Quality Control Optimization for Minimizing Security Risks Associated with Mesenchymal Stromal Cell-Based Product Development.

Mesenchymal stromal cells (MSCs) have been considered a therapeutic strategy in regenerative medicine because of their regenerative and immunomodulatory properties. The translation of MSC-based products has some challenges, such as regulatory and scientific issues. Quality control should be standardized and optimized to guarantee the reproducibility, safety, and efficacy of MSC-based products to be administered to patients. The aim of this study was to develop MSC-based products for use in clinical practice. Quality control assays include cell characterization, cell viability, immunogenicity, and cell differentiation; safety tests such as procoagulant tissue factor (TF), microbiological, mycoplasma, endotoxin, genomic stability, and tumorigenicity tests; and potency tests. The results confirm that the cells express MSC markers; an average cell viability of 96.9%; a low expression of HLA-DR and costimulatory molecules; differentiation potential; a high expression of TF/CD142; an absence of pathogenic microorganisms; negative endotoxins; an absence of chromosomal abnormalities; an absence of genotoxicity and tumorigenicity; and T-lymphocyte proliferation inhibition potential. This study shows the relevance of standardizing the manufacturing process and quality controls to reduce variability due to the heterogeneity between donors. The results might also be useful for the implementation and optimization of new analytical techniques and automated methods to improve safety, which are the major concerns related to MSC-based therapy.

Biological, biochemical and genotoxic effects of Sb in the midge Chironomus sancticaroli Strixino and Strixino, 1981 (Diptera: Chironomidae).

In aquatic systems, antimony (Sb) is found in the water column and associated with sediment particles being bioavailable to organisms. Consequently, toxic effects have been detected in benthic invertebrates, but the toxicity after Sb exposure in Chironomidae have not been investigated. Were investigated DNA damage, activities of cholinesterase (ChE), alpha and beta esterase (EST-α, EST-ß), glutathione S-transferase (GST), catalase (CAT), and superoxide dismutase (SOD) and lipid peroxidation after acute (48 h) and subchronic exposure (8 d). We also investigated the effects of subchronic (8 d) on development of larvae and chronic (25 d) Sb exposure on emergence and size of adults of Chironomus sancticaroli. Were analyze Sb nominal concentrations ranged from 0.5 to 800 µg.L-1. Genotoxic effects occurred at higher concentrations upon acute (50, 800 µg.L-1) and subchronic exposure (50 µg.L-1). Acute exposure increased ChE, EST-α, EST-ß, and GST activities. Subchronic Sb exposure increased EST-α activity at 0.2 µg.L-1 and GST activity at 5 µg.L-1. CAT activity increased at all concentrations while increasing lipid peroxidation levels were observed (1 µg.L-1, 5 µg.L-1 and 50 µg.L-1), indicating oxidative stress. All concentrations of Sb delayed larval development and decreased the number of emerging adults. At high concentrations (50, 500, 800 µg.L-1), the emerging adults were smaller. In conclusion, these varying genotoxic, biochemical and biological effects of Sb make a notable impact on the reproduction and population dynamics of C. sancticaroli.

Genotoxic, metabolic, and biological responses of Chironomus sancticaroli Strixino & Strixino, 1981 (Diptera: Chironomidae) after exposure to BBP.

Benzyl butyl phthalate (BBP), which is widely used in industrial production, reaches the aquatic environment, mainly owing to improper disposal of plastic products. In the water, it remains adsorbed to sedimentary particles causing toxic effects in aquatic invertebrates such as Chironomidae, which are important in maintaining ecosystem dynamics and are an important link in the food chain. However, the effects of BBP on Chironomidae are still poorly known. Thus, the toxic effects of BBP on Chironomus sancticaroli at acute (48 h), subchronic (8 d), and chronic (25 d) exposures of concentrations between 0.1 and 2000 µg·L-1 were determined. Genotoxicity effects, changes in the oxidative stress pathway, and development and emergence of organisms were evaluated. Biochemical markers showed a reduction in cholinesterase (ChE) activity, indicating a neurotoxic effect on acute exposure (1-1000 µg·L-1). The antioxidant pathway, glutathione S-transferase (GST) activity showed reduction on acute (0.1; 1-2000 µg·L-1) and subchronic (1-2000 µg·L-1) exposures and reduction in superoxide dismutase (SOD) activity at all evaluated concentrations, suggesting oxidative stress. In contrast, lipid peroxidation was not observed. DNA damage occurred on acute (10 µg·L-1) and subchronic (10-2000 µg·L-1) exposures, indicating genotoxic effects. At concentrations above 10 µg·L-1, no emergence of adults occurred, while lower concentrations (0.1 and 1 µg·L-1) showed a reduction in the number of adults, mainly males. The observed effects indicate that BBP is genotoxic and causes biochemical alterations presenting high toxicity at the population level.