Botucatu - CEVAP - Centro de Estudos de Venenos e Animais Peçonhentos

URI Permanente para esta coleçãohttps://hdl.handle.net/11449/253747

Navegar

Agora exibindo 1 - 20 de 284

Morphofunctional Improvement of the Facial Nerve and Muscles with Repair Using Heterologous Fibrin Biopolymer and Photobiomodulation
(2023-05-01) Bueno, Cleuber Rodrigo de Souza ; Tonin, Maria Clara Cassola ; Buchaim, Daniela Vieira ; Barraviera, Benedito ; Junior, Rui Seabra Ferreira ; Santos, Paulo Sérgio da Silva ; Reis, Carlos Henrique Bertoni ; Pastori, Cláudio Maldonado ; Pereira, Eliana de Souza Bastos Mazuqueli ; Nogueira, Dayane Maria Braz ; Cini, Marcelo Augusto ; Rosa Junior, Geraldo Marco ; Buchaim, Rogerio Leone ; Universidade de São Paulo (USP) ; University Center of Adamantina (UNIFAI) ; University of Marilia (UNIMAR ; Universidade Estadual Paulista (UNESP) ; University of Marilia (UNIMAR) ; University of West Paulista (UNOESTE) ; Faculty of the Midwest Paulista (FACOP)
Peripheral nerve injuries impair the patient’s functional capacity, including those occurring in the facial nerve, which require effective medical treatment. Thus, we investigated the use of heterologous fibrin biopolymer (HFB) in the repair of the buccal branch of the facial nerve (BBFN) associated with photobiomodulation (PBM), using a low-level laser (LLLT), analyzing the effects on axons, muscles facials, and functional recovery. This experimental study used twenty-one rats randomly divided into three groups of seven animals, using the BBFN bilaterally (the left nerve was used for LLLT): Control group—normal and laser (CGn and CGl); Denervated group—normal and laser (DGn and DGl); Experimental Repair Group—normal and laser (ERGn and ERGl). The photobiomodulation protocol began in the immediate postoperative period and continued for 5 weeks with a weekly application. After 6 weeks of the experiment, the BBFN and the perioral muscles were collected. A significant difference (p < 0.05) was observed in nerve fiber diameter (7.10 ± 0.25 µm and 8.00 ± 0.36 µm, respectively) and axon diameter (3.31 ± 0.19 µm and 4.07 ± 0.27 µm, respectively) between ERGn and ERGl. In the area of muscle fibers, ERGl was similar to GC. In the functional analysis, the ERGn and the ERGI (4.38 ± 0.10) and the ERGI (4.56 ± 0.11) showed parameters of normality. We show that HFB and PBM had positive effects on the morphological and functional stimulation of the buccal branch of the facial nerve, being an alternative and favorable for the regeneration of severe injuries.
Challenges and Opportunities in Clinical Diagnostic Routine of Envenomation Using Blood Plasma Proteomics
(2023-03-01) Cavalcante, Joeliton dos Santos ; de Almeida, Denis Emanuel Garcia ; Moraes, Micael Saggion ; Santos, Sophia Ribeiro ; Pincinato, Pedro Moriel ; Riciopo, Pedro Marques ; de Oliveira, Laís Lacerda B. ; Monteiro, Wuelton Marcelo ; Ferreira-Junior, Rui Seabra ; Universidade Estadual Paulista (UNESP) ; Federal University of Ceará (UFC) ; Amazonas State University
Specific and sensitive tools for the diagnosis and monitoring of accidents by venomous animals are urgently needed. Several diagnostic and monitoring assays have been developed; however, they have not yet reached the clinic. This has resulted in late diagnoses, which represents one of the main causes of progression from mild to severe disease. Human blood is a protein-rich biological fluid that is routinely collected in hospital settings for diagnostic purposes, which can translate research progress from the laboratory to the clinic. Although it is a limited view, blood plasma proteins provide information about the clinical picture of envenomation. Proteome disturbances in response to envenomation by venomous animals have been identified, allowing mass spectrometry (MS)-based plasma proteomics to emerge as a tool in a range of clinical diagnostics and disease management that can be applied to cases of venomous animal envenomation. Here, we provide a review of the state of the art on routine laboratory diagnoses of envenomation by snakes, scorpions, bees, and spiders, as well as a review of the diagnostic methods and the challenges encountered. We present the state of the art on clinical proteomics as the standardization of procedures to be performed within and between research laboratories, favoring a more excellent peptide coverage of candidate proteins for biomarkers. Therefore, the selection of a sample type and method of preparation should be very specific and based on the discovery of biomarkers in specific approaches. However, the sample collection protocol (e.g., collection tube type) and the processing procedure of the sample (e.g., clotting temperature, time allowed for clotting, and anticoagulant used) are equally important to eliminate any bias.
