Platelet Concentration In Platelet-rich Plasma Research Articles

Comparative characterization between autologous serum and platelet lysate under different temperatures and storage times

Therapies using autologous serum and platelet lysate have shown promise among blood and biological products in the treatment of various diseases. The autologous serum has been shown to be a superior alternative to traditional eye drops in treating eye diseases in ophthalmology. Figurelet lysate (PL) has recently been considered a more interesting alternative for the treatment of multiple tissues, as it does not have the unfavorable reactions seen with traditional platelet-rich plasma (PRP), making it a valuable blood derivative for use in ocular therapy. There is no definitive comparison in veterinary medicine between PL and autologous serum in terms of the content of Transforming Growth Factor beta 1 (TGF-1), which is known to have chemotactic, mitogenic, matrix formation, and angiogenesis effects on tissues, and beneficial proteins in ocular tissue. This study aimed to estimate the concentrations of TGF-1, total protein, and albumin, as well as autologous serum and platelet lysate, in horses over an 8-day storage period at temperatures of 4 °C and 37 °C. To produce autologous serum, 63 ml of blood was collected from each animal in seven 9 ml tubes without anticoagulant. For platelet lysate, 180 ml of blood was collected in 50 tubes of 3.6 ml with 3.2% sodium citrate. The most significant findings were the positive relationship between the baseline platelet count in the blood and the final platelet concentration in PRP. Specifically, we found a correlation (R = 0.9) with a p - value of 0.005 between the average baseline platelet level of seven animals and their corresponding PRP results, both on an individual level and as a group. Additionally, there was a correlation between growth factor concentration and PRP platelets, with the highest growth factor concentration in PL. The temperature storage group exhibited higher concentrations of total protein and serum albumin, as well as the maximum amount of growth factor for both products at a temperature of 37 °C.

Composition of Platelet-Rich Plasma Prepared From Knee Osteoarthritic Patients: Platelets, Leukocytes, and Subtypes of Leukocyte.

Platelet-rich plasma (PRP) has gained increasing popularity in the orthopedic field. There has been still no consensus on PRP preparation technique, thus providing a variety of final PRP products. Different preparation techniques lead to different compositions of PRP, which include platelet concentration, the number of leukocytes, and their subtypes. Here, we studied those compositions of PRP compared to whole blood samples. There were 335 participants who met the inclusion and exclusion criteria. Each participant underwent a blood drawing process to prepare PRP for their knee osteoarthritis treatment. We categorized baseline platelet concentration in whole blood samples into three groups: less than 2 x 105/µL (Group 1), between 2 x 105/µL and 2.99 x 105/µL (Group 2), and greater than 2.99 x 105/µL (Group 3). The primary outcome was reported as the platelet concentration in PRP and the ratio of platelet concentration in PRP compared to baseline whole blood samples. The secondary outcome was reported as the ratios of leukocyte, lymphocyte, and neutrophil in PRP compared to the baseline whole blood samples. The average platelet concentration in PRP was 1.26 x 106/µL (6.3 times higher compared to baseline whole blood samples). The mean platelet concentration of PRP in Group 1, Group 2, and Group 3 was 1.08 x 106/µL, 1.38 x 106/µL,and 1.71 x 106/µL, respectively (p-value =0.0001). The platelet concentration ratio of PRP condition to the baseline whole blood was 6.9, 5.8, and 4.2 in Group 1, Group 2, and Group 3, respectively (p-value = 0.0018). The average ratio of leukocytes in PRP to whole blood was 1.5. The average ratio of lymphocytes and neutrophils in PRP to whole blood was 2.0 and 0.5, respectively. Conclusion: Different baseline platelet concentrations in whole blood samples provided significantly different platelet concentrations in PRP. The baseline platelet concentration in whole blood also provided an inverse relation to the fold change of the platelet concentration in PRP. Subtypes of leukocytes changed from neutrophil-predominated in the baseline whole blood samples to lymphocyte-predominated in PRP.

