In Vitro and In Vivo Preparation Evaluations of Bleomycin Implants and Microspheres Prepared with DL-Poly (Lactide-Co-Glycolide)

ABSTRACT

In this investigation, poly(lactide-co-glycolide) (PLGA) gel implants and microspheric depot systems of bleomycin (BLM) were formulated and evaluated in vivo in mice bearing transplantable solid tumor (fibrosarcoma). The pharmacodynamic studies showed that both the formulations retarded tumor growth significantly (p < 0.05) when compared to the control animals (without any drug treatment). Preliminary pharmacokinetic studies illustrated controlled release of the drug into the systemic circulation to elicit the anti-neoplastic action. The gel implants showed better release characteristics and greater pharmacodynamic action when compared to the microspheres, thus demonstrating the feasibility of employing biodegradable depot polymer gel matrix for chronic cancer therapy.     

Development and In Vitro Evaluation of Surface Modified Poly(lactide-co-glycolide) Nanoparticles with Chitosan-4-Thiobutylamidine

ABSTRACT

The objective of this study was to develop a nanoparticulate drug delivery system based on the surface modification of poly(lactide-co-glycolide) (PLGA) nanoparticles with a thiolated chitosan. PLGA nanoparticles were prepared by the emulsification-solvent evaporation method. Immobilization of chitosan to the surface of PLGA nanoparticles via amide bonds was mediated by a carbodiimide. Thiol groups were covalently bound to the chitosan surface of particles by reaction with 2-iminothiolane. Obtained nanoparticles were characterized in vitro regarding size, zeta potential, thiol group content, stability at different pH values, mucoadhesion, and drug release. Results demonstrated that the surface modification of PLGA nanoparticles with thiolated chitosan (chitosan-TBA) leads to nanoparticles of a mean diameter of 889.5 ± 72 nm and positive zeta potential of + 24.74 mV. The modified nanoparticles contained 7.32 ± 0.24 μmol thiol groups per gram nanoparticles. The size of nanoparticles was strongly influenced by the pH of the surrounding medium, being 925.0 ± 76.3 nm at pH 2 and 577.8 ± 66.7 nm at pH 7.4. Thiolated nanoparticles showed a 3.3-fold prolonged residence time on the mucosa and an unchanged release profile in comparison to unmodified PLGA nanoparticles. These data suggest that surface modified chitosan-TBA conjugate PLGA nanoparticles have the potential to be used as mucoadhesive drug delivery system.     

In Vitro and Ex Vivo Permeation Studies of Chlorpheniramine Maleate Gels Prepared by Carbomer Derivatives

The antihistaminic chlorpheniramine maleate (CPM) is used for symptomatic relief of hypersensitivity reactions and in pruritic skin disorders. At present, the drug is marketed in tablet, capsule, syrup, cream, and injectable dosage forms. Chlorpheniramine maleate has some side effects when taken orally. Due to its first pass effect, only 25%–45% of the orally administered dose reaches the blood circulation. To bypass these disadvantages, we aimed to investigate percutaneous absorption of CPM from gel formulations prepared with different carbomer derivatives (Carbopol 934, 940, 941, 2984, 980, and 981; main differences are related to presence of a comonomer and cross‐link density). Cellulose membrane was used as the diffusion barrier for all the formulations' drug‐release studies. The release of active substance from carbopol derivatives, which have the least cross‐linking density (Carbopol 941 and 981) was found to be numerically higher than the others. The formulation (F8; 1% Carbopol 941) that exhibited the maximum drug release through the cellulose membrane was further studied for drug release by using polyurethane membrane, excised rat skin, and human skin. The penetration of the active substance through different diffusion barriers was found to be statistically different (p?<?0.05) when compared. Of all the different diffusion barriers, rat skin gave the closest results to human skin. Thus topical application of CPM in the carbomer gel may be of potential use for local activity. The type and concentration of carbomers can affect drug release. The synthetic membranes are useful in assessments of formulations in quality assurance but they do not give definite indication of how a formulation will behave when it is used on skin.     

