Platelet Count In Platelet-rich Plasma Research Articles

The growth factor content as an indicator of platelet counts in platelet-rich plasma

BackgroundPlatelet contains growth factors that enhance tissue repair mechanisms, including epidermal growth factor (EGF), platelet-derived growth factor (PDGF-AA and −AB), and transforming growth factor (TGF)-β. Autologous platelet-rich plasma (PRP) has been shown to significantly improve the treatment of tendon injuries compared with hyaluronic acid and placebo. The topic of agreement between platelet concentrations and growth factors has been covered in some previous studies, but growth factor levels did not correlate well with platelet concentrations. MethodIn this study, autologous PRP was prepared by concentrating platelets through a J6-MI centrifuge. The automatic hematology analyzer Sysmex XN-20 was used to analyze the platelet concentration in PRP, and the PRP growth factors were determined by ELISA, including PDGF, transforming growth factor- β1 (TGF-β1), and EGF. Statistical analysis was conducted on data from 107 patients who received autologous PRP using Pearson correlation analysis. ResultsPearson correlation analysis revealed PDGF, TGF, and EGF had a strong positive correlation with the platelet concentration of the final PRP product (r = 0.697, p < 0.0001; r = 0.488, p < 0.0001; r = 0.572, p < 0.0001, respectively) ConclusionsThere was a strong positive correlation between the concentration of platelets in the final PRP product and the levels of PDGF-AB, TGF-β, and EGF. These results suggested straightforward and cost-effective growth factor tests can provide valuable information about platelet content in PRP.

Effect of nanofat combined with platelet-rich plasma for treatment of pressure injury wounds in rats

To investigate the effect of allogeneic nanofat combined with platelet-rich plasma (PRP) for treatment of pressure injury wounds in rats. Forty SD rat models with pressure injury wounds were randomly divided into 4 groups (n=10) for treatment with nanofat, PRP (platelet count about 6.2 times that of normal whole blood), nanofat combined with PRP, or PBS only (control group).Wound healing was observed on days 1, 3, 5, 7, 10 and 14 and wound healing rate was calculated. On days 5, 10 and 14, tissue samples were taken from the wounds for HE staining, Masson staining, immunohistochemistry, and Western blotting to observe inflammatory cell infiltration, collagen formation, deposition and arrangement, angiogenesis, inflammatory factor expression and VEGF expression in the wounds. The combined treatment with nanofat and PRP achieved the highest wound healing rates at all the time points of observation (P < 0.05), enhanced inflammatory cell infiltration on day 5, and accelerated dermal and epidermal growth compared with the other treatments (P < 0.05). The combined treatment also more effectively promoted collagen expression and its regular arrangement (P < 0.05) and enhanced angiogenesis in the wounds than nanofat and PRP alone, without significant difference between the latter two treatments (P > 0.05). Immunohistochemistry and Western blotting showed that the expressions of MCP-1 and VEGF in the wounds were the highest in the combined treatment group (P < 0.05) and were higher in nanofat and PRP treatment groups than in the control group (P < 0.05). Nanofat combined with PRP can significantly promote healing of pressure injury wounds in rats, and their synergistic effect sheds light on a new strategy for treatment of pressure injury wounds.

No impact of polymorphism in the phosphodiesterase 5A gene in Cavalier King Charles Spaniels on pimobendan-induced inhibition of platelet aggregation response.

A variant in the canine phosphodiesterase (PDE) 5A gene (PDE5A:E90K) is associated with decreased concentrations of circulating cyclic guanosine monophosphate (cGMP) and response to PDE5 inhibitor treatment. Pimobendan is a PDE inhibitor recommended for medical treatment of certain stages of myxomatous mitral valve disease (MMVD) in dogs. PDE5A:E90K polymorphism attenuates the inhibitory effect of pimobendan on in vitro platelet aggregation and increases basal platelet aggregation in Cavalier King Charles Spaniels (CKCS). Selected clinical variables (MMVD severity, sex, age, hematocrit, platelet count in platelet-rich plasma [PRP], and echocardiographic left ventricular fractional shortening [LV FS]) will not show an association with results. Fifty-two privately owned CKCS with no or preclinical MMVD. Using blood samples, we prospectively assessed PDE5A genotype using Sanger sequencing and adenosine diphosphate-induced platelet aggregation response (area under the curve [AUC], maximal aggregation [MaxA], and velocity [Vel]) with and without pimobendan using light transmission aggregometry. Dogs also underwent echocardiography. Pimobendan inhibited platelet function as measured by AUC, MaxA, and Vel at a concentration of 10 μM (P < .0001) and Vel at 0.03 μM (P < .001). PDE5A:E90K polymorphism did not influence the inhibitory effect of pimobendan or basal platelet aggregation response. The PDE5A:E90K polymorphism did not influence in vitro basal platelet aggregation response or the inhibitory effect of pimobendan on platelet aggregation in CKCS. Dogs with the PDE5A:E90K polymorphism did not appear to have altered platelet function or response to pimobendan treatment.

Factors affecting the features of platelet-rich plasma in patients with knee osteoarthritis.

The aim of this study was to present an analysis of platelet-rich plasma obtained from patients with knee osteoarthritis and reveal the factors affecting its features. A total of 62 patients (mean age: 56.68 ± 7.13 years) with symptomatic knee osteoarthritis were included in this study. Age (years), gender, height (m), weight (kg), body mass index (kg/m2), duration of symptoms, smoking status, smoking index, general health status, and physical activity scores were recorded. Whole blood and platelet-rich plasma cell counts were performed with a hematology analyzer. White blood cell, red blood cell, and platelet counts were recorded. According to the dose of injected platelets, efficiency of the procedure, purity of platelet-rich plasma, and activation classification, dose of platelets, efficiency of the procedure (platelet recovery rate, %), and purity of the obtained platelet-rich plasma product (relative composition in platelets, %) were calculated. Correlation analysis between the features of platelet-rich plasma and the patient-related variables, including age, gender, body mass index, smoking status, smoking index, presence of other health conditions, physical activity scores, duration of symptoms, and pain levels, was performed. Dose of injected platelets, efficiency of the procedure, purity of platelet-rich plasma, and activation analysis showed that the dose of injected platelets was 3.25 billion, the efficiency of the process was 77%, and the purity rate of the platelet-rich plasma was 98.4%. Platelet-rich plasma platelet count was correlated with whole blood platelet count (r = 0.81, P < .001), whole blood white blood cell count (r = 0.39, P = .002), smoking status (r = 0.56, P = .03), smoking index (r = -0.63, P = .002), and the presence of hypertension (r = -0.31, P=.04). Platelet-rich plasma white blood cell and purity of platelet-rich plasma were correlated with the smoking status of the patients (r = 0.52, P = .01; r = 0.64, P = .003, respectively). This study has demonstrated that high dose and very pure platelet-rich plasma with medium efficiency was yielded with this platelet-rich plasma preparation procedure; whole blood platelet count, the presence of hypertension, and the smoking status of patients affect the features of the obtained platelet-rich plasma. Level IV, Diagnostic Study.

Substantial Variability in Platelet-Rich Plasma Composition Is Based on Patient Age and Baseline Platelet Count

