Optimal conditions for mouse follicle culture.

Mammalian ovaries contain a large number of immature follicles. Follicular culture can contribute to the production of fertile oocytes from latent immature follicles, providing a useful tool for exploring the developmental competencies and related factors that oocytes acquire during growth. However, the potential of oocytes produced by follicular culture is limited. Herein, the optimal follicular culture conditions for the addition of polyvinylpyrrolidone to the medium and oxygen concentration were investigated. Polyvinylpyrrolidone with a high molecular weight (≥ 360,000) and a 7% oxygen concentration were found to increase the blastocyst formation rate by more than 20% compared with conventional culture conditions. Although the developmental ability of oocytes produced by follicular culture remained inferior to that of in vivo-derived oocytes, these findings may pave the way for enhanced production of fertile oocytes in vitro and for studying the process of full developmental potency acquisition by oocytes.

Influence of gamma radiation on Amphotericin B incorporated in PVP hydrogel as an alternative treatment for cutaneous leishmaniosis.

Amphotericin B (Amph-B) is an antifungal drug used intravenously for the treatment of leishmaniasis. Side-effects from Amph-B treatment can arise such as cardiac arrhythmia and renal dysfunctions, which will lead to discontinuation of treatment. Unfortunately, patients in endemic countries do not have access to alternative therapies. The objective of this study was to analyze the effects of Cobalt-60 gamma irradiation on crosslinking polymeric hydrogels (Hydg) and the incorporation of Amph-B into the gel as a controlled-release drug delivery alternative. Polyvinylpyrrolidone (PVP)/Amph-B solutions were irradiated with 15 kGy at 0 °C and 25 °C. The drug's stability was ascertained by UV-visible spectrometry, liquid chromatography/mass spectrometry and proton nuclear magnetic resonance. Irradiated Hydg/Amph-B achieved similar stability to the standard Amph-B solution and was enough to promote hydrogel crosslinking. In vitro trials were carried out to ensure Amph-B was still biologically active after irradiation. The results from flow cytometry and MTT assay show that Amph-B had an IC50 = 16.7 nM. A combination of Hydg at 1.324 gmL-1 and Amph-B at 25.1 nM for 24 h lead to the greatest inhibition of L. amazonensis promastigotes, and could be used as an alternative treatment method for cutaneous leishmaniosis.

Treatment with Hyaluronic Acid and Collagen-Polyvinylpyrrolidone Improves Extracellular Matrix Assembly for Scarring after Tracheal Resection.

Treatment of tracheal stenosis is occasionally performed in combination with wound healing modulators to manipulate new extracellular matrix (ECM) formation and prevent fibrosis. Hyaluronic acid (HA) and collagen-polyvinylpyrrolidone (collagen-PVP) decrease fibrosis in experimental tracheal healing. However, they have not been used clinically as their effect on ECM components, which modify tracheal scarring, has not been described. Objective. To evaluate the effect of the application of HA, collagen-PVP, a mixture of HA and collagen-PVP (HA+collagen-PVP), and mitomycin C on the expression of decorin, matrix metalloproteinase 1 (MMP1), and MMP9, as well as the type of collagen and deposits formed in the scar after resection and end-to-end anastomosis (REEA) of the cervical trachea using an experimental model. Materials and Methods. Thirty dogs underwent REEA of the cervical trachea and were treated with different wound healing modulators: group I (n = 6), control; group II (n = 6), HA; group III (n = 6), collagen-PVP; group IV (n = 6), HA+collagen-PVP; and group V (n = 6), mitomycin C. The dogs were evaluated clinically and endoscopically for 4 weeks. Subsequently, macroscopic and microscopic changes, expression of ECM proteins, and collagen deposition in tracheal scars were analysed. Results. Groups II, III, and IV showed reduced endoscopic, macroscopic, and microscopic inflammation, improved neovascularization, high decorin expression (p < 0.01, analysis of variance (ANOVA)), and moderate expression of MMP1 (p < 0.003, ANOVA) and type I and III collagen (p < 0.05, Kruskal-Wallis). Groups IV and V developed fewer collagen deposits (p < 0.001, ANOVA). Conclusion. Treatment with HA and collagen-PVP improved post-REEA healing by increasing neovascularization, stimulating the expression of decorin, and regulating the expression of MMP1, as well as type I and III collagen and their deposition.

