Development and in vitro–in vivo evaluation of a water-in-oil microemulsion formulation for the oral delivery of troxerutin

Objective: The main objective of this study was to develop and evaluate a W/O microemulsion formulation of troxerutin to improve its oral bioavailability.

Methods: The W/O microemulsion was optimized using a pseudo-ternary phase diagram and evaluated for physical properties. In vitro MDCK cell permeability studies were carried out to evaluate the permeability enhancement effect of microemulsion, and in vivo absorption of troxerutin microemulsion in the intestine was compared with that of solution after single-dose administration (56.7?mg/kg) in male Wistar rats.

Results: The optimal formulation consisted of lecithin, ethanol, isopropyl myristate and water (23.30/11.67/52.45/12.59 w/w) was physicochemical stable and the mean droplet size was about 50.20?nm. In vitro study, the troxerutin-loaded microemulsion showed higher intestinal membrane permeability across MDCK monolayer when compared with the control solution. The W/O microemulsion can significantly promote the intestinal absorption of troxerutin in rats in vivo, and the relative bioavailability of the microemulsion was about 205.55% compared to control solution.

Conclusion: These results suggest that novel W/O microemulsion could be used as an effective formulation for improving the oral bioavailability of troxerutin.     

Design of oral drug delivery systems – Past,present and future

Abstract

This paper reviews the past and the present thinking about peroral delivery research and development and how future approaches may be significantly different, both qualitatively and quantitatively. The future should involve the use of comprehensive models capable of incorporating physico-chemical data and biological information such as gastrointestinal flow, how and where drug absorption occurs, and whether and where metabolism of the drug occurs during gastrointestinal transit. Special challenges would involve the use of such models in research protocols in the optimization of drug delivery systems.     

Development of β-cyclodextrin-based sustained release microparticles for oral insulin delivery

Polymeric microparticles have been previously demonstrated to deliver various therapeutic agents efficiently to targeted regions by protecting the drug from harsh gastric milieu of the gastrointestinal tract. In this study, we investigated the hypoglycemic effect of β-cyclodextrin polymeric insulin microparticles in diabetic rats via the oral route of administration. β-cyclodextrin microparticles were prepared by a unique one-step spray-drying technique and stabilized by incorporating enteric retardant polymers in the formulation. The insulin-loaded microparticles had a mean size of 0.8?±?0.25?μm with a zeta potential of 3.57?+?0.62?mV. As seen with the chromatographic analysis, the drug content in the microparticles was determined to be 94.9?±?2.77%. RAW macrophage cells showed greater than 80% viability after 24?h of incubation with the insulin and blank microparticles. For the in vitro release study, the microparticles were able to protect the insulin in gastric fluid where no significant release was detected, followed by only 50% release in intestinal fluid for the first 8?h of the study. This was seen to correlate with the in vivo data where 50% glucose inhibition was seen after 8?h of oral administration in diabetic rats. This data suggest that the oral insulin microparticles were able to reduce glucose levels in disease conditions and would be a favorable route of administration to patients as an alternative to daily subcutaneous injections.     

Adhesion of oral streptococci to all-ceramics dental restorative materials in vitro

In recent years, patients have benefited from the development of better and more esthetic materials, including all-ceramics dental restorative materials. Dental plaque formation on teeth and restorative materials plays an important role in the pathogenesis of oral diseases. This study investigates initial adhesion of stationary phase streptococcal species to different all-ceramics dental restorative materials. The saliva-coated materials were incubated with the bacteria for 1 h in an in vitro flow chamber which mimics environmental conditions in the oral cavity. Number and vitality of adhering bacteria were determined microscopically after staining. Surface roughness and the composition of the materials had no distinctive influence on bacterial adhesion. However, S. mutans and S. sobrinus adhered about tenfold less numerous to all materials than the other streptococcal species. Further, there was a correlation between bacterial vitality and materials' glass content. The results showed that early plaque formation was influenced predominantly by the presence of the salivary pellicle rather than by material dependent parameters whereas the composition of the all-ceramics appeared to have influenced the percentage of viable cells during the adhesion process. This presented in vitro technique may provide a useful model to study the influence of different parameters on adherence of oral streptococcal species.     

