Neural Crest-Derived Chondrocytes Isolation for Tissue Engineering in Regenerative Medicine.

Chondrocyte transplantation has been successfully tested and proposed as a clinical procedure aiming to repair articular cartilage defects. However, the isolation of chondrocytes and the optimization of the enzymatic digestion process, as well as their successful in vitro expansion, remain the main challenges in cartilage tissue engineering. In order to address these issues, we investigated the performance of recombinant collagenases in tissue dissociation assays with the aim of isolating chondrocytes from bovine nasal cartilage in order to establish the optimal enzyme blend to ensure the best outcomes of the overall procedure. We show, for the first time, that collagenase H activity alone is required for effective cartilage digestion, resulting in an improvement in the yield of viable cells. The extracted chondrocytes proved able to grow and activate differentiation/dedifferentiation programs, as assessed by morphological and gene expression analyses.

Assessment and characterization of the bacterial community structure in advanced activated sludge systems.

The present study is aimed to assess and characterize the structure of bacterial community in advanced activated sludge systems. In particular, activated sludge samples were collected from an Integrated Fixed-film Activated Sludge - Membrane Bioreactor pilot plant under a University of Cape Town configuration with in-series anaerobic (Noair)/anoxic (Anox)/aerobic (Oxy) reactors - and further analyzed. The achieved results - based on Next Generation Sequencing (NGS) of 16S rDNA amplicons - revealed that the bacterial biofilm (bf) communities on plastic carriers of Oxy and Anox reactors had a greater diversity compared to suspended (sp) bacterial flocs of Oxy, Anox and Noair. Indeed, the Shannon diversity indices of both biofilm communities were higher than those of suspended growth samples (Oxy-bf = 4.1 and Anox-bf = 4.2 vs. Oxy-sp = 3.4, Anox-sp = 3.5 and Noair-sp = 3.4). The most striking differences have been reported in Rhodobacteraceae being more abundant in biofilm specimens than in suspended biomass samples. The vast majority of the identified bacteria differs from those obtained by culture dependent method, thus suggesting that NGS-based method is really suitable to analyze the bacterial community composition, even in advanced systems for wastewater treatment.

Bacterial community structure and removal performances in IFAS-MBRs: A pilot plant case study.

The paper reports the results of an experimental campaign carried out on a University of Cape Town (UCT) integrated fixed-film activated sludge (IFAS) membrane bioreactor (MBR) pilot plant. The pilot plant was analysed in terms of chemical oxygen demand (COD) and nutrients removal, kinetic/stoichiometric parameters, membrane fouling and sludge dewaterability. Moreover, the cultivable bacterial community structure was also analysed. The pilot plant showed excellent COD removal efficiency throughout experiments, with average value higher than 98%, despite the slight variations of the influent wastewater. The achieved nitrification efficiency was close to 98% for most of the experiments, suggesting that the biofilm in the aerobic compartment might have sustained the complete nitrification of the influent ammonia, even for concentrations higher than 100 mg L-1. The irreversible resistance due to superficial cake deposition was the mechanism that mostly affected the membrane fouling. Moreover, it was noticed an increase of the resistance due pore blocking likely due to the increase of the EPSBound fraction that could derive by biofilm detachment. The bacterial strains isolated from aerobic tank are wastewater bacteria known for exhibiting efficient heterotrophic nitrification-aerobic denitrification and producing biofilm.