Polyvinylpyrrolidone microneedles for localized delivery of sinomenine hydrochloride: preparation, release behavior of in vitro & in vivo, and penetration mechanism.

Sinomenine (SIN) is an anti-inflammatory alkaloid derived from Sinomenium acutum, and the products sinomenine hydrochloride (SH) tablets and injections have been marketed in China to treat rheumatoid arthritis (RA). Oral administration of SH has shortcomings of gastrointestinal irritation and low bioavailability. The injection may require professional training and higher cost. It is of interest to develop an alternative form that is easier to administer and avoids the first-pass metabolism. In this study, SH-loaded dissolving microneedles (SH-MN) were fabricated using polyvinyl pyrrolidone and chondroitin sulfate with a casting method. In percutaneous permeation studies of In vitro, the cumulative permeation and permeation rate of SH-MN were 5.31 and 5.06 times higher than that of SH-gel (SH-G). In percutaneous pharmacokinetic studies, the values of the area under the curve after administration of SH-MN in the skin and blood were 1.43- and 1.63-fold higher than that of SH-G, respectively. In percutaneous absorption studies, SH-MN could absorb into tissue fluid; and dissolve after skin penetration. The drug was released along the channel and spread to surrounding skin tissue. After 4 h, the needle tip was almost completely dissolved, and the drug could penetrate to a depth of 200 µm under the skin. These results demonstrate that the SH-MN is an effective, safe, and simple strategy for transdermal SH delivery.

Efficacy of Divinylbenzenic Resin in Removing Indoxyl Sulfate and P-Cresol Sulfate in Hemodialysis Patients: Results From an In Vitro Study and An In Vivo Pilot Trial (xuanro4-Nature 3.2).

High serum levels of microbiota-derived uremic toxins, indoxyl sulfate (IS) and p-cresyl sulfate (PCS), are associated with chronic kidney disease (CKD) progression and cardiovascular complications. IS and PCS cannot be efficiently removed by conventional hemodialysis (HD), due to their high binding affinity for albumin. This study evaluates the efficacy of a divinylbenzene-polyvinylpyrrolidone (DVB-PVP) cartridge and a synbiotic to reduce uremic toxins in HD patients. First, the in vitro efficacy of DVB-PVP in adsorbing IS and PCS was evaluated. Second, a randomized, placebo-controlled pilot study in HD patients was carried out to establish whether the administration of a synbiotic, either individually and in association with DVB-PVP-HD, could reduce the production of uremic toxins. In vitro data showed that DVB-PVP resin removed a mean of 56% PCS and around 54% IS, after 6 h of perfusion. While, in the in vivo study, the DVB-PVP cartridge showed its adsorbing efficacy only for IS plasma levels. The combination of synbiotic treatment with DVB-PVP HD decreased IS and PCS both at pre- and post-dialysis levels. In conclusion, this study provides the first line of evidence on the synergistic action of gut microbiota modulation and an innovative absorption-based approach in HD patients, aimed at reducing plasma levels of IS and PCS.

Carbon dot cross-linked polyvinylpyrrolidone hybrid hydrogel for simultaneous dye adsorption, photodegradation and bacterial elimination from waste water.

The creation of a polymeric hydrogel from polyvinylpyrrolidone (PVP) cross-linked by Carbon Quantum Dots (CD) for the adsorption and photocatalytic degradation of both cationic and anionic dyes. PVP, an important biocompatible constituent and often surplus in cosmetic industry, was carboxylated through NaOH refluxing and covalently conjugated to surface amine functionality of CD derived from lemon juice and Cysteamine. The hybrid hydrogel was obtained from PVP-CD covalent conjugate by careful manipulation of pH and found to possess better rheological properties than only carboxylate-PVP. The monolayer physisorption of the dyes on the hydrogel was affected by hydrogen bonding, dispersion or inductive effect, and π-π interaction with the polymer backbone as well as the CD that followed pseudo-second-order kinetics. Degradation of the adsorbed dyes was instated by the unique Reactive Oxygen Species (ROS) generating ability of the CD embedded in the hydrogel matrix upon exposure to sunlight, the mechanism of which is also unveiled. The same CD-induced ROS was found to effectively annihilate both gram-positive and gram-negative bacteria in real polluted water in less than 10 min of photoexcitation of the hydrogel. The hydrogel was restored by mild acid wash that is able to perform dye adsorption and photo-degradation upto four cycles.