PurposeTo evaluate the effect of age, sex, body mass index (BMI), and baseline blood count on the final composition of platelet rich-plasma (PRP) and to evaluate the variability of PRP applied in the same patient at 2 different times.MethodsPotential subjects treated with PRP between January 2019 and December 2021 were identified in an institutional registry. Patient demographics and baseline blood count were prospectively recorded in a consecutive series of patients treated with PRP for a musculoskeletal condition in our Institution. The influence of sex, BMI, age, and baseline blood count on final platelet concentrations in PRP was evaluated. Finally, intrapersonal variability was evaluated.ResultsA total of 403 PRP injections from 357 patients were analyzed from an institutional prospective registry of PRP between January 2019 and December 2021. A directly proportional variation in PRP platelet count of 3.8× was observed for each unit increase in baseline blood platelet count. For every decade increase, we observed an approximate decrease of 32,666 platelets. When the first dose of PRP platelet counts was compared with the second dose of PRP platelet counts between the same patients, significant differences were found. A mean of 890,018 platelets in the first PRP and a mean of 1,244,467 in the second PRP with a mean difference of 354,448 was found (P = .008). We did not find differences in the final concentration of platelets regarding sex, BMI, or PRP protocol.ConclusionsOverall the final composition of PRP (platelet count) was significantly influenced by patient’s age and baseline platelet count. In contrast, BMI, sex and the rest of the components of the baseline blood count did not have a significant influence on final PRP. Furthermore, in patients who received 2 doses of PRP, the final concentration of platelets varied significantly between the 2 preparations.Level of EvidenceLevel IV, prognostic case series. To evaluate the effect of age, sex, body mass index (BMI), and baseline blood count on the final composition of platelet rich-plasma (PRP) and to evaluate the variability of PRP applied in the same patient at 2 different times. Potential subjects treated with PRP between January 2019 and December 2021 were identified in an institutional registry. Patient demographics and baseline blood count were prospectively recorded in a consecutive series of patients treated with PRP for a musculoskeletal condition in our Institution. The influence of sex, BMI, age, and baseline blood count on final platelet concentrations in PRP was evaluated. Finally, intrapersonal variability was evaluated. A total of 403 PRP injections from 357 patients were analyzed from an institutional prospective registry of PRP between January 2019 and December 2021. A directly proportional variation in PRP platelet count of 3.8× was observed for each unit increase in baseline blood platelet count. For every decade increase, we observed an approximate decrease of 32,666 platelets. When the first dose of PRP platelet counts was compared with the second dose of PRP platelet counts between the same patients, significant differences were found. A mean of 890,018 platelets in the first PRP and a mean of 1,244,467 in the second PRP with a mean difference of 354,448 was found (P = .008). We did not find differences in the final concentration of platelets regarding sex, BMI, or PRP protocol. Overall the final composition of PRP (platelet count) was significantly influenced by patient’s age and baseline platelet count. In contrast, BMI, sex and the rest of the components of the baseline blood count did not have a significant influence on final PRP. Furthermore, in patients who received 2 doses of PRP, the final concentration of platelets varied significantly between the 2 preparations.

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

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. 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. 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. 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.

Effect of platelet counts and activator in platelet-rich plasma on the treatment of androgenetic alopecia, split-head comparison: A randomised, double-blind study.

Background Androgenetic alopecia is a common, chronic, non-scarring alopecia. It is characterised by stepwise miniaturisation of the hair follicles, due to alteration in the hair cycle dynamics, leading to the transformation of terminal hair follicles into a vellus ones. Oral finasteride and topical minoxidil are the only approved drugs for treating this condition. Due to a limited number of effective therapies for androgenetic alopecia, platelet-rich plasma may be an effective alternative treatment. Aims To study the effect of activator in platelet-rich plasma and baseline platelet count in platelet-rich plasma on the treatment of androgenetic alopecia. Methods A randomised, double-blind split-head comparative study. The sample size was calculated and randomisation was done. Patients with androgenetic alopecia were allocated into two groups; in the first group, autologous activated platelet-rich plasma was injected in the right half of the affected scalp and autologous non-activated platelet-rich plasma was injected in the left half of the affected scalp and vice versa in the second group. Patients were also categorised on the basis of platelet counts in their platelet-rich plasma in three groups; group A (6-8 lakh/mm3), group B (8.1-10 lakh/mm3) and group C (>10 lakh/mm3). Interventions were done monthly for three months and followed up for the next three months. Effects of interventions were assessed by hair density, hair thickness, patient self-assessment and clinical photography. Results A total of 80 patients were included in the study. Activated platelet-rich plasma produced significant improvement of hair density after four months and hair thickness at 6 months. An increase in platelet count led to a significant increase in hair density and hair thickness after three and four months respectively and a highly significant increase in both parameters at the end of the study. Limitations Long-term follow-up of cases was not done and no measurement of vellus hair count was done. Conclusion There is a significant effect of activator and platelet count of the platelet-rich plasma on hair density as well as hair thickness.

Effect of Mean Platelet Volume on Platelet Rich Plasma Based Injection Therapy in Patients with Osteoarthritis of the Knee JointA Longitudinal Study

