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We constructed and optimized the plasmid DNA (pDNA) Opt-S encoding the gene of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein, using poly (lactic-co-glycolic acid) copolymer (PLGA) as a delivery carrier for pDNA. PLGA-pDNA NPs were loaded by nanoprecipitation and its properties in vitro were preliminary evaluated. The results showed that the prepared PLGA-pDNA NPs were regular morphology, clear edges, with an average particle size of (184.2 ± 2.4) nm, polydisperse index (PDI) of 0.093 ± 0.013, zeta potential of (-68.10 ± 0.36) mV, and encapsulation rate of (98.92 ± 0.22)%. The PLGA-pDNA NPs were stable at -20 ℃ for 7 months and could protect pDNA against nuclease degradation. And they also exhibited sustained release of pDNA in vitro. The PLGA-pDNA NPs have low cytotoxicity and high safety. In addition, in vitro transfection experiments showed that the SARS-CoV-2 S gene could enter cells and be expressed. These results indicate that PLGA-pDNA NPs non-viral gene vector have simple preparation process and good performance, which are expected to provide a new idea for the research and development of SARS-CoV-2 vaccine.
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@#In this study, the conjugate of eicosapentaenoic acid (EPA) and hyaluronic acid (HA) was synthesized and the anti-hepatoma activities in vitro were evaluated.The hyaluronic acid-eicosapentaenoic acid (HA-EPA)nanoparticle was synthesized by linking eicosapentaenoic acid with hyaluronic acid with cystamine.The structure of HA-EPA was characterized by nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy (FT-IR).Laser particle sizer and Zeta potential analyzer were used to detect the size and potential of HA-EPA.MTT assay was used to detect the anti-proliferative effect of HA-EPA on HepG2, Huh-7 and LX-2 cells in vitro.The effects of HA-EPA nanoparticles on the proliferation and apoptosis of HepG2 cells in vitro were investigated by EdU staining and TUNEL staining. The apoptosis was further confirmed by flow cytometry.The effect of HA-EPA nanoparticles on the migration and invasion of HepG2 cells was demonstrated by transwell and invasion experiments.The results of 1H NMR showed that HA-EPA was successfully synthesized, and the grafting rate of EPA on HA was (40 ± 5) %. The structure of HA-EPA was further confirmed by FT-IR.The particle size was (162.5 ± 10.2) nm, and the potential was -(4.47 ± 0.31) mV.MTT results showed that, with the prolongation of drug treatment time, HA-EPA showed a better inhibitory effect on the activity of HepG2 and Huh-7 cells than EPA under the same EPA content.After treated for 48 hours, the toxicity of HA-EPA to LX-2 cells was less than that of EPA.The results of 24-hour proliferation, apoptosis, migration and invasion of HepG2 showed that, the graft of hyaluronic acid improved the ability of EPA to inhibit proliferation, promote apoptosis, migration and invasion of HepG2 cells (P < 0.001), indicating that grafting of HA can significantly enhance the inhibitory effect of EPA on liver cancer with some role in reducing toxicity.
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Objective:To conduct theoretical analysis and experimental research on peripherally inserted venous catheters, establish theoretical models of interaction between different materials catheters and simulated skin tissues, and test different catheters at the same time to provide theoretical basis and experimental basis for the optimization design.Methods:According to the mechanical properties of the catheter at 25 ℃ and 37 ℃, a finite element model of the catheter and simulate skin tissue was established. The relationship between catheter folds and material and structure during puncture was analyzed, and the stiffness, radiographic properties, etc. were tested experimentally.Results:The performance of the catheter at different temperatures is closely related to its material. The wrinkle situation of the catheter is related to the catheter material and the inclination of the wedge surface. The elastic modulus of the polyurethane (PU) catheter is about 500 MPa and 250 MPa, respectively at room temperature (25 ℃) and body temperature (37 ℃), which meets the clinical needs of high rigidity during puncture and soft material during indwelling. When the catheter structure is the same, the PU catheter is less prone to wrinkles than the fluorinated ethylene propylene copolymer (FEP) catheter. When the catheter material is the same, the smaller the inclination of the wedge surface, the less likely the catheter to wrinkle.Conclusions:Appropriately reducing the inclination of the wedge-shaped surface of the needle of peripherally inserted venous catheters can improve the success rate of puncture. The PU catheters have good mechanical properties, they are not prone to wrinkles during puncture, and their stiffness can be reduced at body temperature, which can not only increase the success rate of puncture, but also reduce the occurrence of complications. Therefore, PU catheters have a better clinical application prospect.
