Enhancing Oil Solubility of BCS Class II Drug Phenytoin Through Hydrophobic Ion Pairing to Enable High Drug Load in Injectable Nanoemulsion to Prevent Precipitation at Physiological pH With a Potential to Prevent Phlebitis.

This work investigates the micellar titration of phenytoin (a weakly acidic drug) with cetyltrimethylammonium hydroxide (CTAH) to form a hydrophobic ion-pair to enhance oil solubility of phenytoin, followed by an effort to formulate nanoemulsion that could potentially prevent precipitation of phenytoin at physiological pH. The ion-pair formulated in nanoemulsion was evaluated for in vitro precipitation during serial dilution at physiological pH. The formation of ion-pair during titration was explained in context of pH-solubility data. The mathematical model successfully integrated ionization and micellization equilibria to reflect on dominant mechanisms for solubilization. The micellar phenomenon during titration was confirmed using Dynamic Light Scattering (DLS). The phase changes of the excess undissolved solids during titration were evident from X-Ray Powder Diffraction (XRPD) and Fourier Transform Infrared Spectroscopy (FTIR). This analysis confirmed the conversion of phenytoin into ionized state and its subsequent ionic interaction with CTAH forming hydrophobic ion-pair complex (HIP). The complete ion pair formation was evident at pHmax (8.8 to 9.2), and its 1:1 stoichiometry was confirmed using HPLC (Phenytoin and CTAH) and H1 NMR, hence could also be called as a lipophilic salt. The ion-pair (salt) was insoluble in water and showed remarkably high partition coefficient (log P) in octanol/water. As characterized by Hot Stage Microscopy (HSM), the melting point of the ion-pair complex was lowered to 150.8°C compared to the free acid (> 300οC), this was even further lowered to 81.1 °C when evaluated in castor oil. This led to approximately eight-fold higher solubility of hydrophobic ion pair (HIP) in castor oil compared to the free acid form. The high miscibility in castor oil was suitable to formulate a high drug load injectable dispersed system. This was successfully achieved with lecithin and polysorbate as emulsifiers without leaching drug into continuous phase at pH 7.4. This nanoemulsion (<300 nm, and > +30 mV zeta potential) remain stable when evaluated over a period of one month. A serial dilution study of the nanoemulsion was performed in PBS buffer, microscopic observations suggested no birefringence despite incubation at 25°C for several hours. This result indicated that Phenytoin remained strongly partitioned within dispersed oily phase with a higher drug loading when ion-paired phenytoin was used. The higher drug load could enable a small volume slow bolus injection to meet 50 mg/min or lower delivery rate criteria for Phenytoin in the clinical set up. This provided a pathway to further explore potential injectable nano-emulsion formulations that could alleviate typical phlebitis issue associated with the injectable phenytoin solution administration at physiological pH.

Single dose topical inserts containing tenofovir alafenamide fumarate and elvitegravir provide pre- and post-exposure protection against vaginal SHIV infection in macaques.

