Effectiveness of two methods for preparation of autologous platelet-rich plasma: an experimental study in rabbits

The purpose of this study was to compare the quantity and quality of platelets in platelet-rich plasma (PRP) samples prepared using either the single- or the double-centrifugation protocol. Ten adult white New Zealand rabbits were used. Ten ml of blood were drawn from each animal via cardiac puncture. Each blood sample was divided into two equal parts for PRP preparation: 5 ml of blood were centrifuged according to a single-centrifugation protocol (Group I), and 5 ml were centrifuged according to a double-centrifugation protocol (Group II). Manual platelet counts were performed on the whole blood and PRP samples of each group. Smears were also done on all samples in order to see the morphology of the platelets. The data obtained in the manual platelet count were submitted to statistical analysis (repeated measures ANOVA, Tukey, P<.05). The average whole blood platelet count was 446,389/μl. The PRP samples in Group II presented an average platelet amount significantly higher than that of Group I (1,986,875 ± 685,020/μl and 781,875 ± 217,693/μl, respectively). The PRP smears from Group II were the only one to present platelets with altered morphology (75% of the smears). A few lymphocytes with increased cytoplasm were observed in the PRP smears of both Groups I (25% of the smears) and II (62.5% of the smears). Within the limits of this study, it can be concluded that the double-centrifugation protocol resulted in higher platelet concentrations than did the single-centrifugation protocol. However, the double-centrifugation protocol caused alterations in platelet morphology and was more sensitive to small processing errors.

Human Platelet Polymorphism can be a genetic marker associated with HIV/HCV coinfection

To evaluate the associations of HPA polymorphisms -1, -3, and -5 with HIV/HCV coinfection were included in this study 60 HIV/HCV-coinfected patients from the Sao Paulo State health service centers. Data reported by Verdichio-Moraes et al. (2009: J. Med Virol 81:757-759) were used as the non-infected and HCV monoinfected groups. Human Platelet Polymorphism genotyping was performed in 60 Patients co-infected with HIV/HCV by PCR-SSP or PCR-RFLP. HIV subtyping and HCV genotyping was performed by RT-PCR followed sequencing. The data analyses were performed using the χ2 test or Fisher's Exact Test and the logistic regression model. Patients coinfected with HIV/HCV presented HCV either genotype 1 (78.3%) or non-1 (21.7%) and HIV either subtype B (85.0%) or non-B (15%). The Human Platelet Polymorphism-1a/1b genotype was more frequent (P < 0.05) in HIV/HCV coinfection than in HCV monoinfection and the allelic frequency of Human Platelet Polymorphism-5b in the Patients coinfected with HIV/HCV was higher (P < 0.05) than in HCV monoinfected cases and non-infected individuals. These data suggest that the presence of specific HPA allele on platelets could favor the existence of coinfection. On the other hand, Human Platelet Polymorphism-5a/5b was more frequent (P < 0.05) in HIV/HCV coinfected and HCV monoinfected groups than in the non-infected individuals, suggesting that this platelet genotype is related to HCV infection, regardless of HIV presence. Results suggest that the Human Platelet Polymorphism profile in HIV/HCV coinfected individuals differs from the one of both HCV monoinfected and non-infected population. So, the Human Platelet Polymorphism can be a genetic marker associated with HIV/HCV coinfection.

Thrombocytopenia and platelet hypoaggregation induced by Bothrops asper snake venom Toxins involved and their contribution to metalloproteinase-induced pulmonary hemorrhage

