Platelet-rich Plasma Preparation Research Articles

The Efficacy of Platelet-Rich Plasma for Ligament Injuries: A Systematic Review of Basic Science Literature With Protocol Quality Assessment.

Background:Despite the existence of many clinical studies on platelet-rich plasma (PRP) interventions for ligamentous pathology, basic science consensus regarding the indications, mechanisms, and optimal composition of PRP for treating ligament injuries is lacking.Purpose:To (1) compare the efficacy of PRP in animal models of ligament injury with placebo and (2) describe the potential variability in PRP preparation using accepted classification systems.Study Design:Systematic review.Methods:The Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, PubMed, Embase, and Ovid MEDLINE were queried in April 2020 for in vivo and in vitro basic science studies regarding PRP use for ligament injury. Study design, results, PRP composition, and analyzed cellular and molecular markers were extracted, and outcomes relative to control models were documented. Bias was assessed using the SYRCLE risk-of-bias tool.Results:Included were 43 articles (31 in vivo and 12 in vitro studies) investigating the anterior cruciate ligament/cranial cruciate ligament (n = 32), medial collateral ligament (n = 6), suspensory ligament (n = 3), patellar ligament (n = 1), and Hock ligament (n = 1). Platelet concentration was reported in 34 studies (77.3%); leukocyte composition, in 12 (27.3%); and red blood cell counts, in 7 (15.9%). With PRP treatment, 5 of 12 in vitro studies demonstrated significant increases in cell viability, 6 of 12 in gene expression, 14 of 32 in vivo studies reported superior ligament repair via histological evaluation, and 13 in vivo studies reported superior mechanical properties. Variability in PRP preparation methods was observed across all articles, and only 1 study reported all necessary information to be classified by the 4 schemes we used to evaluate reporting. Among the in vivo studies, detection and performance bias were consistently high, whereas selection, attrition, reporting, and other biases were consistently low.Conclusion:Conflicting data on the cellular and molecular effects of PRP for ligament injuries were observed secondary to the finding that included studies were heterogeneous, limiting interpretation across studies and the ability to draw meaningful conclusions. Clinical trials and any causal relationship between PRP use in ligament injuries and its potential for regeneration and healing should be pursued with caution if based solely on basic science data.

An Evaluation of the Effect of Activation Methods on the Release of Growth Factors from Platelet-Rich Plasma.

Activation of platelets in platelet-rich plasma may improve growth factor release, thus enhancing regenerative properties. The authors investigated whether different methods of platelet-rich plasma activation affected growth factor release kinetics over time. Platelet-rich plasma from 20 healthy volunteers was processed by six different methods: (1) control (nonactivated); (2) activation with calcium chloride; (3) activation with calcium chloride and ethanol; (4) activation with calcium chloride and ethanol at 4°C; (5) activation with calcium chloride and ethanol with vitamin C; (6) activation with calcium chloride and ethanol with vitamin C at 4°C. Concentration of secreted vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and insulin-like growth factor over 24 hours was measured by immunoassay. Calcium chloride-activated platelet-rich plasma produced significantly more insulin-like growth factor at 1 hour compared to cold and vitamin C platelet-rich plasma, and calcium chloride plus ethanol produced significantly more at 24 hours compared to vitamin C platelet-rich plasma. The addition of vitamin C reduced release of PDGF over time. Activation with calcium chloride and ethanol with or without cold temperature produced a gradual PDGF release as opposed to calcium chloride alone, which caused higher PDGF within 4 hours. There were no significant differences between groups for VEGF, although calcium chloride and cooled platelet-rich plasma approached significance for producing more than vitamin C platelet-rich plasma. Activation of platelet-rich plasma does not significantly improve growth factor secretion, which is made worse by the addition of vitamin C, a platelet inhibitor. Ethanol does not negatively impact growth factor production and may offer a more gradual release. These findings will help guide platelet-rich plasma preparation methods where therapeutic growth factors are used. Therapeutic, V.