Neuroregeneration and immune response after neurorrhaphy are improved with the use of heterologous fibrin biopolymer in addition to suture repair alone
(2023-06-01) Tibúrcio, Felipe Cantore ; Muller, Kevin Silva ; Leite, Ana Paula Silveira ; de Oliveira, Igor Rodrigues Araújo ; Barraviera, Benedito ; Ferreira, Rui Seabra ; Padovani, Carlos Roberto ; Pinto, Carina Guidi ; Matheus, Selma Maria Michelin ; Universidade Estadual Paulista (UNESP)
Introduction/Aims: Peripheral nerve injuries result in impaired neuromuscular interactions, leading to morphological and functional alterations. Adjuvant suture repair methods have been used to improve nerve regeneration and modulate the immune response. Heterologous fibrin biopolymer (HFB), a scaffold with adhesive properties, plays a critical role in tissue repair. The aim of this study is to evaluate neuroregeneration and immune response focusing on neuromuscular recovery, using suture-associated HFB for sciatic nerve repair. Methods: Forty adult male Wistar rats were distributed into four groups (n = 10): C (control), only sciatic nerve location; D (denervated), neurotmesis and 6-mm gap removal and fixation stumps in subcutaneous tissue; S (suture), neurotmesis followed by suture; and SB (suture + HFB), neurotmesis followed by suture and HFB. Analysis of M2 macrophages (CD206+), as well as the morphology and morphometry of nerves, soleus muscle, and neuromuscular junctions (NMJs), were performed at 7 and 30 days after surgery. Results: The SB group had the highest M2 macrophage area in both periods. After 7 days, SB was the only group similar to the C group regarding the number of axons; furthermore, after 30 days, the SB group was closer to the C group concerning blood vessel and central myonuclear numbers, NMJ angle, and connective tissue volume. After 7 days, increases in nerve area, as well as the number and area of blood vessels, were also observed in SB. Discussion: HFB potentiates the immune response, increases axonal regeneration, induces angiogenesis, prevents severe muscle degeneration, and assists in NMJ recovery. In conclusion, suture-associated HFB has major implications for improved peripheral nerve repair.
LED photobiomodulation therapy associated with heterologous fibrin biopolymer optimizes wound repair in rats
(2023-06-01) Silva, Naldiana Cerqueira ; De Alexandria, Francisco Eugênio Deusdará ; Assis, Livia ; Filho, Antonio Luiz Martins Maia ; Ferreira, Rui Seabra ; Barraviera, Benedito ; Parizotto, Nivaldo Antonio ; Silva, José Figueredo ; Tim, Carla Roberta ; University Brazil ; UESPI ; Facid Wyden ; Universidade Estadual Paulista (UNESP)
Purpose: The skin is a complex organ that covers the entire surface of the body. Any situation that interrupts the integrity of the skin, whether by accidents, trauma, disease, or surgery, is defined as a skin wound. This study aimed to evaluate the effect of LED photobiomodulation therapy associated or not with heterologous fibrin biopolymer to repair skin wounds in rats. Methods: Full-thickness skin wounds were induced on the back of 84 Wistar rats and they were randomly divided into 4 groups: control group (CG), LED group (LED), heterologous fibrin biopolymer group (HFB), and LED + heterologous fibrin biopolymer group (LED + HFB). The animals were euthanized at 7, 14, and 21 days after surgery. In the macroscopic findings, it was observed in the control group, mild local inflammation, evidenced between the 5th and 6th day after the injury, not manifested in the other groups. The analysis of the wound reduction index showed that the treated groups presented greater wound reduction in the three periods evaluated. Results: The results of the degree of inflammation showed that the LED and LED + HFB groups showed a predominance of chronic inflammation, while the control group showed a predominance of diffuse acute inflammation. The quantity of type I collagen was bigger after 7 days in the LED and LED + HFB groups, and at 14 and 21 days this type of collagen appears more intensely in the group that used only HFB. For the traction force at 7 days of the experiment, the treated groups showed better performance, while at 14 and 21 days the group that used only the biopolymer obtained greater strength in the suture region. Conclusion: Treatments using HFB and LED, either isolated or associated, stimulated the wound repair process in rats. HFB promote collagen maturation.