Introduction: There are different types of arthritis seen in elderly patients, of which Osteoarthritis (OA) is the most common. Platelet Rich Plasma (PRP), which is derived from blood, contains many growth factors. Platelet count and indices in PRP has association with pain score of OA, further study will prove its significance. Aim: To find the correlation of platelet parameters like platelet count, Mean Platelet Volume (MPV) and Plateletcrit (PCT) with clinical OA improvement score in PRP-treated patients of OA. Materials and Methods: The longitudinal study was done in the Department of Pathology, PDU Civil Hospital, Rajkot, Gujarat, India, from January 2021 to October 2021. Study was conducted on 40 patients, who were administered intra-articular injection of PRP prepared using double spin method. It was injected at the site of maximum pain in the knee joint. Platelet count, MPV and PCT were noted in the pre and post intervention. Patients were followed-up at two months and six months. The data was analysed using Friedman Analysis of Variance (ANOVA) and Pearson’s correlation test. Results: The mean age of patients was 58.57±4.95 years and there were 26 (65%) females and 14 (35%) males. Increase in platelet count, MPV, PCT in PRP prepared by double spin method and improvement in Visual Analog Scale (VAS) (Before8.38±0.63; After-4.28±0.85), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) (Before-63.30±4.99; After- 43.67±5.22) and International Knee Documentation Committee (IKDC) (Before-45.04±6.06; After-66.14±5.78) score were found to be statistically significant. Correlation of platelet parameters WOMAC with Platelet count (p-value=0.052, R-value=-0.310), VAS with MPV (p-value=0.005, R-value=0.436) and IKDC with PCT (p-value=0.032, R-value=0.339) were also statistically significant at six months follow-up. Conclusion: Single dose intra-articular injection PRP in OA knee with KL Grade 1 and 2 is valuable, efficient and costeffective treatment to reduce or delay in knee replacement. The study showed the significant correlation of platelet parameters with OA improvement scores.

Bacteriostasis of Platelet-Rich Plasma and Its Derivatives in Vitro and Its Relationship with PF4

To investigate the bacteriostatic effect of platelet-rich plasma (PRP) and its derivative platelet gel (PG) supernatant on Escherichia coli in vitro and its relationship with platelet factor 4 (PF4). Apheresis platelets donated by healthy volunteers were obtained from the Blood station of Lu an Blood Center as the source of PRP. The counts of platelet, white blood cell and red blood cell in PRP and its derivative PG supernatant were detected by automatic hematology analyzer. Bacterial growth of PRP and PG supernatants co-cultured with bacteria for different time was observed by plate coating culture method, and the contents of PF4 in PRP and PG supernatants were detected by ELISA. Apheresis platelets were collected from 28 healthy volunteers with a median age of 33 (21-56) years old. PRP can inhibit the growth of escherichia coli, but there were individual differences in antibacterial effect within 24 hours. PRP of 13 healthy volunteers had strong antibacterial effect at 24 hours, 7 cases had weak antibacterial effect at 24 hours, and 8 cases had no antibacterial effect at 24 hours. PG supernatant showed no significant individual difference, and all of them had bacteriostatic effect within 12 hours, but no bacteriostatic effect after 12 hours. There was no statistical difference in the bacteriostatic effect of PRP at 24 hours between healthy volunteers aged ≤30 years and >30 years (P>0.05), and there was no statistical difference between the white blood cell count ≤0.1×109/L and (0.1-1) ×109/L groups (P>0.05). There was significant difference in the bacteriostatic effect of PRP between the two groups with platelet content ≤1 000×109/L and >1 000×109/L (P<0.05). The platelet count in PRP was higher than that in PG supernatant ［(911.57±160.52) ×109/L vs 0］. The PF4 level in PRP was higher than that in PG supernatant (23623.34±9822.14 vs 6664.74±4065.83, P<0.05). Both PRP and PG supernatant have antibacterial effects in Escherichia coli. The bacteriostatic effect of PRP was better than that of PG supernatant, and the platelet and PF4 contents in PRP were higher than those in PG supernatant, suggesting that the platelet and PF4 levels play an important role in bacteriostasis.

Factors Affecting Platelet Count in Platelet-Rich Plasma

Introduction: The amount of platelet count in platelet-rich plasma (PRP) is very important for the concentration of growth factors, which play a major role in the effectiveness of PRP.&#x0D; Aim: The aim of this study was to investigate the effects of demographic and clinical parameters on the number of platelets in PRP.&#x0D; Materials And Methods: The data of 50 patients who received PRP in our stem cell center were scanned retrospectively in electronic environment. Demographic and clinical parameters (age, gender, body mass index, blood group, comorbidities, hypertension, coronary heart disease and diabetes, alcohol and cigarette usage) of the patients were recorded.&#x0D; &#x0D; Results: The results of the correlation analysis performed to evaluate whether there was a relationship between age and BMI with regards to platelet count in PRP revealed no statistical significance (p&gt;0.05). Analysis of Mann Whitney-U and Kruskal Wallis-H showed that there was a statistically significant difference in terms of platelet count, gender, presence of hypertension and coronary artery disease (p

A Simple Double-Spin Closed Method for Preparing Platelet-Rich Plasma.

