Biochemical and biological evaluation of gyroxin isolated from Crotalus durissus terrificus venom

Barros, Luciana Curtolo de; Soares, Andreimar Martins; Costa, Fabíola Medeiros da; Rodrigues, Veridiana M.; Fuly, André Lopes; Giglio, José Roberto; Gallacci, Márcia; Thomazini-Santos, I. A.; Barraviera, Sílvia Regina Catharino Sartori; Barraviera, Benedito; Ferreira, Rui Seabra

doi:10.1590/s1678-91992011000100004

Cited by 52 publications

(39 citation statements)

References 40 publications

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“…At the time of use, the components were previously thawed, reconstituted, mixed, and applied according to the following protocol: The first bottle had fibrinogen obtained from buffalo blood (5 μl), the second contained calcium chloride (2 μl), and the last bottle had a thrombin-like fraction (1 μl), totaling 8 μl [16,[23][24][25][26].…”

Section: Fibrin Sealantmentioning

confidence: 99%

The new heterologous fibrin sealant in combination with low-level laser therapy (LLLT) in the repair of the buccal branch of the facial nerve

et al. 2016

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This study aimed to evaluate the effects of low-level laser therapy (LLLT) in the repair of the buccal branch of the facial nerve with two surgical techniques: end-to-end epineural suture and coaptation with heterologous fibrin sealant. Forty-two male Wistar rats were randomly divided into five groups: control group (CG) in which the buccal branch of the facial nerve was collected without injury; (2) experimental group with suture (EGS) and experimental group with fibrin (EGF): The buccal branch of the facial nerve was transected on both sides of the face. End-to-end suture was performed on the right side and fibrin sealant on the left side; (3) Experimental group with suture and laser (EGSL) and experimental group with fibrin and laser (EGFL). All animals underwent the same surgical procedures in the EGS and EGF groups, in combination with the application of LLLT (wavelength of 830 nm, 30 mW optical power output of potency, and energy density of 6 J/cm(2)). The animals of the five groups were euthanized at 5 weeks post-surgery and 10 weeks post-surgery. Axonal sprouting was observed in the distal stump of the facial nerve in all experimental groups. The observed morphology was similar to the fibers of the control group, with a predominance of myelinated fibers. In the final period of the experiment, the EGSL presented the closest results to the CG, in all variables measured, except in the axon area. Both surgical techniques analyzed were effective in the treatment of peripheral nerve injuries, where the use of fibrin sealant allowed the manipulation of the nerve stumps without trauma. LLLT exhibited satisfactory results on facial nerve regeneration, being therefore a useful technique to stimulate axonal regeneration process.

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Section: Fibrin Sealantmentioning

confidence: 99%

The new heterologous fibrin sealant in combination with low-level laser therapy (LLLT) in the repair of the buccal branch of the facial nerve

et al. 2016

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“…8a, b). Several reports have demonstrated that SVSPs are resistant to heat and/or pH denaturation (Costa et al 2009;Barros et al 2011;Menaldo et al 2012). However, none of these studies has reported the full maintenance of the enzymatic activity after pre-treating the enzymes on extreme pH and temperature conditions.…”

Section: Discussionmentioning

confidence: 99%

“…Similar results were reported for gyroxin from Crotalus durissus terrificus, which is partially and completely inhibited by Ba 2+ and Cu 2+ , respectively. Gyroxin has also undergone the influence of Mn 2+ , which was not tested for collinein-1 ( Barros et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Expression of a new serine protease from Crotalus durissus collilineatus venom in Pichia pastoris and functional comparison with the native enzyme