Human dental pulp stem cell monolayer and spheroid therapy after spinal motor root avulsion in adult rats
(2023-03-01) Paes, Sabrina Moreira ; Castro, Mateus Vidigal de ; Barbosa, Rafael Maza ; Politti Cartarozzi, Luciana ; Coser, Lilian de Oliveira ; Kempe, Paula Regina Gelinski ; Decarli, Monize Caiado ; Moraes, Ângela Maria ; Barraviera, Benedito ; Ferreira Júnior, Rui Seabra ; Oliveira, Alexandre Leite Rodrigues de ; Universidade Estadual de Campinas (UNICAMP) ; Universidade Estadual Paulista (UNESP)
Spinal cord injuries result in severe neurological deficits and neuronal loss, with poor functional recovery. Mesenchymal stem cells have shown promising results; therefore the present objective of this work was to compare motor recovery after treatment with human dental pulp stem cells (hDPSC) cultivated in monolayer (2D) or as spheroids (3D), following avulsion and reimplantation of spinal motor roots in adult rats. Thus, 72 adult female Lewis rats were divided into 4 groups: avulsion (AV); avulsion followed by reimplantation (AR); avulsion associated with reimplant and 2D cell therapy (AR + 2D), and avulsion associated with reimplant and 3D cell therapy (AR + 3D). The application of the cells in 2D and 3D was performed by microsurgery, with subsequent functional assessment using a walking track test (Catwalk system), immunohistochemistry, neuronal survival, and qRT-PCR in 1-, 4-, and 12-weeks post-injury. The animals in the AR + 2D and AR + 3D groups showed the highest neuronal survival rates, and immunofluorescence revealed downregulation of GFAP, and Iba-1, with preservation of synaptophysin, indicating a reduction in glial reactivity, combined with the maintenance of pre-synaptic inputs. There was an increase in anti-inflammatory (IL-4, TGFβ) and a reduction of pro-inflammatory factors (IL-6, TNFα) in animals treated with reimplantation and hDPSC. As for the functional recovery, in all analyzed parameters, the AR + 2D group performed better and was superior to the avulsion alone. Overall, our results indicate that the 2D and 3D cell therapy approaches provide successful immunomodulation and motor recovery, consistent with advanced therapies after spinal cord injury.
Tissue Bioengineering with Fibrin Scaffolds and Deproteinized Bone Matrix Associated or Not with the Transoperative Laser Photobiomodulation Protocol
(2023-01-01) Pomini, Karina Torres ; Buchaim, Daniela Vieira ; Bighetti, Ana Carolina Cestari ; Hamzé, Abdul Latif ; Reis, Carlos Henrique Bertoni ; Duarte, Marco Antonio Húngaro ; Alcalde, Murilo Priori ; Barraviera, Benedito ; Júnior, Rui Seabra Ferreira ; de Souza, Alexandre Teixeira ; da Silva Santos, Paulo Sérgio ; Pilon, João Paulo Galletti ; de Marchi, Miguel Ângelo ; Nogueira, Dayane Maria Braz ; de Souza Bueno, Cleuber Rodrigo ; Soares, Wendel Cleber ; Buchaim, Rogerio Leone ; Universidade de São Paulo (USP) ; University of Marilia (UNIMAR) ; University Center of Adamantina (UNIFAI) ; Universidade Estadual Paulista (UNESP) ; UNINOVE University
Extending the range of use of the heterologous fibrin biopolymer, this pre-clinical study showed a new proportionality of its components directed to the formation of scaffold with a lower density of the resulting mesh to facilitate the infiltration of bone cells, and combined with therapy by laser photobiomodulation, in order to accelerate the repair process and decrease the morphofunctional recovery time. Thus, a transoperative protocol of laser photobiomodulation (L) was evaluated in critical bone defects filled with deproteinized bovine bone particles (P) associated with heterologous fibrin biopolymer (HF). The groups were: BCL (blood clot + laser); HF; HFL; PHF (P+HF); PHFL (P+HF+L). Microtomographically, bone volume (BV) at 14 days, was higher in the PHF and PHFL groups (10.45 ± 3.31 mm3 and 9.94 ± 1.51 mm3), significantly increasing in the BCL, HFL and PHFL groups. Histologically, in all experimental groups, the defects were not reestablished either in the external cortical bone or in the epidural, occurring only in partial bone repair. At 42 days, the bone area (BA) increased in all groups, being significantly higher in the laser-treated groups. The quantification of bone collagen fibers showed that the percentage of collagen fibers in the bone tissue was similar between the groups for each experimental period, but significantly higher at 42 days (35.71 ± 6.89%) compared to 14 days (18.94 ± 6.86%). It can be concluded that the results of the present study denote potential effects of laser radiation capable of inducing functional bone regeneration, through the synergistic combination of biomaterials and the new ratio of heterologous fibrin biopolymer components (1:1:1) was able to make the resulting fibrin mesh less dense and susceptible to cellular permeability. Thus, the best fibrinogen concentration should be evaluated to find the ideal heterologous fibrin scaffold.
Antineoplastic properties and pharmacological applications of Crotalus durissus terrificus snake venom
(2022-01-01) Alves, Brunna Fernanda Arraez ; Ferreira, Rui Seabra ; Universidade de São Paulo (USP) ; Universidade Estadual Paulista (UNESP)
Snake toxins are widely studied owing to their importance in snakebite accidents, a serious public health issue in tropical countries, and their broad therapeutic potential. Isolated fractions from venom produced by snakes of the genus Crotalus sp. present a wide variety of pharmacological uses such as antifungal, antiviral, antibacterial, and antitumor properties, among other therapeutic potentialities. Given the direct effect of this venom on tumor cells, isolation of its compounds is important for the characterization of its anticarcinogenic actions. Crotalus durissus terrificus venom and its toxins have been widely evaluated as potential candidates for the development of new antineoplastic therapies that are efficient against different tumor lines and cellular targets. This review highlights the venom toxins of this species, with a focus on their antineoplastic properties.