BACKGROUND: Vaginal products for HIV prevention that can be used on-demand before or after sex may be a preferable option for women with low frequency or unplanned sexual activity or who prefer not to use daily or long-acting pre-exposure prophylaxis (PrEP). We performed dose ranging pharmacokinetics (PK) and efficacy studies of a vaginally applied insert containing tenofovir alafenamide fumarate (TAF) and elvitegravir (EVG) in macaques under PrEP or post-exposure prophylaxis (PEP) modalities. METHODS: PK studies were performed in 3 groups of pigtailed macaques receiving inserts with different fixed-dose combinations of TAF and EVG (10/8, 20/16 and 40/24 mg). PrEP and PEP efficacy of a selected insert was investigated in a repeat exposure vaginal SHIV transmission model. Inserts were administered 4 h before (n = 6) or after (n = 6) repeated weekly SHIV exposures. Infection outcome was compared with macaques receiving placebo inserts (n = 12). FINDINGS: Dose ranging studies showed rapid and sustained high drug concentrations in vaginal fluids and tissues across insert formulations with minimal dose proportionality. TAF/EVG (20/16 mg) inserts were selected for efficacy evaluation. Five of the 6 animals receiving these inserts 4 h before and 6/6 animals receiving inserts 4 h after SHIV exposure were protected after 13 challenges (p = 0.0088 and 0.0077 compared to placebo, respectively). The calculated PrEP and PEP efficacy was 91.0% (95% CI = 32.2%-98.8%) and 100% (95% CI = undefined), respectively. INTERPRETATION: Inserts containing TAF/EVG provided high protection against vaginal SHIV infection when administered within a 4 h window before or after SHIV exposure. Our results support the clinical development of TAF/EVG inserts for on-demand PrEP and PEP in women. FUNDING: Funded by CDC intramural funds, an interagency agreement between CDC and USAID (USAID/CDC IAA AID-GH-T-15-00002), and by the U.S. President's Emergency Plan for AIDS Relief (PEPFAR) through the U.S. Agency for International Development (USAID) under a Cooperative Agreement (AID-OAA-A-14-00010) with CONRAD/Eastern Virginia Medical School.

Levonorgestrel loaded biodegradable microparticles for injectable contraception: Preparation, characterization and modelling of drug release.

The integration of mechanistic modeling and machine learning facilitates the understanding and engineering of drug release from controlled release systems. Here, we present hybrid models to predict the effect of drug loading on levonorgestrel release from spray-dried poly(L-lactic acid) microparticles. We developed three Monte Carlo methods that differ in the consideration of polymer's degradability and crystallinity, to simulate drug release from the matrices using the Python programming language. To build each method, we utilized data from the characterization of the particles, such as the actual drug content (ranges from 6% to 52%), size (Dv(50) âˆ¼ 5 µm), and polymer crystallinity (ranges from 0% to 15%). We trained each method using drug release data from particles of 4 batches and derived appropriate machine learning models through regression analysis. Results indicate the contribution of drug diffusion and polymer degradation to drug release for particles of lower drug content (<20 %w/w). At higher drug loadings, particles encountered a combination of burst and diffusional release. We validated the predictive powers of the machine learning models by testing them against experimental data. This paper specifically highlights the power of hybrid modeling to engineer drug release for long-term contraception.

Use of Flory-Huggins Interaction Parameter and Contact Angle Values to Predict the Suitability of the Drug-Polymer System for the Production and Stability of Nanosuspensions.

PURPOSE: Use of Flory-Huggins interaction parameter and contact angle values to predict the suitability of the drug-polymer system for the production and stability of nanosuspensions. MATERIAL AND METHODS: Melting point depression of the drug was measured using differential scanning calorimetry. Interaction parameter, χ, was calculated using the melting point depression data to elucidate the drug-polymer interaction strength to predict the suitability of the drug-polymer system for the production and stability of nanosuspensions. Contact angle of the drug films were measured with purified water and 0.1%w/w polymer solutions to predict polymer's suitability for the production and stability of nanosuspension. Nanosuspensions were manufactured to validate the application of the melting point depression approach along with surface property information. RESULTS: All three polymers, HPMC, Soluplus®, and poloxamer exhibited a negative interaction parameter with naproxen and budesonide. Higher negative interaction parameter values for the naproxen-polymer system indicated stronger drug-polymer interactions, while smaller negative interaction parameter values for the budesonide-polymer system indicated weaker drug-polymer interactions. Interaction parameter was not obtained for fenofibrate with HPMC and Soluplus®, and similarly, no interaction parameter was obtained for carvedilol with HPMC, most likely due to weaker drug-polymer interactions. All three polymers provided lower equilibrium contact angle values when compared to purified water, indicating an affinity for polymers. CONCLUSIONS: Successful production and stability of several nanosuspensions were correlated with Flory-Huggins's interaction parameter and contact angle values. In the absence of melting point depression, contact angle values can also be used predict the agglomeration tendencies as we have shown for this study.