Poly(lactide-co-glycolide) encapsulated hydroxyapatite microspheres for sustained release of doxycycline

The purpose of this study was to prepare a poly(lactide-co-glycolide) (PLGA) encapsulated hydroxyapatite microspheres (HAP-MSs) as injectable depot for sustained delivery of Doxycycline (Doxy). Doxy loaded HAP-MSs (Doxy-HAP-MSs) were encapsulated with PLGA by solid-in-oil-in-water (S/O/W) emulsion-solvent evaporation technique, the effects of the PLGA used (various intrinsic viscosity and LA/GA ratio) and ratio of PLGA/HAP-MSs on the formation of Doxy-HAP-MSs and in vitro release of Doxy were studied. The results showed that sustained drug release without obvious burst was obtained by using PLGA encapsulated HAP-MSs as the carrier, also the drug release rate could be tailored by changing the ratio of PLGA/HAP-MSs, or PLGA of various intrinsic viscosities or LA/GA ratio. Lower ratio of PLGA/HAP-MSs corresponded faster Doxy release, e.g. for the microspheres of PLGA/HAP-MSs ratio of 8 and 0.25, the in vitro Doxy release percents at the end of 7days were about 23% and 76%, respectively. Higher hydrophilicity (higher ratio of GA to LA) and lower molecular weight of PLGA corresponded to higher Doxy release rates. For in vivo release study, PLGA encapsulated HAP-MSs were subcutaneously injected to the back of mice, and the results showed good correlation between the in vivo and in vitro drug release. Meanwhile, the plasma Doxy levels after subcutaneous administration of PLGA encapsulated Doxy-HAP-MSs were relatively lower and steady compared to that of the un-encapsulated microspheres. In conclusion, PLGA encapsulated HAP-MSs may be a potential vehicle for the sustained delivery of Doxy.     

In Vitro and In Vivo Interrogation of Bio-sprayed Cells

Bio-sprays can directly form pre-organized cell-bearing structures for applications ranging from engineering functional tissues to the forming of cultures, most useful for modeling disease, to the discovery and development of drugs. Bio-electrosprays and aerodynamically assisted bio-jets, are leading approaches that have been demonstrated as having far-reaching ramifications for regenerative biology and medicine.     

Multiphoton Microscopy of Nonfluorescent Nanoparticles In Vitro and In Vivo

Nanotechnology holds great promise for a plethora of potential applications. The interaction of engineered nanomaterials with living cells, tissues, and organisms is, however, only partly understood. Microscopic investigations of nano‐bio interactions are mostly performed with a few model nanoparticles (NPs) which are easy to visualize, such as fluorescent quantum dots. Here the possibility to visualize nonfluorescent NPs with multiphoton excitation is investigated. Signals from silver (Ag), titanium dioxide (TiO₂), and silica (SiO₂) NPs in nonbiological environments are characterized to determine signal dependency on excitation wavelength and intensity as well as their signal stability over time. Ag NPs generate plasmon‐induced luminescence decaying over time. TiO₂ NPs induce photoluminescent signals of variable intensities and in addition strong third harmonic generation (THG). Optimal settings for microscopic detection are determined and then applied for visualization of these two particle types in living cells, in murine muscle tissue, and in the murine blood stream. Silica NPs produce a THG signal, but in living cells it cannot be discriminated sufficiently from endogenous cellular structures. It is concluded that multiphoton excitation is a viable option for studies of nano‐bio interactions not only for fluorescent but also for some types of nonfluorescent NPs.     

Lonidamine Solid Dispersions: In Vitro and In Vivo Evaluation

ABSTRACT

Solid dispersions of lonidamine in PEG 4000 and PVP K 29/32 were prepared by the spray-drying method. Then, the binary systems were studied and characterized using differential scanning calorimetry, hot stage microscopy, and x-ray diffractometry. In vitro dissolution studies of the solid dispersed powders were performed to verify if any lonidamine dissolution rate or water solubility improvement occurred. In vivo tests were carried out on the solid dispersions and on the cyclodextrin inclusion complexes to verify if this lonidamine water solubility increase was really able to improve the in vivo drug plasma levels. Drug water solubility was increased by the solid dispersion formation, and the extent of increase depended on the polymer content of the powder. The greater increase of solubility corresponded to the highest content of polymer. Both the solid dispersions and the cyclodextrin complexes were able to improve the in vivo bioavailability of the lonidamine when administered per os. Particularly, the AUC of the drug plasma levels was increased from 1.5 to 1.9-fold depending on the type of carrier.     