Acetylcholine receptors of the neuromuscular junctions present normal distribution after peripheral nerve injury and repair through nerve guidance associated with fibrin biopolymer
(2023-02-01) Leite, Ana Paula Silveira ; Pinto, Carina Guidi ; Tibúrcio, Felipe Cantore ; Muller, Kevin Silva ; Padovani, Carlos Roberto ; Barraviera, Benedito ; Junior, Rui Seabra Ferreira ; Leal, Claudenete Vieira ; Matsumura, Cintia Yuri ; Matheus, Selma Maria Michelin ; Universidade Estadual Paulista (UNESP) ; Universidade Estadual de Campinas (UNICAMP)
Peripheral nerve injuries (PNI) lead to alterations in the Agrin-LRP4-MuSK pathway. This results in disaggregation of AChRs and change from epsilon (mature, innervated) to gamma (immature, denervated) subunit. Tubulization technique has been shown to be effective for PNI repair and it also allows the use of adjuvants, such as fibrin biopolymer (FB). This study evaluated the effect of the association of tubulization with FB after PNI on AChRs and associated proteins. Fifty-two adults male Wistar rats were used, distributed in 4 experimental groups: Sham Control (S), Denervated Control (D); Tubulization (TB) and Tubulization + Fibrin Biopolymer (TB+FB). Catwalk was performed every 15 days. Ninety days after surgery the right soleus muscles and ischiatic nerves were submitted to the following analyses: (a) morphological and morphometric analysis of AChRs by confocal microscopy; (b) morphological and morphometric analysis of the ischiatic nerve; (c) protein quantification of AChRs: alpha, gama, and epsilon, of Schwann cells, agrin, LRP4, MuSK, rapsyn, MMP3, MyoD, myogenin, MURF1 and atrogin-1. The main results were about the NMJs that in the TB+FB group presented morphological and morphometric approximation (compactness index; area of the AChRs and motor plate) to the S group. In addition, there were also an increase of S100 and AChRε protein expression and a decrease of MyoD. These positive association resulted in AChRs stabilization that potentiate the neuromuscular regeneration, which strengthens the use of TB for severe injuries repair and the beneficial effect of FB, along with tubulization technique.
Effects of cilostazol, a Phosphodiesterase-3 inhibitor, on kidney function and redox imbalance in acute kidney injury caused by Bothrops alternatus venom
(2022-12-01) Marinho, Aline Diogo ; Coelho Jorge, Antônio Rafael ; Nogueira Junior, Francisco Assis ; Alison de Moraes Silveira, João ; Rocha, Danilo Galvão ; Negreiros Nunes Alves, Ana Paula ; Ferreira, Rui Seabra ; Bezerra Jorge, Roberta Jeane ; Azul Monteiro, Helena Serra ; Federal University of Ceara ; Universidade Estadual Paulista (UNESP)
The mechanisms of pathogenesis of acute kidney injury (AKI) in snakebites is multifactorial and involves hemodynamic disturbances, with release of free radical causing cytotoxic effects. The phosphodiesterase-3 (PDE3) inhibitor, Cilostazol, has been reported to provide protection against renal oxidative stress. Objective: We evaluated the protective effects of cilostazol against Bothrops alternatus snake venom (BaV)-induced nephrotoxicity. Methods: Wistar rat kidneys (n = 6, 260–300 g) were isolated and perfused with Krebs-Henseleit solution containing 6 g/100 mL of bovine serum albumin. After 30 min, the kidneys were perfused with BaV to a final concentration of 1 and 3 μg/mL, and subsequently evaluated for perfusion pressure (PP), renal vascular resistance (RVR), urinary flow (UF), glomerular filtration rate (GFR), and percentage of electrolyte tubular sodium and chloride transport (%TNa+, %TCl–). Oxidative stress and renal histological analyses were performed. Results: BaV caused a reduction in all the evaluated renal parameters (PP, RVR, GFR, UF, %TNa+, and %TCl–). Although only the effects on PP and UF were reversed with cilostazol treatment, the decrease in the malondialdehyde levels, without changes in glutathione levels, further reduced the venom-induced renal tissue changes. Conclusion: Our data suggest that PDE3 is involved in BaV-induced nephrotoxicity, as cilostazol administration significantly ameliorated these effects.