Thrombocytopenia and platelet dysfunction occur in patients bitten by Bothrops sp snakes in Latin America. An experimental model was developed in mice to study the effects of B. asper venom in platelet numbers and function. Intravenous administration of this venom induces rapid and prominent thrombocytopenia and ex vivo platelet hypoaggregation. The drop in platelet numbers was primarily due to aspercetin, a protein of the C-type lectin family which induces von Willebrand factor-mediated platelet aggregation/agglutination. In addition, the effect of class P-III hemorrhagic metalloproteinases on the microvessel wall also contributes to thrombocytopenia since jararhagin, a P-III metalloproteinase, reduced platelet counts. Hypoaggregation was associated with the action of procoagulant and defibrin(ogen)ating proteinases jararacussin-1 (a thrombin-like serine proteinase) and basparin A (a prothrombin activating metalloproteinase). At the doses which induced hypoaggregation, these enzymes caused defibrin(ogen)ation, increments in fibrin(ogen) degradation products and D-dimer and prolongation of the bleeding time. Incubation of B. asper venom with batimastat and α 2-macroglobulin abrogated the hypoaggregating activity, confirming the role of venom proteinases in this effect. Neither aspercetin nor the defibrin(ogen)ating and hypoaggregating components induced hemorrhage upon intravenous injection. However, aspercetin, but not the thrombin-like or the prothrombin-activating proteinases, potentiated the hemorrhagic activity of two hemorrhagic metalloproteinases in the lungs. © 2005 Schattauer GmbH, Stuttgart.

Efeitos do treinamento aeróbio no fator de crescimento endotelial vascular (VEGF), funções cognitivas e funcionalidade motora na Doença de Alzheimer

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aspectos radiográficos e densitométricos de fraturas experimentais do radio de cães tratadas com plasma rico em plaquetas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A standartized research protocol for platelet-rich plasma (PRP) preparation in rats

Introduction: The urgent need for studies using standardized protocols to evaluate the real biological effects of PRP has been emphasized by several authors. Objective: The purpose of this study was to standardize a methodology for autologous Platelet-Rich Plasma (PRP) preparation in rats. Material and methods: Twentyfour, 5 to 6-month-old, male rats, weighing 450 to 500 g were used. After general anesthesia, 3.15 ml of blood was collected from each animal, via cannulation of the jugular vein. A standardized technique of double centrifugation was used to prepare PRP. PRP samples and peripheral blood platelets were then manually counted using a Neubauer chamber. Student’s t-test was used to compare the differences between the number of platelets in peripheral blood and PRP samples (p < 0.05). In addition, PRP and peripheral blood smears were stained to see platelets’ morphology. Results: All surgical procedures were well tolerated by the animals and they were healthy during the entire experimental period. PRP samples showed higher significantly platelet concentrations than peripheral blood samples (2,677,583 and 683,680 respectively). Conclusion: Within the limits of this study, it can be concluded that the method used produced autologous PRP with appropriated platelet quantity and quality, in rats.

A proporção de plasma rico em plaquetas/enxerto ósseo autógeno pode influenciar o reparo ósseo de defeito de tamanho crítico: estudo radiográfico em calvária de ratos

Introduction: It has been suggested that bone grafts combined with platelet rich plasma (PRP) present greater density than bone grafts without PRP after healing process. However, this result may depend on proportion PRP/autogenous bone (AB) graft used. Objective: The purpose of this study was to analyze radiographically the influence of the proportion PRP/particulate autogenous bone (AB) graft on bone healing in surgically created criticalsize defects (CSD) in rat calvaria. Material and method: 50 rats were divided into 5 groups: C, AB, AB/PRP-50, AB/PRP-100 and AB/PRP-150. A 5 mm diameter CSD was created in the calvarium of each animal. In Group C the defect was filled by blood clot only. In Group AB the defect was filled with 0.01 mL of AB graft. In groups AB/ RP-50, AB/PRP-100 and AB/PRP-150 the defects were filled with 0.01 mL of AB graft combined with 50, 100 and 150 µL of PRP, respectively. All animals were euthanized at 30 days post-operative. Standardized radiographic images of the rat calvaria were obtained using Digora System (Soredex, Finland). The images were evaluated by three examiners using scores that indicated the percentage of radiopacity of the defect. The obtained data were subjected to statistical analysis (Kruskal-Wallis test, p < 0.05). Result: Group C presented radiopacity significantly lower than groups AB, AB/PRP-50, AB/PRP-100 and AB/PRP-150. Group AB/PRP-50 showed radiopacity significantly greater than groups AB/PRP-100 and AB. Conclusion: It can be concluded that the proportion PRP/AB graft influences bone healing in CSD in rat calvaria.