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The beneficial or deleterious effects of nanomedicines emerge from their complex interactions with intracellular pathways and their subcellular fate. Moreover, the dynamic nature of plasma membrane accounts for the movement of these nanocarriers within the cell towards different organelles thereby not only influencing their pharmacokinetic and pharmacodynamic properties but also bioavailability, therapeutic efficacy and toxicity. Therefore, an in-depth understanding of underlying parameters controlling nanocarrier endocytosis and intracellular fate is essential. In order to direct nanoparticles towards specific sub-cellular organelles the physicochemical attributes of nanocarriers can be manipulated. These include particle size, shape and surface charge/chemistry. Restricting the particle size of nanocarriers below 200 nm contributes to internalization
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After entering the physiological environment, proteins and other biomolecules bind to the nanoparticles' surface, called protein corona. The corona establishes a new bio-interface that affects its physicochemical properties and biological behaviors. Variations in types and contents of human plasma proteins during the different physiological states can substantially change the composition and effects of the corona. With folic acid (FA)-modified polylactic acid-polyglycolic acid copolymer (PLGA) nanoparticles, the formation of protein coronas and their influence on the targeting capability are studied in healthy and ovarian human plasma. All human plasma samples were collected at the Peking University Third Hospital and this study protocol has been approved by Peking University Third Hospital Medical Science Research Ethics Committee (2019-409-1). Dynamic light scattering measurements demonstrated a 10-40 nm increase in their size distributions and a 30 mV decreased in their absolute zeta-potential since protein corona-coated PLGA-PEG and PLGA-FA were formed. The SDS-PAGE analysis showed the composition of the protein coronas from ovarian and healthy plasma in PLGA-FA were markedly distinct, particularly for proteins with molecular weight of 45, 110 and >180 kDa. Flow cytometry indicated that the absorption of ovarian plasma in PLGA-FA led to a lower cellular uptake by SKOV3 cells. Our results suggest that in vitro formed ovarian plasma protein corona could shield targeting molecules and reduced receptor-mediated internalization. The results of this pilot study will provide evidence of the effectiveness of active targeting nanoparticles under pathologic conditions. Additionally, the protein corona in different diseases is emerging as a key point; thus, a comprehensive understanding could accelerate clinical translation of functionalized nanoparticles.
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To prepare the mimetic exosomes and co-delivery proteins and nucleic acids, and achieve efficient and safe co-delivery of multi-component drugs, an optimized formulation was designed by modifying a polylactic acid-glycolic acid copolymer (PLGA) matrix with a cationic lipid excipient dioleyl trimethylammonium propane (DOTAP), and a PLGA/DOTAP nanoparticles packaged protein and nucleic acid was prepared by double emulsion method, and the outermost membrane structure prepared by reverse phase evaporation method and consists of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and membrane proteins. The structure of the mimetic exosomes is formed by ultrasonic dispersion and extrusion, and analyzed its characteristics and nature of the transfer effect. The size of mimetic exosomes was about 156.13 nm, with negative charge (-18.23 ± 0.57 mV), and it could efficiently co-transfer protein and siRNA, and siRNA could effectively inhibit the expression of target gene Trim28. The mimetic exosomes simulate the structure of exosomes and achieve safe and efficient co-delivery of multi-component drugs.
