Evaluation of polyethersulfone highflux hemodialysis membrane in vitro and in vivo

A new hemodialysis membrane manufactured by a blend of polyethersulfone (PES) and polyvinylpyrrolidone (PVP) was evaluated in vitro and in vivo. Goat was selected as the experimental animal. The clearance and the reduction ratio after the hemodialysis of small molecules (urea, creatinine, phosphate) for the PES membrane were higher in vitro than that in vivo. The reduction ratio of β₂-microglobulin was about 50% after the treatment for 4 h. The biocompatibility profiles of the membranes indicated slight neutropenia and platelet adhesion at the initial stage of the hemodialysis. Electrolyte, blood gas, and blood biochemistry were also analyzed before and after the treatment. The results indicated that PES hollow fiber membrane had a potential widely use for hemodialysis.     

Biocompatibility of polyhydroxybutyrate microspheres: in vitro and in vivo evaluation

Microspheres have been prepared from the resorbable linear polyester of β-hydroxybutyric acid (polyhydroxybutyrate, PHB) by the solvent evaporation technique and investigated in vitro and in vivo. Biocompatibility of the microspheres has been proved in tests in the culture of mouse fibroblast cell line NIH 3T3 and in experiments on intramuscular implantation of the microspheres to Wistar rats for 3 months. Tissue response to the implantation of polymeric microspheres has been found to consist in a mild inflammatory reaction, pronounced macrophage infiltration that increases over time, involving mono- and poly-nuclear foreign body giant cells that resorb the polymeric matrix. No fibrous capsules were formed around polymeric microparticles; neither necrosis nor any other adverse morphological changes and tissue transformation in response to the implantation of the PHB microparticles were recorded. The results of the study suggest that polyhydroxybutyrate is a good candidate for fabricating prolonged-action drugs in the form of microparticles intended for intramuscular injection.     

The controlling biodegradation of chitosan fibers by N-acetylation in vitro and in vivo

Aim In the present study, we investigated the biodegradation of the fibers of chitosan and its acetylated derivatives in vitro and in vivo. Methods A series of chitosan fibers, with acetylation degrees of 7.7%, 21.6%, 40.9%, 61.2%, 82.5% and 93.4%, were obtained by acetylating chitosan filament with acetic anhydride, and were investigated by FT-IR analysis, elemental analysis and scanning electron microscopy analysis. Results The in vitro experimental data indicated that the degradation rate of chitosan fiber was strongly dependent on the degree of acetylation, and the degradation rate increased with an enhancement of the acetylation degree of chitosan fibers. In vivo degradation experiment evaluated by light microscopy as well as scanning electron microscopy, was studied by implanting the fibers between the two nerve stumps of the rat sciatic nerve gap (6 months). The findings demonstrated that acetylation degree could influence the degradation rate of chitosan fibers in vivo. Conclusion These results suggested that acetylated chitosan (chitin) fibers were more biodegradable than chitosan and the biodegradation rate of chitin fiber can be controlled to desirable extent by the variation of acetylation degree.     

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.     

Hyaluronan-based heparin-incorporated hydrogels for generation of axially vascularized bioartificial bone tissues: in vitro and in vivo evaluation in a PLDLLA–TCP–PCL-composite system

Smart matrices are required in bone tissue-engineered grafts that provide an optimal environment for cells and retain osteo-inductive factors for sustained biological activity. We hypothesized that a slow-degrading heparin-incorporated hyaluronan (HA) hydrogel can preserve BMP-2; while an arterio–venous (A–V) loop can support axial vascularization to provide nutrition for a bio-artificial bone graft. HA was evaluated for osteoblast growth and BMP-2 release. Porous PLDLLA–TCP–PCL scaffolds were produced by rapid prototyping technology and applied in vivo along with HA-hydrogel, loaded with either primary osteoblasts or BMP-2. A microsurgically created A–V loop was placed around the scaffold, encased in an isolation chamber in Lewis rats. HA-hydrogel supported growth of osteoblasts over 8 weeks and allowed sustained release of BMP-2 over 35 days. The A–V loop provided an angiogenic stimulus with the formation of vascularized tissue in the scaffolds. Bone-specific genes were detected by real time RT-PCR after 8 weeks. However, no significant amount of bone was observed histologically. The heterotopic isolation chamber in combination with absent biomechanical stimulation might explain the insufficient bone formation despite adequate expression of bone-related genes. Optimization of the interplay of osteogenic cells and osteo-inductive factors might eventually generate sufficient amounts of axially vascularized bone grafts for reconstructive surgery.     