Method to obtain PRP and L-PRP for the use in routine medical practice of orthopedic traumatologist

The paper presents the results of experimental study of two methods for blood processing to obtain platelet-rich plasma (PRP). Due to the widespread clinical use of platelet-rich plasma in orthopedics and traumatology, a differential approach to the use of PRP and leukocyte- and platelet-rich plasma (L-PRP) has become relevant. Most commercial kits for PRP preparation significantly increase the cost of treatment. In addition, the main attention is focused on the concentration of platelets in the final product and the marketing features of a particular kit (separating gel, tube shape, anticoagulant, activator, etc.). The role of leukocytes in the features of the regenerative effect of platelet-rich plasma stays out of focus. The blood of volunteers was processed in two different ways and the cellular composition of the obtained products was analyzed. Sample No. 1 was used as a control for the baseline blood cell composition. Sample No. 2 was centrifuged in a sterile plastic tube, platelet rich plasma was collected manually. Sample No. 3 was centrifuged in a sterile modified syringe, platelet rich plasma was collected using a closed technique by means of connectors together with a rich layer. The results of statistical analysis showed that there were obtained totally different biological products with different concentrations of platelets and leukocytes. The first sample (control) had platelet concentration of (228.69 ± 39.15) × 109/l and leukocyte concentration of (5.18 ± 1.32) × 109/l. In the second sample (tube + manual sampling), platelet concentration was (429.38 ± 79.92) × 109/l and leukocyte concentration — (0.85 ± 0.34) × 109/l. The third sample (syringe plunger + closed vacuum collection) had platelet concentration of (541.15 ± 85.49) × 109/l and leukocytes of (6.56±1.92)×109/l. The data are given without regard to –12.5% deviation on dilution with citrate dextrose. As the result of the work, orthopedic traumatologists can use the simplest non-commercial methods for obtaining PRP and L-PRP in their practice depending on nosology and clinical tasks.

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.

Experimental assessment of regenerative properties of platelet rich plasma on the human skin - a review

Several studies demonstrated the favorable effects of platelet rich plasma (PRP) on the skin and promoted its wide use in clinical practice. The growth factors stored in platelet alfa-granules allow for the tissue regeneration and the main fields of application of PRP in current clinical practice are the cartilage and musculoskeletal defects, osteoarthritis and other bone disorders, chronic and difficult to heal wounds, and aesthetic procedures. The relevant number of different PRP preparation protocols may explain the inconsistency of the different clinical outcomes reported in the literature. Despite the technological advances in PRP preparation, the objective assessment of the clinical efficacy of PRP from the literature reports still is difficult due to the low homogeneity of the samples in terms of both inclusion criteria and size. Therefore, it might be useful to establish standardized and reproducible experimental models to confirm and objectively measure the effectiveness of the available clinical results. Many experimental investigations have been carried out to objectively assess the effectiveness of PRP and platelet gel on several tissues. As far as the skin is concerned, the studies carried out to date are limited to fibroblasts in in-vitro culture models or to collagen, vascular supply, epithelium, and hair follicle in in-vivo models. The skin, however, is a very complex organ, where different cell lines coexist and feature complex mutual interaction. A model that combines the advantages of both in-vitro and in-vivo cultures is the ex-vivo model. The demonstration of the platelet derived growth factors effects through the ex-vivo human full-thickness skin culture model is a keystone to support the evidence of the PRP effectiveness, as it represents an objective, fast, reproducible, and ethical investigational method.

THE INFLUENCE BETWEEN INJECTABLE PLATELET-RICH FIBRIN AND PLATELET-RICH PLASMA TOWARDS GINGIVAL FIBROBLAST CELL PROLIFERATION

ABSTRACTBackground: Gingiva is the outermost periodontal tissue that acts as a mechanical and biological barrier to the root of the teeth and alveolar bone. The main cellular elements in the gingiva are fibroblasts. Fibroblast cell proliferation is an important process in tissue regeneration. Growth factors that can stimulate fibroblast cell proliferation can be found in regenerative agents, such as injectable platelet-rich fibrin (i-PRF) and platelet-rich plasma (PRP). The aim of this study was to examine the influence between i-PRF and PRP on the gingival fibroblast cell proliferation in vitro study on primary cell culture.Method: Gingival fibroblast cell were obtained from primary cell culture derived from healthy gingiva. Ten mL of peripheral blood were centrifuged for i-PRF and PRP preparation. The samples were divided into three groups: i-PRF, PRP, and fibroblast cells without treatment. Cell proliferation were observed at day 1, day 3, day 5 using MTT assay at 550 nm. The data were analyzed by Two-Way ANOVA test, followed by Post Hoc test.Result: The results showed that the cell proliferation increased from day 1, 3, and 5 in all groups. The absorbance value of the cell proliferation in order from highest to lowest: i-PRF, PRP, and cell control.Conclusion: i-PRF and PRP increased the gingival fibroblast cell proliferation. i-PRF increased the cell proliferation higher than PRP.