Evaluating supersaturation in vitro and predicting its performance in vivo with Biphasic gastrointestinal Simulator: A case study of a BCS IIB drug.

This study aimed to develop an evaluation approach for supersaturation by employing an in vitro bio-mimicking apparatus designed to predict in vivo performance. The Biphasic Gastrointestinal Simulator (BGIS) is composed of three chambers with absorption phases that represent the stomach, duodenum, and jejunum, respectively. The concentration of apatinib in each chamber was detected by fiber optical probes in situ. The dissolution data and the pharmacokinetic data were correlated by GastroplusTM. The precipitates were characterized by polarizing microscope, Scanning Electron Microscopy, Powder X-ray diffraction and Differential scanning calorimetry. According to the results, Vinylpyrrolidone-vinyl acetate copolymer (CoPVP) prolonged supersaturation by improving solubility and inhibiting crystallization, while Hydroxypropyl methylcellulose (HPMC) prolonged supersaturation by inhibiting crystallization alone. Furthermore, a predictive in vitro-in vivo correlation was established, which confirmed the anti-precipitation effect of CoPVP and HPMC on in vitro performance and in vivo behavior. In conclusion, CoPVP and HPMC increased and prolonged the supersaturation of apatinib, and then improved its bioavailability. Moreover, BGIS was demonstrated to be a significant approach for simulating in vivo conditions for in vitro-in vivo correlation in a supersaturation study. This study presents a promising approach for evaluating supersaturation, screening precipitation inhibitors in vitro, and predicting their performances in vivo.

Novel method for delayed primary closure and incisional hernia prevention in open abdomen: COmbined and MOdified Definitive Abdominal wall closure (COMODA).

BACKGROUND: Intended open abdomen is an option in cases of trauma and non-trauma patients. Nevertheless, after primary closure, incisional hernia rate is high. We describe a novel method, called COmbined and MOdified Definitive Abdominal closure (COMODA), a delayed primary closure which prevents incisional hernia. METHODS: A negative pressure wound therapy system is combined with a condensed polytetrafluoroethylene (cPTFE) mesh. TRIAL REGISTRATION: ISRCTN72678033. RESULTS: Ten male patients with a median age of 68.8 (43-87) years were included. Primary closure rate was 100% per protocol. The median number of procedures per patient was 5.7 (5-9). Primary closure was obtained in 20.8 (10-32) days and median hospital stay was 36.3 (18-52) days. Only one patient developed incisional hernia during a median follow-up of 27 (8-60) months. CONCLUSION: COMODA method allows for a high rate of delayed primary closure. It is safe and decreases the risk for developing an incisional hernia. However, a large number of patients are needed to support this conclusion.

A novel core-shell nanofiber drug delivery system intended for the synergistic treatment of melanoma.

Here, we introduce core-shell nanofibers based on chitosan (CS)-loaded poly (ε-caprolactone) (PCL) shell and 5-fluorouracil (5-FU)-loaded Poly(N-vinyl-2-pyrrolidone) (PVP) core for synergistic therapy of melanoma skin cancer. The yielded nanofibers exhibited an average diameter of 503 nm with high drug-encapsulating efficiency and good mechanical properties. Moreover, the burst release of 5-FU significantly inhibited melanoma skin cancer cells (B16F10 cells), and the sustained release of CS exhibited "remedying effects" on normal skin cells (L929 cells) after suffering adverse effects from 5-FU treatment. For the B16F10 cells, the early apoptosis cells increased from 0.8% to 62.2% after being treated with blended films loaded with 5-FU (2 wt%) for 24 h; for the L929 cells, the vital cells increased from 68.9% to 77.0%, and the early apoptosis of stage cells decreased from 12.3% to 10.9% after being treated with blended films with CS (8 wt%) for 24 h. In conclusion, the results introduced in this work can be a promising strategy for cancer treatment and possesses synergism potential to broaden an avenue for chemotherapeutic therapy with minimum adverse effects on normal cells.