Correlation of in vitro and in vivo results of vacuum plasma sprayed titanium implants with different surface topography

Research has proven that rough surfaces improve both biologic and biomechanical responses to titanium (Ti) implants. The purpose of this study was to evaluate the correlation between the expression of bone cell-associated proteins to Vacuum Plasma-Sprayed Titanium implants (VPS-Ti) with different surface textures in vitro and the bone integration in vivo. The biological performances of the surfaces were evaluated over a period of 8 weeks using human bone marrow cell cultures and Göttinger mini pigs. Cells were cultured on VPS-Ti with two respectively different surface-roughnesses (Ra). The level of Osteoprotegerin (OPG), Osteocalcin (OC) and alkaline phosphatase activity (ALP) were evaluated. The bone integration in vivo was evaluated by histomorphological analyses. A cancellous structured titanium (CS-Ti) construct was used as reference material in both study designs. Comparison of data was conducted using the Scheffé tests and the paired t-test with Bonferroni’s correction. A comparative analysis was done to measure the degree of association between the in vitro and in vivo data. A total amount of OC was significantly increased for VPS-Ti for cells cultured on both VPS-Ti and CS-Ti, while OPG was only detectable after 8 weeks without any significant differences. The ALP activity on all surfaces was not statistically increased. For VPS-Ti with Ra ranging from 0.025 mm up to 0.059 mm, bone integration response was increased, but there was no statistical difference between the VPS-Ti. Expression of OPG, OC and ALP correlated with the histomorphological data over the 8-week period. The in vitro data suggest the superiority of VPS-Ti over CS-Ti, but more importantly, the biocompatibility of testing an in vitro model to predict the outcome and possible integration of implants in vivo.     

Preparation,mechanical properties and in vitro degradability of wollastonite/tricalcium phosphate macroporous scaffolds from nanocomposite powders

A new class of scaffolds with a gain size of 200 nm was prepared from wollastonite/tricalcium phosphate (WT) nanocomposite powders (termed “nano-sintered scaffolds”) through a two-step chemical precipitation and porogen burnout techniques. For a comparison, WT scaffolds with a grain size of 2 μm were also fabricated from submicron composite powders (termed “submicron-sintered scaffolds”) under the same condition. The resultant scaffolds showed porosities between 50 ± 1.0% and 65 ± 1.0% with a pore size ranging from 100 μm to 300 μm. The WT nano-sintered scaffolds exhibited compressive strength and elastic modulus values that were about twice that of their submicron-sintered counterparts. The in vitro degradation tests demonstrated that the degradability could be regulated by the grain size of bioceramics. The decreased specific surface area of pores in the nano-sintered scaffolds led to their reduced degradation rate. The mechanical properties of the nano-sintered scaffolds exhibited less strength loss during the degradation process. The WT macroporous nano-sintered scaffolds are a promising and potential candidate for bone reconstruction applications.     

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.     

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.     

Human tissue allograft processing: impact on in vitro and in vivo biocompatibility

This work investigates the impact of chemical and physical treatments on biocompatibility for human bone/tendon tissues. Nontreated and treated tissues were compared. In vitro testing assessed indirect and direct cytotoxicity. Tissues were subcutaneously implanted in rats to assess the immunological, recolonization, and revascularization processes at 2–4 weeks postimplantation. No significant cytotoxicity was found for freeze-dried treated bones and tendons in comparison to control. The cellular adhesion was significantly reduced for cells seeded on these treated tissues after 24 h of direct contact. A significant cytotoxicity was found for frozen treated bones in comparison to freeze-dried treated bones. Tissue remodeling with graft stability, no harmful inflammation, and neo-vascularization was observed for freeze-dried chemically treated bones and tendons. Frozen-treated bones were characterized by a lack of matrix recolonization at 4 weeks postimplantation. In conclusion, chemical processing with freeze-drying of human tissues maintains in vitro biocompatibility and in vivo tissue remodeling for clinical application.     

