to mouse skin, Hyal-Gels significantly extended the biological half-life of EVs from 1.37 d to 3.75 d. In the dermis region, EVHyal-Gels induced the overexpression of CD301b on macrophages, resulting in enhanced proliferation of fibroblasts. It was found that miRNAs, such as let-7b-5p and miR-24-3p, were significantly involved in the change of macrophages toward the CD301bhi phenotype. The area of the collagen layer in EVHyal-Gel-treated dermis was 2.4-fold higher than that in Hyal-Gel-treated dermis 4 weeks after a single treatment, and the collagen generated by EVHyal-Gels was maintained for 24 weeks in the dermis. Overall, EVHyal-Gels have the potential as an antiaging dermal filler for reprogramming the dermis microenvironment.
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Preenchedores Dérmicos , Vesículas Extracelulares , Camundongos , Animais , Preenchedores Dérmicos/farmacologia , Derme , Ácido Hialurônico/farmacologia , Fibroblastos , Colágeno/farmacologia , Hidrogéis/farmacologia , Células-Tronco , MacrófagosRESUMO
Hepatocellular carcinoma is the most common malignancy with a high incidence rate and is the leading cause of cancer-related deaths. Herein, we developed a thermo-responsive hydrogel comprising poly(ε-caprolactone-co-lactide)-b-poly(ethylene glycol)-b-poly(ε-caprolactone-co-lactide (PCLA) that exhibits acidity-accelerated delivery of the tumor-targeting glucuronic acid-bearing doxorubicin (DOX-pH-GA) conjugate into tumor tissues. The PCLA copolymer was post-modified with boronic acid (BA-PCLA) to covalently cross-link with the pH-responsive DOX-pH-GA conjugate. The BA-PCLA copolymer effectively coordinated with the DOX-pH-GA conjugate through the boronate ester formation and showed a lower critical gelation temperature. The DOX conjugated via boronate ester exhibited a sustained release in vitro. Subcutaneous administration of PCLA copolymers formed in situ gels in the subcutaneous layers of Sprague-Dawley rats and degraded after 6 weeks. Similarly, BA-PCLA copolymers coordinated with DOX-pH-GA formed a stable in situ gel in vivo. In vivo imaging studies demonstrated that DOX-pH-GA was released in a sustained manner. The anti-tumor activity of the DOX releasing injectable hydrogel was examined using a HepG2 liver cancer xenograft model. The in vivo antitumor effect demonstrated that the DOX releasing hydrogel depot remarkably suppresses the tumor growth. These results demonstrate that the pH-responsive DOX releasing thermo-responsive hydrogel depot has great potential for application in localized anticancer therapy.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Animais , Carcinoma Hepatocelular/tratamento farmacológico , Ésteres , Hidrogéis , Concentração de Íons de Hidrogênio , Neoplasias Hepáticas/tratamento farmacológico , Ratos , Ratos Sprague-DawleyRESUMO
Immune checkpoint therapy (ICT), which reinvigorates cytotoxic T cells, provides clinical benefits as an alternative to conventional cancer therapies. However, its clinical response rate is too low to treat an immune-excluded tumor, owing to the presence of abundant stromal elements impeding the penetration of immune cells. Here, we report that macitentan, a dual endothelin receptor antagonist approved by the FDA to treat pulmonary arterial hypertension, can be repositioned to modulate the desmoplastic tumor microenvironment (TME). In the 4T1 orthotopic tumor model, the polymeric nanoparticles bearing macitentan (M-NPs) prevent fibrotic progression by regulating the function of cancer-associated fibroblasts, attenuate the biogenesis of cancer cell-derived exosomes, and modulate the T cell subsets and distribution in TME. These results demonstrate that the M-NPs effectively reorganize the immunosuppressive TME by targeting the endothelin-1 axis and consequently exhibit synergistic antitumor effects in combination with ICT.
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Nanopartículas , Microambiente Tumoral , Inibidores de Checkpoint Imunológico , Pirimidinas , Sulfonamidas/farmacologiaRESUMO
Osteoporosis is one of the most common skeletal disorders caused by the imbalance between bone formation and resorption, resulting in quantitative loss of bone tissue. Since stem cell-derived extracellular vesicles (EVs) are growing attention as novel cell-free therapeutics that have advantages over parental stem cells, the therapeutic effects of EVs from adipose tissue-derived stem cells (ASC-EVs) on osteoporosis pathogenesis were investigated. ASC-EVs were isolated by a multi-filtration system based on the tangential flow filtration (TFF) system and characterized using transmission electron microscopy, dynamic light scattering, zeta potential, flow cytometry, cytokine arrays, and enzyme-linked immunosorbent assay. EVs are rich in growth factors and cytokines related to bone metabolism and mesenchymal stem cell (MSC) migration. In particular, osteoprotegerin (OPG), a natural inhibitor of receptor activator of nuclear factor-κB ligand (RANKL), was highly enriched in ASC-EVs. We found that the intravenous administration of ASC-EVs attenuated bone loss in osteoporosis mice. Also, ASC-EVs significantly inhibited osteoclast differentiation of macrophages and promoted the migration of bone marrow-derived MSCs (BM-MSCs). However, OPG-depleted ASC-EVs did not show anti-osteoclastogenesis effects, demonstrating that OPG is critical for the therapeutic effects of ASC-EVs. Additionally, small RNA sequencing data were analysed to identify miRNA candidates related to anti-osteoporosis effects. miR-21-5p in ASC-EVs inhibited osteoclast differentiation through Acvr2a down-regulation. Also, let-7b-5p in ASC-EVs significantly reduced the expression of genes related to osteoclastogenesis. Finally, ASC-EVs reached the bone tissue after they were injected intravenously, and they remained longer. OPG, miR-21-5p, and let-7b-5p in ASC-EVs inhibit osteoclast differentiation and reduce gene expression related to bone resorption, suggesting that ASC-EVs are highly promising as cell-free therapeutic agents for osteoporosis treatment.