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BACKGROUND: Polyisobutylene and block copolymer and its crosslinked product are a kind of novel thermoplastic elastomer. They have unique properties and excellent biocompatibility, which is a promising medical biomaterial and applied extensively. OBJECTIVE: To review the research progress and applications of polyisobutylene and its thermoplastic elastomers, and to discuss the application prospect of polyisobutylene-based polymers as medical implant materials. METHODS: A computer-based retrieval of PubMed, Web of Science, CNKI and WanFang databases was conducted for the articles about polyisobutylene published from 1958 to 2019. The key words were “polyisobutylene and block copolymer, polyisobutylene and thermoplastic elastomer, polyisobutylene and biomaterials, polyisobutylene and modification, polyisobutylene and medical application” in English and Chinese, respectively. In accordance with the inclusion and exclusion criteria, 65 eligible articles were included for review. RESULTS AND CONCLUSION: Polyisobutylene and block copolymer and its crosslinked products have favorable biocompatibility and stability. By making full use of polyisobutylene-based materials’ advantages, with the combination of other biomaterials and usage of new technology for surface modification, the copolymer will be more competitive in the field of medical implant in the future, including eye implant materials, soft biomaterials and drug delivery systems.
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Objective: To investigate the applicability of the spray-dried microspheres of vinyl pyrrolidone/vinyl acetate copolymer VA64-Soluplus for inclusion of cinnamon oil (CO) and compare with traditional inclusion technology of β-cyclodextrin. Methods: HPLC was used to determine the encapsulation rate of inclusion complex. Transmission electron microscopy (TEM), scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and X-ray powder diffraction (XRD) were used to characterize the inclusion complex; The dissolution and stability of the inclusion complex was investigated by in vitro release test and accelerated stability test; The pharmacokinetic and analgesic efficacy tests were used to examine the bioavailability and efficacy of the inclusion complex. Results: The encapsulation rate of microsphere inclusion complex and β-cyclodextrin inclusion complex was (98.38 ± 0.30)% and (86.51 ± 0.52)%, respectively. Observation of the inclusion complex under TEM showed a uniform spherical-like structure with uniform dispersion; Observation under SEM showed that the inclusion complex was spherical with a concave surface; The endothermic peak of volatile oil of cinnamon in DSC and the diffraction peak in XRD disappeared. The cinnamon volatile oil was dispersed in theinclusion complex in the form of non-aggregation; The cumulative release rates of cinnamon volatile oil, microsphere inclusion complex and β-cyclodextrin inclusion complex in in vitro dissolution experiments were 97.05%, 93.36% and 80.26%, respectively; Accelerated stability test at 60 ℃ showed that the loss rate of volatile oil of microsphere inclusion complex was significantly lower than that of cinnamon volatile oil and β-cyclodextrin inclusion complex; Pharmacokinetics showed that the AUC0-∞ of cinnamon essential oil, microsphere inclusion complex and β-cyclodextrin inclusion complex were basically the same; Pharmacodynamics showed that the analgesic rates of cinnamon volatile oil in the three groups were 53.0%, 47.5% and 21.1%, respectively. Conclusion: The stability of cinnamon volatile oil was enhanced by the combination of spray-dried microspheres of vinyl pyrrolidone/vinyl acetate copolymer VA64-Soluplus. The in vitro release, bioavailability and analgesic efficacy of microsphere were basically consistent with the volatile oil of cinnamon volatile oil, and it was superior to the inclusion compound of β-cyclodextrin. The vinyl pyrrolidone/vinyl acetate copolymer VA64-Soluplus microsphere inclusion compound had better water solubility. This study provides a new method for the inclusion of volatile oil.
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Inhibition of bitterness is a significant measure to improve the compliance and clinical efficacy of traditional Chinese medicine(TCM) decoction. According to the characteristics of TCM decoction, such as high dispersion of bitterness components, multi-component bitterness superposition and strong instantaneous stimulation, the research group put forward a new strategy to inhibit bitterness in the early stage based on the self-assembly characteristics of amphiphilic substances in aqueous solution, in order to reduce the distribution of bitterness components in real solution and achieve the purpose of bitter-masking. It was found that the bitter-masking effect of amphiphilic substances was different on the bitter compounds of various structures. Therefore, it was speculated that there might be a certain relationship between the bitter inhibition effect and the substrate structure. In this paper, the interaction between mPEG-PLLA and five bitter alkaloids(bamatine, jatrorrhizine, berberine, epiberberine and coptisine) in Coptidis Rhizoma was studied to explore the effect of substrate structure on the inhibition of bitterness. The sensory test of volunteers was used to determine the bitter-masking effect of mPEG-PLLA on the decoction of Coptidis Rhizoma and its main bitter alkaloids. The molecular docking and molecular force field were applied to locate the bitter groups and the bitter-masking parts. The relationship between the bitter strength and the structure was analyzed by the surface electrostatic potential of the bitter alkaloids, and the correlation between the bitter-masking effect and the structural parameters of the bitter components was explored by factor analysis, so as to clarify the structure-activity relationship of mPEG-PLLA in masking the bitterness of coptis alkaloids. It was found that mPEG-PLLA had significant taste masking effect on the decoction of Coptidis Rhizoma and five alkaloids. The masking effect was obviously related to the structure of different alkaloids: the effect increased with the increase of the number of hydrogen donors, rotatable bonds, molecular weight, and hydrophobicity, and decreased with the increase of surface electrostatic potential, electrophilicity and binding energy with bitter receptors. In this study, the influence of alkaloid structure of Coptidis Rhizoma on the butter-masking effect of mPEG-PLLA was preliminarily elucidated, providing a scientific basis for better exerting the bitter-masking effect of amphiphilic block copolymers.