Objective: To describe and analyze a new protocol for the extraction of platelet-rich plasma (PRP) for use in clinical practice and compare this technique with methods that have been previously described in the medical literature.Methods: Sixteen blood samples from healthy volunteers were collected. PRP was prepared using our new double-spin technique, consisting of successive centrifugation of blood samples with two different spins, without opening the container. Descriptive analysis of cell counts in baseline and PRP samples was undertaken. Comparison between cell and platelet count in baseline and PRP samples, as well as the statistical analysis, were done.Results: The mean platelet concentration ratio was 3.47 (SD: 0.85; 95% CI: 3.01-3.92; range: 2.48-5.71). The baseline whole blood platelet count correlated positively to the PRP platelet count (rP = 0.56; 95% CI: 0.09-0.88; P = 0.023). The PRP was enriched for lymphocytes and monocytes but presented significantly lower counts of neutrophils and eosinophils in comparison to baseline.Conclusion: Results show a safe and easily reproducible method to obtain PRP for use in clinical daily practice.

Different platelet-rich plasma preparation protocols in Female pattern hair loss: Does it affect the outcome? A pilot study.

Platelet-rich plasma (PRP) injection is a promising modality for hair regeneration in female pattern hair loss (FPHL). A standard protocol on best methods for PRP preparation has not been established. To optimize standard PRP preparation protocols and evaluate its clinical efficacy in FPHL. Comparative study enrolled 40 female patients with FPHL divided randomly into 4 equal groups. Each group received 3 sessions of monthly intradermal injection of PRP prepared by different methods regarding number of spins, centrifugation speeds, type of the centrifuge, and the size of PRP tube. Patients were evaluated by trichoscan before and 1month after the 3rd session for number of terminal, vellus hair, and average hair width. A statistically significant increase in platelet count in PRP prepared by combination of digital centrifuge, large-sized sodium citrate tube, and low centrifugation speed (900rpm). All patients showed statistically significant increase in percentage of terminal hair and average width of hair after treatment as assessed by trichoscan, without statistically significant difference between studied groups. Digital centrifuge, large-sized sodium citrate tubes, and a single spin with low centrifugation speed (900rpm) were ideal for PRP preparation. PRP is an effective and safe modality in FPHL therapy.

Two or four injections of platelet-rich plasma for osteoarthritic knee did not change synovial biomarkers but similarly improved clinical outcomes

We compared two and four intra-articular injections of platelet-rich plasma (PRP) in terms of changes of synovial cytokines and clinical outcomes. One hundred twenty-five patients having knee osteoarthritis (OA) underwent PRP injections at a 6-week interval. Before each PRP injection, synovial fluid aspiration was collected for investigation. Patients were divided into two or four intra-articular PRP injections (group A and B, respectively). Changes in synovial biomarkers were compared with the baseline levels of both groups, and clinical outcomes were evaluated until one year. Ninety-four patients who had completed synovial fluid collection were included for final evaluation, 51 in group A and 43 in group B. There were no differences in mean age, gender, body mass index (BMI), and radiographic OA grading. The average platelet count and white blood cell count in PRP were 430,000/µL and 200/ µL, respectively. There were no changes of synovial inflammatory cytokines (IL-1β, IL-6, IA-17A, and TNF-alpha), anti-inflammatory cytokines (IL-4, IL-10, IL-13, and IL-1RA), and growth factors (TGF-B1, VEGF, PDGF-AA, and PDGF-BB) between baseline levels and six weeks in group A, and 18 weeks in group B. Both groups had significantly improved clinical outcomes from six weeks including visual analog scale (VAS), patient-reported outcome measures [PROMs; Western Ontario and McMaster Universities Osteoarthritis (WOMAC) Index and Short Form-12 (SF-12)], with a significant delayed improvement of performance-based measures [PBMs; time up and go (TUG), 5-time sit to stand test (5 × SST), and 3-min walk test (3-min WT)]. In conclusion, two- or four-PRP intra-articular injection at a 6-week interval for knee OA demonstrated no changes of synovial cytokines and growth factors but similarly improved clinical outcomes from 6 weeks until 1 year.