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Tecido Adiposo/metabolismo , Vesículas Extracelulares/metabolismo , Osteoporose/terapia , Osteoprotegerina/genética , Células-Tronco/metabolismo , Animais , Modelos Animais de Doenças , Feminino , Humanos , Camundongos , Osteoporose/patologiaRESUMO
Allogeneic transplantation of mesenchymal stem cell-derived extracellular vesicles (EVs) offers great potential for treating liver fibrosis. However, owing to their intrinsic surface characteristics, bare EVs are non-specifically distributed in the liver tissue after systemic administration, leading to limited therapeutic efficacy. To target activated hepatic stellate cells (HSCs), which are responsible for hepatic fibrogenesis, vitamin A-coupled small EVs (V-EVs) were prepared by incorporating vitamin A derivative into the membrane of bare EVs. No significant differences were found in the particle size and morphology between bare and V-EVs. In addition, surface engineering of EVs did not affect the expression of surface marker proteins (e.g., CD63 and CD9), as demonstrated by flow cytometry. Owing to the surface incorporation of vitamin A, V-EVs were selectively taken up by activated HSCs via receptor-mediated endocytosis. When systemically administered to mice with liver fibrosis, V-EVs effectively targeted activated HSCs in the liver tissue, resulting in reversal of the fibrotic cascade. Consequently, even at a 10-fold lower dose, V-EVs exhibited comparable anti-fibrotic effects to those of bare EVs, substantiating their therapeutic potential for liver fibrosis.
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Vesículas Extracelulares , Células-Tronco Mesenquimais , Animais , Células Estreladas do Fígado , Cirrose Hepática/tratamento farmacológico , Camundongos , Vitamina ARESUMO
Therapeutic proteins are attractive candidates for the treatment of human diseases. However, their short half-life often limits their clinical application. To overcome this problem, injectable hydrogels have been developed as depots for controlled release of therapeutic proteins, but these systems have not yet achieved the desired extended, sustained drug release profile. Our strategy herein was to implement selective and strong interactions between the hydrogels and therapeutic proteins. Specifically, we investigated whether strong and specific interactions between human serum albumin (HSA) and albumin-binding peptide (ABP) can be used to achieve extended release of urate oxidase (Uox), a therapeutic protein for hyperuricemia treatment, from pH- and temperature-sensitive injectable hydrogels consisting of poly(ethylene glycol)-poly(ß-amino ester urethane) (PEG-PAEU) copolymer. Thus, HSA was conjugated to Uox (Uox-HSA) and ABP was introduced in PEG-PAEU (PEG-PAEU-ABP). Polymers, conjugates, and hydrogels were extensively characterized for their physicochemical characteristics and in vivo efficacy in a hyperuricemia mouse model. Briefly, the hydrogels exhibited good injectability, in vitro biocompatibility and extended drug release, and in vivo gel formation and degradability. The serum half-life of the Uox-HSA loaded in PEG-PAEU-ABP hydrogels was ~96 h in mice, which was ~88, ~5.5, and ~2 times longer than that of free native Uox, free Uox-HSA, and Uox-HSA loaded in PEG-PAEU hydrogels, respectively. In the hyperuricemia mouse model, Uox-HSA loaded in PEG-PAEU-ABP hydrogels exhibited a substantially extended period of uric acid-lowering efficacy. These results clearly show that by applying ABP-HSA strong interaction to injectable hydrogels and therapeutic protein, the concentration of the therapeutic protein can be maintained for a long period in vivo, prolonging its therapeutic effect. Further, our approach can be tailored to accommodate other therapeutic proteins, which potentially expands the clinical applicability range of these systems.
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Hiperuricemia , Urato Oxidase , Animais , Preparações de Ação Retardada , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Hidrogéis , Hiperuricemia/tratamento farmacológico , Camundongos , Polietilenoglicóis , Albumina Sérica HumanaRESUMO
Mito-TEMPO is a well-known mitochondria-specific superoxide scavenger. However, the effect of Mito-TEMPO on porcine embryo development, to our knowledge, has not been studied yet. In the present study, porcine embryos were classified into two groups (G1 and G2) based on the cytoplasm lipid contents at the zygote stage. The development of blastocysts derived from G2 zygotes was reduced (G2:16.2 ± 7.9% vs G1: 26.5 ± 5.9%; 1.6-fold, p < 0.05) compared to those from G1 zygotes. In G2 embryos, the proportion of TUNEL-positive cells was also higher than that of G1 embryos. Superoxide in G2 embryos was significantly increased compared to that in G1 embryos. Mitochondrial membrane potential and ATP production were lower in G2 embryos than in G1 embryos. Phosphorylation of Drp1 at Ser 616 increased in G1 embryos during the cleavage stages compared to that in the zygote but was not significantly different in G2 embryos. Then, the effects of Mito-TEMPO were investigated in G2 embryos. Blastocyst formation rate (G2: 19.1 ± 5.1% vs G2 + Mito-TEMPO: 28.8 ± 4.0%; 1.5-fold, p < 0.05) and mitochondrial aggregation were recovered after superoxide reduction by Mito-TEMPO treatment. Thus, we showed that Mito-TEMPO improves blastocyst development by superoxide reduction in porcine embryos in vitro.