Rapidly dissolving bilayer microneedle arrays - A minimally invasive transdermal drug delivery system for vitamin B12.

Vitamin B12 plays an essential role in one-carbon metabolism in the human body. A deficiency in this vitamin can lead to severe haematopoietic and neuropsychiatric disorders and is currently treated by oral or parenteral administration of exogenous vitamin. Unfortunately, the absorption of orally taken vitamin B12 is low and highly variable, while injections can cause pain and anxiety. Thus, an efficient alternative drug delivery system for overcoming these shortcomings is highly desirable. Novel polymeric microneedle (MN) arrays have the potential for minimally invasive transdermal treatment of vitamin B12 deficiency. Bilayer dissolving MN arrays (19 × 19 needles, 600 µm height) containing 135 µg vitamin B12 were cast using two different aqueous polymer blends. MN arrays showed sufficient mechanical strength for skin insertion, dissolved rapidly and delivered 72.92% of their drug load in vitro over 5 h. Ultimately, the potential of delivering a therapeutically relevant dose of vitamin B12 transdermally was demonstrated in vivo in Sprague-Dawley rats by comparison to subcutaneous injections. Maximum plasma levels of 0.37 µg/mL occurred 30 min post-MN application, highlighting the ability of fabricated MN arrays to rapidly deliver vitamin B12 transdermally.

Development of sorafenib loaded nanoparticles to improve oral bioavailability using a quality by design approach.

Sorafenib, a potent anticancer drug, has low absorption in the gastrointestinal tract due to its poor aqueous solubility. The main purpose of this investigation was to design sorafenib nanoparticle using a newly developed technique, nanoparticulation using fat and supercritical fluid (NUFS™) to improve the absorption of sorafenib. The quality by design (QbD) tool was adopted to define the optimal formulation variables: hydroxypropyl methyl cellulose (HPMC), polyvinyl pyrrolidone K30 (PVP), and poloxamer. The studied response variables were particle size of nanoparticle, dissolution (5, 60, and 180 min), drug concentration time profile of nanoparticle formulations, and maximum drug concentration. The result of particle size revealed that an increase in concentration of poloxamer and HPMC decreased the particle size of nanoparticles (p < 0.05). Likewise, the concentration of drug release at different time point (5, 60, and 180 min) showed HPMC and poloxamer had positive effects on drug dissolution while PVP had negative effects on it. The design space was built in accordance with the particle size of nanoparticle (target < 500 nm) and dissolution of sorafenib (target > 7 µm/mL), following failure probability analysis using Monte Carlo simulations. In vivo pharmacokinetics studies in beagle dogs demonstrated that optimized formulation of sorafenib (F3 and F4 tablets) exhibited higher blood drug profiles indicating better absorption compared to the reference tablet (Nexavar®). In conclusion, this study showed the importance of systematic formulation design for understanding the effect of formulation variables on the characteristics of nanoparticles of the poorly soluble drug.

Interaction Studies Between Levodopa and Different Excipients to Develop Coground Binary Mixtures for Intranasal Application.

Levodopa (LEVO) as the gold standard in the treatment of Parkinson's disease is usually administrated per os but its bioavailability is low. The intranasal administration is a potential alternative route to increase bioavailability of the drug and treat the off period. Our aim was to develop LEVO-containing binary nasal powders with different excipients by dry cogrinding process. The interactions between the components were examined. The optimized cogrinding process parameters (LEVO:excipient ratio and grinding time) resulted in the desired particle size range (5-40 µm). The α-cyclodextrin and poly(vinylpyrrolidone) (PVP) had an intensive crystallinity degree reducing effect on LEVO measured by XRPD, and they functioned as cogrinding agents. Hydroxypropyl methylcellulose, poly (vinyl alcohol) (PVA), and D-mannitol associate around the LEVO crystals preventing its crystalline structure. Hydrogen bonding was detected only for LEVO-PVP and LEVO-D-mannitol used Fourier-transformed infrared spectroscopy. Chemical degradation of LEVO in the products was not detected even after the accelerated stability test. The dissolution profile of the products can be characterized by the first-order kinetic model with different dissolution rate. The dissolution rate of LEVO was increased with α-cyclodextrin and PVP, and the drug release decreased in the case of hydroxypropyl methylcellulose, PVA, and D-mannitol compared to the LEVO powder.