DETERMINATION OF THE OPTIMUM PLATELET-RICH PLASMA METHOD FOR THE ESSENTIAL AMOUNT OF PLATELETS

Objective: Platelet-rich plasma (PRP) is the cellular plasma component containing a higher platelet concentration obtained by centrifugation of autologous peripheral blood. PRP has an auxiliary role in regenerative medicine treatment. Current treatment protocols for tissue repair strategies may be globally insufficient. PRP treatment via Platelet-released growth factors (GF) supports inflammation, proliferation, and wound healing. PRP studies have progressed and have proposed different formulations in recent years. Although various methods and devices are used, there are few and different standardized protocols to obtain PRP with the highest platelet concentration. Although anticoagulants containing citrate are primarily preferred in PRP production To provide effective platelet concentration in PRP, we noticed differences in the literature regarding PRP tube volumes, anticoagulant-citrate, and blood amounts. We aimed to obtain the most appropriate platelet concentration in PRP treatment by using different concentration ratios of Acid Citrate dextrose (ACD) solution A and different blood amounts and introducing a new treatment method to the literature. Methods: The blood of 30 individuals aged between 30 and 75 was taken into 10cc laboratory tubes, and 15cc and 18cc hourglass-shaped PRP tubes containing 8% ACD-solution A. Blood fractions were separated by centrifugation, buffy coat area was collected, and count platelet amount by Hematology Analyzer. Results: The platelet amount per µl was statistically significantly higher in 18cc blood compared to 15cc and 10cc. Platelet amounts obtained from 15 and 18cc peripheral blood with 8% ACD-solution A were above the effective treatment amount recommended in the literature. Conclusion: It was observed that the amount of used blood volume was correlated with the amount of obtained platelets.

Effect of platelet-rich plasma scaffolding combined with osteochondral autograft transfer for full-thickness articular cartilage defects of the femoral condyle

We aimed to investigate the local application methods of platelet-rich plasma (PRP) and the effect and safety of PRP scaffolding combined with osteochondral autograft transfer (OAT) in the treatment of full-thickness articular cartilage defects of the femoral condyle. Patients with cartilage defects of the femoral condyle were treated with OAT combined with PRP scaffolding between July 2017 and December 2020. Preoperative magnetic resonance imaging (MRI) and computed tomography were utilized to assess the size, location, and severity of the osteochondral defects. X-ray and MRI images of the knee were obtained at the final follow-up to assess the osseointegration and integrity of the implanted articular cartilage. Osteoarthritic changes in the knee joint were evaluated using the Kellgren–Lawrence grading system. Clinical status was assessed using the visual analog scale (VAS), International Knee Documentation Committee (IKDC), and Lysholm scores before the treatment and at the final follow-up. Complications and patient satisfaction were recorded to assess the safety of this combination therapy. Twenty-one patients were recruited, with a mean follow-up duration of 18.23 ± 6.84 months. The mean lesion size was 2.3 ± 0.59 cm2. The mean platelet concentration in PRP at baseline was 6.27 ± 0.63 times greater than that in the peripheral blood. The VAS, IKDC, and Lysholm scores had improved significantly at the final follow-up (P < 0.001). No serious complications such as joint infection, deep venous thrombosis, or hematoma were observed. Eighteen patients (85.72%) were satisfied with their knee function and quality of life at the final follow-up. Three patients (14.28%) complained of mild anterior knee pain, which was relieved by oral administration of nonsteroidal anti-inflammatory drugs. MRI examinations of all patients showed bony consolidation and the defect surface was covered with cartilage-like tissue. X-ray evaluations indicated that osteoarthritis in two knees (9.5%) had progressed from grade 1 to grade 2 at the final follow-up. The preliminary results showed that OAT combined with PRP may be a safe and effective technique for the treatment of full-thickness articular cartilage defects in the knee.

P-799 To evaluate the effect of Intra-ovarian platelet rich plasma instillation on the clinical outcome of women with diminished ovarian reserve: A prospective interventional study