Experimental Bothrops atrox Envenomation: Blood Plasma Proteome Effects after Local Tissue Damage and Perspectives on Thromboinflammation
(2022-09-01) Cavalcante, Joeliton S. ; Brito, Ingrid Mayara da Cunha ; De Oliveira, Laudicéia Alves ; De Barros, Luciana Curtolo ; Almeida, Cayo ; Rossini, Bruno Cesar ; Sousa, Duaran Lopes ; Alves, Renata Sousa ; Jorge, Roberta Jeane Bezerra ; Santos, Lucilene Delazari dos ; Universidade Estadual Paulista (UNESP) ; Federal University of ABC ; Federal University of Ceará ; Federal University of Ceará (UFC)
The clinical manifestations of Bothrops atrox envenoming involve local and systemic changes, among which edema requires substantial attention due to its ability to progress to compartmental syndromes and sometimes cause tissue loss and amputations. However, the impact of edema on the poisoned body’s system has not been explored. Thus, the present study aimed to explore the systemic pathological and inflammatory events that are altered by intraplantar injection of B. atrox venom in a mouse model through hematologic, lipidic, and shotgun proteomics analysis. Plasma samples collected showed a greater abundance of proteins related to complement, coagulation, lipid system, platelet and neutrophil degranulation, and pathways related to cell death and ischemic tolerance. Interestingly, some proteins, in particular, Prdx2 (peroxiredoxin 2), Hba (hemoglobin subunit alpha), and F9 (Factor IX), increased according to the amount of venom injected. Our findings support that B. atrox venom activates multiple blood systems that are involved in thromboinflammation, an observation that may have implications for the pathophysiological progression of envenomations. Furthermore, we report for the first time a potential role of Prdx2, Hba, and F9 as potential markers of the severity of edema/inflammation in mice caused by B. atrox.
Proteomic study of facial melasma
(2022-11-01) Schaefer, Luiza Vasconcelos ; Pontes, Leticia Gomes de ; Cavassan, Nayara Rodrigues Vieira ; Santos, Lucilene Delazari dos ; Miot, Hélio Amante ; Universidade Estadual Paulista (UNESP) ; Universidade do Oeste Paulista
Launching a CDMO in Brazil aiming to develop biopharmaceuticals for clinical trials
(2022-01-01) Ferreira Junior, Rui Seabra ; Morales, Marcelo Marcos ; Barretti, Pasqual ; Barraviera, Benedito ; Universidade Estadual Paulista (UNESP) ; Universidade Federal do Rio de Janeiro (UFRJ) ; Ministry of Science Technology and Innovation (MCTI)
The innovation timeline is expensive, risky, competitive, time-consuming, and laborintensive. In order to overcome such challenges and optimize financial resources, pharmaceutical companies nowadays hire contract development and manufacturing organizations (CDMO) to help them. Based on the experience acquired first from the development of two biopharmaceuticals, the Heterologous Fibrin Sealant and the Apilic Antivenom, and more recently, during their respective clinical trials; the Center for the Study of Venoms and Venomous Animals (CEVAP) proposed to the Ministry of Health the creation of the first Brazilian CDMO. This groundbreaking venture will assist in converting a candidate molecule - from its discovery, proof of concept, product development, up to pilot batch production - into a product. The CDMO impact and legacy will be immense, offering service provision to the public and private sector by producing validated samples for clinical trials and academic training on translational research for those seeking a position in pharmaceutical industries and manufacturing platforms.
Effects of a Biocomplex Formed by Two Scaffold Biomaterials, Hydroxyapatite/Tricalcium Phosphate Ceramic and Fibrin Biopolymer, with Photobiomodulation, on Bone Repair
(2022-05-01) Reis, Carlos Henrique Bertoni ; Buchaim, Rogerio Leone ; Pomini, Karina Torres ; Hamzé, Abdul Latif ; Zattiti, Isabella Vasconcelos ; Duarte, Marco Antonio Hungaro ; Alcalde, Murilo Priori ; Barraviera, Benedito ; Ferreira Júnior, Rui Seabra ; Pontes, Fenelon Martinho Lima ; Grandini, Carlos Roberto ; Ortiz, Adriana de Cássia ; Fideles, Simone Ortiz Moura ; Eugênio, Renata Maria de Camargo ; Rosa Junior, Geraldo Marco ; Teixeira, Daniel de Bortoli ; Pereira, Eliana de Souza Bastos Mazuqueli ; Pilon, João Paulo Galletti ; Miglino, Maria Angelica ; Buchaim, Daniela Vieira ; University of Marilia (UNIMAR) ; Universidade de São Paulo (USP) ; Unisagrado University Center ; Universidade Estadual Paulista (UNESP) ; FIASP ; University Center of Adamantina (UniFAI)
There are several treatment methods available for bone repair, although the effectiveness becomes limited in cases of large defects. The objective of this pre-clinical protocol was to evalu-ate the grafting of hydroxyapatite/tricalcium phosphate (BCP) ceramic biomaterial (B; QualyBone BCP®, QualyLive, Amadora, Portugal) together with the heterologous fibrin biopolymer (FB; CE-VAP/UNESP Botucatu, Brazil) and with photobiomodulation (PBM; Laserpulse®, Ibramed, Amparo, Brazil) in the repair process of bone defects. Fifty-six rats were randomly divided into four groups of seven animals each: the biomaterial group (G1/B), the biomaterial plus FB group (G2/BFB); the bioma-terial plus PBM group (G3/B + PBM), and the biomaterial plus FB plus PBM group (G4/BFB + PBM). After anesthesia, a critical defect was performed in the center of the rats’ parietal bones, then filled and treated according to their respective groups. The rats were euthanized at 14 and 42 postopera-tive days. Histomorphologically, at 42 days, the G4/BFB + PBM group showed a more advanced maturation transition, with more organized and mature bone areas forming concentric lamellae. A birefringence analysis of collagen fibers also showed a more advanced degree of maturation for the G4/BFB + PBM group. In the comparison between the groups, in the two experimental periods (14 and 42 days), in relation to the percentage of formation of new bone tissue, a significant difference was found between all groups (G1/B (5.42 ± 1.12; 21.49 ± 4.74), G2/BFB (5.00 ± 0.94; 21.77 ± 2.83), G3/B + PBM (12.65 ± 1.78; 29.29 ± 2.93), and G4/BFB + PBM (12.65 ± 2.32; 31.38 ± 2.89)). It was concluded that the use of PBM with low-level laser therapy (LLLT) positively interfered in the repair process of bone defects previously filled with the biocomplex formed by the heterologous fibrin biopolymer associated with the synthetic ceramic of hydroxyapatite and tricalcium phosphate.