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Humans , Alkaloids , Coptis , Drugs, Chinese Herbal , Molecular Docking Simulation , Structure-Activity Relationship , TasteABSTRACT
Objectives: Compare tensile and transverse strength of new copolymers for denture base. Materials and methods: The specimens were prepared from heat cured acrylic resin with three types of additives: Acryester B, Ethoxycarbonylethylene, and Propenoic acid at a percentage of 5% and 10%. The tensile and transverse strains were tested, recorded and compared. Results: The analysis of variance display statistically significant difference. The p-value was 0.001 for each of tensile and transverse strain tests. Conclusions: The tensile strength of the novel copolymers increased. The transverse strength of some of the novel copolymers increased.
Subject(s)
Humans , Resins, Synthetic/chemistry , Tensile Strength , Denture Bases , Polymethyl MethacrylateABSTRACT
The surface hydrophobicity of nanoparticles plays an important role in drug delivery process. The aim of this study was to verify the feasibility of using self-assembly method to prepare drug-loaded nanoparticles with tunable surface hydrophobicity. Here, Soluplus was selected as the polymeric carrier to prepare panobinostat (PNB) loaded micelles. Three different monoglycerides, glycerly monooleate (GMO), glycerly linoleate (GML) and glycerly linolenate (GMLO), were used to modify the surface of PNB-Soluplus micelles to prepare polymer-lipid hybrid nanoparticles (PLHNs). The effect of monoglyceride type and amount on the physico-chemical properties of PNB-loaded PLHNs was investigated, and the surface hydrophobicity of PLHNs was characterized by Rose Bengal (RB) binding method and mucin particle method. The results suggested that compared with the PNB-Soluplus micelles (particle size 77.97 ± 0.78 nm, zeta potential 0.44 ± 0.29 mV, entrapment efficiency 99.45% ± 1.47%, the RB binding constant (K) value 0.008 ± 0.002, the increased particle size after mixing with mucin particles 7.90 ± 1.41 nm), surface hydrophobicity of the PLHNs increased significantly when modified by GMO, GML, GMLO, with K values of 0.055 ± 0.010, 0.050 ± 0.011 and 0.058 ± 0.008, respectively. The increased particle sizes after mixing with mucin particles were 17.37 ± 4.48 nm, 22.60 ± 2.10 nm and 25.13 ± 3.89 nm, respectively. Among them, the physico-chemical properties of the GMLO modified PNB-loaded PLHNs (particle size 81.60 ± 4.52 nm, zeta potential 0.77 ± 0.03 mV, entrapment efficiency 99.59% ± 0.20%) kept constant, thus GMLO was selected to further investigate the effect of GMLO mass ratio (1%-3%) to Soluplus on the properties of the nanoparticles. While no statistical significant difference in particle size, zeta potential, entrapment efficiency or in vitro release behavior was found when GMLO ratio increased, the surface lipophilicity of the PLHNs, as characterized by K values and the increased particle sizes after mixing with mucin particles, increased almost linearly with the increase of GMLO amount. In conclusion, we demonstrated that drug-loaded PLHNs based on Soluplus and GMLO can be prepared by self-assembly method, and the surface hydrophobicity was tunable by modifying the mass ratio of GMLO to Soluplus. This approach could be used for related basic science research aiming to elucidate the effect of surface hydrophobicity on in vivo behavior of drug-loaded system.