Design and characterization of 3D printed, neomycin-eluting poly-L-lactide mats for wound-healing applications.

This study evaluates the suitability of 3D printed biodegradable mats to load and deliver the topical antibiotic, neomycin, for up to 3 weeks in vitro. A 3D printer equipped with a hot melt extruder was used to print bandage-like wound coverings with porous sizes appropriate for cellular attachment and viability. The semicrystalline polyester, poly-l-lactic acid (PLLA) was used as the base polymer, coated (post-printing) with polyethylene glycols (PEGs) of MWs 400 Da, 6 kDa, or 20 kDa to enable manipulation of physicochemical and biological properties to suit intended applications. The mats were further loaded with a topical antibiotic (neomycin sulfate), and cumulative drug-release monitored for 3 weeks in vitro. Microscopic imaging as well as Scanning Electron Microscopy (SEM) studies showed pore dimensions of 100 × 400 µm. These pore dimensions were achieved without compromising mechanical strength; because of the "tough" individual fibers constituting the mat (Young's Moduli of 50 ± 20 MPa and Elastic Elongation of 10 ± 5%). The in vitro dissolution study showed first-order release kinetics for neomycin during the first 20 h, followed by diffusion-controlled (Fickian) release for the remaining duration of the study. The release of neomycin suggested that the ability to load neomycin on to PLLA mats increases threefold, as the MW of the applied PEG coating is lowered from 20 kDa to 400 Da. Overall, this study demonstrates a successful approach to using a 3D printer to prepare porous degradable mats for antibiotic delivery with potential applications to dermal regeneration and tissue engineering. Illustration of the process used to create and characterize 3D printed PLLA mats.

Advances in controlled release hormonal technologies for contraception: A review of existing devices, underlying mechanisms, and future directions.

This article presents a comprehensive review on controlled release hormonal contraceptive systems that include transdermal patches, intravaginal rings (IVRs), intrauterine devices (IUDs), injectables and subdermal implants. These systems represent a substantial advance from traditional oral contraceptive pills, to improve upon safety, efficiency, and compliance among women. The widespread use of controlled release systems is hindered by limitations, which are discussed in this review. Biodegradable polymers such as poly (lactic-co-glycolic acid) and polycaprolactone have been used to formulate subdermal implants and injectable microspheres to eliminate the need for implant removal and reduce provider intervention. To address low permeability in transdermal patches, permeation enhancers such as alkanols, fatty acids, prodrugs, and vesicular delivery for steroids have been investigated. Local anesthetics in the form of creams, gels and sprays have been evaluated to alleviate the pain associated with IUD insertion. Among methods for device fabrication, 3D printing has emerged as a potential approach for fabricating customizable IVRs and IUDs. Several other modified delivery systems such as transdermal microneedle patches, in situ forming injectable implants, microspheres embedded in implants and IVRs addressing multiple clinical conditions have been investigated for controlled release of contraceptive hormones. To ensure drug release at zero-order rates, empirical and theoretical modelling have been extensively employed and evaluated. The limitations of low predictive power associated with empirical modelling may be overcome through theoretical modelling and simulation that consider underlying mechanisms. Newer approaches such as Monte Carlo based simulation and deep learning models based on artificial neural networks can prove highly beneficial in developing precise contraceptive delivery system, to enhance the quality of life for women worldwide.

Lipid Nanoparticles Improve the Uptake of α-Asarone Into the Brain Parenchyma: Formulation, Characterization, In Vivo Pharmacokinetics, and Brain Delivery.