Platelet Concentrates and Their Clinical Applications in Oral and Maxillofacial Surgery, Periodontal, Dentoalveolar and Reconstructive Surgery and Endodontics. Current Review.

During the last two decades a new technology appeared called platelet rich plasma preparations (PRP) [1], suggesting the main benefit in both hard and soft tissue wound healing for surgical use in regenerative medicine [2], as well as aiding in decreasing: rate of postoperative infections, pain and blood loss [3]. The PRP clinical applications include periodontal surgery, Oral and Maxillofacial surgery, aesthetic plastic surgery, spinal fusion, heart by-pass surgery, treatment of soft tissue ulcers, etc. [3]. However in the recent times there have been described different types of PRP concentrates and different clinical indications for every of them. The aim of this review is to enlist and describe the different types of PRP concentrates and their current clinical use in Dentistry and Oral and Maxillofacial surgery.

Effect of topical application of platelet-rich plasma on chronic venous leg ulcerations.

Wound products that reliably support healing of chronic leg ulcers remain a huge unmet need in clinical practice. Due to the lack of standardized comparable protocols and different systems for platelet-rich plasma (PRP) preparation, there is limited data on healing rates in chronic venous ulcers. In our case series with a total of seven chronic leg ulcers in four patients, we investigated the healing rates based on standardized digital photographs of chronic venous ulcers after application of topical PRP using a digital imaging software. In 5 out of 7 ulcers, the PRP-treated wound half showed faster healing as compared the control half of the wound. In this case series, PRP-treated sides of chronic venous leg ulcers showed a tendency for accelerated healing as compared to nontreated collateral wound side. Our data support the evaluation of topical PRP treatment in the management of chronic venous leg ulcers.

Simple Tube Centrifugation Method for Platelet-Rich Plasma (PRP) Preparation in Catalonian Donkeys as a Treatment of Endometritis-Endometrosis.

Simple SummaryPlatelet-rich plasma (PRP) is used to improve the regenerative capacity of damaged tissues in different species. In equine medicine, PRP is commonly used to treat joint diseases, tendinitis, ligamentous lesions, and persistent endometritis. Jenny endometrium shows a high sensitivity to endometritis. There are important differences between donkey and horse blood characteristics. Several protocols to obtain horse PRP have been reported, but no protocols have yet been reported for obtaining donkey PRP. Our study shows that single-spin tube centrifugation at 133× g for 10 min is appropriate to obtain donkey PRP with therapeutic potential in jenny endometritis-endometriosis.The aim of this study was to standardize a simple, manual platelet-rich plasma (PRP) protocol in Catalonian donkeys using single-spin tube centrifugation as a treatment for jenny endometritis. The objective was to obtain a blood product with a moderate concentration of platelets (2 or 3 times baseline physiologic values) and a low WBC (White Blood Cells) concentration. Blood was drawn from six Catalonian donkeys using acid citrate dextrose (ACD) as an anticoagulant, and then processed by single centrifugation at 133× g for two different centrifugation times (10 and 15 min). The PRP samples were evaluated by flow cytometry, and TGF-β1 (Transforming Growth Factor-Beta1) concentrations were determined by enzyme-linked immunosorbent assay (ELISA). The 10 min centrifugation protocol resulted in a slightly greater release of TGF-β1 (6044.79 ng/mL), a 2.06-fold increase in platelet concentration, and a 15-fold reduction in leukocyte concentration when compared to the initial values. The 15 min centrifugation time resulted in a 2.44-fold increase in baseline platelet concentration, a reduction in WBC count by a factor of 20, and slightly lower TGF levels (5206 ng/mL). We conclude that both protocols are adequate for the obtention of PRP, and both may have an acceptable therapeutic potential for use in this species, although this needs to be further validated.