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The study was designed to synthesize a novel dendritic copolymer composed of polyamidoamine dendrimer G0 as the inner core and poly(L-glutamic acid) grafted low molecular weight polyethylenimine (PGLP) as surrounding arms for gene delivery vector. The molecular structure of PGLP was confirmed by 1H NMR (proton nuclear magnetic resonance spectroscopy). The DNA combination capability of PGLP was examined by gel retardation electrophoresis. The particle sizes and zeta potentials of PGLP/pDNA complexes were determined by dynamic light scattering (DLS). The cytotoxicity of PGLP was evaluated by Cell Counting Kit-8 (CCK-8) and hemolysis assays, which was approved by Research Ethics Committee of the First Affiliated Hospital of Nanchang University. The in vitro transfection efficiency of PGLP was measured by a flow cytometry. The results of physicochemical properties suggested that PGLP could self-assemble with DNA to form complexes with average particle sizes of about 105-200 nm and zeta potentials of about +10 - +28 mV, which could protect DNA from serum degradation. The results of biological properties suggested that PGLP showed more higher transfection efficiencies but lower cytotoxicity than PEI 25K or Lipofectamine 2000 in various cell lines (HEK 293T, HeLa, BEL 7402, RASMC). Importantly, it was found that PGLP/pDNA complexes at w/w = 8 showed more strong serum-resistant capacity than PEI 25K/pDNA complexes. Therefore, PGLP is a promising candidate vector for gene delivery.
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Multidrug resistance (MDR) has been considered as a huge challenge to the effective chemotherapy. Therefore, it is necessary to develop new strategies to effectively overcome MDR. Here, based on the previous research of -(2-hydroxypropyl)methacrylamide (HPMA) polymer-drug conjugates, we designed an effective system that combined drug-efflux circumvention and mitochondria targeting of anticancer drug doxorubicin (Dox). Briefly, Dox was modified with mitochondrial membrane penetrating peptide (MPP) and then attached to (HPMA) copolymers (P-M-Dox). Our study showed that macromolecular HPMA copolymers successfully bypassed drug efflux pumps and escorted Dox into resistant MCF-7/ADR cells endocytic pathway. Subsequently, the mitochondria accumulation of drugs was significantly enhanced with 11.6-fold increase by MPP modification. The excellent mitochondria targeting then resulted in significant enhancement of reactive oxygen species (ROS) as well as reduction of adenosine triphosphate (ATP) production, which could further inhibit drug efflux and resistant cancer cell growth. By reversing Dox resistance, P-M-Dox achieved much better suppression in the growth of 3D MCF-7/ADR tumor spheroids compared with free Dox. Hence, our study provides a promising approach to treat drug-resistant cancer through simultaneous drug efflux circumvention and direct mitochondria delivery.
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Objective: To investigate the preparation process of polylactic acid-glycolic acid copolymer/ hydroxyapatite (PLGA/HA) microcarriers by electrostatic spraying method, and to elucidate the superiority of PLGA/HA microcarriers. Methods: The PLGA/HA microcarriers were prepared by electrostatic spraying method using nano-HA (20 nm, 99.9%) and PLGA (LA/GA = 50/50, Mw30k), and the influence of different concentrations (1%, 3%, 5%) and different voltages (4. 0, 4. 5, > 5. 0 kV) of HA in the morphology of the microcarriers was investigated and the best ball making parameters were obtained. The PLGA microcarriers were used as PLGA group, PLGA+1%HA as 1% PLGA/HA group, PLGA + 3% HA as 3% PLGA/HA group, PLGA+ 5% HA as 5% PLGA/HA group, and the simple cells were used as blank control group. The characteristics of PLGA/HA microcarriers were detected by scanning electron microscope (SEM), cell proliferation test, cell fluorescence staining experiment, and Fourier transform infrared spectroscopy (FTIR). Results: The SEM results showed that the microcarrier particles were uniform, all of them were elliptical or circular, without abnormal shape spheres, with smooth surface, without sharp edges, adhesion between the spheres and aggregation, and there were no significant differences between the different concentrations of microcarriers. The cell proliferation test results showed that the order of adhesion cells was 5% PLGA/HA group > 3% PLGA/HA group > 1% PLGA/HA group > PLGA group > blank control group (P<0. 05); the number of cells was increased with the increasing of HA concentration; the microcarriers in 5% PLGA/HA group had the best cell affinity and the microcarriers had no cytotoxity. The cell fluorescence staining experiment showed that the MC3T3-E1 cells adhered well on the microcarriers. The FTIR analysis results showed that HA characteristic absorption peak was observed, indicating that the composite microcarrier contained PLGA and HA. Conclusion: The preparation process of PLGA/ HA microcarriers is successfully established by electrostatic spraying method. The method is simple and convenient to operate, and has excellent ball-making effect. It has broadly application prospects in bone tissue engineering.