In Vitro and In Vivo Percutaneous Absorption Studies of Ketotifen Patches

Abstract

The purpose of the study is to develop a suitable transdermal delivery system for ketotifen. The physicochemical properties and skin penetration of ketotifen were evaluated for the optimization of the method of patch preparation. The distribution coefficients of ketotifen were determined in octanol/phosphate buffer systems, the partition coefficient and pka value of ketotifen were obtained as 120 and 6.73, respectively. In vitro skin penetration of ketotifen solution was determined at 35°C in the Valia-Chien diffusion cell by using the abdominal skin of nude mouse. The results showed that ketotifen had an optimal skin permeability at pH 7.5. In addition, ketotifen had higher rate of penetration through stripped skin, it was suggested that the main barrier for percutaneous absorption of ketotifen is stratum corneum. Ketotifen patch was fabricated in a stainless mold containing Eudragit S-100 and PEG 400. Other components, tween, span and fatty acids were also incorporated into the patch as penetration enhancers. For animal study, a patch with area of 30 cm² was applied on the dorsal skin of rabbit. The plasma level, after 10 hrs administration were reached 60 ng/mL and maintained a constant level. The results proposed that ketotifen was successfully absorbed through the skin from the applied patch.     

In Vitro and In Vivo Percutaneous Absorption Studies of Ketotifen Patches

The purpose of the study is to develop a suitable transdermal delivery system for ketotifen. The physicochemical properties and skin penetration of ketotifen were evaluated for the optimization of the method of patch preparation. The distribution coefficients of ketotifen were determined in octanol/phosphate buffer systems, the partition coefficient and pka value of ketotifen were obtained as 120 and 6.73, respectively. In vitro skin penetration of ketotifen solution was determined at 35°C in the Valia-Chien diffusion cell by using the abdominal skin of nude mouse. The results showed that ketotifen had an optimal skin permeability at pH 7.5. In addition, ketotifen had higher rate of penetration through stripped skin, it was suggested that the main barrier for percutaneous absorption of ketotifen is stratum corneum. Ketotifen patch was fabricated in a stainless mold containing Eudragit S-100 and PEG 400. Other components, tween, span and fatty acids were also incorporated into the patch as penetration enhancers. For animal study, a patch with area of 30 cm² was applied on the dorsal skin of rabbit. The plasma level, after 10 hrs administration were reached 60 ng/mL and maintained a constant level. The results proposed that ketotifen was successfully absorbed through the skin from the applied patch.     

Fluorescent Polymer Nanoparticles for Cell Barcoding In Vitro and In Vivo

Fluorescent polymer nanoparticles for long‐term labeling and tracking of living cells with any desired color code are developed. They are built from biodegradable poly(lactic‐co‐glycolic acid) polymer loaded with cyanine dyes (DiO, DiI, and DiD) with the help of bulky fluorinated counterions, which minimize aggregation‐caused quenching. At the single particle level, these particles are ≈20‐fold brighter than quantum dots of similar color. Due to their identical 40 nm size and surface properties, these nanoparticles are endocytosed equally well by living cells. Mixing nanoparticles of three colors in different proportions generates a homogeneous RGB (red, green, and blue) barcode in cells, which is transmitted through many cell generations. Cell barcoding is validated on 7 cell lines (HeLa, KB, embryonic kidney (293T), Chinese hamster ovary, rat basophilic leucemia, U97, and D2A1), 13 color codes, and it enables simultaneous tracking of co‐cultured barcoded cell populations for >2 weeks. It is also applied to studying competition among drug‐treated cell populations. This technology enabled six‐color imaging in vivo for (1) tracking xenografted cancer cells and (2) monitoring morphogenesis after microinjection in zebrafish embryos. In addition to a robust method of multicolor cell labeling and tracking, this work suggests that multiple functions can be co‐localized inside cells by combining structurally close nanoparticles carrying different functions.     

