ABSTRACT
In the current global emergency due to SARS-CoV-2 outbreak, passive immunotherapy emerges as a promising treatment for COVID-19. Among animal-derived products, equine formulations are still the cornerstone therapy for treating envenomations due to animal bites and stings. Therefore, drawing upon decades of experience in manufacturing snake antivenom, we developed and preclinically evaluated two anti-SARS-CoV-2 polyclonal equine formulations as potential alternative therapy for COVID-19. We immunized two groups of horses with either S1 (anti-S1) or a mixture of S1, N, and SEM mosaic (anti-Mix) viral recombinant proteins. Horses reached a maximum anti-viral antibody level at 7 weeks following priming, and showed no major adverse acute or chronic clinical alterations. Two whole-IgG formulations were prepared via hyperimmune plasma precipitation with caprylic acid and then formulated for parenteral use. Both preparations had similar physicochemical and microbiological quality and showed ELISA immunoreactivity towards S1 protein and the receptor binding domain (RBD). The anti-Mix formulation also presented immunoreactivity against N protein. Due to high anti-S1 and anti-RBD antibody content, final products exhibited high in vitro neutralizing capacity of SARS-CoV-2 infection, 80 times higher than a pool of human convalescent plasma. Pre-clinical quality profiles were similar among both products, but clinical efficacy and safety must be tested in clinical trials. The technological strategy we describe here can be adapted by other producers, particularly in low- and middle-income countries.
ABSTRACT
The aim of this study was to evaluate the effect of Er:YAG laser for selective removal of carious lesion, followed by biomodification with chitosan gel where the subsurface microhardness, chemical composition, and morphological changes of the residual caries-affected dentin were examined. Artificial dentinal lesions were created by pH-cycling method (14 days) in 104 bovine specimens (5 × 5 mm). Specimens were randomly divided according to the carious removal method: bur (low-speed handpiece) or Er:YAG laser (250 mJ/4 Hz). Specimens were treated with 35% phosphoric acid and were subdivided into two groups according to dentin biomodification: without chitosan (control) and 2.5% chitosan. Forty specimens were restored with an adhesive system and composite resin. Subsurface microhardness tests were performed in sound dentin, caries-affected dentin, residual caries-affected dentin, and after the restoration. The other 64 specimens were subjected to SEM-EDS atomic analysis. Data were statistically analyzed (p < 0.05). After the Er:YAG laser excavation, the microhardness value of residual caries-affected dentin was higher (p < 0.05) than bur-treated dentin. A significant decrease in the amount of Ca, P, and Ca/P ratio was found after the removal of carious lesions with Er:YAG laser (p < 0.05). The biomodification with chitosan did not influence the microhardness and atomic percentage of Ca, P, and Ca/P ratio of residual caries-affected dentin (p > 0.05). SEM analysis showed morphological changes on residual caries-affected dentin (p > 0.05). The selective removal of carious dentin with Er:YAG laser increased microhardness of residual caries-affected dentin, changing its surface morphology and chemical composition. The biomodification with chitosan did not influence the structural and chemical composition of residual caries-affected dentin.