Development and evaluation of a novel drug delivery: Soluplus®/TPGS mixed micelles loaded with piperine in vitro and in vivo

Background: Although piperine can inhibit cells of tumors, the poor water solubility restricted its clinical application. This paper aimed to develop mixed micelles based on Soluplus^® and D-α-tocopherol polyethylene glycol succinate (TPGS) to improve the aqueous solubility and anti-cancer effect.

Methods: Piperine-loaded mixed micelles were prepared using a thin-film hydration method, and their physicochemical properties were characterized. The cellular uptake of the micelles was confirmed by confocal laser scanning microscopy in A549 lung cancer cells and HepG₂ liver cancer cells. In addition, cytotoxicity of the piperine mixed micelles was studied in A549 lung cancer cells and HepG₂ liver cancer cells. Free piperine or piperine-loaded Soluplus^®/TPGS mixed micelles were administered at an equivalent dose of piperine at 3.2?mg/kg via a single intravenous injection in the tail vain for the pharmacokinetic study in vivo.

Results: The diameter of piperine-loaded Soluplus^®/TPGS (4:1) mixed micelles was about 61.9?nm and the zeta potential –1.16?±?1.06?mV with 90.9% of drug encapsulation efficiency and 4.67% of drug-loading efficiency. Differential scanning calorimetry (DSC) studies confirmed that piperine is encapsulated by the Soluplus^®/TPGS. The release results in vitro showed that the piperine-loaded Soluplus^®/TPGS mixed micelles presented sustained release behavior compared to the free piperine. The mixed micelles exhibited better antitumor efficacy compared to free piperine and physical mixture against in A549 and HepG₂ cells by MTT assay. The pharmacokinetic study revealed that the AUC of piperine-loaded mixed micelles was 2.56 times higher than that of piperine and the MRT for piperine-loaded mixed micelles was 1.2-fold higher than piperine (p?Conclusion: The results of the study suggested that the piperine-loaded mixed micelles developed might be a potential nano-drug delivery system for cancer chemotherapy. These results demonstrated that piperine-loaded Soluplus^®/TPGS mixed micelles are an effective strategy to deliver piperine for cancer therapy.     

Evaluation of in vitro degradation of PCL scaffolds fabricated via BioExtrusion – Part 2: Influence of pore size and geometry

The in vivo degradation processes by which scaffolds degrade and are replaced by neo-tissue are complex and may be influenced by many factors, including environmental conditions, material properties, porosity and 3D architecture. The present study is focused on the influence of design parameters, filament distance (FD) and lay-down pattern, on the degradation kinetics of Polycaprolactone (PCL) scaffolds obtained via BioExtrusion. Through the variation of design parameters it was possible to obtain two groups of scaffolds with distinct pore geometry and size. The in vitro degradation was performed in simulated body fluid (SBF) and in phosphate buffer solution (PBS) for six months. Our results highlight a more complex degradation pattern of the scaffolds in SBF than in PBS, probably related to a mineral deposition. Significant statistical differences in weight loss values at month 6, allowed us to conclude that degradation kinetics of PCL scaffolds is strongly influenced by the pore size.     

Lipid nanoparticles of zaleplon for improved oral delivery by Box–Behnken design: optimization,in vitro and in vivo evaluation

Purpose: Zaleplon (ZL) is a hypnotic drug prescribed for the management of insomnia and convulsions. The oral bioavailability of ZL was low (～30%) owing to poor water solubility and hepatic first-pass metabolism. The cornerstone of this investigation is to develop and optimize solid lipid nanoparticles (SLNs) of ZL with the aid of Box–Behnken design (BBD) to improve the oral bioavailability.