Design and characterisation of electrospun shellac-polyvinylpyrrolidone blended micro/nanofibres loaded with monolaurin for application in wound healing.

Due to their ultrafine network structures, electrospun nanofibres have been potentially used for wound application. In order to develop a desired wound dressing material, shellac (SHL) was blended with polyvinyl pyrrolidone (PVP). Monolaurin (ML), which is a natural antimicrobial lipid, was incorporated into the SHL-PVP blended fibres to prevent delayed wound healing resulting from microbial infection. A full factorial design with three replicated centre points was employed in order to determine the main and interaction effects of various factors including SHL ratio in SHL-PVP blended solution, ML content and applied voltage on the multiple responses such as morphology, surface wettability, absorbency and mechanical properties. According to the results, an increase in the PVP content could lead to a significant increase in tensile strength and elongation. In addition, the presence of PVP contributed to an improvement in the drug loading capacity and dissolution rate. The fabricated fibres loaded with ML exhibited an excellent activity against Staphylococcus aureus and Candida albicans, and also provided an enhanced ability in the cell adhesion. Therefore, SHL-PVP blended fibres loaded with ML might be effectively used for application in wound healing.

Translocation, biotransformation-related degradation, and toxicity assessment of polyvinylpyrrolidone-modified 2H-phase nano-MoS2.

Nano-MoS2 has been extensively investigated in materials science and biomedicine. However, the effects of different methods of exposure on their translocation, biosafety, and biotransformation-related degradability remain unclear. In this study, we combined the advantages of synchrotron radiation (SR) X-ray absorption near-edge structure (XANES) and high-resolution single-cell SR transmission X-ray microscopy (SR-TXM) with traditional analytical techniques to investigate translocation, precise degraded species/ratio, and correlation between the degradation and toxicity levels of polyvinylpyrrolidone-modified 2H-phase MoS2 nanosheets (MoS2-PVP NSs). These NSs demonstrated different biodegradability levels in biomicroenvironments with H2O2, catalase, and human myeloperoxidase (hMPO) (H2O2 < catalase < hMPO). The effects of NSs and their biodegraded byproducts on cell viability and 3D translocation at the single-cell level were also assessed. Toxicity and translocation in mice via intravenous (i.v.), intraperitoneal (i.p.), and intragastric (i.g.) administration routes guided by fluorescence (FL) imaging were investigated within the tested dosage. After i.g. administration, NSs accumulated in the gastrointestinal organs and were excreted from feces within 48 h. After i.v. injection, NSs showed noticeable clearance due to their decreased accumulation in the liver and spleen within 30 days when compared with that in the i.p. group, which exhibited slight accumulation in the spleen. This work paves the way for understanding the biological behaviors of nano-MoS2 using SR techniques that provide more opportunities for future applications.

A Combined Utilization of Plasdone-S630 and HPMCAS-HF in Ziprasidone Hydrochloride Solid Dispersion by Hot-Melt Extrusion to Enhance the Oral Bioavailability and No Food Effect.

The purpose of this study was to research a novel combination of Plasdone-S630 and HPMCAS-HF as hot-melt carrier used in ziprasidone hydrochloride for enhanced oral bioavailability and dismissed food effect. Ziprasidone hydrochloride solid dispersion (ZH-SD) was prepared by hot-melt extrusion technique, and its optimized formulation was selected by the central composite design (CCD), which was characterized for powder X-ray diffraction (PXRD), fourier transform infrared spectroscopy (FTIR), in vitro dissolution study, and stability study. Finally, the in vivo study in fasted/fed state was carried out in beagle dogs. Based on PXRD analysis, HME technique successfully dispersed ziprasidone with a low crystallinity hydrochloride form in the polymers. According to the analysis of FTIR, hydrogen bonds were formed between drug and polymers during the process of HME. Without any noticeable bulk, crystalline could be found from the micrograph of ZH-SD when analyzed the result of scanning electron microscope (SEM). Pharmacokinetics studies indicated that the bioavailability of ZH-SD formulation had no significant difference in fasted and fed state, and the Cmax and AUC of ZH-SD were two fold higher than Zeldox® in fasted state. This result indicated that ziprasidone has achieved a desired oral bioavailability in fasted state and no food effect.