Using of Fibrin Sealant on Treatment for Tendon Lesion: Study in Vivo
(2022-01-01) Dutra Junior, Enéas de Freitas ; Hidd, S. M.C.M. ; Amaral, M. M. ; Maia Filho, A. L.M. ; Assis, L. ; Ferreira, R. S. ; Barraviera, B. ; Tim, C. R. ; Universidade Brasil ; UESPI ; Universidade Estadual Paulista (UNESP)
Tendon injuries are among the most common orthopaedic problems with long-term disability as a frequent consequence due to its prolonged healing time. Thus, the study has investigated the effect a new fibrin sealant (FS) derived from the venom of crotalus durissus terrificus in tendon repair. Therefore, 12 animals (±185.8 g) from Wistar lineage, showed average weight ± 185.8 g. All animals received the partial transected tendons and then they were separated in two group a random way: Lesion group (GL) and fibrin sealant group (GS). Immediately after the tendon injuries, 9 µL of fibrin sealant was applicated in each transected tendon, in order to form a stable clot with a dense fibrin network. The methodology of edema analysis was made in three stages: before the partial transected tendons, 24 h after tendon injuries induction and after 21 days of fibrin sealant treatment. The quantification of collagen was by slide colored with picrosirius red. We applied the Kolmogorov–Smirnov test to verify the normality between the groups. For the comparison between the groups the Student’s teste was applied for parametric samples and Mann–Whitney test for non-parametric samples with significance level of p < 0.05. The results showed that there were not significant difference between the groups in the first 24 h before the lesion, but the fibrin sealant treatment for tendon lesion can be promote the reduction of edemas after 21 days. The fibrin sealant group showed similar collagen deposition with the injury group, but with the collagen fibers more organized. The results suggest that the fibrin sealant is effectiveness to treat tendon injuries.
The Heterologous Fibrin Sealant and Aquatic Exercise Treatment of Tendon Injury in Rats
(2022-01-01) Hidd, S. M.C.M. ; Dutra, E. F. ; Tim, C. R. ; Filho, A. L.M.M. ; Assis, L. ; Ferreira, R. S. ; Barraviera, B. ; Amaral, M. M. ; Universidade Brasil ; UESPI ; Universidade Estadual Paulista (UNESP)
Acute rupture of the calcaneus tendon is relatively common, usually related to sports practice. In recent years, the number of researches in search of more efficient techniques, which induce the healing process, has been growing. The Fibrin Sealant Derived from Snake Venom (FSDSV) or Heterologous Sealant has been standing out in animal and human application for accelerating the repair of lesions, reducing the likelihood of hemorrhage and infectious diseases and having low production cost. Aquatic exercise also presents itself as an efficient strategy for rehabilitation, reducing pain and edemas, improving muscle properties and enhancing the repair process due to the numerous beneficial effects provided by the liquid medium. The aim of this research is to evaluate the use of fibrin sealant derived from snake venom associated with aquatic exercise in tendon repair. We used 84 rats of the Wistar strain, weighing between 170 and 250 g of weight who underwent surgery to induce partial rupture of the calcaneus tendon. The animals were randomly separated into four experimental groups. The technique used was the application of fibrin sealant and aquatic exercises according to the studied group. There was a greater reduction in the edema of the animals’ paws from the seventh day on in all treatments (p < 0.002), when compared to the control group. After 7 and 14 days of treatment, LE showed a greater reduction in the volume of edema (p = 0.03041) compared to the control. After 21 days, the (LS) showed a greater reduction in edema compared to the control group. It was possible to verify a higher collagen to LSE ratio in the evaluated period after 21 days of treatment. Thus, the heterologous fibrin sealant associated or not with aquatic exercise has a beneficial influence on tendon repair, becoming a propitious technique for future clinical applications.