Methods: A design space with three formulation variables at three levels were evaluated in BBD. Amount of lipid (A₁), amount of surfactant (A₂) and concentration of co-surfactant (%) (A₃) were selected as independent variables, whereas, particle size (B₁), entrapment efficiency (B₂) and zeta potential (ZP, B₃) as responses. ZL-SLNs were prepared by hot homogenization with ultrasonication method and evaluated for responses to obtain optimized formulation. Morphology of nanoparticles was observed under SEM. DSC and XRD studies were examined to understand the native crystalline behavior of drug in SLN formulations. Further, in vivo studies were performed in Wistar rats.

Results: The optimized formulation with 132.89?mg of lipid, 106.7?mg of surfactant and 0.2% w/v of co-surfactant ensued in the nanoparticles with 219.9?±?3.7?nm of size, ?25.66?±?2.83?mV surface charge and 86.83?±?2.65% of entrapment efficiency. SEM studies confirmed the spherical shape of SLN formulations. The DSC and XRD studies revealed the transformation of crystalline drug to amorphous form in SLN formulation. In conclusion, in vivo studies in male Wistar rats demonstrated an improvement in the oral bioavailability of ZL from SLN over control ZL suspension.

Conclusions: The enhancement in the oral bioavailability of ZL from SLNs, developed with the aid of BBD, explicated the potential of lipid-based nanoparticles as a potential carrier in improving the oral delivery of this poorly soluble drug.     

Comparative study of human monocyte and platelet adhesion to hydrogels in vitro – effect of polymer structure

Blood platelet and monocyte adhesion was studied in vitro with respect to the influence of hydrophilic polymer chemical functional groups and their charge. The results showed that the strongest adhesion of human monocytes was to coverslips covered with cationic polymer. Platelet adhesion to all tested polymers proved to be negligible; no differences related to the charge of the polymers used were observed. These results show the obvious difference between the biocompatibility and haemocompatibility in vitro which must be taken into consideration during polymer biological properties testing before clinical trials.     

Synthesis and in vitro characterization of a novel PAA–ATP conjugate

The objective of this study was to improve the multifunctional properties of poly(acrylic acid) (PAA) by covalent attachment of 4-aminothiophenol (ATP) to its backbone. The permeation enhancing effect of PAA–ATP together with glutathione was evaluated in Ussing-type chambers using fluorescein isothiocyanate dextran as model compound. The mucoadhesive properties were evaluated in vitro on freshly excised porcine intestinal mucosa through the rotating cylinder method. The resulting conjugates PAA–ATP1 and PAA–ATP2 displayed 168 ± 35 and 426 ± 55 μmol immobilized free thiol groups per gram polymer, respectively. In addition, 279 ± 28 and 139 ± 22 μmol disulfide bonds per gram polymer, respectively, were identified on PAA–ATP1 and PAA–ATP2. Within disintegration studies in aqueous buffer solution, the modified polymers showed improved cohesive properties. Because of the immobilization of ATP, the swelling of PAA–ATP1 and PAA–ATP2 improved 12.0- and 17.8-fold, respectively. The adhesion times of the conjugates PAA–ATP1 and PAA–ATP2 were more than 20- and 30-fold increased in comparison to unmodified PAA. Furthermore, conjugates PAA–ATP1 and PAA–ATP2 exhibited a 1.86- and 2.07-fold higher permeation enhancing effect, respectively, over unmodified PAA. According to these results, PAA–ATP conjugates represent a very promising novel type of thiomer for the development of various mucoadhesive drug delivery systems.     

Melt dispersion granules: formulation and evaluation to improve oral delivery of poorly soluble drugs – a case study with valsartan

