poly(D,L-lactic-co-glycolic) Acid Research Articles

The influence of extracellular matrix on the generation of vascularized, engineered, transplantable tissue.

In a recently described model for tissue engineering, an arteriovenous loop comprising the femoral artery and vein with interposed vein graft is fabricated in the groin of an adult male rat, placed inside a polycarbonate chamber, and incubated subcutaneously. New vascularized granulation tissue will generate on this loop for up to 12 weeks. In the study described in this paper three different extracellular matrices were investigated for their ability to accelerate the amount of tissue generated compared with a no-matrix control. Poly-D,L-lactic-co-glycolic acid (PLGA) produced the maximal weight of new tissue and vascularization and this peaked at two weeks, but regressed by four weeks. Matrigel was next best. It peaked at four weeks but by eight weeks it also had regressed. Fibrin (20 and 80 mg/ml), by contrast, did not integrate with the generating vascularized tissue and produced less weight and volume of tissue than controls without matrix. The limiting factors to growth appear to be the chamber size and the capacity of the neotissue to integrate with the matrix. Once the sides of the chamber are reached or tissue fails to integrate, encapsulation and regression follow. The intrinsic position of the blood supply within the neotissue has many advantages for tissue and organ engineering, such as ability to seed the construct with stem cells and microsurgically transfer new tissue to another site within the individual. In conclusion, this study has found that PLGA and Matrigel are the best matrices for the rapid growth of new vascularized tissue suitable for replantation or transplantation.

Morphology, drug distribution, and in vitro release profiles of biodegradable polymeric microspheres containing protein fabricated by double-emulsion solvent extraction/evaporation method

The surface and internal morphology, drug distribution and release kinetics at 22°C of polyesters such as PCL (polycaprolactone) and PLGA (poly( dl-lactic- co-glycolic acid)) 65 : 35 microspheres containing BSA (bovine serum albumin) have been investigated in order to understand the relationship amongst morphology, drug distribution and in vitro release profiles and to develop controlled release devices for marine fishes in tropical area. CLSM (confocal laser scanning microscope) micrographs reveal that the polyvinylalcohol (PVA as an emulsifier) concentration in the external water phase strongly influences drug distribution within microspheres and release profiles. The presence of PVA in the internal water phase enhances the stabilization of inner water droplets against coalescence. This results in a more uniform drug distribution and a slower BSA release. Different oil-phase volumes and polymer concentrations yield different solvent exchange and precipitation mechanisms, which lead to different morphologies. A low oil-phase volume yields microspheres with a porous matrix and defective skin surface, which gives a high initial BSA burst as well as a fast release profile. Microspheres fabricated from a low polymer concentration have less defective skin surface, but with a less tortuous inner matrix which results in a more rapid BSA release. A higher BSA loading yields a larger concentration gradient between the emulsion droplet and the continuous water phase as well as between the microspheres and the in vitro medium. The former results in a lower encapsulation efficiency, whereas the latter yields a faster initial burst and a more rapid release profile. High stirring speed can reduce microsphere size, but decreases the yield of microspheres.

Effect of preparation conditions on morphology and release profiles of biodegradable polymeric microspheres containing protein fabricated by double-emulsion method

We have investigated the key parameters to fabricate PDLLA (Poly( dl-lactic acid)), PDLLGA (Poly( dl-lactic-co-glycolic acid) 65 : 35 and blends of PDLLGA 65 : 35 and PEG (Poly(ethylene glycol)) microspheres containing bovine serum albumin (BSA) as a model protein using the double-emulsion (water-in-oil-in-water) solvent extraction/evaporation method. The release profiles of microspheres were investigated at 22°C in order to develop controlled release devices for marine fishes in tropical area. Various factors that influence the size of microspheres, encapsulation efficiency, initial release, morphology and release profiles of microspheres, and BSA distribution within microspheres have been investigated. These factors include preparation temperature, solvent removal rate, volume ratio of oil phase to internal water phase, and polymer concentration. Microspheres fabricated at a low volume ratio of oil phase to internal water phase and a low polymer concentration tend to have a large surface area, a low bulk density, resulting in a high initial burst and a fast release of BSA. Fabrication temperature heavily affects solvent extraction/evaporation and mechanism of phase-inversion. The microspheres fabricated at 4 and 38°C yield the highest encapsulation efficiency (52.0–48.0%) and lowest initial BSA release (18.8–20.0%), while microspheres produced at 22°C have the lowest encapsulation efficiency and highest initial burst. This interesting phenomenon is due to the fact that different phase-inversion paths occur when preparation temperature varies. Nucleation growth and spinodal decomposition dominate the skin formation at low preparation temperatures, while evaporation-driven skin formation takes place at high preparation temperatures. The relationship between the release profile and the rate of continuous water-phase addition is extremely complicated. Slow demixing dominates the interface skin formation at low continuous water-phase addition rates and results in fine porous skin structure, while rapid demixing dominates at high continuous water-phase addition rates and also leads to microspheres with a porous skin. Thus both have high initial bursts and fast release rates. A continuous water-phase addition of 3 ml/min may yield the microspheres having a low initial burst and a slow release rate.

Encapsulation and release of rhodium(II) citrate and its association complex with hydroxypropyl-β-cyclodextrin from biodegradable polymer microspheres

Rhodium(II) carboxylates and their derivatives constitute a promising class of second-generation transition metal compounds with anticancer properties. While most transition metal anticancer compounds chelate DNA and cause extensive chromosomal damage, rhodium(II) carboxylates act on the enzyme DNA polymerase R and hence cause minimal chromosomal damage. Rhodium(II) citrate, a recent member of the rhodium(II) carboxylate family is highly promising as an antitumor agent. However, due to its high water solubility, a high systemic dose is necessary to achieve efficacy. In this paper, we have explored the complexation of rhodium(II) citrate with hydroxypropyl-β-cyclodextrin as a means to improve encapsulation and release kinetics from poly(dl-lactic-co-glycolic) acid (PLGA) and poly(anhydride) microspheres. We observed that complexation of rhodium(II) citrate with hydroxypropyl-β-cyclodextrin significantly increased both the encapsulation efficiency and duration of release in both polymer systems.

