T. K. Makhina

Effect of growth conditions on the molecular weight of poly-3-hydroxybutyrate produced by Azotobacter chroococcum 7B

It has been shown that poly-3-hydroxybutyrate (PHB) of predetermined molecular weight can be obtained by varying the growth conditions of the producer strain, Azotobacter chroococcum 7B: pH, temperature, aeration, presence of sodium acetate as an additional carbon source, or growth on crude complex carbon sources (molasses, vinasse, or starch). High-molecular-weight polymer can be obtained at pH 7.0, optimal for the culture (1485 kDa), temperature 30–37°C (1600–1450 kDa, respectively), and low aeration (2215 kDa). The following factors decrease PHB MW: pH deviation to the acidic (pH 6.0, 476 kDa) or alkaline (pH 8.0, 354 kDa) range or lower temperature (20°C, 897 kDa). Introduction of additional carbon source (sodium acetate) at concentrations in the medium varying from 0 to 5 g/l provides an original method of production of PHB with predetermined MW in a wide range, from 270 to 1515 kDa, with high PHB content in the cell.

Hydrolytic Degradation of Poly(3-hydroxybutyrate), Polylactide and their Derivatives: Kinetics, Crystallinity, and Surface Morphology

Hydrolytic degradations of biodegradable poly(3-hydroxybutyrate) (PHB), polylactide (PLA) and their derivatives were explored by kinetic and structure methods at 37 and 70°C in phosphate buffer. It was revealed the kinetic profiles for copolymer PHBV (20% of 3-hydroxyvalerate) and the blend PHB-PLA (1:1 wt. ratio). The intensity of biopolymer hydrolysis depending on temperature is characterized by total weight loss and the viscosity-averaged molecular weight decrement (ΔMW) as well as by WAXS and AMF techniques. Characterization of PHB and PHBV includes both ΔMW and crystallinity evolution (x-ray diffraction) as well as the AFM analysis of PHB film surfaces before and after aggressive medium exposition. The degradation is enhanced in the series PHBV < PHB < PHB-PLA blend < PLA. The impact of MW on the biopolymer hydrolysis is shown.

New poly(3-hydroxybutyrate)-based systems for controlled release of dipyridamole and indomethacin

New poly(3-hydroxybutyrate)-based systems for controlled release of anti-inflammatory and anti-thrombogenic drugs have been studied. The release occurs via two mechanisms (diffusion and degradation) operating simultaneously. Dipyridamole and indomethacin diffusion processes determine the rate of the release at the early stages of the contact of the system with the environment (the first 6–8 h). The coefficient of the release diffusion of a drug depends on its nature, the thickness of the poly(3-hydroxybutyrate) films containing the drug, the concentrations of dipyridamole and indomethacin, and the molecular weight of the poly(3-hydroxybutyrate). The results obtained are critical for developing systems of release of diverse drugs, thus, enabling the attainment of the requisite physiological effects on tissues and organs of humans.

Biodegradation kinetics of poly(3-hydroxybutyrate)-based biopolymer systems

The aim of this study was to evaluate and to compare the long-term kinetics curves of biodegradation of poly(3-hydroxybutyrate) (PHB), its copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate), and a PHB/polylactic acid composite. The total weight loss and the change of average viscosity molecular weight were used as the parameters reflecting the biodegradation degree. The rate of biodegradation was analyzed in vitro in the presence of lipase and in vivo after film implantation in animal tissues. The morphology of the PHB film surface was studied by the atomic force microscopy technique. It was shown that PHB biodegradation involves both polymer hydrolysis and its enzymatic biodegradation. The results obtained in this study can be used for the development of various PHB-based medical devices.

Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer by Azotobacter chroococcum strain 7B

The ability of Azotobacter chroococcum strain 7B, producer of poly(3-hydroxybutyrate) (PHB), to synthesize its copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(3HB-co-3HV)) was studied. It was demonstrated, for the first time, that A. chroococcum strain 7B was able to synthesize P(3HB-co-3HV) with various molar rates of HV in the polymer chain when cultivated on medium with sucrose and carboxylic acids as precursors of HV elements in the PHB chain, namely, valeric (13.1–21.6 mol %), propanoic (3.1 mol %), and hexanoic (2.1 mol %) acids. Qualitative and functional differences between PHB and P(3HB-co-3HV) were demonstrated by example of the release kinetic of methyl red from films made of synthesized polymers. Maximal HV incorporation into the polymer chain (28.8mol %) was recorded when the nutrient medium was supplemented with 0.1% peptone on the background of 20 mM valerate. These results suggest that that the studied strain can be regarded as a potential producer of not only PHB but also P(3HB-co-3HV).