A biocomplex to repair experimental critical size defects associated with photobiomodulation therapy
(2022-01-01) Buchaim, Daniela Vieira ; Andreo, Jesus Carlos ; Pomini, Karina Torres ; Barraviera, Benedito ; Ferreira, Rui Seabra ; Duarte, Marco Antonio Hungaro ; Alcalde, Murilo Priori ; Reis, Carlos Henrique Bertoni ; de Bortoli Teixeira, Daniel ; de Souza Bueno, Cleuber Rodrigo ; Detregiachi, Cláudia Rucco Penteado ; Araujo, Adriano Cressoni ; Buchaim, Rogério Leone ; University of Marilia (UNIMAR) ; University Center of Adamantina (UniFAI) ; Universidade Estadual Paulista (UNESP) ; Universidade de São Paulo (USP) ; Unisagrado University Center
Background: The association of scaffolds to repair extensive bone defects can contribute to their evolution and morphophysiological recomposition. The incorporation of particulate biomaterials into three-dimensional fibrin bioproducts together with photobiomodulation therapy (PBM) has potential and can improve regenerative medicine procedures. The objective of this experiment was to evaluate the effects of PBM therapy on critical size defects filled with xenogenic bone substitute associated with fibrin biopolymer. Methods: A critical defect of 8 mm was performed in 36 Wistar male adult rats that were divided into four groups. Groups BC and BC-PBM were defined as controls with defects filled by a clot (without or with PBM, respectively) and groups XS and XS-PBM that comprised those filled with biocomplex Bio-OssTM in association with fibrin biopolymer. PBM was applied immediately after the surgery and three times a week every other day, with the parameters: wavelength of 830 nm, energy density 6.2 J/cm2, output power 30 mW, beam area of 0.116 cm2, irradiance 0.258,62 W/cm2, energy/point 0.72 J, total energy 2.88 J. Fourteen and 42 days after the surgery, animals were euthanatized and subjected to microtomography, qualitative and quantitative histological analysis. Results: The BC-PBM and XS-PBM groups had a similar evolution in the tissue repair process, with a higher density of the volume of new formed bone in relation to the groups without PBM (p = 0.04086; p = 0.07093, respectively). Intense vascular proliferation and bone deposition around the biomaterial particles were observed in the animals of the groups in which biocomplex was applied (XS and XS-PBM). Conclusion: PBM therapy allowed an improvement in the formation of new bone, with a more organized deposition of collagen fibers in the defect area. Biocomplex favored the insertion and permanence of the particulate material in bone defects, creating a favorable microenvironment for accelerate repair process.
A fingerprint of plasma proteome alteration after local tissue damage induced by Bothrops leucurus snake venom in mice
(2022-02-20) Cavalcante, Joeliton dos Santos ; de Almeida, Cayo Antônio Soares ; Clasen, Milan Avila ; da Silva, Emerson Lucena ; de Barros, Luciana Curtolo ; Marinho, Aline Diogo ; Rossini, Bruno Cesar ; Marino, Celso Luís ; Carvalho, Paulo Costa ; Jorge, Roberta Jeane Bezerra ; dos Santos, Lucilene Delazari ; Universidade Estadual Paulista (UNESP) ; Federal University of ABC ; Oswaldo Cruz Foundation (FIOCRUZ) ; Federal University of Ceará (UFC)
Bothrops spp. is responsible for about 70% of snakebites in Brazil, causing a diverse and complex pathophysiological condition. Bothrops leucurus is the main species of medical relevance found in the Atlantic coast in the Brazilian Northeast region. The pathophysiological effects involved B. leucurus snakebite as well as the organism's reaction in response to this envenoming, it has not been explored yet. Thus, edema was induced in mice paw using 1.2, 2.5, and 5.0 μg of B. leucurus venom, the percentage of edema was measured 30 min after injection and the blood plasma was collected and analyzed by shotgun proteomic strategy. We identified 80 common plasma proteins with differential abundance among the experimental groups and we can understand the early aspects of this snake envenomation, regardless of the suggestive severity of an ophidian accident. The results showed B. leucurus venom triggers a thromboinflammation scenario where family's proteins of the Serpins, Apolipoproteins, Complement factors and Component subunits, Cathepsins, Kinases, Oxidoreductases, Proteases inhibitors, Proteases, Collagens, Growth factors are related to inflammation, complement and coagulation systems, modulators platelets and neutrophils, lipid and retinoid metabolism, oxidative stress and tissue repair. Our findings set precedents for future studies in the area of early diagnosis and/or treatment of snakebites. Significance: The physiopathological effects that the snake venoms can cause have been investigated through classical and reductionist tools, which allowed, so far, the identification of action mechanisms of individual components associated with specific tissue damage. The currently incomplete limitations of this knowledge must be expanded through new approaches, such as proteomics, which may represent a big leap in understanding the venom-modulated pathological process. The exploration of the complete protein set that suffer modifications by the simultaneous action of multiple toxins, provides a map of the establishment of physiopathological phenotypes, which favors the identification of multiple toxin targets, that may or may not act in synergy, as well as favoring the discovery of biomarkers and therapeutic targets for manifestations that are not neutralized by the antivenom.