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Objective: To observe the effect of vascular endothelial growth factor/polylactide-polyethyleneglycol-polylactic acid copolymer/basic fibroblast growth factor (VEGF/PELA/bFGF) mixed microcapsules in promoting the angiogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods: The BMSCs were isolated by the method of whole bone marrow adherent, and sub-cultured. The passage 3 BMSCs were identified by Wright-Giemsa staining and flow cytometry, and used for subsequent experiments. VEGF/PELA/bFGF (group A), PELA/bFGF (group B), VEGF/PELA (group C), and PELA (group D) microcapsules were prepared. The biodegradable ability and cytotoxicity of PELA microcapsule were determined，and the slow-released ability of VEGF/PELA/bFGF mixed microcapsules was measured. The passage 3 BMSCs were co-cultured with the extracts of groups A, B, C, and D, separately. At 1, 3, 7, 14, and 20 days after being cultured, the morphological changes of induced BMSCs were recorded. At 21 days, the induced BMSCs were tested for DiI-labeled acetylated low density lipoprotein (Dil-ac-LDL) and FITC-labeled ulex europaeus agglutinin I (FITC-UEA-I) uptake ability. The tube-forming ability of the induced cells on Matrigel was also verified. The differences of the vascularize indexes in nodes, master junctions, master segments, and tot.master segments length in 4 groups were summarized and analyzed. Results: The isolated and cultured cells were identified as BMSCs. The degradation time of PELA was more than 20 days. There was no significant effect on cell viability under co-culture conditions. At 20 days, the cumulative release of VEGF in the mixed microcapsules exceeded 95%, and the quantity of bFGF exceeded 80%. The morphology of cells in groups A, B, and C were changed. The cells in groups A and B showed the typical change of cobble-stone morphology. The numbers of double fluorescent labeled cells observed by fluorescence microscope were the most in group A, and decreases from group B and group C, with the lowest in group D. The cells in groups A and B formed a grid-like structure on Matrigel. Quantitative analysis showed that the differences in the number of nodes, master junctions, master segments, and tot.master segments length between groups A, B and groups C, D were significant ( P0.05). Conclusion: VEGF/PELA/bFGF mixed microcapsules have significantly ability to promote the angiogenic differentiation of rat BMSCs in vitro.
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Objective@#To investigate whether poly (lactic-co-glycolic acid) (PLGA) as protein delivery vehicles that encapsulate CC chemokine receptor 5 antibody (anti-CCR5) has more suppressive function on macrophages than single anti-CCR5 in mouse endometriosis model.@*Methods@#The PLGA/anti-CCR5 nanoparticles were synthesized. The cumulative release of anti-CCR5 from PLGA/anti-CCR5 nanoparticles was evaluated. The mouse endometriosis model was established and divided into control group, anti-CCR5 group and PLGA/anti-CCR5 group. Meanwhile, ectopic endometrial cells (EEC) and macrophages isolated from peritoneal fluid were cultured in vitro. Flow cytometry was used to detect the proportion of macrophages in the peritoneal fluid of each group. The secretion of interleukin 10 (IL-10) and transforming growth factor β (TGF-β) in each group were determined by ELISA. The proliferation and infiltration of EEC were detected by 5-bromodeoxyuridine proliferation kit and matrigel invasion kit.@*Results@#The PLGA/anti-CCR5 nanoparticles were successfully synthesized. The mouse endometriosis model was established and the EEC and macrophages were cultured. Compared with the anti-CCR5 without nanoparticles, the bioconjugate PLGA/anti-CCR5 nanoparticles could control the release of anti-CCR5 from day 3 to day 24. The proportion of macrophages in PLGA/anti-CCR5 group were gradually reduced compared with those in anti-CCR5 group (P<0.01), the ratios of day 7 [(4.5±1.5)%] and day 3 [(6.3±0.6)%], day 14 [(2.6±0.7)%] and day 7 were significantly different (P<0.01 and P<0.05). PLGA/anti-CCR5 reduced IL-10 and TGF-β levels relative to anti-CCR5 (P<0.01),and decreased gradually on day 3, day 7, and day 14 (P<0.01). Anti-IL-10+anti-TGF-β could reduce the proliferation [(70.8±7.6)%] and invasion ability [(50.2±9.1)%] of EEC (P<0.05).@*Conclusions@#In mouse endometriosis model, PLGA/anti-CCR5 may inhibit the proliferation and invasion of EEC by inhibiting the secretion of IL-10 and TGF-β by macrophages, suggesting that it provide a new idea for the treatment of clinical endometriosis.