In vitro and in vivo biological performance of collagen-chitosan/silicone membrane bilayer dermal equivalent

Skin loss or damage affects severely the life quality of human being and can even cause death in many cases. We report here a bilayer dermal equivalent (BDE) composed of collagen-chitosan porous scaffold and silicone membrane, which can effectively induce the regeneration of dermis in an animal model of full thickness skin loss. The in vitro biosecurity test showed that the BDE had no cytotoxicity, and no remarkable sensitization and irritability. In vitro cell culture proved that the BDE had good biocompatibility to support the proliferation of fibroblasts. Animal test was performed on Bama miniature pig skin. Gross view and histological sections found plenty of fibroblasts and extracellular matrix in the regenerative scaffold after transplantation of the BDE for 4 weeks. Immunohistochemistry results proved that the BDE has the ability to support the angiogenesis of the regenerated dermis. All these results indicate that the BDE might be a promising equivalent in treating dermal loss.     

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.     

Fabrication and properties of novel composites in the system Al–Zr–C

Composite bodies in the system Al–Zr–C, with about 95% relative density, were obtained by heating the compact body of powder mixture consisting of Al and ZrC (5 : 1 mol %) in Ar at 1100–1500°C for various lengths of time. Components of the material heated at more than 1200°C were Al, Al3Zr, ZrC and AlZrC2. The Al3Zr exhibited plate-like aggregation, and its size increased with increasing temperature. In the material heated at 1500°C for 1 h, the largest plate-like Al3Zr aggregation was 2000 m long and 133 m thick. Then the AlZrC2 was present as well-proportioned hexagonal platelet particles with a 8–9 m diameter and a 1–2 m thickness in the interior of the plate-like Al3Zr aggregation and Al matrix phase. The average three-point bending strength of the bodies was 140–190 MPa, and the maximum strength was 203 MPa in the body heated at 1300°C for 1 h. The body heated at 1500°C for 1 h showed high oxidation resistivity to air up to 1000°C.     

In vivo degradation of poly(DTE carbonate) membranes. Analysis of the tissue reactions and mechanical properties

Different bioabsorbable polymers and their co-polymers have been used to construct an optimal material for guided bone regeneration applications. Our aim was to evaluate a novel bioabsorbable material in a soft tissue environment. In this study, a poly(DTE carbonate) membrane (0.2–0.3 mm) was implanted into 20 NZW rabbits’ subcutaneous pouches for 6, 12, 24 and 52 weeks. The material was evaluated by means of histological reactions to the material and mechanical properties of the membrane. Based on this study, it can be concluded that poly(DTE carbonate) elicited a very modest foreign body reaction in the soft tissues. This reaction was uniform throughout the study. Varying amounts of calcification was seen in the fibrous capsule surrounding the implant. The number of calcified bodies did not correlate to healing time.     

In vitro and in vivo sustained release of exenatide from vesicular phospholipid gels for type II diabetes

Diabetes is a chronic disease that requires daily treatment to maintain a stable blood glucose level. Sustained-release formulations can thus benefit the treatment of diabetes by reducing the repeated administration of therapeutics. Our study aimed to develop a sustained-release platform for exenatide that is biocompatible and capable of mass production. Vesicular phospholipid gels (VPGs) are semisolid phospholipid dispersions with controlled release profiles. Exenatide-VPGs prepared via simple magnetic stirring showed excellent biocompatibility with an average particle size of about 15?μm after redispersion. VPGs were shown to achieve sustained release for up to 21 days in vitro with no obvious burst effect. The in vivo release study showed that VPGs sustained the release of the exenatide for up to 11 days. Moreover, after subcutaneous injection of the exenatide-VPGs in the diabetic rats, the hypoglycemic effect lasted for 10 days compared with exenatide solution. In sum, the exenatide-VPGs system represents a promising sustained-release formulation for exenatide with a long-acting therapeutic efficacy in vivo.     

Biocompatibility evaluation of a novel hydroxyapatite-polymer coating for medical implants (in vitro tests)

Nanocomposites consisting of hydroxyapatite (HA) and a sodium maleate copolymer (maleic polyelectrolyte), synthesized by hydrothermal method and deposited on titanium substrates by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique were tested for the biological properties. Coating bioanalysis was carried out by triple staining of actin, microtubules and nuclei followed by immunofluorescence microscopy. Within 24 h cells that occupied the biomaterial surface displayed the morphology and cytoskeleton pattern similar to the controls. Cells grown on nanocomposite coated surfaces had a higher proliferation rate than their counterparts grown on Ti coated with HA alone, indicating that maleic polyelectrolyte improved surface bio-adhesive characteristics. The capacity to induce cell attachment, spreading and proliferation demonstrated the potential of Ti coated with HA-polymer nanocomposites to be used as scaffolds in dental or orthopedic implantology.     

Preparation and evaluation of a timolol maleate drug–resin ophthalmic suspension as a sustained-release formulation in vitro and in vivo

Abstract

The aim of this work was to assess the performance of resin as an ocular delivery system. Timolol maleate (TM) was chosen as the model drug and an ion exchange resin (IER) as the carrier. The drug–resin complex was prepared using an oscillation method and then characterized regarding particle size, zeta potential, morphology, and drug content. After in vitro drug release study and corneal permeation study were performed, in vivo studies were performed in New Zealand albino rabbits using a suspension with particles sized 4.8?±?1.2?μm and drug loading at 43.00?±?0.09 %. The results indicate that drug released from the drug–resin ophthalmic suspension permeated the cornea and displayed a sustained-release behavior. Drug levels in the ocular tissues after administration of the drug–resin ophthalmic suspension were significantly higher than after treatment with an eye drop formulation but were lower in body tissues and in the plasma. In conclusion, resins have great potential as effective ocular drug delivery carriers to increase ocular bioavailability of timolol while simultaneously reducing systemic drug absorption.     

Microstructures, mechanical properties and in vitro corrosion behaviour of biodegradable Mg–Zr–Ca alloys

The microstructures, mechanical properties, corrosion behaviour and biocompatibility of the Mg–Zr–Ca alloys have been investigated for potential use in orthopaedic applications. The microstructures of the alloys were examined using X-ray diffraction analysis, optical microscopy and scanning electron microscopy. The mechanical properties of Mg–Zr–Ca alloys were determined from compressive tests. The corrosion behaviour has been investigated using an immersion test and electrochemical measurement. The biocompatibility was evaluated by cell growth factor using osteoblast-like SaOS2 cell. The experimental results indicate that the hot-rolled Mg–Zr–Ca alloys exhibit much finer microstructures than the as-cast Mg–Zr–Ca alloys which show coarse microstructures. The compressive strength of the hot-rolled alloys is much higher than that of the as-cast alloys and the human bone, which would offer appropriate mechanical properties for orthopaedic applications. The corrosion resistance of the alloys can be enhanced significantly by hot-rolling process. Hot-rolled Mg–0.5Zr–1Ca alloy (wt %) exhibits the lowest corrosion rate among all alloys studied in this paper. The hot-rolled Mg–0.5Zr–1Ca and Mg–1Zr–1Ca alloys exhibit better biocompatibility than other studied alloys and possess advanced mechanical properties, corrosion resistance and biocompatibility, suggesting that they have a great potential to be good candidates for orthopaedic applications.     

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.     

In vivo and in vitro response to electrochemically anodized Ti-6Al-4V alloy

Tissues’ reactions to metals depend on a variety of properties of the metal, most notably surface structure. Anodizing has been shown to alter the surface properties of metal, thus eliciting a change in the biocompatibility of the metal. In order to evaluate the biocompatibility of unoxidized titanium alloy (Ti-6Al-4V) and anodized titanium alloy samples, the samples were implanted in murine abdominal subcutaneous tissues, and maintained for 2 and 4 weeks. The reaction of the abdominal subcutaneous connective tissues to the samples was then assessed. Fibrous connective tissue capsules were observed around the vicinity of the sample, and these capsules were shown to harbor fibroblasts, fibrocytes, and other cells, including neutrophils, macrophages, and giant multinucleated cells. The average thickness of the fibrous capsules observed around the anodized alloy samples was less than that of the capsules seen around samples of the unoxidized titanium alloy. Blood was obtained from the tails of the experimental mice, and blood cell analyses were conducted in order to assess the levels of leukocytes, red blood cells, and thrombocytes. The blood analysis results of the unoxidized control group and treatment group were all within normal ranges. In addition, the biocompatibility of the titanium alloy samples was evaluated using cell culture techniques. The numbers of MG-63 cells cultured on oxidized samples tended to be greater than those in the controls; however, these increases were not statistically significant. The alkaline phosphatase activity of the sample oxidized at 310 V evidenced significantly higher activity than was observed in the control group. These results indicate that the anodized Ti-6Al-4V alloy will be of considerable utility in biomedical applications.