Variant expression signatures of microRNAs and protein related to growth in a crossbreed between two strains of Nile tilapia (Oreochromis niloticus)
(2021-11-01) Herkenhoff, Marcos Edgar ; Bovolenta, Luiz A. ; Broedel, Oliver ; dos Santos, Lucilene D. ; de Oliveira, Arthur C. ; Chuffa, Luiz G.A. ; Ribeiro, Amanda de O. ; Lupi, Luiz A. ; Dias, Marco A.D. ; Hilsdorf, Alexandre W.S. ; Frohme, Marcus ; Pinhal, Danillo ; Universidade Estadual Paulista (UNESP) ; Technical University of Applied Sciences Wildau ; University of Mogi das Cruzes ; Federal University of Lavras
Nile tilapia (Oreochromis niloticus) is a species of worldwide importance for aquaculture. A crossbred lineage was developed through introgressive backcross breeding techniques and combines the high growth performance of the Chitralada (CHIT) lwith attractive reddish color of the Red Stirling (REDS) strains. Since the crossbreed has an unknown genetically improved background, the objective of this work was to characterize expression signatures that portray the advantageous phenotype of the crossbreeds. We characterized the microRNA transcriptome by high throughput sequencing (RNA-seq) and the proteome through mass spectrometry (ESI-Q-TOF-MS) and applied bioinformatics for the comparative analysis of such molecular data on the three strains. Crossbreed expressed a distinct set of miRNAs and proteins compared to the parents. They comprised several microRNAs regulate traits of economic interest. Proteomic profiles revealed differences between parental and crossbreed in expression of proteins associated with glycolisis. Distinctive miRNA and protein signatures contribute to the phenotype of crossbreed.
Phosphodiesterase-5 inhibitor sildenafil attenuates kidney injury induced by Bothrops alternatus snake venom
(2021-10-30) Jorge, Antônio Rafael Coelho ; Marinho, Aline Diogo ; Silveira, João Alison de Moraes ; Nogueira Junior, Francisco Assis ; de Aquino, Pedro Everson Alexandre ; Alves, Ana Paula Negreiros Nunes ; Jorge, Roberta Jeane Bezerra ; Ferreira Junior, Rui Seabra ; Monteiro, Helena Serra Azul ; Federal University of Ceara ; Universidade Estadual Paulista (UNESP)
Acute kidney injury pathogenesis in envenoming by snakes is multifactorial and involves immunologic reactions, hemodynamic disturbances, and direct nephrotoxicity. Sildenafil (SFC), a phosphodiesterase 5 inhibitor, has been reported to protect against pathological kidney changes. Objective: This study aimed to investigate the protective effect of sildenafil against Bothrops alternatus snake venom (BaV)-induced nephrotoxicity. Methods: Kidneys from Wistar rats (n = 6, weighing 260–300 g) were isolated and divided into four groups: (1) perfused with a modified Krebs-Henseleit solution (MKHS) containing 6 g% of bovine serum albumin; (2) administered 3 μg/mL SFC; (3) perfused with 3 μg/mL BaV; and (4) administered SFC + BaV, both at 3 μg/mL. Subsequently, the perfusion pressure (PP), renal vascular resistance (RVR), urinary flow (UF), glomerular filtration rate (GFR), and percentage of electrolyte tubular sodium and chloride transport (%TNa+, %TCl−, respectively) were evaluated. The cyclic guanosine monophosphate (cGMP) levels were analyzed in the perfusate, and the kidneys were removed to perform oxidative stress and histopathological analyses. Results: All renal parameters evaluated were reduced with BaV. In the SFC + BaV group, SFC restored PP to normal values and promoted a significant increase in %TNa+ and %TCl−. cGMP levels were increased in the SFC + BaV group. The oxidative stress biomarkers, malondialdehyde (MDA) and glutathione (GSH), were reduced by BaV. In the SFC + BaV group, a decrease in MDA without an increase in GSH was observed. These findings were confirmed by histological analysis, which showed improvement mainly in tubulis. Conclusion: Our data suggest the involvement of phosphodiesterase-5 and cGMP in BaV-induced nephrotoxicity since its effects were attenuated by the administration of SFC.
Crotoxin: A novel allergen to occupational anaphylaxis
(2016-06-01) De Pontes, Leticia Gomes ; Cavassan, Nayara Rodrigues Vieira ; Creste, Camila Fernanda Zorzella ; Junior, Airton Lourenço ; Arcuri, Helen Andrade ; Ferreira, Rui Seabra ; Barraviera, Benedito ; Gagete, Elaine ; Dos Santos, Lucilene Delazari ; Universidade Estadual Paulista (UNESP) ; Anaphylaxis Committee of the Brazilian Association of Allergy and Immunology ; National Institutes for Science and Technology

Navegar

Submissões Recentes