Differential effect of polyvinylpyrrolidone-coated superparamagnetic iron oxide nanoparticles on BT-474 human breast cancer cell viability.

Polyvinylpyrrolidone superparamagnetic iron oxide nanoparticles (PVP-SPIONs) have unique properties. Due to these characteristics, PVP-SPIONs have been used in several medical applications such as magnetic resonance imaging (MRI) contrast agent or drug delivery system. However, a more comprehensive understanding of the environmental safety of PVP-SPIONs is vital for consumption of these nanomaterials. In this study, we describe the effects of PVP-SPIONs on cell viability of the BT-474 human breast cancer cells. Cell viability of the BT-474 cells treated with PVP-SPIONs (10-800 µg/ml) was assessed by MTT assay. MRC-5 cell line was used as a control. Quantitative real-time PCR was performed to investigate the mRNA expression levels of apoptotic (caspase 3) and anti-apoptotic (BCL2) genes Confluent BT-474 monolayers exposed to PVP-SPIONs showed biphasic effects on cell proliferation. PVP-SPIONs at 10-100 µg /ml promote proliferation of BT-474 cells but not the MRC-5 cells. At higher dosage, PVP-SPIONs have toxicity on BT-474 cells. The results of real-time PCR was in line with MTT assay. The increase of cell proliferation at low PVP-SPIONs concentrations is different from what would be expected for these nanoparticles. Our results suggest that more attentions are needed to ensure the safer use of SPION in nanomedicine.

Novel Orally Disintegrating Tablets Produced Using a High-Pressure Carbon Dioxides Process.

It is well known that high-pressure carbon dioxide (CO2) lowers the glass transition temperature (Tg) of polymers. We therefore investigated whether Tg depression of high-pressure CO2 results in interparticle bridging of a polymer and the tablet characteristics that makes the manufacture of an orally disintegrating tablet (ODT) possible. Copolyvidone (Kollidon®) and polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus®) were examined and found to exhibit a large Tg depression. Placebo ODTs were prepared and hardness, disintegration rate, porosity, and change in thickness and appearance were evaluated before and after the high-pressure CO2 treatment. This enabled the establishment of the optimal conditions for pressure, temperature, and treatment time under pressure. Experimental results showed that it was possible to manufacture ODTs comprising Kollidon® as a water-soluble polymer with CO2 treatment under the suitable conditions such as temperature at 45°C, pressure lower than 8 MPa, and a treatment time shorter than 30 min, which is a new ODT manufacturing process called "Carbon Dioxide Assisted Tablet Formation Scheme" (CATS). In comparison to the conventional processes, which require high temperatures or humidity, CATS is expected to be applicable to drugs that are unstable at high temperature and humidity, and to functional drug particles used for bitter taste masking, sustained release, and other uses.

Modelling the in-vitro dissolution and release of sumatriptan succinate from polyvinylpyrrolidone-based microneedles.

A mathematical model was developed to predict the transport of sumatriptan molecules across the skin followed by absorption into the bloodstream. The drug was encapsulated in dissolving polyvinylpyrrolidone-based microneedles shaped in the form of pyramids. Mass balance equations were derived to simulate the dissolution and transport of the pharmaceutical ingredient. The theoretical framework made it possible to assess and predict the effects of key parameters on the release profile. The skin concentration increased with the loading dose and the height of the microneedle. An inverse relationship was noted between the amount of drug released in the dermal layer and the pitch width. These results were validated with in-vitro diffusion studies previously conducted using Göttingen minipig skin. The new mathematical approach successfully explained the in-vitro permeation of three different sumatriptan-containing formulations.

A comparative study of dissolving hyaluronic acid microneedles with trehalose and poly(vinyl pyrrolidone) for efficient peptide drug delivery.

We studied the role of the additives trehalose and poly(vinyl pyrrolidone) in the physical and pharmacokinetic properties of peptide drug incorporated hyaluronic acid microneedles. Poly(vinyl pyrrolidone) increases the mechanical strength of microneedles and ameliorates drug bioavailability in vivo, suggesting that poly(vinyl pyrrolidone) can be a promising additive in the fabrication of peptide drug-encapsulated fully dissolving microneedles.