M. Vert

The preparation of sub-200 nm poly(lactide-co-glycolide) microspheres for site-specific drug delivery

Abstract The biodistribution of injected colloidal carriers for targeted delivery will be highly dependent upon their size and surface properties. This paper describes investigations on the preparation of sub-200 nm poly(lactide-co-glycolide) (PLGA) microspheres by the variation of processing parameters using the solvent evaporation technique. Smaller particle sizes were found to be favoured by a two stage emulsification process, a low PLGA concentration and an increased surfactant concentration. In the latter case, it would appear that the viscosity of the continuous phase is a crucial factor. The process of particle size reduction could further be complimented by using a surfactant of lower molecular weight. Employing such procedures, microspheres as low as 90 nm in diameter of low polydispersity were prepared in a reproducible manner.

More about the polymerization of lactides in the presence of stannous octoate

The ring-opening polymerization of lactide cyclic monomers in the bulk in the presence of tin(II) 2-ethylhexanoate (stannous octoate or SnOct2) was reexamined under conditions allowing for the end group characterization of growing chains by high-resolution 1H-NMR. Data collected for low values of the monomer/initiator (M/I) ratio showed that the DL-lactide ring was opened to yield lactyl octoate-terminated short chains. A cationic-type mechanism involving co-initiation by octanoic acid was proposed to account for experimental findings. The formation of a side product, hydroxytin(II) lactate (HTL), was found which appeared able to initiate lactide polymerization and to yield a high molecular weight PLA50 polymer. However the polymerization with stannous octoate was faster than the HTL one. Anyhow, data suggested that both SnOct2 and HTL are likely to act simultaneously as initiators during the polymerization of lactides in the presence of SnOct2. A complete reaction scheme was proposed to account for the presence of the various compounds likely to be formed under these conditions.

Something new in the field of PLA/GA bioresorbable polymers?

Polymers issued from glycolic acid and lactic acids (PLAGA) are now used worldwide as bioresorbable devices in surgery and in pharmacology. Their abiotic hydrolytic degradation has been shown to depend on diffusion-reaction phenomena and to proceed homogeneously or heterogeneously, depending on many factors. Two initiators are presently used industrially to make PLAGA polymers by ring opening polymerisation of lactide and/or glycolide in the bulk, namely Sn octanoate and zinc metal. In this contribution, attention is paid to the differences generated by the use of these two initiator systems in the case of the polymerisation of DL-lactide. Various poly(DL-lactide)s were prepared and characterised by size-exclusion chromatography (SEC), differential scanning calorimetry (DSC) and nuclear magnetic resonance spectroscopy (NMR). These polymers were allowed to age in pH=7.4 isoosmolar phosphate buffer at 37 degrees C. Under these conditions, polymers prepared by the two initiator systems showed dramatic differences when the fates of parallel sided specimens of rather large dimensions were considered. These differences were related to the esterification of some of the OH chain ends by octanoic acid and to the presence of rather hydrophobic low molecular weight by-products which were insoluble in the solvent generally used to purify the crude PLAGA polymers. These new findings should be of great interest in the case of PLAGA based matrices aimed at drug delivery.

Detection and determination of surface levels of poloxamer and PVA surfactant on biodegradable nanospheres using SSIMS and XPS

The surface chemical characterisation of sub-200 nm poly(DL-lactide co-glycolide) nanospheres has been carried out using the complementary analytical techniques of static secondary ion mass spectrometry (SSIMS) and X-ray photoelectron spectroscopy (XPS). The nanospheres, which are of interest for site-specific drug delivery, were prepared using an emulsification-solvent evaporation technique with poly(vinyl alcohol), Poloxamer 407 and Poloxamine 908 respectively as stabilisers. The presence of surfactant molecules on the surface of cleaned biodegradable colloids was confirmed and identified on a qualitative molecular level (SSIMS) and from a quantitative elemental and functional group analysis (XPS) perspective. SSIMS and XPS data were also used in combination with electron microscopy to monitor the effectiveness of cleaning procedures in removing poorly bound surfactant molecules from the surface of nanospheres. The findings are discussed with respect to the development of nanoparticle delivery systems, particularly the composition of the surface for extending blood circulation times and achieving site-specific deposition.

More about the stereodependence of DD and LL pair linkages during the ring‐opening polymerization of racemic lactide

For the last two decades, mixtures of D- and L-lactides, the DD and LL cyclic dimers of lactic acid enantiomers, were polymerized under various conditions of temperature, time, and initiators. Consequences of the dimeric structure of lactide molecules on the distribution of chiral units and on the properties of resulting PLAs were discussed occasionally. In the meantime, NMR techniques have grown up and are now powerful tools to investigate configurational structures from stereosensitive resonances. Thus far, correlation between experimental data and theoretical n-ades were based on Bernoullian-type enchainments of DD and LL pairs of lactyl units present in PLA stereocopolymer chains obtained by ring-opening polymerization. Analysis of NMR spectra of racemic PLAs of different origins, including literature data, was reconsidered on the basis of the probabilities to form iso- and hetero-dyads during chain propagation. Consideration of equireactivity and stereodependent reactivity between the DD and LL pairs generated from a racemic mixture of D- and L-lactide diastereoisomers led us to the conclusion that DD/LL and LL/DD heterotactic junctions are preferentially formed when one uses Sn octoate and Zn metal as initiator systems, even though completed macromolecules remain predominantly isotactic because of the isodyads inherently present in lactide diastereoisomers. A simple method is proposed to determine whether homo- or cross-addition of DD and LL pairs is favorized in any poly(racemic lactide) in the absence of transesterification rearrangements. This method is based on the comparison of experimental relative weights of the three distinct groups of resonances due to carbonyl carbon atoms with relative weights calculated according to the Bernoullian statistics. It allows the determination of a coefficient of stereoselectivity Pi from which one can easily evaluate the average length of isotactic blocks Li without any computation approximations.