Abstract Study question Does intra-ovarian instillation of platelet rich plasma (PRP) improves the clinical outcome of IVF cycles in women with Diminished Ovarian reserve (DOR)? Summary answer PRP instillation leads to consistent improvement in Antral follicle count (AFC), thus achieving clinical pregnancy rate of 33.3% per cycle in women with DOR. What is known already There is rising incidence of females with diminished ovarian reserve (DOR) especially among Asian ethnicity. With the emergence of regenerative medicine, multiple studies have evaluated the role of intra-ovarian PRP, demonstrating a beneficial role in improving ovarian reserve parameters (serum Follicular stimulating hormone (FSH), serum anti-Mullerian hormone (AMH), AFC). Despite its’ favorable effects on biochemical markers and AFC, data regarding improvement in clinical outcome remains elusive and led to inception of this study. Study design, size, duration A prospective interventional study was conducted at Division of Reproductive Medicine of a tertiary care institute. 41 infertile females aged 20-39 years with DOR (AMH &amp;lt;1.2 ng /ml; AFC&amp;lt;5) were enrolled in the study during a 6-month period beginning from August 2021. Participants/materials, setting, methods After informed consent, patients received fresh autologous PRP, prepared from 30 ml venous blood. 1.5ml of PRP instilled in each ovarian stroma between day 7-10 of menstrual cycle under sedation. Patients were followed up for three-consecutive months to assess ovarian reserve parameters including serum FSH, AMH and AFC. Patients showing significant improvement in parameters were recruited for fresh IVF cycles using Antagonist protocol with 1% transdermal testosterone. Outcomes were analysed using linear mix effect model. Main results and the role of chance The average platelet concentration in PRP was ∼10,00,000 platelets/µL. The mean age of enrolled patients was 31.22±4.16 years. Linear improvement in AFC (3.63 vs 6.98 vs 7.97 vs 6.90, p&amp;lt;0.001) was observed from baseline to three consecutive follow-up months with maximal response witnessed in second month in 57.1% of those undergoing IVF cycle. However, there was no significant difference in Serum FSH (p=0.11) and AMH (p=0.16) from the baseline post intervention. Of the 41 patients, 35 (85.3%) responded to the treatment and underwent IVF antagonist cycle. 5 out of 35 IVF cycles were cancelled mid-cycle due to poor ovarian response. The mean dose of gonadotropin requirement was 2667.5±281.1 IU (Follicular stimulating hormone) and 1400±337.3 IU (Human menopausal gonadotropin). The average number of oocytes retrieved was 5.7±2.2 whereas mean number of MII oocytes was 4.63±1.85. The fertilization rate and the cleavage rate were 92.4% and 74.1% respectively. Of the thirty patients, eight patients underwent day 2 transfer due to poor grade of embryos. Mean number of grade 1- day 3 embryos was 1.25±0.55 with surplus embryos available for cryopreservation in 14 patients. The overall clinical pregnancy rate per transfer was 33.33%. No adverse events were reported. Limitations, reasons for caution This was a prospective single arm study. A randomized controlled trial comprising a “no-treatment” arm would establish a Level-I evidence. However, “no-treatment” arm in a developing country like ours, imposes financial burden on the couple with no guaranteed clinical success and thus raising ethical concern and need for ovarian rejuvenation. Wider implications of the findings With the impetus to provide a biological child to these DOR women, intra-ovarian PRP instillation as a method of ovarian rejuvenation holds promising results. Evidently, PRP is not only effective in improving ovarian reserve but this translates into an improved reproductive outcome in a population, previously limited to oocyte donation. Trial registration number REF/2022/01/051033

Efficacy of Platelet-Rich Plasma Injection in the Management of Rotator Cuff Tendinopathy: A Review of the Current Literature.

Platelet-rich plasma (PRP) is being projected as a newer and superior treatment option for rotator cuff tendinopathy. With the first use of PRP in trauma and orthopedics dating back only to 1998, its advent into the field is relatively recent. Thus, data on long-term efficacy, large cohort studies, or large randomized controlled trials are fairly limited. Moreover, owing to the absence of standardized methods of platelet extraction and lack of consensus on the ideal concentration of platelets in PRP, data comparison from different studies is difficult. Things are complicated by the fact that it is also not clear whether a single injection of PRP is therapeutic or if multiple serial injections are needed to produce the desired effects. The literature on efficacy and pain relief is also obtained from studies with low sample sizes and short follow-ups. The dropout rate and noncompliance rate were also found to be high in some studies. Thus, the data is often not clinically significant and may also be biased due to the non-standardized inclusion and exclusion criteria of the studies. Though multiple studies have suggested good pain control with PRP injection, many studies have found that PRP injection therapy may not be any better than the physical therapy regimen prescribed to patients with rotator cuff tendinopathy. Also, the data on the efficacy of PRP on shoulder function and shoulder range of motion is at variance. This submission aims to evaluate the efficacy and use of PRP in the management of rotator cuff tendinopathy.