Evaluation of Factors Affecting the Quantity and Quality of Platelet-Rich Plasma and Platelet-Derived Growth Factor-BB

Context: Platelet-rich plasma (PRP) releases growth factors (GFs) upon activation which accelerates the healing and regeneration in the target tissue. Wider applications of PRP warrant product standardization. PRP compositions may vary according to the patient's demographics and the constituents of PRP. Aims: Our study aimed to assess the correlations between age, gender, platelet counts, platelet indices, platelet yield, platelet dose, GF dose, and platelet-derived GF-BB (PDGF-BB) concentration. Settings and Design: This was an experimental study done in the Department of Pathology along with the Department of Dentistry at a tertiary care hospital in North India. Subjects and Methods: PRP was prepared from 40 ml of whole blood. Platelet counts, platelet indices (LH 750 coulter analyzer, Florida, USA), and PDGF-BB levels (enzyme-linked immunosorbent assay, Qayee Bio., Ltd, China) were measured. Platelet yield (%), platelet dose (×106), and GF dose (×1012) in PRP were also calculated. Statistical Analysis Used: All the parameters were analyzed using Pearson's correlation coefficient and linear regression. Paired t-test and Mann–Whitney test were used for two-group analysis. Results: Mean platelet count, PDGF-BB concentration, platelet yield, platelet dose, and GF dose in PRP were 1317 × 103/mm3, 30 ± 9.89 ng/ml, 71.62 ± 28.34%, 6.5 ± 3.5 × 109, and 159.62 ± 52.39 ng/ml, respectively. PRP platelet counts were positively correlated with PDGF-BB concentration, platelet yield, platelet dose, and GF dose. Conclusions: Of all the parameters, PDGF-BB concentration, platelet yield, platelet dose, and GF dose showed a positive correlation with PRP platelet count. Age, gender, and platelet indices had no significant influence.

COMPARISON OF DIFFERENT METHODS OF CENTRIFUGATION FOR OPTIMUM PREPARATION OF PLATELET RICH PLASMA (PRP)&amp; FRAMING A CUSTOMIZED PRP PROTOCOL-AN INNOVATIVE APPROACH

Introduction:The rationale behind the therapeutic potential of the high concentration of platelets in PRP (Platelet Rich Plasma) for intervention procedures of soft tissue injuries is often challenged by the ability of the centrifugation methods followed in extracting the high quality PRP with prompt contented quantity too.This study aims to unravel the basic science of PRP preparation involved in centrifugation protocols, spin methods, speed and time duration of centrifugation to obtain consistent therapeutic platelets yield. Materials and Methods:30 participants were subjected to intervention procedures using PRP. For therapeutic purpose 3 centrifugation protocol types are followed. Ø Protocol 1-Conventional Old protocol Ø Protocol 2-Customised New protocol Ø Protocol 3-Spin Reversal of the routine -hard spin followed by soft spin The Platelets and WBC concentration and composition in the final autologous PRP sample harvested as a result of double centrifugation method are compared for quantity and efficiency. Results:The maximum PCF-3.21 times above baseline and PRR -64.21% are through new customized spin proving its efficiency. As determined by one-way ANOVA, there is statistically significant difference between the PROTOCOL groups (1,2,3) for the variables PRP platelet count, PRP Leucocytes, PCF, PRR. A Tukey post hoc test revealed that there is statistically significant value for Protocol 2 and Protocol 3 compared to the Protocol 1 for PRP platelet count, PCF and PRR whereas for PRP Leucocytes, there was a statistically significant difference between the Protocol 2 and Protocol 3 which again implicates the necessity of spin reversal in yielding better quantity of leucocytes. Conclusion:To produce PRP samples with consistent and reproducible compositionsof platelets and leucocytes with better quality control standard through a detailed, precise and stepwise description of the manufacturing protocol has been explained in our study. Our perspective is in standardizing a safe, simple protocol that can be followed to obtain an optimal consistent platelet yield without the use of commercial kits, which has been proved statistically too.