Solid dispersion (SD) technique is a promising strategy to improve the solubility and dissolution of BCS class II drugs. However, only few products are marketed till today based on SD technology due to poor flow properties and stability. The present work was intended to solve these problems by using combination approach, melt dispersion and surface adsorption technologies. The main aim of the present work is to improve the absorption in the stomach (at lower pH) where the absorption window exists for the drug by improving the dissolution, resulting in the enhancement of oral bioavailability of poorly soluble, weakly acidic drug with pH dependant solubility, i.e. valsartan. Melt dispersion granules were prepared in different ratios using different carriers (Gelucire 50/13, PEG 8000 and Pluronic F-68) and lactose as an adsorbent. Similarly, physical mixtures were also prepared at corresponding ratios. The prepared dispersion granules and physical mixtures were characterized by FTIR, DSC and in vitro dissolution studies. DSC studies revealed reduction in the crystallinity with a possibility of presence of amorphous character of drug in the dispersion granules. From dissolution studies, valsartan Gelucire dispersion (GSD4; 1:4 ratio) showed complete drug release in 30?min against the plain drug which showed only 11.31% of drug release in 30?min. Pharmacokinetic studies of optimized formulation in male Wistar rats showed 2.65-fold higher bioavailability and 1.47-fold higher C_max compared to pure drug. The melt dispersion technology has the potential to improve dissolution and the bioavailability of BCS class II drugs.     

In vitro and in vivo degradation of poly(l-lactide-co-glycolide) films and scaffolds

Poly(l-lactide-co-glycolide) (PLGA) was synthesized using a biocompatible initiator, zirconium acetylacetonate. In vitro and in vivo degradation properties of PLGA films (produced by solvent casting, 180 μm thick) and PLGA scaffolds (produced by an innovated solvent casting and particulate leaching, 3 mm thick) were evaluated. The samples were either submitted for degradation in phosphate buffered saline (PBS) at 37 °C for 30 weeks, or implanted into rat skeletal muscles for 1, 4, 12, 22 and 30 weeks. The degradation was monitored by scanning electron microscopy, atomic force microscopy, weight loss, and molecular weight changes (in vitro), and by microscopic observations of the materials’ morphology after histological staining with May-Grunwald-Giemsa (in vivo). The results show that the films in both conditions degraded much faster than the scaffolds. The scaffolds were dimensionally stable for 23 weeks, while the films lost their integrity after 7 weeks in vitro. The films’ degradation was heterogenous—degradation in their central parts was faster than in the surface and subsurface regions due to the increased concentration of the acidic degradation products inside. In the scaffolds, having much thinner pore walls, heterogenous degradation due to the autocatalytic effect was not observed.     

Development and comparative evaluation of in vitro,in vivo properties of novel controlled release compositions of paroxetine hydrochloride hemihydrate as against Geomatrix™ platform technology

The objective of this study is to develop, in vitro and in vivo evaluation of novel approaches for controlled release of paroxetine hydrochloride hemihydrate (PHH) in comparison to patented formulation PAXIL CR^® tablets of GlaxoSmithKline (Geomatrix? technology). In one of the approaches, hydrophilic core matrix tablets containing 85% of the dose were prepared and further coated with methacrylic acid copolymer to delay the release. An immediate release coating of 15% was given as top coat. The tablets were further optionally coated using ethyl cellulose. In the second approach, hydrophobic matrix core tablets containing metharylic acid copolymer were prepared. In the third approach, PHH was granulated with enteric polymer and further hydrophobic matrix core tablets were prepared. The effect of polymer concentration, level of enteric coating on drug release was evaluated by in vitro dissolution study by varying dissolution apparatus and the rotation speeds. It was found that increase in concentration of high viscosity hydroxypropylmethylcellulose (HPMC) resulted in reduction of the release rate. The drug release was observed to be dependent on the level of enteric coating and ethyl cellulose coating, being slower at increased coating. The release mechanism of PHH followed zero-order shifting to dissolution dependent by the increase of HPMC content. The formulation was stable without change in drug release rate. In vivo study in human volunteers confirmed the similarity between test and innovator formulations. In conclusion, HPMC-based matrix tablets, which were further coated using methacrylic acid copolymer, were found to be suitable for the formulation of single layer-controlled release PHH.     

