Biodegradable Porous Silk Microtubes for Tissue Vascularization.

Cardiovascular diseases are the leading cause of mortality around the globe, and microvasculature replacements to help stem these diseases are not available. Additionally, some vascular surgeries needing small diameter vascular grafts present different performance requirements. In this work silk fibroin scaffolds based on silk/polyethylene oxide blends were developed as microtubes for vasculature needs and for different tissue regeneration times, mechanical properties and structural designs. Systems with 13, 14 and 15% silk alone or blended with 1 or 2% of polyethylene oxide (PEO) were used to generate porous microtubes using gel-spinning. Microtubes with inner diameters (ID) of 150-300 µm and 100 µm wall thickness were fabricated. The systems were assessed for porosity, mechanical properties, enzymatic degradability, and in vitro vascular endothelial cell attachment and metabolic activity. After 14 days all tubes supported the proliferation of cells and cell attachment increased with porosity. The silk tubes with PEO had similar crystallinity but higher elastic modulus compared with the systems without PEO. The silk (13%)/PEO (1%) system showed the highest porosity (20 µm pore diameters on average), highest cell attachment and fastest degradation profile. There was a good correlation between these parameters with silk concentration and the presence of PEO. The results demonstrate the ability to generate versatile and tunable tubular biomaterials based on silk-PEO-blends with potential for microvascular grafts.

Development and characterization of new enzymatic modified hybrid calcium carbonate microparticles to obtain nano-architectured surfaces for enhanced drug loading.

HYPOTHESIS: Biopolymer-CaCO3 hybrid microparticles exposed to hydrolytic enzymes can provide new surface tailorable architectures. Soluble Alginate Lyase hydrolyzed alginate chains exposed on microparticle surface are generating considerable matrix changes. The change of porosity and surface to volume ratio is expected to influence absorption of drugs, thereby affecting controlled release profiles. The developed hybrid system potentially shows interesting properties for lung drug administration. EXPERIMENTAL: Hybrid microparticles were developed by colloidal co-precipitation of CaCO3 in presence of biopolymers: alginate (Alg) or Alg-High Methoxylated Pectin (HMP), followed by treatment with Alginate Lyase (AL). Surface architectures were observed by SEM. The increase in area to volume ratio was confirmed by BET isotherms. Also, enzymatic changes were elucidated by biophysical methods (EDAX, DSC, FTIR, XRD) and determination of the total carbohydrates content. Levofloxacin (a fluoroquinolone antibiotic) as model drug was incorporated by absorption. The drug release profile and the antimicrobial activity of the microparticles were tested against Pseudomonas aeruginosa. FINDINGS: After enzyme treatment, microspheres showed 4µm diameter and increased porosity. While CaCO3-Alg microspheres resulted in a rougher surface, CaCO3-Alg-HMP ones exhibited "nano-balloon" patterns on surface. Both AL-treated microparticles showed up to 3 and 7 times higher Levofloxacin encapsulation than no treated ones. Microparticles showed controlled drug release profiles and enhanced antimicrobial effect. The present work demonstrates a significant progress in the development of new carriers with potential application for lung infections treatment.

Effects of organic solvents on immobilized lipase in pectin microspheres.

Lipase from Brevibacillus agri 52 was found stable up to 90% diethylenglycol (DEG), glycerol (GLY), and 1,2 propanediol (1,2 PRO) at 37 degrees C for 1 h and the stability was reduced only approximately 20% after 12 h incubation, but in 40% dimethylsulfoxide (DMSO), lipase activity was stable only for 1 h. Inhibition of the biocatalysts with dimethylformamide (DMF) was detected at 20% solvent concentration. In water immiscible systems, the stability of lipase in n-hexane, n-tetradecane and n-heptane resembles the water activity, but in the presence of isobutanol, 1-hexanol, and butylbutirate, the stability was significantly reduced. Lipase 52 precipitates in the presence of 50% acetone or ethanol/water mixtures, but enzymatic activity was partially recovered by adding 20% GLY, DEG, 1,2 PRO, or DMSO to the reaction mixture. Furthermore, by increasing DEG in 70% DMF/DEG mixtures, the lipase activity was protected. Encapsulation of lipase in pectin gels cross-linked with calcium ions brings three to four times more enzymatic activity in 70% water miscible organic solvents compared to aqueous systems.