Comparison of Process Parameters for Microencapsulation of Plasmid DNA in Poly(D,L-Lactic-co-Glycolic) Acid Microspheres

Poly(D,L-lactic-co-glycolic) acid (PLGA) microspheres containing plasmid DNA encoding the firefly luciferase gene were prepared using the water-in-oil-in-water (w/o/w) double emulsion and solvent evaporation method. In this study, we investigated the effects of three process parameters on DNA microencapsulation: (1) emulsification method used to generate the primary emulsion, (2) water/oil ratio during formation of the first emulsion, and (3) surfactant concentration used in the preparation of the second emulsion. The resulting formulations were also analyzed for microsphere size, encapsulation efficiency, and kinetics of DNA release. We found that although each process alteration resulted in encapsulation of biologically active, structurally intact DNA, the surfactant and water/oil ratio significantly affected the size, release kinetics and encapsulation efficiency of plasmid DNA.

Repair of ulnar segmental defect by recombinant human bone morphogenetic protein-2 in dogs.

The efficacy of recombinant human bone morphogenetic protein-2 (rhBMP-2) combined with poly D, L lactic-co-glycolic acid (PLGA)/gelatin sponge complex (PGS) as a carrier on the repair of segmental long-bone defects was evaluated using an ulnar model in dogs. The defect was 2 cm in length and was fixed with bone plating. After implantation of PGS with or without rhBMP-2, the repair process of the defect was evaluated by serial radiography until 16 postoperative weeks. All defects treated with 160 micrograms or 640 micrograms of rhBMP-2/PGS revealed bone union radiographically by 12 postoperative weeks, whereas all defects treated with PGS alone revealed no radiographic evidence of healing throughout the experimental period. In defects treated with 40 micrograms of rhBMP-2/PGS, new bone appeared partially at the defects but did not accomplish union. Bone mineral contents at the defect sites after harvest at 16 weeks postoperatively were significantly (p < 0.05) higher in those treated with 160 micrograms or 640 micrograms of rhBMP-2 than in those treated with 40 micrograms of rhBMP-2 or PGS alone. Histologically, defects radiographically diagnosed as having achieved union showed the appearance of cortical bone and bone marrow cells. These findings suggest the use of rhBMP-2/PGS as a potential bone graft substitute in reconstructive surgery in dogs.

A Theoretical Model of Erosion and Macromolecular Drug Release from Biodegrading Microspheres

□ A theoretical model is outlined for predicting the time evolution of total mass, mean molecular weight, and drug release for the case of a spherical bulk-eroding microsphere, prepared by a double emulsification procedure and containing a hydrophilic drug, such as a protein or peptide. Explicit analytical formulae are derived for calculating the time evolution of measurable macroscopic characteristics, such as drug release or mean molecular weight. Microsphere hydration, polymer erosion, and drug release phases are each described. Results indicate that polymer degradation by only random-chain scission or only end scission (or unzipping) cannot explain experimentally observed kinetics of particle mass loss and molecular weight change; thus, a combined model (incorporating both random and end scission) is proposed. A general methodology for determining the microscopic transport coefficients (such as polymer degradation rate constant or drug diffusion coefficient) from erosion and release data is outlined. This paradigm is applied to the specific case of 50:50 poly(D,L-lactic-co-glycolic acid (PLGA) microspheres encapsulating glycoprotein 120 (gp 120), a candidate AIDS vaccine. Predictions permit comparisons with experimental data for mean weight- and number- averaged molecular weights, as well as for mass loss and protein release. Other comparisons are made with data appearing in the literature for release of tetanus toxoid from PLA and PLGA microspheres of variable molecular weight. Agreement between theory and experiment is observed.

Biodegradation and tissue reaction to intravitreous biodegradable poly(D,L-lactic-co-glycolic)acid microspheres.

We studied the biodegradation of and the tissue reaction to microspheres of 50:50 poly(D,L-lactic-co-glycolic)acid (PLGA) (viscosity-average MW: 3000 d), injected intravitreous in rabbits. These microspheres are under investigation as injectable devices for intravitreous sustained drug delivery. The rate of intravitreous degradation of PLGA microspheres has not been well documented in the literature. Twenty two pigmented rabbits underwent gas vitrectomy in one eye: 19 eyes received 2.5 mg of PLGA microspheres in 1 ml of balanced salt solution (BSS) and 3 control eyes received 1 ml of BSS only. Slit lamp exam and indirect ophthalmoscopy were performed periodically from day 1 to 6 months after surgery. The eyes were enucleated and studied by light microscopy and immunohistochemistry at various time points. The electroretinogram (ERG) was recorded in a subgroup of rabbits before injection and after 1 and 6 months. The amount of microspheres in the vitreous cavity progressively decreased. At 6 months microspheres were found in 1/4 rabbits at indirect ophthalmoscopy and in 4/4 rabbits histopathologically. A mild localized, non progressive foreign body reaction was observed. The cell reaction was composed mostly of vimentin and glial fibrillary acidic protein positive cells which probably represent glial cells and fibroblasts. The choroid and retina were normal. The ERG showed no abnormalities. No clinical inflammatory signs were observed 4 days postoperatively and thereafter.