Evaluation of the analytical performance of the PC100 platelet counter

IntroductionPlatelet count can be altered in various diseases and treatments and measuring it may provide better insight into the expected outcome. So far, quantification of platelet count is done within laboratory conditions by using established hematology analyzers, whereas a point-of-care device could be used for this purpose outside of the clinical laboratories.AimOur aim was to assess the closeness of agreement between a newly developed point-of-care PC100 platelet counter and two reference methods (Sysmex® XP-300, Sysmex® XN-9000) in measuring platelet counts in whole blood and platelet-rich-plasma (PRP).MethodWhole blood was obtained from 119 individuals, of which 74 were used to prepare PRP samples. Whole blood platelet count was measured by the two reference methods and the PC100 platelet counter. PRP was prepared from the whole blood and platelet count was adjusted to the range of 250–3600 × 103/μl and measured with the PC100 platelet counter and Sysmex® XP-300.ResultsA median difference of − 1.35% and − 2.98% occurred in whole blood platelet count between the PC100 platelet counter and the Sysmex® XP-300 and Sysmex® XN-9000, respectively. A strong linear correlation (r ≥ 0.98) was seen in both cases and regression equations indicated neither a constant nor a proportional bias between the methods. Direct comparison of the two reference methods revealed a median difference of − 1.15% and a strongly linear relationship (r = 0.99). Platelet count in PRP resulted in a median difference of 1.42% between the PC100 platelet counter and the reference method, Sysmex® XP-300. While the difference between two methods increased with concentration of platelets in PRP, a strong linear relationship remained throughout the whole measuring interval indicated by the high correlation coefficient (r = 0.99). Assessment of the predicted bias at predefined platelet counts showed that the bias in platelet counts falls within the acceptance criterion for both whole blood and PRP measurements.ConclusionsOur results show that the PC100 platelet counter can be used interchangeably with the reference methods for determining platelet counts.

Comparing novel versus conventional technique of platelet-rich plasma therapy in periorbital hyperpigmentation: A randomized prospective split-face study.

Platelet-rich plasma (PRP) has been found to be effective in treating periorbital hyperpigmentation (POH). PRP prepared by double-spin (DS) method and activated by calcium has been used conventionally. PRP can be prepared by single spin (SS) and activated at low temperature (novel method), but the evidence is limited. To compare the novel and conventional PRP in the treatment of periorbital hyperpigmentation. We selected 21 patients of POH and randomly divided the face into two halves. One-half of the face (group A) was treated with novel PRP (SS and low-temperature activation). The other half (group B) was treated with conventional PRP (DS and calcium activation). A total of 3 PRP injections were given at 4 weekly intervals. Patients were observed and assessed on 12th week by photography, dermoscopy, visual analog scale (VAS) score, and Dermatology life quality index (DLQI). Platelet counts and growth factors were assessed in PRP. Mean platelet count in novel and conventional PRP was 7.41±1.76 lacs and 8.17±2.23 lacs (p=0.348). Mean photographic and dermoscopic assessment at the end of the study in group A and group B was 52.33±6.468 and 53.14±6.99 (p=0.151). Change in VAS in groups A and B was 3.85±1.27 and 3.90±1.04 (p=0.895). Levels of various growth factors assessed by ELISA did not differ significantly. There was significant decline in DLQI. The novel method is not inferior to conventional method of PRP in the treatment of periorbital hyperpigmentation.

Factors affecting the quantity and quality of platelet-rich plasma and platelet-derived growth factor-BB: an observational study