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Objective To investigate whether poly (lactic?co?glycolic acid) (PLGA) as protein delivery vehicles that encapsulate CC chemokine receptor 5 antibody (anti?CCR5) has more suppressive function on macrophages than single anti?CCR5 in mouse endometriosis model. Methods The PLGA/anti?CCR5 nanoparticles were synthesized. The cumulative release of anti?CCR5 from PLGA/anti?CCR5 nanoparticles was evaluated. The mouse endometriosis model was established and divided into control group, anti?CCR5 group and PLGA/anti?CCR5 group. Meanwhile, ectopic endometrial cells (EEC) and macrophages isolated from peritoneal fluid were cultured in vitro. Flow cytometry was used to detect the proportion of macrophages in the peritoneal fluid of each group. The secretion of interleukin 10 (IL?10) and transforming growth factor β (TGF?β) in each group were determined by ELISA. The proliferation and infiltration of EEC were detected by 5?bromodeoxyuridine proliferation kit and matrigel invasion kit. Results The PLGA/anti?CCR5 nanoparticles were successfully synthesized. The mouse endometriosis model was established and the EEC and macrophages were cultured. Compared with the anti?CCR5 without nanoparticles, the bioconjugate PLGA/anti?CCR5 nanoparticles could control the release of anti?CCR5 from day 3 to day 24. The proportion of macrophages in PLGA/anti?CCR5 group were gradually reduced compared with those in anti?CCR5 group (P<0.01), the ratios of day 7 [(4.5±1.5)%] and day 3 [(6.3±0.6)%], day 14 [(2.6±0.7)%] and day 7 were significantly different (P<0.01 and P<0.05). PLGA/anti?CCR5 reduced IL?10 and TGF?β levels relative to anti?CCR5 (P<0.01),and decreased gradually on day 3, day 7, and day 14 (P<0.01). Anti?IL?10+anti?TGF?β could reduce the proliferation [(70.8 ± 7.6)% ] and invasion ability [(50.2 ± 9.1)% ] of EEC (P<0.05). Conclusions In mouse endometriosis model, PLGA/anti?CCR5 may inhibit the proliferation and invasion of EEC by inhibiting the secretion of IL?10 and TGF?β by macrophages, suggesting that it provide a new idea for the treatment of clinical endometriosis.
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Aim: To investigate the in vitro release behavior and pharmacokinetic characteristics of quercetinloaded mixed micelles composed of Pluronic P123/ Poloxamer 188 in rats. Methods: Polymer micelles were prepared by the thin-film hydration method. The quercetin release rate of quercetin mixed micelles was investigated by dynamic membrane dialysis technique in the physiological saline contained 1% Tween-80. The drug release curves were drawn. The results were evaluated by DD Solver. Ultra performance liquid chromatography (UPLC) method was employed to measure the plasma concentration of quercetin in rats after tail intravenous administration. The plasma concentration-time curves were drawn, and the pharmacokinetic parameters were calculated with PK Solver. Results: The entrapment efficiency (EE%) and loading efficiency (DL%) of mixed micelles was (94. 25 ± 2. 13)% and (8.61 ± 0. 18)%, respectively. The quercetin release rate of quercetin mixed micelles in the physiological saline contained 1% Tween-80 was (52.90 ± 1.08)%. The quercetin mixed micelles could be described by Higuchi model and expressed by the equation; F = 6. 735 t0.5 (Rsqr-adj = 0. 960 4, AlC = 75. 584 0). T1/2β, AUC0.4 and MRT0-inf of quercetin mixed micelles were calculated to be 1.2,1.5 and 2. 4 times of free quercetin, respectively. Conclusions: The quercetin-loaded mixed micelles have high entrapment efficiency and loading efficiency, which can improve the release behavior in vitro, prolong the time of quercetin, and significantly enhance the fraction of bioavailability of quercetin in rats.