The Effect of a Single Freeze-Thaw Cycle on Matrix Metalloproteinases in Different Human Platelet-Rich Plasma Formulations.

Storing platelet-rich plasma (PRP) for future use is a compelling approach, presuming the retention of biological properties is maintained. However, certain factors in PRP preparations have deleterious effects for the treatment of certain musculoskeletal conditions. The purpose of this study was to measure and compare matrix metalloproteinase protein (MMP) concentrations between fresh and freeze-thawed leukocyte-rich PRP (LR-PRP) inactivated (LR-I) and activated (LR-A) preparations, and leukocyte-poor PRP (LP-PRP) inactivated (LP-I) and activated (LP-A) preparations. A volume of 60 mL of whole blood was drawn from 19 healthy donors. LP-I and LR-I samples were processed using a manual extraction and centrifugation methodology. LP-A and LR-A products were activated with 10% CaCl2 and recombinant thrombin. Blood fractions were either immediately assayed and analyzed or stored at −80 °C for 24, 72 and 160 h. Multiplex immunoassay was used to measure MMP-1, MMP-2, MMP-3, MMP-9, MMP-10, and MMP-12. MMP-1 concentrations increased in LR-A (p < 0.05) and MMP-9 significantly increased in LR-I (p < 0.05), while MMP-2 significantly decreased in LR-I (p < 0.05) and MMP-3 concentrations significantly decreased in LR-A (p < 0.05). MMP-12 concentrations also significantly decreased in LR-I (p < 0.05) from baseline concentrations. There were no significant differences between LP-A and LP-I preparations and MMP concentrations. MMP-10 concentrations in all PRP samples compared to each freezing time point were also not significantly different. MMPs regulate components of the extracellular matrix (ECM) in the remodeling phase of musculoskeletal injury. In this study, we observed a significant increase and decrease in MMP concentrations in response to a single freeze–thaw cycle in inactivated PRP and activated PRP preparations. This evidence contributes to the growing body of literature on the optimization of PRP preparation and storage strategies prior to delivery. Our findings suggest that specific PRP preparations after a single freeze–thaw may be more advantageous for certain musculoskeletal applications based on the presence of MMP concentrations.

A Split Scalp Study of Single versus Double Spin Platelet-rich plasma Injections in Treatment of Female Pattern Hair Loss: Clinical Effect and Relation to Vascular Endothelial Growth Factor in PRP

Abstract Background Female pattern hair loss (FPHL) is the most common hair loss disorder in women. Platelet-rich plasma (PRP) provides an option for patients recalcitrant to the currently approved therapies. As there are many methods for the preparation of PRP, the most suitable for AGA therapy is not yet clear. Aims To compare the efficacy between single and double spin PRP injections in treatment of FPHL and to assess the relationship between the concentrations of vascular endothelial growth factor (VEGF) in the PRP preparation and the treatment results obtained. Patients and Methods 15 female patients with FPHL were enrolled. Each patient was subjected to intradermal injection of PRP (prepared by double-spin method) into the right half of the scalp, in addition to intradermal injection of PRP (prepared by singlespin method) into the left half of the scalp in the form of three treatment sessions three weeks apart. Evaluation of treatment response was done through comparing patients' global photographs, assessment of patients' satisfaction and folliscopic assessment before and after treatment. Results Our results showed clinically significant improvement by comparing patients' photographs in most of the treated patients. In addition, folliscopy revealed significant change of the terminal hair density in the right half of the scalp following treatment sessions.VEGF concentration didn't differ significantly between double-spin and single-spin prepared PRP, also it didn't differ significantly upon calcium activation. Conclusions PRP is an effective treatment modality for FPHL. Double-spin prepared PRP could yield better results in the treatment of FPHL than the single-spin method. However, adding Calcium gluconate prior to PRP injection is of no benefit as it didn't significantly increase the release of VEGF. Yet, further studies discussing different PRP preparation protocols and involving a larger number of cases are needed to minimize statistical errors and to detect the most accurate PRP preparation method with the greatest yield of growth factors.

Platelet-rich plasma in the treatment of chronic inflammatory diseases of lower extremities

Objectives to evaluate the effectivenes of platelet-rich plasma preparations in the treatment of chronic inflammatory diseases of lower extremities.&#x0D; Material and methods. The study involved patients hospitalized in the Research Institute S. V. Ochapovsky Regional Clinic Hospital No.1, Krasnodar, with trophic ulcers and chronic osteomyelitis. The patients received injections of platelet-rich plasma as platelet lysate and/or platelet gel.&#x0D; Results. The study involved 19 patients with trophic ulcers of the lower extremities with a duration of more than 2 months. Previously, the patients had underwent therapy without effect. In all cases, the patients received a complex of therapeutic measures, including debridement, dressing, conservative treatment, physiotherapy, antibiotic therapy. In the presence of large ulcers, 8 patients underwent autodermoplasty with complete engraftment. It has been noted that during platelet-rich plasma use the average healing time was 6-7 weeks.&#x0D; Four patients were observed with chronic osteomyelitis. During operation, the bone cavity was filled with platelet gel. The postoperative period was uneventful in all cases. The maximum observation period was 1.5 years. No recurrence of osteomyelitis was detected.&#x0D; Conclusion. The results of the study demonstrate the positive effects of platelet-rich plasma products use.

Platelet-rich Plasma and Its Application in Clinical Trial

Platelet-rich Plasma and Its Application in Clinical Trial

Закономірності зміни концентрацій ІЛ-17А, ІЛ-10 та ІЛ-2 у крові людей із гніздовою алопецією до та після лікування збагаченою тромбоцитами плазмою

Alopecia areata is a heterogeneous disease characterized by nonscarring hair loss. It affects 1-2% of the population of both genders and occurs in all age groups. According to the recent studies, helper T cells 1, helper T cells 2, helper T cells 17, and regulatory T cells may be the main possible initiators of alopecia areata. The purpose of the study was to determine the concentrations of serum interleukin-17A, interleukin-10 and interleukin-2 in patients with alopecia areata, and to study the relationship between their levels before and after treatment with platelet-rich plasma. Materials and methods. The study involved 37 patients with alopecia areata aged (35.7±8.9) years and 30 people of the control group aged (37.4±7.9) years. People with alopecia areata were injected with platelet-rich plasma. For this, immediately before administration, venous blood was taken from the patients from the cubital vein into a special tube for the preparation of platelet-rich plasma. Platelet-rich plasma was injected into the lesions at the rate of 0.2 ml/cm2 using an insulin syringe and a 30 G needle, once every 2 weeks for 16 weeks. The patients were examined before the treatment and in 3, 6 and 17 months. The concentrations of interleukin-17A, interleukin-2 and interleukin-10 were determined in blood serum before treatment and after 3, 6 and 17 months of observation by the Sandwich-ELISA method (enzyme-linked immunosorbent assay). Interleukin-17A, interleukin-2 and interleukin-10 in the blood serum in the control group were also determined. The software program SPSS 22.0 was used for the statistical analysis. The normality distribution was tested by Shapiro-Wilk’s method. Independent Student’s t-test for unpaired samples was used to compare mean values between groups. The p-value &lt; 0.05 was considered significant. Results and discussion. The results obtained in our study showed that the concentrations of the pro-inflammatory cytokines interleukin-17A and interleukin-2 were significantly higher in persons with alopecia areata before treatment. After 17 months of therapy with platelet-rich plasma, serum interleukin-17A and interleukin-2 levels approached the control level. Conclusion. There was no difference in the concentration of anti-inflammatory interleukin-10 between patients with alopecia areata and healthy people. In the course of treatment, the concentration of interleukin-10 increased after 3 and 17 months of therapy

Platelet HMGB1 in Platelet-Rich Plasma (PRP) promotes tendon wound healing.

Platelet-rich plasma (PRP) is a widely used autologous treatment for tendon injuries in clinics. Platelets (PLTs) are a major source of high mobility group box1 (HMGB1) that is gaining attention as a chemoattractant that can recruit stem cells to the wound area to enhance healing of injured tissues; however, the contribution of PLT HMGB1 in wounded tendon healing remains unexplored. This study investigated the effect of PLT HMGB1 within PRP on tendon healing using PLT HMGB1 knockout (KO) and GFP mice. A window defect was created in the patellar tendons of both groups of mice, and wounds were treated with either saline, PRP isolated from PLT HMGB1-KO mice, or PRP isolated from GFP mice. Seven days post-treatment, animals were sacrificed and analyzed by gross inspection, histology, and immunostaining for characteristic signs of tendon healing and repair. Our results showed that in comparison to mice treated with PRP from PLT HMGB1-KO mice, wounds treated with PRP from GFP mice healed faster and exhibited a better organization in tendon structure. Mice treated with PRP from PLT HMGB1-KO mice produced tendon tissue with large premature wound areas and low cell densities. However, wounds of PLT HMGB1-KO mice showed better healing with PRP from HMGB1-KO mice compared to saline treatment. Moreover, wounds treated with PRP from GFP mice had increased extracellular HMGB1, decreased CD68, increased stem cell markers CD146 and CD73, and increased collagen III protein expression levels compared to those treated with PRP from PLT HMGB1-KO mice. Thus, PLT HMGB1 within PRP plays an important role in tendon wound healing by decreasing inflammation, increasing local HMGB1 levels, and recruiting stem cells to the wound area in the tendon. Our findings also suggest that the efficacy of PRP treatment for tendon injuries in clinics may depend on PLT HMGB1 within PRP preparations.

Heritable platelet disorders: an enigma even guidelines can't unravel.

Heritable platelet disorders: an enigma even guidelines can't unravel.

P-050 The effectiveness of the platelet-rich plasma treatment of men with severe oligoasthenoteratozoospermia

Abstract Study question To evaluate the effect of platelet-rich plasma (PRP) testicular injections on spermogram parameters of men with severe oligoasthenoteratozoospermia (OAT). Summary answer The PRP testicular injections have beneficial effects on spermatogenesis and enhance sperm concentration and motility in infertile men with OAT. What is known already The use of PRP therapy in assisted reproductive technologies is debatable. Despite the recent evidence of its positive effects in promoting endometrial and follicular growth, data from clinical studies are limited. There are only a few papers on the effectiveness of PRP therapy in the treatment of male infertility and sexual dysfunction. In more detail, the influence of PRP on spermatogenesis was carried out only on experimental animals. Although the mechanisms of its action have not yet been clarified, it is assumed that PRP, containing many biologically active molecules, realizes its effect through the tissue regeneration and cell proliferation. Study design, size, duration This prospective study included 68 men (34.6±5.2) years old with severe OAT (≤4 million/ml, motility ≤30%, normal sperm morphology ≤1%) receiving hormonal and antioxidant (AO) therapy during 6 months before in vitro fertilization cycles. 33 of them were injected once with autologous PRP (0.5 ml in each testicle). Spermogram and testosterone level were analyzed before the treatment and in 3, 4 and 6 months after it. Participants/materials, setting, methods Sperm concentration, motility and morphology in ejaculate of 33 men of PRP group were compared with those in the group of 35 men without PRP within 6 months of starting the treatment. Total and free testosterone level were measured in blood serum. PRP was prepared by centrifuging the patient’s own blood in the anticoagulant-containing tubes. The final concentration of platelets in the obtained sample was 950.000 – 1.250 000 cells in 1 ml. Main results and the role of chance 4 months after the PRP injection, sperm concentration and motility increased in 18 of 33 men of the PRP group compared with the baseline (before the treatment) – 4.2 (1.0;6.9) vs 1.4 (0.1;3.4) mln/ml (p &amp;lt; 0.05) and 36.7 (30.6;45.8) vs 17.7 (6.7;28.2) % respectively (p &amp;lt; 0.05).The maximum increase in sperm motility (but not in sperm concentration!) was observed in 24 men in 6 months – 49.6 (39.6;56.4) % (p &amp;lt; 0.05). Percent of morphologically normal spermatozoa in ejaculate slightly increased only in 12 men in that time period from 0-1 % to 1-2%. The total testosterone level was 2.4 times higher than the baseline (31.6±7.2 vs 13.2±4.3 nmol/l, p &amp;lt; 0.05), the free testosterone level was 1.8 times higher (14.5±3.5 vs 7.9±3.0 pgl/ml, p &amp;lt; 0.05). Unlike the PRP group, in the group of men without PRP treatment, the sperm parameters did not changed compared with the baseline in 4 months after the starting hormonal and AO treatment. A significant increase of sperin concentration was observed only in 17 of 35 patients in 6 months. Sperm motility and percent of morphologically normal spermatozoa after the treatment did not differ from the baseline. Changes in the testosterone levels were similar to changes in PRP group. Limitations, reasons for caution Only young and middle-aged men were considered in the study. Large randomized controlled studies are required to confirm the PRP therapy efficacy and safety of f various fertility disorders. There are also no standardized protocols for PRP preparation. Wider implications of the findings PRP therapy may have great potential for the treatment of male infertility and improving spermatogenesis. Optimization of methods of PRP preparation and dosage of testicular injections can enhance reproductive outcomes in assisted reproductive technologies. Trial registration number not applicable

P–050 The effectiveness of the platelet-rich plasma treatment of men with severe oligoasthenoteratozoospermia

Abstract Study question To evaluate the effect of platelet-rich plasma (PRP) testicular injections on spermogram parameters of men with severe oligoasthenoteratozoospermia (OAT). Summary answer The PRP testicular injections have beneficial effects on spermatogenesis and enhance sperm concentration and motility in infertile men with OAT. What is known already The use of PRP therapy in assisted reproductive technologies is debatable. Despite the recent evidence of its positive effects in promoting endometrial and follicular growth, data from clinical studies are limited. There are only a few papers on the effectiveness of PRP therapy in the treatment of male infertility and sexual dysfunction. In more detail, the influence of PRP on spermatogenesis was carried out only on experimental animals. Although the mechanisms of its action have not yet been clarified, it is assumed that PRP, containing many biologically active molecules, realizes its effect through the tissue regeneration and cell proliferation. Study design, size, duration This prospective study included 68 men (34.6±5.2) years old with severe OAT (≤4 million/ml, motility ≤30%, normal sperm morphology ≤1%) receiving hormonal and antioxidant (AO) therapy during 6 months before in vitro fertilization cycles. 33 of them were injected once with autologous PRP (0.5 ml in each testicle). Spermogram and testosterone level were analyzed before the treatment and in 3, 4 and 6 months after it. Participants/materials, setting, methods: Sperm concentration, motility and morphology in ejaculate of 33 men of PRP group were compared with those in the group of 35 men without PRP within 6 months of starting the treatment. Total and free testosterone level were measured in blood serum. PRP was prepared by centrifuging the patient’s own blood in the anticoagulant-containing tubes. The final concentration of platelets in the obtained sample was 950.000 – 1.250 000 cells in 1 ml. Main results and the role of chance 4 months after the PRP injection, sperm concentration and motility increased in 18 of 33 men of the PRP group compared with the baseline (before the treatment) – 4.2 (1.0; 6.9) vs 1.4 (0.1; 3.4) mln/ml (p &amp;lt; 0.05) and 36.7 (30.6; 45.8) vs 17.7 (6.7; 28.2)% respectively (p &amp;lt; 0.05).The maximum increase in sperm motility (but not in sperm concentration!) was observed in 24 men in 6 months – 49.6 (39.6; 56.4)% (p &amp;lt; 0.05). Percent of morphologically normal spermatozoa in ejaculate slightly increased only in 12 men in that time period from 0–1% to 1–2%. The total testosterone level was 2.4 times higher than the baseline (31.6±7.2 vs 13.2±4.3 nmol/l, p &amp;lt; 0.05), the free testosterone level was 1.8 times higher (14.5±3.5 vs 7.9±3.0 pgl/ml, p &amp;lt; 0.05). Unlike the PRP group, in the group of men without PRP treatment, the sperm parameters did not changed compared with the baseline in 4 months after the starting hormonal and AO treatment. A significant increase of sperin concentration was observed only in 17 of 35 patients in 6 months. Sperm motility and percent of morphologically normal spermatozoa after the treatment did not differ from the baseline. Changes in the testosterone levels were similar to changes in PRP group. Limitations, reasons for caution Only young and middle-aged men were considered in the study. Large randomized controlled studies are required to confirm the PRP therapy efficacy and safety of f various fertility disorders. There are also no standardized protocols for PRP preparation. Wider implications of the findings: PRP therapy may have great potential for the treatment of male infertility and improving spermatogenesis. Optimization of methods of PRP preparation and dosage of testicular injections can enhance reproductive outcomes in assisted reproductive technologies. Trial registration number Not applicable

