Chondrogenic differentiation of mesenchymal stem/stromal cells on 3D porous poly (ε-caprolactone) scaffolds: Effects of material alkaline treatment and chondroitin sulfate supplementation.

Cartilage defects resultant from trauma or degenerative diseases (e.g., osteoarthritis) can potentially be repaired using tissue engineering (TE) strategies combining progenitor cells, biomaterial scaffolds and bio-physical/chemical cues. This work examines promoting chondrogenic differentiation of human bone marrow mesenchymal stem/stromal cells (BM-MSCs) by combining the effects of modified poly (ε-caprolactone) (PCL) scaffolds hydrophilicity and chondroitin sulfate (CS) supplementation in a hypoxic 5% oxygen atmosphere. 3D-extruded PCL scaffolds, characterized by µCT, featured a 21 mm-1 surface area to volume ratio, 390 µm pore size and approximately 100% pore interconnectivity. Scaffold immersion in sodium hydroxide solutions for different periods of time had major effects in scaffold surface morphology, wettability and mechanical properties, but without improvements on cell adhesion. In-situ chondrogenic differentiation of BM-MSC seeded in 3D-extruded PCL scaffolds resulted in higher cell populations and ECM deposition along all scaffold structure, when chondrogenesis was preceded by an expansion phase. Additionally, CS supplementation during BM-MSC expansion was crucial to enhance aggrecan gene expression, known as a hallmark of chondrogenesis. Overall, this study presents an approach to tailor the wettability and mechanical properties of PCL scaffolds and supports the use of CS-supplementation as a biochemical cue in integrated TE strategies for cartilage regeneration.

Modification of the activity of an alpha-amylase from Bacillus licheniformis by several surfactants

The influence of different commercial surfactants on the enzymatic activity of a commercial alpha-amylase from Bacillus licheniformis (Termamyl 300 L) has been studied. As non-ionic surfactants, alkyl polyglycosides (Glucopon® 215, Glucopon® 600 and Glucopon® 650) were studied, as were fatty alcohol ethoxylates (Findet 1214N/23 and Findet 10/15), and nonyl phenol ethoxylate (Findet 9Q/21.5NF). Also, an anionic surfactant, linear alkyl benzene sulfonate (LAS) was assayed. In general, none of the non-ionic surfactants studied, except Findet 10/15, vary substantially the enzymatic activity. Findet 10/15 has the strongest hydrophobic character and reduces the enzymatic activity more significantly the greater its concentration. Regarding LAS, this surfactant significantly depressed enzymatic activity, presumably due to the electrostatic interactions caused by its anionic character.

Solvent tolerance in bacteria: role of efflux pumps and cross-resistance with antibiotics.

Microorganisms have mechanisms that enable them to tolerate lethal concentrations of toxic compounds. This feature has been exploited in a wide range of bioprocesses that range from bioremediation applications to production of fine chemicals in two-phase reaction media. The ability to modify the physical properties of cellular membranes has long been put forward as a protection mechanism that enables microorganisms to tolerate solvents. More recently, efflux pumps have been shown to extrude deleterious compounds, such as antibiotics, drugs and solvents. An understanding of the mechanism of solvent tolerance and its relationship to cross-resistance of pathogenic organisms to antibiotics has major impact on the type and use of disinfectants and disinfecting procedures. The presence of solvents in the growth environment may lead to the emergence of solvent resistant strains and, therefore, overuse may propagate resistant microbial variants. In this paper, mechanisms that lead to solvent tolerance of microbes and accompanying specific antibiotic resistance are reviewed.