Combination of β-cyclodextrin inclusion complex and self-microemulsifying drug delivery system for photostability and enhanced oral bioavailability of methotrexate: novel technique

In the present study, we prepared an inclusion complex of methotrexate (MTX) with β-cyclodextrin (β-CD) in order to decrease its photosensitivity and enhance its aqueous solubility. Then we incorporated this inclusion complex in a self-microemulsifying drug delivery system (SMEDDS) overall to increase its oral bioavailability. The inclusion complex has been prepared by freeze drying method and characterized by differential scanning calorimetry (DSC), ultraviolet (UV), and infrared (IR) spectroscopy assays. The proper molecular ratio of MTX/β-CD was found to be of 1:7, and the water-solubility of MTX was increased in an average of 10-fold. The photostability studies showed that the MTX became stable on exposure to light. Construction of pseudoternary diagrams were investigated to prepare a MTX/β-CD inclusion complex loaded SMEDDS which was characterized by measuring the particle size and the zeta-potential. The optimum formulation of SMEDDS was a system consisting of ethyl oleate, tween 80, and propylene glycol with a mean droplet size of 39.42?nm. In vitro drug release in different pH media showed that the release profile of MTX from the MTX/β-CD loaded SMEDDS was influenced by the pH of the release medium and presented the characteristics of a sustained release profile. Finally, in-vivo studies showed an enhancement of the bioavailability of MTX from the MTX/β-CD loaded SMEDDS form of 1.57-fold. We concluded that the β-CD inclusion complex loaded SMEDDS improved the chemical and physiological properties of MTX and could be a promising means for the delivery of MTX and other unstable and lipophilic drugs by oral route.     

Effect of polymerization degree of calcium polyphosphate on its microstructure and in vitro degradation performance

Preparation, characterization and in vitro study of a series of calcium polyphosphate (CPP) with different polymerization degree were reported. A series of CPP with different polymerization degree were prepared by controlling calcining time. Average polymerization degree was analyzed by liquid state ³¹P nuclear magnetic resonance (NMR). The microstructure was observed by scanning electric microscope (SEM). X-ray diffraction (XRD) analysis was used to demonstrate that polymerization degree would not affect the crystal system and space group of CPP. The results showed that polymerization degree increased with the increase of calcining time. Degradation studies were performed during 32 days in physiological saline solution (aqueous solution, 0.9 wt.%NaCl) to assess the effect of polymerization degree on the degradation velocity of the samples. It was also shown that the degradation velocity of CPP (polymerization degree = 13) doubles than another two samples (polymerization degree = 9,19). The results in the present study may be able to provide some fundamental data for controlling CPP degradation.     

Bioactive borosilicate glass scaffolds: in vitro degradation and bioactivity behaviors

Bioactive borosilicate glass scaffolds with the pores of several hundred micrometers and a competent compressive strength were prepared through replication method. The in vitro degradation and bioactivity behaviors of the scaffolds have been investigated by immersing the scaffolds statically in diluted phosphate solution at 37°C, up to 360 h. To monitor the degradation progress of the scaffolds, the amount of leaching elements from the scaffolds were determined by ICP-AES. The XRD and SEM results reveal that, during the degradation of scaffolds, the borosilicate scaffolds converted to hydroxyapatite. The compressive strength of the scaffolds decreased during degradation, in the way that can be well predicted by the degradation products, or the leachates, from the scaffolds. MTT assay results demonstrate that the degradation products have little, if any, inhibition effect on the cell proliferation, when diluted to a certain concentration ([B] <2.690 and pH value at neutral level). The study shows that borosilicate glass scaffold could be a promising candidate for bone tissue engineering material.     

Formulation,characterization and in vitro–in vivo evaluation of flurbiprofen-loaded nanostructured lipid carriers for transdermal delivery