Importância da avaliação hematológica e sorológica (dot-blot ELISA) no diagnóstico de erliquiose em cães

This study compared the hematological and serological analysis of diagnosis of canine ehrlichiosis. The survey of Ehrlichia canis was performed through the evaluation of blood smears from 150 dogs. The serological test was performed on 12 samples selected by the platelet count (less than 170,000 platelets / uL). Serologic testing was performed with the Imunocomb kit - Dot-blot-ELISA. No cytoplasmatic inclusion characteristic of morula of E. canis was found in blood smears. In serologic testing, eight samples were positive for Ehrlichia canis, concluding that thrombocytopenia is an important hematological finding of ehrlichiosis diagnosis and the detection of Ehrlichia canis morulae is uncommon. The serological evaluation Dot-blot ELISA is an accurate and brief diagnosis method of canine ehrlichiosis, been the most appropriate to be used in veterinary practice routine.

Púrpura trombocitopênica idiopática: importância do cirurgião-dentista no diagnóstico

This paper presents a case report of a patient with idiopathic thrombocytopenic purpura (ITP) diagnosed in routine dental care. ITP is defined as a decrease in the number of circulating platelets and generally shows oral manifestations. In this light, we show that the general dental practitioner must be ready to detect ITP signs and symptoms, especially oral changes that are not of dental origin, so as to immediately prescribe the proper treatment. Without this treatment, this pathology may well be irreversible and fatal.

Acidose ruminal subaguda em ovinos Santa Inês: estudo clínico, laboratorial e avaliação da laminite por termografia infravermelha e radiologia digital

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação clínico-laboratorial e da expressão de metaloproteinases 2 e 9 de equinos com obstrução experimental do cólon menor

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação das alterações hemorrágicas e tromboembólicas em cães com doença renal crônica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da eficácia dos biocurativos em ratos wistar com lesões de pele causadas por queimaduras

Accidents caused by thermal, chemical, electrical or radioactive agents cause skin lesions causing burns of varying degrees. The therapeutic approach aims to restore damaged tissues and involves a wide range of products on the market. This study aims to evaluate the use of biological dressing, biotech product developed at the Blood Center of Botucatu / UNESP obtained from fresh frozen plasma or platelet concentrate with in vitro addition of thrombin and calcium gluconate. This addition in the platelet concentrate, intended to release the active growth factors of the platelets granules on the healing process. The study of the effectiveness of Platelet Gel home made in Wistar rats was established, in agreement with scald burns, comparing efficacy and cost of Platelet Gel with usual hospital -based treatment collagenase + chloramphenicol plus cost analysis through pharmacoeconomics. We used 25 Wistar rats were divided into 3 treatment groups: Group A, Collagenase + Chloramphenicol; Group B, Platelet Gel and C, control. The products were applied every other day for 30 days in animals. In group A, there was the presence of erythema and crust in all animals. The exudates was indentified 2/10 animals. For the Group B, we observed the presence of erythema and crust at all and no presence of exudates. In group C all the animals showed erythema with no presence of exudates and scab occurred in 1/10. Statistical analysis showed significant difference ( p < 0.0 ) for crust formation between Groups B and C. In the histological analysis, group A showed a slight amount of blood vessels and collagen fibers, moderate amounts of macrophages and fibroblasts was observed while B and C groups showed moderate amounts of blood vessels, macrophages and fibroblasts and discreet presence of collagen fibers. The re-epithelialization occurred in most animals of all groups without significant statistical differences. For the aspects of pharmacoeconomics, the platelet gel presented a better cost - effectiveness in relation to treatment based on collagenase / chloramphenicol. In light of the ethical aspects of the raw material is the result of spontaneous blood donation, the proposal should have biological dressings productions the responsibility of public blood transfusion centers for free distribution. This may point to the production chain of Brazilian blood banks like special blood components for use no intravenous.

Alterações no fenômeno da coagulação na vigência do mastocitoma cutâneo em cães

Pós-graduação em Medicina Veterinária - FCAV

Perfil de mutações e resistência do Vírus da Imunodeficiência Humana (HIV) em pacientes coinfectados com o Vírus da Hepatite B (VHB)

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB