Comparison of single and double centrifugation methods for preparation of Platelet-Rich Plasma (PRP).

Objective: To compare the platelet count, platelet concentration/yield, residual Red blood cells (RBCs) and White blood cells (WBCs) counts in platelet-rich plasma (PRP) samples prepared from the single- and the double-centrifugation protocols. Methods: It was a Cross-Sectional study, conducted at the Department of Hematology & Transfusion Medicine, The Children's Hospital and UCHS, Lahore from October 2021 to January 2022 including 50 voluntary, healthy individuals of age 20-45 years of both genders, after taking informed consent. Complete blood count analysis of all participants was done initially by drawing 3ml blood in EDTA vial. From all the participants, 20 ml venous blood sample was taken in syringes containing tri-sodium citrate and then shifted to harvest tubes. Group-I comprised of PRP samples prepared by single- centrifugation method. While Group-II samples were prepared by Double-centrifugation method consisting of soft and hard spin. The platelet, RBC and WBC counts in prepared PRP samples were determined by using automated SYSMEX XP-100 hematology analyzer. Platelet yield or Platelet concentration (%) was calculated for samples using formula. The data analysis was done using SPSS version 23. Results: The mean PRP platelet count in Group-I was 594.6±157.4×103/µl whereas in Group-II was 923.06 ± 127.58×103/µl. In Group-I, the mean platelet concentration/yield in PRP was 175.75 ± 55.08% while in Group-II, it was 276.78 ± 112.7%. Significant difference was observed between the platelet counts and platelet concentration/yields from the PRP samples of two Group-s (p < 0.01). Significant difference between the WBCs count was also observed (p < 0.01) with higher WBCs in Group- I PRP. Residual RBCs were almost same among two Group-s. Conclusions: The double centrifugation protocol resulted in higher platelet quantity and yield with less contamination by red and white blood cells than did the single centrifugation protocol for PRP preparation. So, double centrifugation method is beneficial in preparation of autologous as well as allogenic PRP.

Intradiscal Platelet-Rich Plasma Injection for Discogenic Low Back Pain and Correlation with Platelet Concentration: A Prospective Clinical Trial.

OBJECTIVE: Discogenic pain is common cause of low back ache and may result in significant morbidity. Platelet-rich plasma (PRP) is an upcoming regenerative therapy that has treatment potential for this condition. The objective of this study was to correlate platelet concentration in intradiscal PRP injection with improvement in low back pain and functional status at three and six months. DESIGN: Prospective single-arm interventional study. SETTING: Outpatient pain clinic and operation theater. SUBJECTS: Twenty-five patients with discogenic pain diagnosed by clinical means and imaging with confirmation by provocative discography were recruited. METHODS: The patients received PRP injection at a single or multiple disc levels. Preprocedure numerical rating scale (NRS) pain scores and Oswestry Disability Index (ODI) scores were calculated. Platelet counts of patients and PRP samples were measured. At three and six months postprocedure, NRS and ODI scores were measured, and improvement in these scores was correlated with platelet concentrations in the PRP sample. RESULTS: Twenty patients completed the study. The improvement in NRS and ODI scores positively correlated with platelet concentrations in the PRP sample. We determined the correlation coefficient (r) of platelet concentrations with a reduction in NRS at three months (r = 0.65) and six months (r = 0.73) and in ODI score at three months (r = 0.72) and six months (r = 0.7). CONCLUSIONS: This study supports the use of intradiscal PRP for treatment of discogenic pain with preferably higher platelet counts to elicit a favorable response.

Platelet-Rich Plasma Therapy: An Effective Approach for Managing Knee Osteoarthritis.

Platelet-rich plasma (PRP) is a promising non-invasive therapeutic intervention for knee osteoarthritis (KOA) that has generated significant interest due to anecdotal accounts of its efficacy, resulting in reduced recovery time in various orthopedic interventions. This systematic review examines the effectiveness of PRP in managing KOA. Specifically, it seeks to determine the extent to which PRP can treat KOA patients effectively, alleviate KOA symptoms, and improve patient outcomes. Additionally, the review aims to identify the optimal concentration and composition of PRP required to achieve therapeutic results in KOA. Furthermore, the review investigates whether PRP can modify the synovial environment structurally and immunologically to improve outcomes in KOA patients. We conducted a comprehensive literature search on PubMed, Orthogate, Clinicaltrials.gov, and Embase of clinical trials investigating PRP treatment in KOA patients in the last five years. The results indicated that PRP is effective in treating KOA patients. Evidence shows that PRP therapy can alleviate pain, enhance joint function, increase range of motion, and improve mobility in KOA patients. PRP was effective in treating KOA when the mean platelet concentration of PRP treatment was 4.83 to 5.91 times higher than the baseline whole blood platelet concentration. However, studies investigating PRP with a mean platelet concentration of 3.48 to 4.04 times higher than baseline failed to demonstrate statistically significant improvements. PRP therapy slowed down KOA progression, which validates its effectiveness in impeding further structural damage and arresting the degenerative impact of the disease. Nonetheless, further investigation is necessary to examine how PRP therapy can modify the progression of the disease. Furthermore, future research should identify the most effective platelet concentration levels that provide optimal symptom relief. There is a need for further research to identify the specific PRP configuration that is most pertinent in a clinical setting, as there is a lack of standardization in PRP manufacturing protocols, including the variety of experimental setups and dosing schedules utilized in different studies.

Circulating platelet concentration is associated with bone mineral density in women.

In this cross-sectional study, enrollment included 818 female adults undergoing bone mineral density (BMD) assessment during the health examination. Subjects with osteoporosis had the lowest circulating platelet concentrations. The circulating platelet concentration was positively correlated with BMD. A high platelet concentration had independently low odds of osteoporosis. PURPOSE: Platelets play an important role in bone metabolism. However, the association between circulating platelet counts and bone mineral density (BMD) has been inconsistently reported. We aimed to investigate the relationship between platelet counts and osteoporosis in Chinese women. METHODS: In this cross-sectional study, a total of 818 female adults who underwent BMD assessment during the health examination were enrolled. Blood cell counts and biochemistry data were recorded. RESULTS: Subjects with osteoporosis had the lowest platelet counts (238 ± 59 × 109/L) compared with subjects with osteopenia (256 ± 64 × 109/L) and a normal BMD (269 ± 76 × 109/L, P < 0.001). The circulating platelet concentration was positively correlated with the BMD of the lumbar spine (r = 0.195, P < 0.001), left hip (r = 0.145, P < 0.001), and right hip (r = 0.149, P < 0.001). According to the receiver operating characteristic curve, the cutoff platelet concentration for differentiating osteoporosis was 260 × 109/L. A high platelet concentration had significantly low odds of osteoporosis after adjusting for other covariates (odds ratio = 0.574, 95% confidence interval: 0.346‒0.953, P = 0.032). CONCLUSION: The circulating platelet concentration was significantly correlated with BMD in Chinese women.

Evaluation of Platelet and Leukocyte Counts in Canine Platelet-Rich Plasma Obtained After Successive Blood Collections From the Same Patient and the Effects of Freezing on the Concentration of Growth Factors Present in It.

Objective: The purpose of this study was 2-fold: to evaluate whether the timing of collection influences the platelet counts and leukocyte counts of PRP samples, and to evaluate growth factor concentrations in canine PRP after freezing and storage without a preservation agent for 6 months of time. Materials and Methods: Whole blood collection was performed three times over the course of 4 weeks. All PRP samples were evaluated with a CBC analysis. The PRP samples were frozen and stored without a preservation agent for the duration of the 4-week study. Results: The platelet and leukocyte counts were not statistically significant between the timing of blood draws over the course of 4 weeks. All three growth factors were present in measurable quantities after freezing and storage for 6 months without a preservation agent. Clinical Relevance: PDGF, TGF-ß1, and VEGF were all present in measurable quantities. Furthermore, PDGF and TGF-ß1 were correlated with platelet count of the final PRP. VEGF concentrations were able to be quantified. We correctly hypothesized growth factor concentrations would be present and measurable in canine PRP frozen and stored without a preservation agent for 6 months.

Does platelet aggregation have any importance in fetal growth restriction pregnancies?

OBJECTIVES: The aim of the study was to evaluate platelet (PLT) concentration, mean platelet volume (MPV), PLT aggregation and its velocity in pregnancy complicated with fetal growth restriction (FGR) and to analyze the PLT aggregation according to the gestational age and Doppler velocimetry. MATERIAL AND METHODS: The study group included 29 pregnant women diagnosed with FGR. The control group-consisted of 27 females in uncomplicated pregnancy. Then both groups were divided according to the gestational week (< and ≥ 36 weeks) and Doppler velocimetry results. The adenosine diphosphate (ADP) induced PLT aggregation was performed with the help of the electrical impedance. RESULTS: There was a significant positive correlation between gestational age and PLT aggregation and between gestational age and velocity of PLT aggregation in FGR. Patients with FGR ≥ 36 weeks of gestation had 73% higher PLT aggregation than control group. Within the FGR group, the PLT aggregation was 135% higher in pregnancies ≥ 36 weeks as compared to < 36 weeks of gestation. In FGR pregnancies ≥ 36 weeks with impaired flow in both uterine arteries (UtA), 2.3-fold higher PLT aggregation was found as compared to FGR patients with normal flow or abnormal flow in one UtA. CONCLUSIONS: The increased PLT aggregation in FGR is related to gestational week and occurs in pregnancies ≥ 36 weeks of gestation. The PLT hyperaggregability in growth-restricted pregnancies is associated with abnormal Doppler velocimetry in both UtA, comparing to patients with altered blood flow in one UtA or normal pulsatility index in both UtA, suggesting the PLT activation due to impaired uteroplacental circulation.

Therapeutic Efficacy of Autologous Platelet Concentrate Injection on Macular Holes with High Myopia, Large Macular Holes, or Recurrent Macular Holes: A Multicenter Randomized Controlled Trial.

We aimed to evaluate the anatomical and functional outcomes of pars-plana vitrectomy (PPV) with or without autologous platelet concentrate (APC) injection in patients with recurrent macular holes (MHs), large MHs, or MHs with high myopia. This multicenter, prospective, interventional randomized controlled trial was conducted from March 2017 to April 2020. Participants were randomly allocated to a PPV group or a PPV+APC group. All participants underwent standard 25-gauge PPV, and eyes in the PPV+APC group underwent PPV with intravitreal APC injection before air-gas exchange. A total of 117 patients were enrolled (PPV group: n = 59, PPV+APC group: n = 58). Hole closure was achieved in 47 participants (79.7%) in the PPV group and 52 participants (89.7%) in the PPV+APC group. There were no between-group differences in the anatomical closure rate or functional outcomes including best-corrected visual acuity, metamorphopsia, pattern-reversal visual evoked potential, or Visual Function Questionnaire-25 score. The use of APC injection does not improve the anatomical and functional outcomes of surgery for large MHs, recurrent MHs, or MHs with high myopia. The adjunctive use of APC can be considered in selected cases because it is not inferior to conventional MH surgery, is relatively simple to perform, and is not affected by the surgeon's skill.

Optimized centrifugation preparation of the platelet rich plasma: Literature review.

Nowadays, the Platelet Rich Plasma (PRP) is frequently used for its therapeutic effects on wound healing, and this due to secretion of many growth factors. However, no standardized procedure has been set up. The aim of this article is to check the various preparations (centrifugation time and speed). This review recorded all the international articles published between 2007 and 2018, for which the assessment criteria were the platelet concentration and/or the growth factor release rate. A multitude of protocols has been looked at with a simple or double centrifugation. All of them have shown an increase in the platelet concentration allowing a therapeutic effect. However, when the centrifugation force is extended, platelets can possibly be altered. The diversity of methods can be linked to the use of various centrifuges. A procedure with simple centrifugation would be a good compromise for the day-to-day use of the PRP in surgery.

Validation and Characterization of Platelet-Rich Plasma in the Feline: A Prospective Analysis.

Objective: To quantitate key parameters of the platelet-rich plasma (PRP) product from a commercially available system in healthy, adult felines. Materials and methods: A prospective study was performed from January 2019 to April 2019. 11 adult, healthy cats were used to prospectively analyze a commercially available PRP system. A whole blood sample and a PRP sample that was processed immediately following blood draw according to the manufacturer's protocol were collected from each cat. The mean whole blood and PRP product platelet, RBC, WBC, neutrophil, monocyte, and lymphocyte concentrations were determined. The mean PRP product values were compared to the mean whole blood baseline values using a paired t-test with significance established at p = 0.05. Results: Mean platelet concentration was significantly increased (p = 0.0155). Mean RBC concentration was significantly decreased (p < 0.0001). Mean neutrophil concentration was significantly decreased (p < 0.0001). There was no statistically significant difference in mean WBC, monocyte, and lymphocyte concentrations. Clinical Relevance: The analyzed PRP system increased platelet concentration, while significantly reducing the RBC and neutrophil concentrations. Further study is warranted to determine the clinical applications and efficacy of PRP in felines, and the ideal concentrations of and relationships between platelets, red blood cells, and leukocytes needed for therapeutic effect.

Osteogenic capacity of diluted platelet-rich plasma in ectopic bone-forming model: Benefits for bone regeneration.

Platelet-rich plasma (PRP) with normal and below-normal physiological concentrations of platelets is designated as diluted PRP (dPRP). The aims of this study are to evaluate whether bone mineral matrix in combination with dPRP possesses osteogenic capacity; and whether the differences in dynamics and osteogenic process pattern depend on different platelet concentrations, to what extent, and also what could be benefits for bone regeneration in clinical practice. Three types of implants were made: BMM-bone mineral matrix alone; dPRP/10-bone mineral matrix mixed with dPRP (concentration of platelets 10 times lower than physiological level) and dPRP/3-bone mineral matrix mixed with dPRP (concentration of platelets 3 times lower than physiological level). A subcutaneous implantation model in Balb/c mice was used. The implants were analyzed using expression analysis of bone-related genes, histochemical, immunohistochemical and histomorphometrical analysis. All types of implants induced creation of necessary preconditions for supporting osteogenic processes, but did not induce visible young bone growth. Implant types dPRP/10 and dPRP/3 showed very similar and significantly better stimulatory effects on osteogenic processes than bone matrix alone. In this study, significant ectopic osteogenic potential of concentration of platelets in PRP that are lower than physiological level in blood plasma in combination with bone mineral matrix was demonstrated. Diluted platelet-rich plasma could be a promising and useful adjuvant therapeutic agent in bone regeneration.

The significance of changes in platelet concentration during the early phase after severe burn injury in a Chinese mass casualty.

BACKGROUND: Changes in platelet concentration are common in severe burn patients. Platelets play a key role in the course of disease. This study aims to explore the significance of platelet concentration during the course of the disease in victims of a mass burn casualty. METHODS: A total of 180 patients were involved in the "8.2" Kunshan explosion accident in China. The examined data included age, gender, total burn area (% TBSA), third-degree burn area (% TBSA), and platelet concentration within the first 5 days after the burn injury. The patients were divided into two groups according to four indicators (resuscitation, acute respiratory distress syndrome, acute kidney injury, septic shock). We collected several types of data for the patients and divided the patients into a complication group and non-complication group according to the diagnostic criteria. We analyzed the platelet concentration of the two groups using t tests to determine whether significant differences were present. P values < 0.05 were considered statistically significant. RESULTS: The group with successful resuscitation had higher platelet concentration than the failure group on day 3 and day 5. The patients who suffered from acute kidney injury (AKI) and septic shock had a lower platelet concentration than non-sufferers on day 3 and day 5. CONCLUSIONS: The platelet concentration of burn patients can dynamically reflect the pathophysiological changes of the body. It can be used as an early objective indicator of prognosis in mass burn casualty cases.

The influence of environmental variables on platelet concentration in horse platelet-rich plasma.

Platelet-rich plasma (PRP) commonly refers to blood products which contain a higher platelet (PLT) concentration as compared to normal plasma. Autologous PRP has been shown to be safe and effective in promoting the natural processes of soft tissue healing or reconstruction in humans and horses. Variability in PLT concentration has been observed in practice between PRP preparations from different patients or from the same individual under different conditions. A change in PLT concentration could modify PRP efficacy in routine applications. The aim of this study was to test the influence of environmental, individual and agonistic variables on the PLT concentration of PRP in horses. Six healthy Standardbred mares were exposed to six different variables with a one-week washout period between variables, and PRP was subsequently obtained from each horse. The variables were time of withdrawal during the day (morning/evening), hydration status (overhydration/dehydration) treatment with anti-inflammatory drugs and training periods on a treadmill. The platelet concentration was significantly higher in horses treated with a non-steroidal anti-inflammatory drug (P = 0.03). The leukocyte concentration increased 2-9 fold with respect to whole blood in the PRP which was obtained after exposure to all the variable considered. Environmental variation in platelet concentration should be taken into consideration during PRP preparation.

Human embryonic stem cells and macroporous calcium phosphate construct for bone regeneration in cranial defects in rats.

Human embryonic stem cells (hESCs) are an exciting cell source as they offer an unlimited supply of cells that can differentiate into all cell types for regenerative medicine applications. To date, there has been no report on hESCs with calcium phosphate cement (CPC) scaffolds for bone regeneration in vivo. The objectives of this study were to: (i) investigate hESCs for bone regeneration in vivo in critical-sized cranial defects in rats; and (ii) determine the effects of cell seeding and platelets in macroporous CPC on new bone and blood vessel formation. hESCs were cultured to yield mesenchymal stem cells (MSCs), which underwent osteogenic differentiation. Four groups were tested in rats: (i) CPC control without cells; (ii) CPC with hESC-derived MSCs (CPC+hESC-MSC); (iii) CPC with hESC-MSCs and 30% human platelet concentrate (hPC) (CPC+hESC-MSC+30% hPC); and (iv) CPC+hESC-MSC+50% hPC. In vitro, MSCs were derived from embryoid bodies of hESCs. Cells on CPC were differentiated into the osteogenic lineage, with highly elevated alkaline phosphatase and osteocalcin expressions, as well as mineralization. At 12weeks in vivo, the groups with hESC-MSCs and hPC had three times as much new bone as, and twice the blood vessel density of, the CPC control. The new bone in the defects contained osteocytes and blood vessels, and the new bone front was lined with osteoblasts. The group with 30% hPC and hESC-MSCs had a blood vessel density that was 49% greater than the hESC-MSC group without hPC, likely due to the various growth factors in the platelets enhancing both new bone and blood vessel formation. In conclusion, hESCs are promising for bone tissue engineering, and hPC can enhance new bone and blood vessel formation. Macroporous CPC with hESC-MSCs and hPC may be useful for bone regeneration in craniofacial and orthopedic applications.

Enumeration of feline platelets in ethylenediamine tetra-acetic acid anticoagulated blood with the ADVIA 2120 system and two manual methods: Leucoplate and Thrombo-TIC.

A manual method (Thrombo-TIC; Bioanalytic GmbH, Umkirch/Freiburg, Germany) was advertised to disaggregate platelet clumps and to make human platelets spherical to improve platelet enumeration. The current study's hypothesis was that this method would perform better than current methods for feline blood anticoagulated with ethylenediamine tetra-acetic acid (EDTA), which often contains platelet aggregates. Platelet concentrations (PLTs) were determined in 21 feline blood samples by 3 methods. Thrombo-TIC was compared to the manual method (Leucoplate; Sobioda, Montbonnot-Saint-Martin, France) currently used in the authors' laboratory along with an ADVIA 2120 (Siemens AG, Eschborn, Germany) optical platelet concentration. Feline blood samples often contained platelet aggregates. ADVIA flagged for platelet aggregates in 11 of the 21 feline blood samples, and examination of blood smear revealed platelet aggregates in 14 of the 21 samples. The hemocytometers displayed more platelet aggregates with the Thrombo-TIC method than with Leucoplate. The method giving the greatest PLT was considered most accurate. The Leucoplate median PLT (238 × 10(9)/1) was greater than Thrombo-TIC (202 × 10(9)/1) or ADVIA (157 × 10(9)/1). Intra-assay precision was determined for the 3 methods using the 21 feline blood samples. Median Thrombo-TIC and Leucoplate precision (7.4% and 7.3% coefficient of variation [CV], respectively) were similar and not much worse than ADVIA (5.9% CV). The Thrombo-TIC method did not appear to perform better than the current manual method (Leucoplate). Leucoplate appeared least affected by platelet aggregation in feline blood. The ADVIA automated PLT appeared to be most negatively affected by platelet aggregation. The Thrombo-TIC method did not appear to prevent platelet aggregation in feline EDTA blood samples and, thus, is not recommended for such use.

Method to obtain platelet-rich plasma from rabbits (Oryctolagus cuniculus ) / Método de obtenção de plasma rico em plaquetas de coelhos (Oryctolagus cuniculus)

Platelet-rich plasma (PRP) is a product easy and inxpesnsive, and stands out to for its growth factors in tissue repair. To obtain PRP, centrifugation of whole blood is made with specific time and gravitational forces. Thus, the present work aimed to study a method of double centrifugation to obtain PRP in order to evaluate the effective increase of platelet concentration in the final product, the preparation of PRP gel, and to optimize preparation time of the final sample. Fifteen female White New Zealand rabbits underwent blood sampling for the preparation of PRP. Samples were separated in two sterile tubes containing sodium citrate. Tubes were submitted to the double centrifugation protocol, with lid closed and 1600 revolutions per minute (rpm) for 10 minutes, resulting in the separation of red blood cells, plasma with platelets and leucocytes. After were opened and plasma was pipetted and transferred into another sterile tube. Plasma was centrifuged again at 2000rpm for 10 minutes; as a result it was split into two parts: on the top, consisting of platelet-poor plasma (PPP) and at the bottom of the platelet button. Part of the PPP was discarded so that only 1ml remained in the tube along with the platelet button. This material was gently agitated to promote platelets resuspension and activated when added 0.3ml of calcium gluconate, resulting in PRP gel. Double centrifugation protocol was able to make platelet concentration 3 times higher in relation to the initial blood sample. The volume of calcium gluconate used for platelet activation was 0.3ml, and was sufficient to coagulate the sample. Coagulation time ranged from 8 to 20 minutes, with an average of 17.6 minutes. Therefore, time of blood centrifugation until to obtain PRP gel took only 40 minutes...

O plasma rico em plaquetas (PRP) é um produto de fácil obtenção a baixo custo, destacando-se pelos seus fatores de crescimento na reparação tecidual. Para obtenção do PRP, a centrifugação do sangue total é realizada com tempos e forças gravitacionais específicas. Assim, o presente trabalho teve por objetivo estudar o método da dupla centrifugação para obtenção do PRP, a fim de avaliar a eficácia de aumento da concentração de plaquetas no produto final, a preparação de gel de PRP e otimizar o tempo de preparação da amostra final. Quinze coelhos Nova Zelândia Branco, fêmeas, foram submetidos à coleta de sangue para a preparação de PRP. As amostras foram separadas em dois tubos estéreis contendo citrato de sódio. Os tubos foram submetidos ao protoloco de dupla centrifugação, com a tampa fechada a 1600 revoluções por minuto (rpm) durante 10 minutos, resultando na separação dos glóbulos vermelhos, plaquetas e plasma contendo os leucócitos. Na sequência, foram destapados para pipetar o plasma e transferí-lo para outro tubo de estéril. O plasma foi novamente centrifugado a 2000pm durante 10 minutos, resultando em duas partes: a parte superior, que consistia em plasma pobre em plaquetas (PPP) e a parte inferior do botão de plaquetas. Parte PPP foi descartado de modo que apenas 1ml de PPP permaneceu no frasco juntamente com o botão de plaquetas. Este material foi agitado suavemente para promover a ressuspensão das plaquetas, o que resultou na produção de PRP. O protocolo de centrifugação dupla foi capaz de promover a concentração de plaquetas 3 vezes maior em relação à amostra de sangue inicial. O volume de gluconato de cálcio utilizado para a ativação das plaquetas foi de 0,3ml, e foi suficiente para coagular a amostra, e o tempo de coagulação variou de 8 a 20 minutos, com uma média de 17,6 minutos. O tempo da centrifugação do sangue até a obtenção do PRP gel levou apenas 40 minutos...

Plasma rico em plaquetas de ratos wistar / Platelet-rich plasma of wistar rats

Uma das biotecnologias com propósito relevante à saúde humana é o plasma rico em plaquetas (PRP) o qual, não há registro protocolar padronizado para Rattus novergicus (Wistar). De fato, a falta de padronização do método leva pesquisadores à dúvida sob qual protocolo seguir para os melhores resultados referente as concentrações e mudanças morfológicas plaquetárias. Tal indeterminação promove morosidade nas pesquisas científicas. Frente a conjectura, usou-se o mínimo de animais possíveis em experimentação duplo-cego, retirando-se 3,2 mL de sangue de cada animal por pulsão cardíaca o qual posteriormente, foi testado em três protocolos distintos. Cada protocolo foi designado como P1; P2; P3 e o grupo controle designado PSP. Desta forma, i. P1 foi submetido a uma única centrifugação de 160G por 6min resultando em 100 µL de PRP; ii. P2 foi resignado a dupla centrifugação sendo a primeira etapa 160G por 20min e a segunda etapa 400G por 15 min, resultando em 100 µL de PRP e iii. P3 destinado a dupla centrifugação sendo a primeira etapa 300G por 10 min e a segunda etapa 640G por 10 min, resultando em 100 µL de PRP. Obtivemos maiores concentrações plaquetárias dos protocolos P2 e P3 com concentrações superiores a 519%. Quanto a visualização morfológica plaquetária por esfregaço, os três protocolos apresentaram boa visualização, no entanto P3 sobressaiu-se aos demais quanto ao tempo global de preparo e menor inclusão eritrocitária. Assim, padronizamos um modelo animal in vivo para futuras investigações científicas interessadas na utilização do PRP de Rattus novergicus (Wistar)(AU)

One of the biotechnologies with a relevant human health purpose is platelet rich plasma (PRP), which has no standardized protocol record for Rattus novergicus (Wistar). In fact, the lack of standardization of the method leads researchers to doubt which protocol to follow for the best results regarding platelet concentrations and morphological changes. Such indeterminacy promotes slowness in scientific research. In view of the conjecture, how few animals were used in a double-blind experiment and 3.2 mL of blood was drawn from each animal by heartbeat and subsequently tested in three separate protocols. Each protocol was designated as P1; P2; P3 is the control group designated PSP. i. P1 was subjected to a single 160G centrifugation for 6min resulting in 100 µL of PRP, ii. P2 was resigned to double centrifugation with the first stage 160G for 20min and the second stage for 400G for 15 min, resulting in 100 µL of PRP and iii. P3 intended for double centrifugation with the first stage of 300G for 10 min and the second stage of 640G for 10 min, resulting in 100 µL of PRP. We obtained higher platelet concentrations of protocols P2 and P3 with concentrations higher than 519%. Regarding the morphological visualization of platelet smears, the three protocols presented good visualization, but P3 stood out for the others in terms of total preparation time and less erythrocyte invasion. Thus, we standardized an in vivo animal model for future scientific investigations interested in the use of PRP Rattus novergicus (Wistar)(AU)