Flurbiprofen is used in the treatment of arthritis. However, its multiple dosing due to short elimination half life is a concern for such treatment. This work aims to develop nanostructured lipid carriers (NLCs) of flurbiprofen and evaluate their potential for transdermal delivery. The NLCs were prepared by the optimized o/w emulsification-homogenization-sonication technique using coconut oil (liquid lipid). The NLCs were found to be spherical with uniform size (214 nm). The entrapment efficiency and zeta potential were 92.58% and ?30.70 mV, respectively. Differential scanning calorimetry (DSC) showed the amorphous state of flurbiprofen encapsulated in NLCs. The percentage cumulative drug release through the excised rat skin from NLCs was biphasic and significantly prolonged compared with the commercial gel. DSC of the treated skin indicated that the NLCs penetrate into follicles of the skin and accumulate in the dermis. The bioavailability of flurbiprofen from NLCs was more than 1.7-fold that of the commercial gel. The NLCs showed a quick onset and sustained anti-inflammatory effect over period of 24 h for carrageenan-induced rat paw edema than the commercial gel. The stability data revealed that the NLCs were more stable when stored at 5°C. In conclusion, prepared NLCs have potential for skin targeting and sustained drug release.     

Effect of fluoride coating on in vitro dynamic degradation of Mg–Zn alloy

A compact and flat fluoride coating with some pores was prepared on a Mg–Zn alloy in order to control its degradation behavior. The electrochemical tests demonstrated that the real impedance (Z_re) of the fluoride-coated Mg–Zn was approximately 10 times as large as that of the untreated alloy. The free corrosion potential (E_corr), compared to that of the uncoated Mg–Zn alloy, increased 646 mV for the coated metal. The free corrosion current (I_corr) of the Mg–Zn specimen with the fluoride film was about one tenth of that of the uncoated one. The in vitro dynamic degradation tests showed that the average weight loss of the fluoride-coated Mg–Zn was lower than that of the untreated alloy in the initial 4 h of the tests, indicating the film could function as a barrier coating on Mg–Zn matrix. However, the coating cracked and peeled severely after 4 h dynamic tests, which implied that the fluoride coating could not endure the sustaining washing of the modified simulated body fluid.     

Development of zolmitriptan transfersomes by Box–Behnken design for nasal delivery: in vitro and in vivo evaluation

The aim was to prepare an optimized zolmitriptan (ZT)-loaded transfersome formulation using Box–Behnken design for improving the bioavailability by nasal route for quick relief of migraine and further to compare with a marketed nasal spray. Here, three factors were evaluated at three levels. Independent variables include: amount of soya lecithin (X₁), amount of drug (X₂) and amount of tween 80 (X₃). The dependent responses were vesicle size (Y₁), flexibility index (Y₂) and regression coefficient of drug release kinetics (Y₃). Prepared formulations were evaluated for physical characters and an optimal system was identified. Further, in vivo pharmacokinetic study was performed in male wistar rats to compare the amount of drug in systemic circulation after intranasal administration. Optimized ZT-transfersome formulation containing 82.74?mg of lecithin (X₁), 98.37?mg of zolmitriptan (X₂) and 32.2?mg of Tween 80 (X₃) and had vesicle size of 93.3?nm, flexibility index of 20.25 and drug release regression coefficient of 0.992. SEM picture analysis revealed that the vesicles were spherical in morphology and had a size more than 1?µm. The formulations were found to be physically stable upon storage at room temperature up to 2?months period, as there were no significant changes noticed in size and ZP. The nasal bioavailability of optimized transfersome formulation was found to be increased by 1.72 times than that of marketed nasal spray (Zolmist^®). The design and development of zolmitriptan as transfersome provided improved nasal delivery over a conventional nasal spray for a better therapeutic effect.     

Design and in vitro evaluation of novel γ-PGA/hydroxyapatite nanocomposites for bone tissue engineering

In order to improve biocompatibility and cell growth, poly-γ-glutamic acid (γ-PGA) was used as an additive, and a series of γ-PGA/hydroxyapatite (γ-PGA/HAp) nanocomposites were prepared. Then the morphology, water uptake and retention abilities, in vitro degradation properties in the simulated medium, and cytotoxicity of these γ-PGA/HAp nanocomposites were investigated. The results show that the γ-PGA/HAp nanocomposites have homogenous nano-sized grains, hydrophilicity, biocompatibility, and controlled in vitro degradation, suggesting that the γ-PGA/HAp nanocomposites are novel nanostructure composites with great potential application in the field of bone tissue engineering.