Treatment of brain-related diseases is one of the most strenuous challenges in drug delivery research due to numerous hurdles, including poor blood-brain barrier penetration, lack of specificity, and severe systemic toxicities. Our research primarily focuses on the delivery of natural therapeutic compound, α-asarone, for the treatment of brain-related diseases. However, α-asarone has poor aqueous solubility, bioavailability, and stability, all of which are critical issues that need to be addressed. This study aims at formulating a lipid nanoparticulate system of α-asarone (A-LNPs) that could be used as a brain drug delivery system. The physicochemical, solid-state properties, stability, and in vitro and in vivo studies of the A-LNPs were characterized. The release of α-asarone from the A-LNPs was prolonged and sustained. After intravenous administration of A-LNPs or free α-asarone, significantly higher levels of α-asarone from the A-LNPs were detected in murine plasma and brain parenchyma fractions, confirming the ability of A-LNPs to not only maintain a therapeutic concentration of α-asarone in the plasma, but also transport α-asarone across the blood-brain barrier. These findings confirm that lipid nanoparticulate systems enable penetration of natural therapeutic compound α-asarone through the blood-brain barrier and may be a candidate for the treatment of brain-related diseases.

Adaptive Hybrid Molecular Brushes Composed of Chitosan, Polylactide, and Poly(N-vinyl pyrrolidone) for Support and Guiding Human Dermal Fibroblasts.

Hybrid molecular brushes (HMBs) are macromolecular constructs made up of a backbone polymer and side-chain polymers with distinct properties. They adapt to a changing microenvironment via the conformational mechanism and thus may affect mammalian cell proliferation. Two biobenign HMBs were synthesized in this work: (1) polylactide (PLA) grafted to the chitosan (CHI) backbone to form chitosan-graft-polylactide (CHI-g-PLA), a two-component molecular brush, and (2) poly(N-vinyl pyrrolidone) (PNVP) grafted to chitosan moieties of CHI-g-PLA to form a three-component HMB. The molecular brushes were used to fabricate polymer coatings and nanofibers, guiding the attachment and growth of human dermal fibroblasts (HDFs) while silencing the response of macrophages to the scaffolds. The exterior surface composition of the coatings can be switched by exposure to solvents of different polarities: hydrophilic PNVP chains upon exposure to water or hydrophobic PLA chains upon treatment by anisole. Our experiments demonstrate substantial improvement of the HDF cell attachment and proliferation on the surface of the HMBs as compared to the parent polymers CHI, PLA, and PNVP. A Sirius Red assay and immunofluorescence show that HMBs stimulate production of collagen by HDF cells, which propagate on the polymer substrates revealing well-developed focal adhesion structures. On the other hand, a low attachment of macrophages is observed on the HMB surfaces, in particular if HMBs are switched to the hydrophilic state, i.e., PNVP in the top strata.

Determinants of zero-order release kinetics from acetaminophen-layered Suglet® pellets, Wurster-coated with plasticized Aquacoat® ECD (ethyl cellulose dispersion).

The potential for zero-order drug-release was evaluated for ethyl cellulose (EC) coated, acetaminophen-layered sugar pellets (Suglets®) of mesh size 18/20 (850-1000 µm). To determine optimal plasticizer/pore-former concentrations for EC films, solvent-cast Aquacoat® ECD (ethyl cellulose dispersion) films were prepared with 0-50% w/w ratios of two triethyl citrate (TEC) or triacetin (TA). Characterization studies showed that films with excipient concentrations ≥20% were homogenous, mechanically strong at room temperature (Young's modulus of 25-35 MPa), and have a glass transition (Tg) in the range of 20-45 °C. Based on these results, a working range of 20-50% weight concentrations was selected for drug release studies. Suglets® were layered with acetaminophen (APAP) using Wurster Glatt GPCG-3 to yield roughly 10% w/w coating (controls). The Controls were coated using the same Wurster process with Aquacoat® ECD containing 20-50% w/w of TEC or TA. Samples were removed periodically at 3-11% weight gain, to evaluate impact of weight gain, and consequently film-formation, on drug release. Dissolution was monitored over a period of 12 h in a media consisting of simulated gastric fluid (first two hours), followed by simulated intestinal fluid. The controls showed near 100% release within the first 30 min, indicating the value of EC-coating to achieve controlled release. Dissolution release profiles showed that TEC is more effective than TA as a plasticizer and pore-former, as linear profiles were apparent at lower concentrations and % weight gain. For a quantitative evaluation of these results, linear regression was fitted to all cumulative release profiles, and R-squared values examined as a function of excipient concentration and % weight gain. The corresponding response surface plots and the second order regression were shown to aid in optimization. The design space for zero-order release was represented as contour plots between excipient concentration and % weight gain.