A Comparative Study between Fresh and Stored Platelet Concentrate after Transfusion in same ALL patients in a Tertiary Hospital

Background: Platelet transfusion is needed for the treatment of ALL children. Researchers have shown that transfusion of stored platelet concentrates (up to 5days) is as effective as fresh platelet concentrates (24 hours). Objectives: The present study was conducted to access the comparison between fresh and stored platelet concentrate after transfusion in same all patients in a tertiary hospital. Materials and methods: The study was a cross-sectional descriptive study which was conducted in department of Clinical Pathology, Paediatric Haemato-Oncology and transfusion Medicine BSMMU, and Department of Haematology and Paediatric Haemato-Oncology in Dhaka Medical College Hospital. Over a period of March 2010 to February 2011.The sample size was 81. A data sheet with two parts (Part A and Part B) was designed with a view to collect data from the patient to be enrolled in the study .The data were analyzed using the SPSS version 25.0. Results: Out of 81 patients fresh platelet concentrates (FPC) or day-0 platelets were transfused in 47 acute Lymphoblastic leukaemia (ALL) children. Stored platelet concentrates (day 1-5) were transfused in 34 children. In 27cases both fresh and stored platelet concentrates were transfused. Corrected count increment at 1 hour (CCI 1h) in FPC and SPC were 20.5 and 18.9 respectively. Mean corrected count increment at 24 hour (CCI 24h) FPC and SPC were 15.5 and 13.8 respectively. There were no significant differences between FPC and SPC P&gt;0.05. Conclusions: It was observed that 1-5 day's stored platelet-rich-plasma platelet concentrate (PRP-PC) was as effective as fresh PC to obtain acceptable platelet increment in childhood acute lymphoblastic leukemia. Furthermore, corrected count increment and percentage platelet recovery are important tools to evaluate platelet refractoriness.

Platelet-rich Plasma Extract promoting Migration of Mouse Bone Marrow Cells

Platelet-rich plasma (PRP) is a rich source of plateletderived growth factors such as PDGF, TGF and VEGF, making it a potential material in tissue regeneration, especially in regenerative medicine. Due to the limitation of ethical issues for direct research on humans, this study aimed to collect PRP from peripheral blood of the mouse and evaluate its features as platelet concentration, the structure of activated PRP gel, in vitro cytotoxicity and biological characteristics through migration with mouse bone marrow cells. The results showed that the platelet concentration of PRP was five times higher than that in whole blood and PRP has a dense fibrin network after activating by CaCl2 after 10 minutes. PRP is not only safe for cells but also supports bone marrow cells to migrate and close the gap. With these results, PRP can be seen as a material with a considerable potential for wound healing, especially regenerative medicine.

Platelet-rich plasma in the treatment of alopecia

&lt;p&gt;Platelet rich plasma (PRP) is a promising treatment choice for patients with thinning hair. Despite excellent clinical safety and low cost, its clinical standing is still weak. The effectiveness of this method depends on its dosage, number of sessions, their intervals and technique of injection incorporated. PRP can produce particularly some phenomenal effects when applied in cosmetic dermatology. The therapeutic value of PRP is equivalent to stem cells and considered as one of the promising therapeutic agents in regenerative medicine. Harvesting of PRP plays a significant role, which is obtained from the patient's blood after centrifugation of the sample i.e., the platelet concentrates above the baseline which is the plasma fraction of the autologous blood. There are many applications of PRP in the medical field and has an incredibly significant role in dermatologic conditions e.g., tissue regeneration, wound healing, scar revision, skin rejuvenation and alopecia. In this review, we will be analyzing the authenticity of the use of PRP in the treatment of alopecia. PRP, in current scenario, is considered as a novel treatment modality. The efficacy of PRP therapy carries some deficiencies, which include lacking standard in preparation and concentration of platelets in PRP.&lt;/p&gt;&lt;p&gt;&lt;strong&gt; &lt;/strong&gt;&lt;/p&gt;

Platelet rich plasma concentration improves biologic mesh incorporation and decreases multinucleated giant cells in a dose dependent fashion.