The Biodegradation of Poly-β-Hydroxybutyrate (PHB) by a Model Soil Community: The Effect of Cultivation Conditions on the Degradation Rate and the Physicochemical Characteristics of PHB

The biodegradation of films made of poly-β-hydroxybutyrate (PHB) with a molecular mass of 1500 kDa was studied using a model soil community in the presence and absence of nitrate and at different concentrations of oxygen in the gas phase. The biodegradation of PHB was investigated with respect to changes in its molecular mass, crystallinity, and some mechanical properties.

Sustained release of the antitumor drug paclitaxel from poly(3-hydroxybutyrate)-based microspheres

The development of sustained release formulations based on biodegradable polymers is a promising trend in modern pharmacology. Polyhydroxyalkanoates (PHA) attract increasing attention due to their biodegradability and high biocompatibility, which make them suitable for the development of novel drug dosage forms. We have produced poly(3-hydroxybutyrate) (PHB)-based microspheres loaded with the antitumor drug paclitaxel and investigated morphology, drug release kinetics and the effect of these microspheres on tumor cells in vitro. The data on the kinetics of drug release, biocompatibility and biological activity of the biopolymer microspheres in vitro have demonstrated that the studied system of prolonged drug release had lower toxicity and higher efficiency compared to the traditional dosage forms of paclitaxel.

Culturing of Mouse Mesenchymal Stem Cells on Poly-3-Hydroxybutyrate Scaffolds.

We studied the possibility of long-term culturing of mouse mesenchymal stem cells on a porous scaffold made of biocompatible polymer poly-3-hydroxybutyrate. The cells remained viable for at least 2 months and passed more than 65 population doublings in culture. Culturing on the scaffold did not change surface phenotype of cells. 3D poly-3-hydroxybutyrate scaffolds are appropriate substrate for long-term culturing of mesenchymal stem cells.

Prolonged release of chlorambucil and etoposide from poly-3-oxybutyrate-based microspheres

Microspheres were obtained on the basis of poly(3-oxibutyrate) (POB) with the inclusion of the Chlorambucil and Etoposide cytostatic drugs in a polymer matrix, and the morphology, kinetics of drug release from microspheres, and the interaction between microspheres and tumor cells in vitro were studied. Data on the kinetics of drug release suggests that a prolonged release occurs by drug diffusion from the polymer matrix at the initial stage and at the expense of hydrolytic degradation of the polymer at a later stage. A study of the biocompatibility and biological activity of biopolymeric microspheres showed that chlorambucil operates actively and strongly inhibits the growth of cultured cells for a short time (24 h). Etoposide acts weaker (the percentage of cell growth suppression during 48 h does not exceed 50%), but subsequently it has a basis for the creation of new dosage forms with prolonged action of Etoposide and chlorambucil for cancer therapy.

Low molecular weight poly(3-hydroxybutyrate) microparticles synthesized by piezoelectric spray drying for the sustained release of paclitaxel

Biocompatible and biodegradable polymer microparticles are contemporary medicines able to eliminate the side effects and unsatisfactory pharmacokinetics of already existing preparations. In this work we have developed a high-tech scalable method for the synthesis of paclitaxel-loaded poly(3-hydroxybutyrate) (PHB)-based microparticles. These particles were synthesized on a B-90 Buchi nano spray dryer by piezoelectric spray drying in an inert atmosphere. A regular spherical shape, narrow size distribution, and satisfactory results for the release of paclitaxel from the polymeric matric of microparticles in vitro make this polymeric medicinal form promising for its further application in pharmaceutics. Nanoparticles with a similar composition synthesized via the laboratory one-stage emulsification method were used for comparison. This study is the first stage in the creation of a sustained-action anticancer paclitaxel preparation.