Advances in bioprinting using additive manufacturing.

Since its conception in the 1980's, several advances in the field of additive manufacturing have led to exploration of alternate as well as combination biomaterials. These progresses have directed the use of 3D printing in wider applications such as printing of dermal layers, cartilage, bone defects, and surgical implants. Furthermore, the incorporation of live and functional cells with or atop biomaterials has laid the foundation for its use in tissue engineering. The purpose of this review is to summarize the advances in 3D printing and bioprinting of several types of tissues such as skin, cartilage, bones, and cardiac valves. This review will address the current 3D technologies used in tissue construction and study the biomaterials being investigated. There are several requirements that need to be addressed, in order to reconstruct functional tissue such as mechanical strength, porosity of the replicate and cellular incorporation. Researchers have focused their studies to answer questions regarding these requirements.

Design of hybrid molecular brushes with reversible surface adaptability on exposure to specific solvents.

Hybrid molecular brushes (HMBs) are macromolecules made of a linear backbone and polymeric side chains that differ in their chemical nature. The authors developed a new method of synthesis of HMB with chitosan (CHI) backbone. In the first step, chitosan-graft-polylactide (CHI-g-PLA) was synthesized by interfacial ring opening polymerization of lactide initiated from CHI. CHI-g-PLA is characterized for its molecular weight and structure. In the second step, polyvinylpyrrolidone (PNVP) or polyacrylamide (PAAm) is grafted by radical polymerization from the CHI in CHI-g-PLA to form CHI-g-PLA-g-PNVP and CHI-g-PLA-g-PAAm. This results in the formation of HMB, with hydrophobic PLA and hydrophilic PNVP or PAAm side chains grafted to CHI. The chemical structure and thermal behavior of the HMBs are characterized. The morphology of CHI-g-PLA as well as the HMBs is determined using atomic force microscopy (AFM). Both the HMBs tethered to separate surfaces exhibit reversible switching between the hydrophilic and hydrophobic polymers on exposure to specific solvents. This is studied by AFM and water contact angle measurements. Hence, the authors developed a method for synthesis of HMB that can be applied for surface modification.

Physicochemical characterization of spray-dried PLGA/PEG microspheres, and preliminary assessment of biological response.

CONTEXT: The use of spray-drying to prepare blended PLGA:PEG microspheres with lower immune detection. OBJECTIVE: To study physical properties, polymer miscibility and alveolar macrophage response for blended PLGA:PEG microspheres prepared by a laboratory-scale spray-drying process. METHODS: Microspheres were prepared by spray-drying 0-20% w/w ratios of PLGA 65:35 and PEG 3350 in dichloromethane. Particle size and morphology was studied using scanning electron microscopy. Polymer miscibility and residual solvent levels evaluated by thermal analysis (differential scanning calorimetry - DSC and thermogravimetric analysis - TGA). Immunogenicity was assessed in vitro by response of rat alveolar macrophages (NR8383) by the MTT-based cell viability assay and reactive oxygen species (ROS) detection. RESULTS: The spray dried particles were spherical, with a size range of about 2-3 µm and a yield of 16-60%. Highest yield was obtained at 1% PEG concentration. Thermal analysis showed a melting peak at 59 °C (enthalpy: 170.61 J/g) and a degradation-onset of 180 °C for PEG 3350. PLGA 65:35 was amorphous, with a Tg of 43 °C. Blended PLGA:PEG microspheres showed a delayed degradation-onset of 280 °C, and PEG enthalpy-loss corresponding to 15% miscibility of PEG in PLGA. NR8383 viability studies and ROS detection upon exposure to these cells suggested that blended PLGA:PEG microspheres containing 1 and 5% PEG are optimal in controling cell proliferation and activation. CONCLUSION: This research establishes the feasibility of using a spray-drying process to prepare spherical particles (2-3 µm) of molecularly-blended PLGA 65:35 and PEG 3350. A PEG concentration of 1-5% was optimal to maximize process yield, with minimal potential for immune detection.

Enhanced osteogenic potential of human mesenchymal stem cells on electrospun nanofibrous scaffolds prepared from eri-tasar silk fibroin.

This study evaluated the mechanical properties and osteogenic potential of a silk fibroin scaffold prepared from a 70:30 blend of Eri (Philosamia ricini) and Tasar (Antheraea mylitta) silk, respectively (ET scaffolds). An electrospinning process was used to prepare uniformly blended, fibrous scaffolds of nanoscale dimensions, as confirmed by scanning and transmission electron microscopy (fiber diameter < 300 nm). Similarly prepared scaffolds derived from gelatin and Bombyx mori (BM) silk fibroin were used as controls. Mechanical testing and atomic force microscopy showed that the ET scaffolds had significantly higher tensile strength (1.83 ± 0.13 MPa) and surface roughness (0.44 µm) compared with BM (1.47 ± 0.10 MPa; 0.37 µm) and gelatin scaffolds (0.6 ± 0.07 MPa; 0.28 µm). All scaffolds were exposed to mesenchymal stem cells isolated to human chord blood (hMSCs) for up to 28 days in vitro. Alamar blue and alkaline phosphatase assay showed greater attachment and proliferation for both ET and BM scaffolds compared with gelatin. The ET scaffolds also promoted greater differentiation of the attached hMSCs as evidenced by higher expression of RunX2, osteocalcin, and CD29/CD44 expression. ET scaffolds also showed significantly higher mineralization, as evidenced by glycosaminoglycan assay, alizarin red staining, and elemental analysis of crystalline composites isolated from the scaffolds.

A molecular dynamics approach for predicting the glass transition temperature and plasticization effect in amorphous pharmaceuticals.

The objectives of this study were as follows: (i) To develop an in silico technique, based on molecular dynamics (MD) simulations, to predict glass transition temperatures (Tg) of amorphous pharmaceuticals. (ii) To computationally study the effect of plasticizer on Tg. (iii) To investigate the intermolecular interactions using radial distribution function (RDF). Amorphous sucrose and water were selected as the model compound and plasticizer, respectively. MD simulations were performed using COMPASS force field and isothermal-isobaric ensembles. The specific volumes of amorphous cells were computed in the temperature range of 440-265 K. The characteristic "kink" observed in volume-temperature curves, in conjunction with regression analysis, defined the Tg. The MD computed Tg values were 367 K, 352 K and 343 K for amorphous sucrose containing 0%, 3% and 5% w/w water, respectively. The MD technique thus effectively simulated the plasticization effect of water; and the corresponding Tg values were in reasonable agreement with theoretical models and literature reports. The RDF measurements revealed strong hydrogen bond interactions between sucrose hydroxyl oxygens and water oxygen. Steric effects led to weak interactions between sucrose acetal oxygens and water oxygen. MD is thus a powerful predictive tool for probing temperature and water effects on the stability of amorphous systems during drug development.

Development and evaluation of cross-linked collagen-hydroxyapatite scaffolds for tissue engineering.

This study examines the tissue engineering potential of type I collagen cross-linked in the presence of hydroxyapatite (HAp). Scaffolds were prepared by controlled freezing followed by lyophilization of composite mixtures of collagen and HAp in acetic acid, followed by cross-linking with 0.3% glutaraldehyde. Scaffolds of three ratios were prepared, corresponding to collagen/HAp ratios of 1:2, 1:4, and 1:6. The scaffolds were evaluated for their microstructure, chemical and physical properties, swelling behavior, mechanical strength, biodegradability hemocompatability, cytocompatibility, and histopathology following subcutaneous implantation in Sprague Dawley rats. The collagen/HAp matrices showed a smaller pore size of 10-40 µm compared to 50-100 µm for pure collagen scaffolds. Pure collagen showed a mechanical strength of 0.25 MPa, and the value almost doubled for cross-linked composites with collagen/HAp ratio 1:6. The improvement in mechanical strength corresponded to a decrease in swelling and enzymatic degradation (measured by resistance to collagenases). FTIR spectra results in conjunction with scanning electron micrographs showed that cross-linking in the presence of HAp did not significantly alter the structure of collagen. MTT assay and calcein AM staining revealed prominent and healthy growth of mesenchymal stem cells in both the pure collagen as well as collagen:HAp composites of ratio 1:2. In vivo implantation in Sprague Dawley rats showed an initial acute inflammatory response during days 3 and 7, followed by a chronic, macrophage-mediated inflammatory response on days 14 and 28. Overall, a cross-linked collagen/HAp composite scaffold of ratio 1:2 was identified as having potential for further development in tissue engineering.

Preparation and evaluation of sustained release infliximab microspheres.

A sustained release microsphere system for an antibody (infliximab, molecular weight: 140 Kd) was formulated with PLGA 50:50 co-polymer using two methods of preparation: phase separation technique and double emulsion technique. Microspheres were made in triplicate using each technique and varying drug-to-polymer ratios. Drug-to-polymer ratio was maintained at 1:5, 1:10, or 1:20. In vitro release profile of infliximab was studied in phosphate-buffered saline solution at 37 °C. The release profile and encapsulation efficiency was compared between the two methods of preparation. The releases data was modeled by the sum of squares method to isolate the dominant release mechanism. The physical attributes of the microspheres prepared were characterized. The biochemical characteristics of infliximab before and after encapsulation were also evaluated by several analytical techniques such as size exclusion chromatography, isoelectric focusing, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Biological activity of infliximab was also evaluated before and after encapsulation. The phase separation technique showed much higher entrapment efficiency than the double emulsion technique. Microspheres prepared using the double emulsion technique showed a longer release profile (∼5 days) compared to microspheres prepared using the phase separation technique (∼72 h). Physical and biochemical properties of infliximab did not change significantly after encapsulation into microspheres with either methods of preparation. Microspheres prepared using phase separation showed some loss in bioactivity. Based on this research it can be concluded that the microspheres can present an alternative delivery method for infliximab. LAY ABSTRACT: A sustained release microsphere system for an antibody (infliximab, molecular weight: 140 Kd) was formulated using polymers. This antibody is currently in the market for rheumatoid arthritis among various indications. Microspheres were made in triplicate using two techniques and varying drug-to-polymer ratios. Drug-to-polymer ratio was maintained at 1:5, 1:10, or 1:20. The release of drug from microsphere was studied. Biochemical properties of microspheres were also studied before and after encapsulation in microspheres. Various analytical techniques were used to study the biochemical properties of infliximab to ensure that it would be efficacious and safe after encapsulation. Sustained release of drug was observed from the microspheres. Infliximab showed no change in biochemical properties and also showed bioactivity. Based on this research it can be concluded that the microspheres can present an alternative delivery method for infliximab that is safe and efficacious and my result in cost savings for patients.

On-column refolding of bone morphogenetic protein-2 using cation exchange resin.

Refolding is often the bottle-neck step in producing recombinant proteins from inclusion bodies of Escherichia coli, especially for dimer proteins. The refolding process is protein specific, engaging a lot of time and cost to optimize conditions so that the thermodynamics favor protein refolding over competitive aggregation. Bone morphogenetic protein-2 (BMP-2) is a potent osteogenic agent having significant applications in bone regeneration therapy. In this study, we present a novel solid-phase refolding method for rapid and efficient refolding of recombinant BMP-2 dimer from E. coli. We employed a weak cation exchange resin as the adsorbing support, with decreasing gradient of denaturing agent and exposure to oxidizing conditions for adequate disulfide bond formation. Refolded BMP-2 was further purified using size exclusion chromatography and analyzed for its secondary structure and biological activity. The purified BMP-2 dimer showed dose-dependent induction of alkaline phosphatase (ALP) activity in MC3T3 pre-osteoblast cells, thus translating the success of our refolding method. This simple and rapid method can also be applied in refolding and purification of other BMP-2 like dimer proteins.