Boldrini-França

Rodrigues

Santos-Silva

et al. 2015

Appl Microbiol Biotechnol

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Snake venom serine proteases (SVSPs) act primarily on plasma proteins related to blood clotting and are considered promising for the treatment of several hemostatic disorders. We report the heterologous expression of a serine protease from Crotalus durissus collilineatus, named collinein-1, in Pichia pastoris, as well as the enzymatic comparative characterization of the toxin in native and recombinant forms. The complementary DNA (cDNA) encoding collinein-1 was amplified from cDNA library of C. d. collilineatus venom gland and cloned into the pPICZαA vector. The recombinant plasmid was used to transform cells of KM71H P. pastoris. Heterologous expression was induced by methanol and yielded 56 mg of recombinant collinein-1 (rCollinein-1) per liter of culture. The native collinein-1 was purified from C. d. collilineatus venom, and its identity was confirmed by amino acid sequencing. The native and recombinant enzymes showed similar effects upon bovine fibrinogen by releasing preferentially fibrinopeptide A. Although both enzymes have induced plasma coagulation, native Colinein-1 has shown higher coagulant activity. The serine proteases were able to hydrolyze the chromogenic substrates S-2222, S-2238, and S2302. Both enzymes showed high stability on different pH and temperature, and their esterase activities were inhibited in the presence of Zn2+ and Cu2+. The serine proteases showed similar k cat/K m values in enzyme kinetics assays, suggesting no significant differences in efficiency of these proteins to hydrolyze the substrate. These results demonstrated that rCollinein-1 was expressed with functional integrity on the evaluated parameters. The success in producing a functionally active recombinant SVSP may generate perspectives to their future therapeutic applications.

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“…[31][32][33] In contrast, the new heterologous fibrin sealant derived from Crotalus durissus terrificus snake venom contains a thrombin-like protein with an activity similar to other forms of thrombin but about 4000-fold more potent than human thrombin. [34][35] Therefore, our fibrin sealant is produced from buffalo fibrinogen, a species able to donate a large amount of blood-diminishing product costs. 36,37 Together, these findings led us to conclude that fibrin gel made from the venom of the rattlesnake seems to present adequate consistency and stability.…”

mentioning

confidence: 99%

A new heterologous fibrin sealant as a scaffold to cartilage repair—Experimental study and preliminary results

Barros

Yamada

et al. 2015

Exp Biol Med (Maywood)

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Autologous fibrin gel is commonly used as a scaffold for filling defects in articular cartilage. This biomaterial can also be used as a sealant to control small hemorrhages and is especially helpful in situations where tissue reparation capacity is limited. In particular, fibrin can act as a scaffold for various cell types because it can accommodate cell migration, differentiation, and proliferation. Despite knowledge of the advantages of this biomaterial and mastery of the techniques required for its application, the durability of several types of sealant at the site of injury remains questionable. Due to the importance of such data for evaluating the quality and efficiency of fibrin gel formulations on its use as a scaffold, this study sought to analyze the heterologous fibrin sealant developed from the venom of Crotalus durissus terrificus using studies in ovine experimental models. The fibrin gel developed from the venom of this snake was shown to act as a safe, stable, and durable scaffold for up to seven days, without causing adverse side effects. Fibrin gel produced from the venom of the Crotalus durissus terrificus snake possesses many clinical and surgical uses. It presents the potential to be used as a biomaterial to help repair skin lesions or control bleeding, and it may also be used as a scaffold when applied together with various cell types. The intralesional use of the fibrin gel from the venom of this snake may improve surgical and clinical treatments in addition to being inexpensive and adequately consistent, durable, and stable. The new heterologous fibrin sealant is a scaffold candidate to cartilage repair in this study.

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Biochemical and biological evaluation of gyroxin isolated from Crotalus durissus terrificus venom

Cited by 52 publications

References 40 publications

The new heterologous fibrin sealant in combination with low-level laser therapy (LLLT) in the repair of the buccal branch of the facial nerve

The new heterologous fibrin sealant in combination with low-level laser therapy (LLLT) in the repair of the buccal branch of the facial nerve

Expression of a new serine protease from Crotalus durissus collilineatus venom in Pichia pastoris and functional comparison with the native enzyme

A new heterologous fibrin sealant as a scaffold to cartilage repair—Experimental study and preliminary results

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