Abstract
Serine proteases (SPases) isolated from snake venoms present thrombin-like enzymatic activity that acts on the haemostatic system to convert fibrinogen into fibrin. The latter can be polymerized to acquire sealant and adhesive properties via the formation of a stable net. Based on these premises, researchers at CEVAP (Center for the Study of Venoms and Venomous Animals at UNESP) proposed, about 20 years ago, the prospect of a new sealant from the SPase obtained from Crotalus durissus terrificus venom and fibrinogen extracted from large animals. This product enables numerous biotechnological applications without presenting a risk of transmitting infectious diseases; furthermore, being genuinely Brazilian, its production cost is lower.Although it is known from the literature that SPases contribute significantly to tissue recovery this reconstruction has never been monitored dynamically in vivo in a non-invasive manner. We chose to study this phenomenon, for the first time in the world, utilizing near-infrared fluorescent imaging (fluo-IVP) in vivo in murine models of avulsion and re-implantation of motor neuron roots, utilizing the sealant as a biological scaffold in association with mesenchymal stem cells that must act synergetically to stimulate regeneration. This non-invasive strategy is based on the use of stained and synthesized fluorescent probes, of the enzyme-indocyanine (SPase-ICA) type, and of fluo-stained stem cells for in vivo monitoring, with the results observed by the modern imaging apparatus fluo-IVP, which is still unavailable in Brazil. The latter has greater potential for use in basic research as well as translational medicine. Based on these new molecular tools and stained cells, the reconstructive action from associating SPase with stem cells at the nervous system level will be evaluated in a dynamic, integrated and non-invasive manner.To elucidate and execute this translational project, it will be divided into two subprojects, namely: 1-Fluor-staining, by means of chemical synthesis, and evaluation of the bioactivity of the new fluorescent conjugates of SPase (SPase-ICA) as imaging probes in vivo; Fluo-staining the mesenchymal stem cells for in vivo monitoring together with fluor-stained sealant. 2-Imaging in vivo of nerve regeneration of rats avulsed at / from the interface of the central and peripheral nervous systems treated with mesenchymal stem cells and fluor-stained fibrin sealant as biological scaffolding.The present translational project, besides augmenting knowledge on the action mechanisms of the sealant with the stem cells, will have applications both in fundamental research (accompanying the tissue regeneration in vivo in a non-invasive manner), and in the clinical setting, by means of applied biotechnology (imaging in vivo with stained fluorescent enzyme employing the fibrin sealant as a biological scaffold and mesenchymal stem cells). The nerve regeneration subproject, in the case of experimental success, would then be immediately applicable in patients, in phase I clinical trials, after due approval by the Research Ethics Committee, since until the present date, there is still no successful gold standard restorative surgery for these patients. (AU)
Scientific publications
(17)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
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