Abstract Objective: Platelet-rich plasma (PRP) releases growth factors upon activation, which in turn accelerates healing and regeneration of the target tissue. However, PRP composition may vary according to the patient's demographics, and wider applications of PRP warrant product standardization. The current study aimed to examine variables influencing the platelet-derived growth factor BB (PDGF-BB) concentration in PRP. Methods: This observational study was conducted in the Department of Pathology and Dentistry at Swami Rama Himalayan University, a tertiary care hospital in northern India from December 2016 to November 2017. PRP was prepared from 40 mL of whole blood from 35 individuals (22 women, 13 men). Platelet counts, platelet indices (platelet distribution width, mean platelet volume) and PDGF-BB levels were measured, and platelet yield, platelet dose, and growth factor dose in PRP were also calculated. All parameters were analyzed using Pearson's correlation coefficient. The association between PDGF-BB and PRP platelet count was evaluated using logistic regression. This study was approved by the Ethics Committee of Swami Rama Himalayan University (SRHU/HIMS/ETHICS/2016/103) on September 7, 2016. Results: The mean platelet count, PDGF-BB concentration, platelet yield, platelet dose, and growth factor dose in PRP were 1317 × 109/L, 30 ± 9.89ng/mL, 71.62 ± 28.34%, 6.5 ± 3.5 × 109, and 159.62 ± 52.39ng/mL, respectively. Linear regression analysis indicated that PRP platelet counts were a good predictor for PGDF-BB (P &lt; 0.05; adjusted R 2 = 0.96. PRP platelet count was significantly positively correlated with PDGF-BB concentration (r = 0.74, P &lt; 0.001), platelet yield (r = 0.80, P &lt; 0.001), platelet dose (r = 1, P &lt; 0.001), and growth factor dose (r = 0.74, P &lt; 0.001). Conclusions: PRP has wide clinical applications associated with its healing and regenerative properties, and both the quality and quantity of PRP thus need to be standardized as per the requirements. Evaluating variables affecting PRP will thus aid pathologists and clinical practitioners.

Intraoperative collection of autologous platelet-rich plasma from the cardiopulmonary bypass circuit upon initiation of extracorporeal circulation

ObjectivesThe aim of this study is to evaluate the possibility of the autologous platelet-rich plasma (PRP) collection from the cardiopulmonary bypass (CPB) circuit and to evaluate its effect on the aggregative function.MethodsFor seventy-two patients undergoing cardiac surgery with CPB, an autologous PRP was prepared using the Haemonetics Component Collection System® by drawing blood from the CPB circuit immediately after CPB was established. The blood samples were taken at three points for examination, A: beginning of surgery, B: immediately after heparin reversal with protamine following discontinuation of CPB, C: after the collected autologous PRP was returned to the patient. Platelet count and platelet aggregation ability were analyzed.ResultsThe mean platelet count in autologous PRP was 5.5 (range: 3–14) units. Platelet count decreased by 115.0 (±27.3) × 1000/μl from A to B and increased by 27.3 ± 17.2 (× 1000/μl) from B to C.When platelet aggregation was measured by Adenosine Diphosphate (ADP) 3.0 μM, it decreased by 42.6% ± 12.1% from A to B and increased by 8.7% ± 7.4% from B to C.ConclusionsAutologous PRP can be safely collected by drawing blood from the CPB circuit, platelet count and aggregation ability significantly decreased after CPB including autologous PRP collection. Some improvement was detected in the number of the platelets count and platelet aggregation ability by administrating an autologous PRP even if autologous PRP is collected from CPB circuit.Trial registrationUMI-CTR, UMIN000023776. Registered 1 October 2016.

484 FDA cleared devices produce inconsistent platelet-rich plasma product

Platelet-rich plasma (PRP) has emerged as a novel option to treat androgenetic alopecia. However, data focused on quality of product from FDA cleared devices is lacking. Herein, we assessed and compared cellular PRP content produced from FDA cleared devices. Within one academic institution, white blood cell (WBC) counts and platelet counts from whole blood and resultant PRP were retrospectively reviewed. Institutional quality control (QC) procedures were utilized to obtain counts. Of the fifty-two PRP samples assessed, only 2 met the Insitutions QC standard (defined a 3-fold increase in platelet concentration). Paired analysis demonstrated similar platelet counts in whole blood and PRP. Platelet counts from individual devices were significantly different. Within the devices, platelet counts from PRP samples were inconsistent, failing the Shapiro-Wilk test. The Emcyte device demonstrated the highest capacity to concentrate, even though mean enrichment was just 2.9-fold and capture efficiency was found to be 34%. WBC numbers were negligible in Eclipse and Selphyl final PRP samples. However, the Emcyte device enriched PRP with WBCs similar to or greater than whole blood. These results demonstrate inconsistent PRP end product with variable platelet counts among multiple devices and within devices. QC testing standards were not passed by most samples. Efficiency of platelet capture was lower than that claimed by device manufacturers. WBC content varied by device used for preparation. Of note, no patients worsened while receiving these treatments. This study presents an additional variable in PRP preparation – inconsistent PRP product from FDA-cleared devices. Additionally, clinical improvement despite QC failure suggests platelet numbers may not be vital for clinical improvement.