Prediction of solubility parameters and miscibility of pharmaceutical compounds by molecular dynamics simulations.

The objectives of this study were (i) to develop a computational model based on molecular dynamics technique to predict the miscibility of indomethacin in carriers (polyethylene oxide, glucose, and sucrose) and (ii) to experimentally verify the in silico predictions by characterizing the drug-carrier mixtures using thermoanalytical techniques. Molecular dynamics (MD) simulations were performed using the COMPASS force field, and the cohesive energy density and the solubility parameters were determined for the model compounds. The magnitude of difference in the solubility parameters of drug and carrier is indicative of their miscibility. The MD simulations predicted indomethacin to be miscible with polyethylene oxide and to be borderline miscible with sucrose and immiscible with glucose. The solubility parameter values obtained using the MD simulations values were in reasonable agreement with those calculated using group contribution methods. Differential scanning calorimetry showed melting point depression of polyethylene oxide with increasing levels of indomethacin accompanied by peak broadening, confirming miscibility. In contrast, thermal analysis of blends of indomethacin with sucrose and glucose verified general immiscibility. The findings demonstrate that molecular modeling is a powerful technique for determining the solubility parameters and predicting miscibility of pharmaceutical compounds.

The design of flexible ciprofloxacin-loaded PLGA implants using a reversed phase separation/coacervation method.

The purpose of this research is to design and characterize flexible PLGA-based implants for the controlled release of ciprofloxacin hydrochloride for up to 6 weeks in vitro. This research uses a reversed phase separation/coacervation method to fabricate flexible PLA and PLGA: excipient implants with dichloromethane/mineral oil as solvent/non-solvent. Physical characterization was performed using thermal and mechanical analyses. Drug loading and release studies were performed with ciprofloxacin HCl as the model drug. Release kinetics was modeled to elucidate possible mechanisms of drug release. Four polymer-excipient combinations with glass transition temperatures less than 20°C and representing a wide range of Young's moduli were shown to entrap up to 8% of ciprofloxacin HCl that could be released at a controlled rate for 65 days in vitro. The release rate could consistently fit a ternary Gaussian pattern with an R(2)>0.99. It was postulated that these release patterns could be related to ciprofloxacin that was loosely or poorly bound (burst release), trapped within the polymer matrix, or encapsulated by the polymer. These studies show that flexible implants can be fabricated from PLGA-based polymers for the controlled release of ciprofloxacin hydrochloride for up to 6 weeks in vitro.

Novel poly-DL-lactide-polycaprolactone copolymer based flexible drug delivery system for sustained release of ciprofloxacin.

This research evaluated 7525DLPCL for soft flexible drug delivery systems. The effect of ciprofloxacin hydrochloride (CIP) loading at three levels (10, 20, and 30%), on thermo-mechanical properties was studied. CIP release was monitored for 12 weeks. Addition of CIP to 7525DLPCL caused an increase in compressive modulus of 7525DLPCL. CIP release was found to be sigmoidal with two phenomena (apart from a minor burst) contributing to release-diffusion and later diffusion plus erosion. An increased burst was observed with greater CIP loading and the majority of CIP (> 70%) was released as an effect of diffusion plus erosion. Additional factors, like the effect of CIP particle size, had no significant effect on drug release. Change in the implant shape from a cylinder (5 mm diameter; 3 mm thickness) to disc (6 mm diameter, 0.5 mm thickness) also failed to show a significant impact on drug release. Erosion of 7525DLPCL is a major contributing factor towards this release and other factors like shape of implants and particle size of drug have little effect on CIP release. Such flexible drug delivery systems offer new avenues for long-term skeletal drug delivery of antibiotics for conditions like osteomyelitis or periodontitis.