Platelet rich plasma (PRP) has been shown to improve incorporation and reduce inflammation in ventral hernia repair (VHR) with acellular dermal matrix (ADM). The concentration of platelets in PRP varies in clinical studies and an ideal concentration has yet to be defined. The effects of varied concentrations of PRP on ADM incorporation and inflammatory cell infiltration in a rat model of VHR. We hypothesized that increasing concentration of PRP would lead to improved incorporation, decreased CD8+ and multinucleated giant cell (MNGC) infiltrate. Lewis rats underwent ventral hernia creation and repair 30days later with porcine non-crosslinked ADM. PRP was applied to the mesh prior to skin closure at concentrations of 1×104 plt/μL (PRP-LOW), 1×106 plt/μL (PRP-MID), or 1×107 plt/μL (PRP-HIGH) and tissue harvested at 2 and 4weeks. Cellularization, tissue deposition, and mesh thickness using hematoxylin and eosin and Masson's trichrome, and neovascularization was assessed with VVG staining, to establish the relationship of PRP concentration to metrics of incorporation. MNGC and CD8+ T-cell infiltration were quantified to establish the relationship of inflammatory cell infiltration in response to PRP concentration. Lymphocyte infiltration was assessed using immunohistochemical staining for CD8. PRP-HIGH treated had significantly greater tissue deposition at 4weeks. PRP-MID showed increasing mesh thickness at 2weeks. Cell infiltration was significantly higher with PRP-HIGH at both 2 and 4weeks while PRP-LOW showed increased cell infiltration only at 4weeks. At both time points there was a trend towards a dose dependent response in cell infiltration to PRP concentration. Neovascularization was highest with MID-plt at 2weeks, yet no significant differences were noted compared to controls. CD8+ cell infiltrate was significantly decreased at 2 and 4weeks in PRP-LOW and PRP-MID treated groups. PRP at all concentrations significantly decreased MNGC infiltration at 2weeks while only PRP-HIGH and PRP-MID had significant reductions in MNGC at 4weeks. Both MNGC and CD8+ cell infiltration demonstrated dose dependent reduction in relation to PRP concentration. Increasing platelet concentrations of PRP correlated with improved incorporation, tissue deposition, and decreased scaffold degradation. These findings were associated with a blunted foreign body response. These findings suggest PRP reduces inflammation which may be beneficial for ADM incorporation in VHR.

OPTIMIZING A PLATELET-RICH PLASMA CONCENTRATION PROTOCOL FOR SOUTH AMERICAN SEA LIONS (OTARIA FLAVESCENS).

Accelerated healing in wild or captive South American sea lions (Otaria flavescens) is a key tool to help minimize infection and complications associated with open wounds, dental disease, and ocular pathology. Platelet-rich plasma (PRP) is an autogenous source for growth factors based on platelet concentration, which can be obtained by centrifuging whole blood collected in sodium citrate anticoagulant. Currently, there are well-defined PRP concentration protocols for humans and most domestic companion animal species. However, there is no clear centrifugation protocol for obtaining PRP in most marine mammal species. This study aimed to optimize the platelet concentration protocol based on whole blood centrifugation using speeds ranging from 500 to 5,000 rpm and times ranging from 3 to 6 min. Blood was drawn from seven adult South American sea lions, placed into 1-ml sodium citrate tubes, and centrifuged following 12 different centrifugation protocols. PRP was designated as the lower third fraction of the centrifuged plasma. Platelet counts were performed using flow cytometry and statistical analysis was carried out to establish a well-defined protocol for efficient PRP production. Transmission electron microscopy (TEM) analysis was performed to evaluate possible platelet degranulation during the different centrifugation protocols and measure platelet areas. Maximum concentration of platelets in PRP was 4.73-fold higher than the number of platelets in equal volume of whole blood, and significant differences in the concentrations obtained were found between the 12 centrifugation protocols evaluated using different speed and time combinations. The best one-step centrifugation protocol resulted from using 900-rpm speed for 3 min. The highest-fold increase was achieved using a two-step centrifugation protocol, which combined the most efficient one-step centrifugation protocol (900 rpm, 3 min) with a second centrifugation using 2,000-rpm speed for 6 min. TEM analysis confirmed that platelets were complete and maintained integrity after the proposed protocol.

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.