Fabrication using a rapid prototyping system and in vitro characterization of PEG-PCL-PLA scaffolds for tissue engineering

In the field of tissue engineering new polymers are needed to fabricate scaffolds with specific properties depending on the targeted tissue. This work aimed at designing and developing a 3D scaffold with variable mechanical strength, fully interconnected porous network, controllable hydrophilicity and degradability. For this, a desktop-robot-based melt-extrusion rapid prototyping technique was applied to a novel tri-block co-polymer, namely poly(ethylene glycol)-block-poly(ε-caprolactone)-block-poly(DL-lactide), PEG-PCL-P(DL)LA. This co-polymer was melted by electrical heating and directly extruded out using computer-controlled rapid prototyping by means of compressed purified air to build porous scaffolds. Various lay-down patterns (0/30/60/90/120/150°, 0/45/90/135°, 0/60/120° and 0/90°) were produced by using appropriate positioning of the robotic control system. Scanning electron microscopy and micro-computed tomography were used to show that 3D scaffold architectures were honeycomb-like with completely interconnected and controlled channel characteristics. Compression tests were performed and the data obtained agreed well with the typical behavior of a porous material undergoing deformation. Preliminary cell response to the as-fabricated scaffolds has been studied with primary human fibroblasts. The results demonstrated the suitability of the process and the cell biocompatibility of the polymer, two important properties among the many required for effective clinical use and efficient tissue-engineering scaffolding.

Ring opening polymerization of D,L-lactide in the presence of zinc metal and zinc lactate

The polymerization of D,L-lactide in the presence of zinc metal was investigated for low values of the monomer/catalyst ratio in order to allow end-group characterization of the growing chains by high resolution 360MHz 1H NMR. The results showed that the polymerization was moisture sensitive and that only a fraction of zinc was active. Small quantities of a side-product were detected and identified as zinc lactate. This compound appeared to be an efficient initiator of the ring-opening polymerization in the bulk. Zinc lactate is thus proposed as the actual initiator of the polymerization of 1,4-dioxane-2,5-diones in the presence of zinc metal. A polymerization mechanism based on a cationic process co-catalysed by lactic acid is also proposed, which accounted very satisfactorily for experimental data. Initiation by zinc lactate yielded high molecular weight polymers with a high degree of conversion and rather high polymerization rates. Therefore, zinc lactate appears to be an attractive alternative to the initiations by zinc metal and stannous octoate, the latter being suspected of some toxicity.

Attempts to map the structure and degradation characteristics of aliphatic polyesters derived from lactic and glycolic acids.

During the past 5 years, important advances have been accomplished in the understanding of the fate of aliphatic polyesters derived from lactic acid (LA) and glycolic acid (GA) in aqueous media. Hydrolysis of solid LA/GA polymers is now regarded as dependent upon a diffusion-reaction mechanism. Faster central degradation, degradation-induced composition, and morphology changes are three of the most important findings which appeared to be composition-dependent as deduced from the behavior of different LA/GA polymers. An attempt is made to generalize these findings to the whole family and to elaborate a map which could be used to predict degradation characteristics of LA/GA polymers from their initial composition and morphology.

Preparation and degradation of surfactant-free PLAGA microspheres

Abstract PLA oligomers which have an amphiphilic surfactant-like structure with a polar head and a long hydrophobic tail were used to prepare self-stabilized microparticles of the PLAGA-type by the evaporation solvent method. It is shown that PLA50 oligomers prepared by polycondensation of dl -lactic acid were able to emulsify an organic solution of high molecular weight PLA37.5GA25 (37.5% L-LA, 37.5% D-LA and 25% GA units) in salt-containing aqueous media. Resulting microspheres were characterized by SEC, DSC, SEM and by size measurements. In vitro degradation was investigated. The mechanism of degradation is driven by the rapid release of oligomeric materials. However, the degradation rate of the high molecular weight components of the matrix was not greatly affected by the short-term presence of the oligomeric materials introduced as matrix-like surfactant. As an example, surfactant-free progesterone-containing microspheres were prepared by the solvent evaporation method.

Poly(lactic acid) degradation in soil or under controlled conditions

The fate of a racemic lactic acid polymer (PLA50) was investigated by allowing parallelepiped plates of PLA50 to age in liquid medium containing a mixed culture of Fusarium moniliforme and Pseudomonas putida at 30°C. Microbial activity was monitored by measuring pH, lactic acid formation by high-performance liquid chromatography, and esterase activity in supernatant. Degradation of the plates was monitored by weighing, size exclusion chromatography, and visual examination. Under the selected conditions, pH increased up to 8, whereas it remained constant in the microorganism-free control. No significant lactic acid formation or esterase activity was detected in the supernatant during the experimental period (32 weeks). For the first 17 weeks, the water absorption rate of the plates was the same in both the microbial and the control media. Subsequently, plates absorbed more water in microbial than in control media. Typical surface/center differential degradation of plates was observed until 17 weeks, but it reduced progressively. At 32 weeks, specimens appeared completely disintegrated in the microbial medium. PLA50 plates were also buried in the soil for 8 weeks. After recovery, plates were allowed to degrade 8 weeks under controlled conditions. Scanning electron microscopy of these plates showed the development of some filamentous fungi at the surface and into the bulk of plates. Five fungal strains were isolated which appeared to be able to assimilate PLA50 oligomers in mixed cultures. It was assumed that PLA50 plates had been first degraded by chemical hydrolysis, followed by the bioassimilation of degradation by-products in both experiments.