Interactions of Osteoblastic and Other Cells with Bioactive Glasses and Silica In Vitro and In Vivo

Responses of cells to the presence of 90‐150 μm particles of 3 bioactive glasses were compared with their reaction to non‐bioactive borosilicate glass and to a known tissue irritant, quartz/silica. Measurements were made in vitro and in vivo of changes of intra‐ and extracellular ion activities. Ion‐selective microelectrodes were used to determine intracellular pH, [Na⁺], [K⁺] and [Ca²⁺] in cultured rodent osteoblasts, fibroblasts and macrophages in contact with, or exposed to medium containing, Bioglass® 45S5, borosilicate glass and bioactive glasses 77S and 58S. The same materials were implanted in connective tissue in rabbit ear chambers and similar measurements were made in vivo. No significant differences were observed among the 3 cell‐types in their reactions, which were minimal, to the presence of the borosilicate and 58S and 77S glasses either in vivo or in vitro. However all cells were affected by interaction with Bioglass® 45S5 which caused significant alkalinisation at its extracellular contact sites and concomitant but smaller changes in internal pH. The latter were accompanied by significant increases in [K⁺], [Na⁺] and [Ca²⁺]. There was also hyperpolarisation of plasma membrane potential in fibroblasts and osteoblasts. In the presence of silica, macrophages showed marked and significant decreases in pH both at the contact zone and intracellularly. Among the cells used, osteoblasts showed higher and macrophages lower pH than fibroblasts when in contact with the test materials. Despite the known leaching of silicic acid from Bioglass® macrophages were not “activated” by its presence, but were by silica. It is concluded that materials developed as potential prostheses should also be examined for their ability to alter cellular metabolism.     

In Vivo and In Vitro Degradation Behavior of Magnesium Alloys as Biomaterials

The corrosion behavior of pure Mg,AZ31,and AZ91D were evaluated in various in vitro and in vivo environments to investigate the potential application of these metals as biodegradable implant materials.DC polarization tests and immersion tests were performed in different simulated body solutions,such as distilled(DI) water,simulated body fluid(SBF) and phosphate buffered solution(PBS).Mg/Mg alloys were also implanted in different places in a mouse for in vivo weight loss and biocompatibility investigations.The in vivo subcutis bio-corrosion rate was lower than the corrosion rate for all of the in vitro simulated corrosive environments.The Mg/Mg alloys were biocompatible based on histology results for the liver,heart,kidney,skin and lung of the mouse during the two months implantation.Optical microscopy and scanning electron microscopy were carried out to investigate the morphology and topography of Mg/Mg alloys after immersion testing and implantation to understand the corrosion mechanisms.     

In Vitro and In Vivo Evaluation of Glibenclamide in Solid Dispersion Systems

The purpose of this work is to improve the dissolution and bioavailability characteristics of glibenclamide as compared to Daonil® tablets (Hoechst). Solid dispersions of glibenclamide in Gelucire 44/14 (Formula 1) and in polyethylene glycol 6000 (PEG 6000) (Formula 2) were prepared by fusion method. In vitro dissolution studies showed that the dispersing systems containing glibenclamide and Gelucire 44/14 or PEG 6000 gave faster dissolution rates than the reference product Daonil. The in vivo bioavailability study was assessed in six healthy male volunteers in crossover design with a 1‐week washout period. Both formulas were found to be significantly different from Daonil with regard to the extent of absorption as indicated by the area under serum concentration‐time curve. Both formulas are not significantly different from Daonil with respect to time of peak plasma concentration (T_max). It is concluded from this pilot study that the ranking of the in vitro dissolution is similar to the ranking of in vivo availability. The ranking of the three preparations in term of dissolution rate and extent of absorption is as follows: Formula 2?>?Formula 1?>?Daonil.     