A cross-sectional analysis of the effects of various centrifugation speeds and inclusion of the buffy coat in platelet-rich plasma preparation.

Platelet-rich plasma is an autologous blood preparation which is used in various medical specialties because of its regenerative properties. There is a wide variation in platelet-rich plasma preparation protocols and attaining the ideal platelet yield (>1 million platelets/μL) in a clinic setting can be challenging. We aimed at analyzing the centrifuge spin rates at which to attain an ideal platelet-rich plasma yield and also to study the effect of inclusion of the buffy coat after the first spin on the final platelet concentration in platelet-rich plasma. Seventy-five whole blood samples were obtained and divided into two groups - (1) leukocyte-rich platelet-rich plasma group and (2) leukocyte-poor platelet-rich plasma group. Samples in both groups were centrifuged using the dual spin method, at one of three centrifugation speed combinations (initial "soft" spin and second "hard" spin speeds, respectively): (1) 100 g/400 g, (2) 350 g/1350 g and (3) 900 g/1800 g. Platelet, red blood cell (RBC) and white blood cell (WBC) counts in both groups were compared. The 100 g/400 g spin gave a high platelet yield (increase of 395.4 ± 111.1%) in the leukocyte-poor-platelet-rich plasma group, while in the leukocyte-rich platelet-rich plasma group both 100 g/400 g and 350 g/1350 g spins resulted in significantly higher yields with an increase of 691.5 ± 316.3% and 738.6 ± 193.3%, respectively. The study was limited by a smaller sample size in the pure platelet-rich plasma (leukocyte-poor platelet-rich plasma) group. Ideal platelet yields can be achieved with both the 100 g/400 g as well as the 350 g/1350 g spins using the buffy coat inclusion method while the 100 g/400 g spin for "pure" platelet-rich plasma accomplishes a near-ideal platelet count with significantly reduced contamination with other cells.

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.

Qualitative Evaluation of Platelet Concentrates Prepared with Different Methods

Introduction: One of the most important responsibility of Blood Bank is to ensure uninterrupted supply of quality blood products, namely Platelet Concentrate (PC), which are in high demand particularly during pandemic. In India, PCs are of three types prepared by different methods like Platelet Rich Plasma-Platelet Concentrate (PRP-PC), Buffy Coat Platelet Concentrates (BC- PC), and Single Donor Platelets (SDP) units. Hence, establishing optimum quality standards across all the three types of PC remains a challenge in order to maximum benefit to recipients. Aim: To assess and compare the quality parameters of platelet concentrate prepared by PRP-PC, BC-PC and apheresis methods (SDP). Materials and Methods: This laboratory based observational study was conducted at rural, tertiary care, postgraduate, teaching hospital located at South India from December 2018 to November 2019. A total of 156 PCs were studied with 52 units belonging to PRP method, 52 units BC-PC method and the remaining were 52 units of SDP prepared by apheresis method. Quality parameters include volume, pH, swirling, White Blood Cell (WBC) count/bag, platelet count/bag and platelet indices. Platelet indices include Mean Platelet Volume (MPV), Platelet Distribution Width (PDW), and Platelet Large Cell Ratio (PLCR). Arterial Blood Gas (ABG) analysis was also done to assess (pH, partial pressure of Carbon Dioxide (pCO2), plasma bicarbonate (pHCO3-)). The p-value less than 0.05 were considered statistically significant. Statistical Package for the Social Sciences (SPSS) software version 23.0 used for statistical analysis. Results: All the 156 PC units were analysed as per the Director General of Health Services (DGHS) India, criteria. Among the 156 platelet concentrates scored as per DGHS criteria, a score of 5 was obtained for 34.61% [18/52], 30.76 [16/52] and 11.5% [6/52] SDP, BC-PC and PRP-PC units, respectively. All the PC units were sterile and Red Blood Cell (RBC) contamination was within acceptable limits. Conclusion: In this study, quality parameters of SDP was better than BC-PC and PRP-PC but with improvement and more standardisation of procedures and storage, BC-PC units can give a yield similar to that of SDP units.

Preparation of Platelet-Rich Plasma: National IADVL PRP Taskforce Recommendations.