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Introduction: Ostene is a water-soluble wax-like alkylene oxide copolymer preparation for use as a mechanical hemostatic agent. this study aims to evaluate the effects of Ostene on bone healing. materials and methods: twenty albino rabbits were divided into four groups according to post-treatment follow-up (24 hr, 3 days, 7 days, 14 days) with five rabbits in each group. each rabbit in all groups was treated with two study materials (Ostene and Gelfoam). three holes were made in the mandibular bone of each rabbit using 5mm surgical bur; two holes were made on right side: one for testing Ostene and another for Gelfoam. a third hole, on the left side of mandible, was not treated, and was used as a control. finally, the incision was closed. the specimens were collected at different days post-treatment and examined by histopathology. result and discussion: this study showed that there is a significant difference (p-value≤ 0.05) between the Ostene group and the other groups (Gelfoam and control). at 24 hr post intervention, there is a significant difference in osteoblast cell formation (p-value=0.03), and osteoclast cell formation (p-value=0.05). new blood vessel formation, osteoblast and osteoclast cell formation for Ostene group at 3 days post-intervention were also significantly different (p-values = 0.05, 0.03, 0.04, respectively). at 7 days post-intervention p-values were 0.05 for osteoblast formation and 0.04 for osteoclast formation, respectively. after 14 days of healing p-value for osteoblast cell formation in the Ostene group was 0.05 and 0.04 for osteoclast cell formation. conclusions: the bone hemostatic agent Ostene is an effective at enhancing osteogenesis by initiating proliferation of osteoblast and osteoclast cells.
Subject(s)
Animals , Rabbits , Osteogenesis/drug effects , Wound Healing/drug effects , Bone and Bones/drug effects , Hemostatics/pharmacology , Hemostasis , Osteoblasts , Osteoclasts , Disease Models, Animal , Mandible/drug effectsABSTRACT
BACKGROUND: To report the long-term outcomes of endoscopic surgery (ES) in pediatric patients with vesicoureteral reflux in terms of success rate, urinary tract infection, and renal function. METHODS: We retrospectively reviewed the records of 73 pediatric patients (110 ureters) who underwent ES for vesicoureteral reflux. Ultrasonography was performed 1, 3, and 12 months postoperatively. Voiding cystourethrography was performed 3 months postoperatively and repeated after 1 year if vesicoureteral reflux persisted. Success was defined as the absence of reflux at the first voiding cystourethrography. Renal scans were performed at least 12 months postoperatively. Renal function deterioration was defined as a new scar or a greater than 5% decrease in function. RESULTS: The median follow-up duration was 24 (12–118) months. The overall success was 65.6%, while it was 78.9%, 87.0%, 62.5%, 37.5%, 66.7% for grades I, II, III, IV, and V, respectively. In multivariate analyses, significant predictive factors for success were vesicoureteral reflux grade (odds ratio [OR], 0.28; P < 0.001) and mound detection at the first postoperative ultrasonography (OR, 13.53; P < 0.001). Renal function deterioration was found in 8 (15.3%) ureters and was less common in those with successful surgeries than in those with failures (9.5% vs. 40.0%; P = 0.035). No significant predictive factor for renal function deterioration or urinary tract infection was found. CONCLUSION: Successful short-term outcomes of ES are expected in low-grade vesicoureteral reflux, especially when a mound is detected by postoperative ultrasonography. However, unpredictable long-term renal deterioration warrants continued follow-up.