In Vitro and In Vivo Adhesion Testing of Mucoadhesive Drug Delivery Systems

Bioadhesive tablets were prepared by physical mixing of polymers and drug, then granulating and compressing into a tablet. The mucoadhesion was evaluated by shear stress measurement, detachment force measurement, and X-ray photography of the rabbit gastrointestinal tract. The strong interaction between the polymer and the mucous lining of the tissue helps increase contact time and permit localization. Polymers like hydroxypropyl methylcellulose K4M (HPMC K4M), hydroxypropyl methylcellulose 100 cps (HPMC 100 cps), carbopol-934, sodium carboxy methylcellulose (Na CMC), guar gum, and polyvinylpyrrolidone (PVP) were tested by shear stress measurement and detachment force measurement methods. HPMC K4M, showing maximum bioadhesion, was used in further studies. Adhesion was maximum between pH 5 and pH 6. Maximum adhesion was observed in the duodenum, followed by the jejunum and ileum. Barium sulfate (BaSO₄) matrix tablets containing polymer and drug were subjected to X-ray studies in rabbits, and it was found that the tablet was mucoadhesive even after 8 hr. Enteric coating did not show any effect on mucoadhesion after passing from the stomach.     

Development of Copper Nanoclusters for In Vitro and In Vivo Theranostic Applications

Theranostics refers to the incorporation of therapeutic and diagnostic functions into one material system. An important class of nanomaterials exploited for theranostics is metal nanoclusters (NCs). In contrast to gold and silver NCs, copper is an essential trace element for humans. It can be more easily removed from the body. This, along with the low cost of copper that offers potential large-scale nanotechnology applications, means that copper NCs have attracted great interest in recent years. The latest advances in the design, synthesis, surface engineering, and applications of copper NCs in disease diagnosis, monitoring, and treatment are reviewed. Strategies to control and enhance the emission of copper NCs are considered. With this synopsis of the up-to-date development of copper NCs as theranostic agents, it is hoped that insights and directions for translating current advances from the laboratory to the clinic can be further advanced and accelerated.     

In Vitro and In Vivo Release of Naltrexone from Biodegradable Depot Systems

ABSTRACT

The aim of this study was to prepare poly(d, l-lactide) (PLA) microspheres containing naltrexone (NTX) by a solvent evaporation method, and to evaluate both in vitro and in vivo release characteristics and histopathological findings of tissue surrounding an implant formulation in rats.

This method enabled the preparation of microspheres of regular shape and relatively narrow particle size distribution. The in vitro release profiles of NTX from PLA microspheres showed the release of NTX did not follow zero-order kinetics. An initial burst release was observed, subsequently followed by a nearly constant rate of 0.4% per day after ten days. The cumulative amount of NTX released at the end of 60 days was 80%. Compressed microspheres showed near zero-order sustained release of NTX for 360 days. The plasma NTX levels in rats showed that for compressed microspheres NTX concentrations were constant and exceeded 2 ng/mL for 28 days. Throughout the 28 days of study, the implantations cause a minor inflammatory response, which can be regarded as a normal defence mechanism. The sustained release performance of NTX from the biodegradable depot systems may provide a reliable, convenient, and safe mechanism for the administration of NTX for the long-term treatment of opioid dependence.     

乳酸-乙醇酸共聚物/壳聚糖复式微球缓释蛋白

采用两次乳化包埋技术制备了一种载蛋白的乳酸-乙醇酸共聚物/壳聚糖复式微球（Poly（lactide-co-glycolide）/chitosan微球,简称PLGA/chitosan微球）。先以复乳法（W/O/W）制备加载牛血清白蛋白（BSA）的PLGA微球,再以壳聚糖（Chitosan）为基体对PLGA微球进行包埋,用...     

A Molecular Hero Suit for In Vitro and In Vivo DNA Nanostructures

Precise control of DNA base pairing has rapidly developed into a field full of diverse nanoscale structures and devices that are capable of automation, performing molecular analyses, mimicking enzymatic cascades, biosensing, and delivering drugs. This DNA‐based platform has shown the potential of offering novel therapeutics and biomolecular analysis but will ultimately require clever modification to enrich or achieve the needed “properties” and make it whole. These modifications total what are categorized as the molecular hero suit of DNA nanotechnology. Like a hero, DNA nanostructures have the ability to put on a suit equipped with honing mechanisms, molecular flares, encapsulated cargoes, a protective body armor, and an evasive stealth mode.