The goal of these recommendations is to provide a framework to practitioners for implementing useful, evidence-based recommendations for the preparation of platelet-rich plasma (PRP) in various dermatological indications. The Indian Association of Dermatologists, Venereologists and Leprologists (IADVL) assigned the task of preparing these recommendations to its task force on PRP. A comprehensive literature search was done in the English language on the preparation of PRP across multiple databases. The grade of evidence and strength of recommendation was evaluated on the GRADE framework (Grading of Recommendation, Assessment, Development and Evaluation). A draft of clinical recommendations was developed on the best available evidence which was also scrutinized and critically evaluated by the IADVL Academy of Dermatology. Based on the inputs received, this final consensus statement was prepared. A total of 45 articles (meta-analyses, prospective and retrospective studies, reviews [including chapters in books], and case series) were critically evaluated and the evidence thus gathered was used in the preparation of these recommendations. This expert group recommends the use of double-spin manual method for the preparation of PRP. The recommended parameters for centrifuge are 100–300 g for 5–10 min for the first spin and 400–700 g for 10–17 min for the second spin. The recommended platelet concentration in PRP for the treatment of various dermatological conditions is 1–1.5 million platelets/μL. The activation of PRP is not required when it is injected into soft tissues.

Platelet-Rich Plasma in Androgenetic Alopecia.

The goal of these recommendations is to provide a framework to practitioners for implementing useful, evidence-based recommendations for the preparation of platelet-rich plasma (PRP) and its use in androgenetic alopecia (AGA). The Indian Association of Dermatologists, Venereologists and Leprologists (IADVL) assigned the task of preparing these recommendations to its taskforce on PRP. A comprehensive literature search was done in the English language on the PRP across multiple databases. The grade of evidence and strength of recommendation were evaluated on the GRADE (Grading of Recommendation, Assessment, Development, and Evaluation) framework. A draft of clinical recommendations was developed on the best available evidence, which was also scrutinized and critically evaluated by the IADVL Academy of Dermatology. Based on the inputs received, the final consensus statement was prepared. A total of 30 articles (meta-analyses, prospective and retrospective studies, reviews [including chapters in books], and case series) were critically evaluated, and the evidence thus gathered was used in the preparation of these recommendations. This expert group recommends use of manual double-spin method for the preparation of PRP for AGA. Minimum three to five sessions of PRP are recommended for AGA with a gap of 1 month between the two sessions. Patients with Grade II to V Norwood Hamilton classification of AGA are the ideal subset for PRP. A total of 5 to 7 mL of PRP and 0.05 to 0.1 mL/cm2 is the recommended dose of PRP for AGA. Activation of PRP is not required when it is used for AGA. About 1 to 1.5 million platelets/μL of platelets in PRP is the recommended platelet concentration in PRP for the treatment of AGA. I-PRF (injectable platelet-rich fibrin) has also been found to be useful in AGA, although further studies are required to establish its role. PRP can also have an adjunctive role in hair transplantation procedures.

The optimal platelet concentration in platelet-rich plasma for proliferation of human cells in vitro-diversity, biases, and possible basic experimental principles for further research in the field: A review.