Physiological Parameters to Identify Suitable Blood Donor Cows for Preparation of Platelet Rich Plasma.

Simple SummaryPlatelet rich plasma is a biological product obtained from blood and used for regenerative treatments of different pathologies. It is characterized by a high concentration of platelets (at least 3 times the physiological level) containing many growth factors with anti-inflammatory, bactericidal and regenerative properties. In human medicine, PRP is used in an autologous way, it means that the blood donor is also the recipient. In veterinary medicine, PRP is used to treat different diseases or lesions and in bovine species to treat mastitis. In this context, the opportunity to have PRP ready to use, stored from donor cows of the same farm where it will be used, would be very useful in treating this pathology immediately when it occurs. For this purpose, the present research aimed to detect parameters useful to identify the most suitable cows to be used as blood donors to obtain the highest yield of PRP (milliliters of PRP obtained with respect to milliliters of initial blood). Our results showed that blood collection from the mammary vein within three months of parturition, from nonpregnant cows at 5 years of age, but not the blood collection season, were associated with a high yield of PRP.Platelet rich plasma (PRP) has been shown to be beneficial in the treatment of bovine mastitis, with an action comparable to that of antibiotics. Autologous treatment is feasible in experimental conditions but is difficult to apply in field conditions, particularly in acute mastitis. The ideal scenario would be to have heterologous PRP stored on every farm so that it is readily available when needed. In this paper, we analysed data collected during bovine mastitis treatment with heterologous PRP produced by casual donor cows on several farms. We tried to identify parameters which might be useful to identify the most suitable cows to be used as blood donors, to obtain the highest yield of PRP. Variables considered for each animal were the age, the parity, the date of the last parturition, the season of blood collection, the site of blood collection (jugular or mammary vein) and the reproductive status e.g., pregnant or not pregnant. There were statistically significant differences for all the variables considered from the 135 blood cows, except for the blood collection season. The highest yield of PRP was associated with nonpregnancy blood collection within three months of parturition, parity 3 or 4, and blood collection from the mammary vein.

The Effect of Platelet Rich Plasma on Tissue Healing after Mandibular Third Molar Surgery

Introduction: This study aimed, effect of platelet-rich plasma (PRP) on soft tissue healing after mandibular third molar surgery. Materials &amp; Methods: In this semi-blinded clinical trial study,30 selected patients requiring surgical extraction of soft tissue impacted mandibular third molar participated from cases referred to the Department of Surgery, Faculty of Dentistry, Isfahan Azad University 2017.Patients divided into both test and control groups. PRP was placed in the extracted socket of the test group, whereas the control group had no PRP. Arzhangian standard kit utilized for preparation of PRP. The outcome variables in this study were pain, swelling, inter incisal mouth opening, wound dehiscence, dry socket, bleeding, and tissue color. The collected data analyzed using statistical tests followed independent T-test, Mann-Whitney, and fisher (p value &lt; 0.05). Results: The mean postoperative pain score (Visual Analog Scale) lowered for the test group after several wound dehiscence on the third and seventh days after surgery which was statistically significant (p value &lt; 0.001). Although the mean bleeding time on the third day after surgery decreased in the test group, this difference was not statistically significant (p value = 0.59). There were no statistically significant between both groups for The dry socket incidence on the seventh day and the mean of inter incisal mouth opening in the third and seventh days after surgery as well. Conclusion: The usage of topical PRP has beneficial advantages on soft tissue healing after mandibular third molar surgery. Unlike the control group, The PRP group provided reduced pain and better soft tissue healing.