BackgroundIn the last decades, several in vitro studies have tested the effect of plate-rich plasma (PRP) on the proliferation of human cells in search of a wizard for the use of PRP in a clinical setting. However, the literature displays striking differences regarding this question despite the relatively similar experimental design. The aim of this review is twofold: describe and explain this diversity and suggest basic principles for further in vitro studies in the field. The optimal platelet concentration in vivo will also be discussed.MethodsA search in mainly EMBASE and PubMed was performed to identify in vitro studies that investigate the effect of different PRP concentrations on human cell proliferation. The assessment of bias was based on the principles of “Good Cell Culture Practice” and adapted.ResultsIn total, 965 in vitro studies were detected. After the initial screening, 31 studies remained for full-text screening. A total of 16 studies met the criteria of final inclusion and appeared relatively sound. In general, the studies state consistently that PRP stimulates the proliferation of the human cell. Two main types of experimental techniques were detected: 1. The Fixed PRP Concentration Group using a fixed PRP concentration throughout the experiment, which leads to a substantial decrease in nutrition available at higher concentrations. 2. The Fixed PRP Volume Group using a fixed PRP-to-media ratio (Vol/Vol) throughout the experiment. A general tendency was observed in both groups: when the PRP to media ratio increased (Vol/Vol), the proliferation rate decreased. Further, The Low Leukocyte group observed a substantial higher optimal PRP concentration than The High leukocyte group. No prominent tendencies was seen regarding anticoagulants, activation methods, and blood donor (age or sex).DiscussionTwo major biases regarding optimal proliferation in vitro is pointed out: 1. Too high PRP volume. It is speculated that the techniques used by some studies led to an adverse growth condition and even cell starvation at higher concentrations. 2. High leukocyte levels. Reduced proliferation rate due to proinflammatory substances released during degranulation of leukocytes.ConclusionsThe two main biases may explain the bell-shaped effect of PRP and the detrimental effects at higher platelet concentrations observed in several studies. These biases may also explain the low optimal PRP concentration observed in some studies. Even if one universal optimal PRP concentration does not exist, the review indicates that PRP concentrations in the upper parts of the scale is optimal or at least beneficial. Finally, following basic experimental principles are suggested. 1: The PRP/media ratio (Vol/Vol) should be kept as constant. 2: The PRP/media ratio should provide a sufficient nutrition supply, that is, PRP ≤ 10% (Vol/Vol). 3: The cell density per well (cells/mL) should be defined. 4: Leukocyte level should be kept low, preferable depleted (< 0.1 PLT/µL).

Platelet Rich Plasma - Platelet Counts and Application - A Literature Review

Platelet rich plasma (PRP) is a novel method of using plasma concentrated with platelets for wound healing and tissue regeneration. Platelet rich plasma is prepared from the venous blood using a differential centrifugation technique. It involves a separation spin and a concentration spin, yielding platelet rich plasma. PRP products have been classified into 4 types depending upon major cell constituent and fibrin density upon activation. These are as follows: Pure PRP, Leukocyte and PRP, Pure PRF, Leukocyte and PRF. PRF differs from PRP in that it is rich in a high density fibrin network after activation. PRP is abundant in a variety of growth factors such as VEGF, PDGF, TGF, EGF, and Interleukin-1. Literature consists of reports by different authors about the platelet yield of PRP centrifuged by different systems. A number of factors have also been quoted to influence the platelet concentration in platelet rich plasma. Hence, the aim of this review is to discuss the platelet concentration in PRP centrifuged by different systems and to observe for variations if any.

Comparison of Platelet-Rich Plasma-Impregnated Suture Material with Low and High Platelet Concentration to Improve Colonic Anastomotic Wound Healing in Rats.

Objective This study was designed to investigate the impact of using suture material impregnated with platelet-rich plasma (PRP) in different platelet concentrations on colonic anastomotic wound healing in rats. Methods A total of 24 Sprague Dawley female rats were separated into 3 groups (n = 8 for each) including the control group (CON; standard vicryl suture repair), the low platelet concentrate PRP group (L-PRP; suture material impregnated with PRP containing average 2.7-fold (range, 2.0 to 3.1) higher amount of platelets vs. control), and the high platelet concentrate PRP group (H-PRP; suture material impregnated with PRP containing average 5.1-fold (range, 4.8 to 5.4) higher amount of platelets vs. control). Rats were sacrificed on the postoperative 7th day for analysis of colonic anastomosis region including macroscopic observation, measurement of anastomotic bursting pressure (ABP), and the hydroxyproline levels and histopathological findings in colon tissue samples. Results Total injury scores were significantly lower in the L-PRP and H-PRP groups than those in the control group (median (range) 13.00 (7.00) and 11.50 (6.00) vs. 15.50 (4.00), p < 0.05 and p < 0.01, respectively). ABP values (180.00 (49.00) vs. 124.00 (62.00) and 121.00 (57.00) mmHg, p < 0.001 for each) and tissue hydroxyproline levels (0.56 (0.37) vs. 0.25 (0.17) and 0.39 (0.10) μg/mg tissue, p < 0.001 and p < 0.05, respectively) were significantly higher in the L-PRP group as compared with those in the control and H-PRP groups. Conclusion In conclusion, our findings revealed PRP application to colonic anastomosis sutures to promote the anastomotic healing process. The platelet concentration of PRP seems to have a significant impact on the outcome with superior efficacy of L-PRP over H-PRP in terms of bursting pressures and collagen concentration at the anastomotic site.