Pure Platelet-rich Plasma Research Articles

Leukocyte Platelet-Rich Plasma-Derived Exosomes Restrained Macrophages Viability and Induced Apoptosis, NO Generation, and M1 Polarization.

Chronic refractory wounds refer to wounds that cannot be repaired timely. Platelet-rich plasma (PRP) has significant potential in chronic wound healing therapy. The exosomes isolated from PRP were proved to exhibit more effectiveness than PRP. However, the therapeutic potential of exosomes from PRP on chronic refractory wounds remained elusive. Hence, this study aimed to clarify the action of exosomes from PRP on chronic refractory wounds by evaluating the response of macrophages to exosomes. Pure platelet-rich plasma (P-PRP) and leukocyte platelet-rich plasma (L-PRP) were prepared from the fasting venous blood of healthy volunteers. Exosomes were extracted from P-PRP and L-PRP using ultracentrifugation and identified by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and western blot. Macrophages were obtained by inducing THP-1 cells with phorbol-12-myristate-13 acetate (PMA). The internalization of exosomes into macrophages was observed utilizing confocal laser scanning microscopy after being labeled with PKH67. Cell viability was determined by CCK-8 assay. Cell apoptosis was measured utilizing a flow cytometer. The polarization status of M1 and M2 macrophages were evaluated by detecting their markers. Nitric oxide (NO) detection was conducted using the commercial kit. Exosomes from P-PRP and L-PRP were absorbed by macrophages. Exosomes from L-PRP restrained viability and induced apoptosis of macrophages. Besides, exosomes from P-PRP promoted M2 polarization, and exosomes from L-PRP promoted M1 polarization. Furthermore, exosomes from L-PRP promoted NO generation of macrophages. Exosomes from L-PRP restrained viability, induced apoptosis and NO generation of macrophages, and promoted M1 polarization, while exosomes from P-PRP increased M2 polarization. The exosomes from L-PRP presented a more effective effect on macrophages than that from P-PRP, making it a promising strategy for chronic refractory wound management.

Efficacy of Platelet-Rich Plasma Intra-articular Injections in Hip and Knee Osteoarthritis.

Osteoarthritis (OA) is a chronic degenerative disorder that causes significant pain and functional limitations. Platelet-rich plasma (PRP) therapy has gained considerable attention in recent years in the treatment of musculoskeletal injuries. In this narrative review, we aim to investigate the role of intra-articular PRP injections in the treatment of knee and hip OA. The review also discusses different classifications of PRP based on composition. Furthermore, this narrative review also identified various limitations of PRP therapy in OA. PRP is classified into different types based on cell content and fibrin architecture, including pure platelet-rich plasma (P-PRP), leukocyte- and platelet-rich plasma (L-PRP),pure platelet-rich fibrin (P-PRF), and leukocyte- andplatelet-rich fibrin (L-PRF).Various clinical trials have shown that PRP is an effective option for the treatment of knee and hip OA. However, the superiority of PRP over hyaluronic acid has been reported inconsistently. This variability can be attributed to PRP preparation techniques. The safety profiles of PRP are generally favorable and the adverse effects are generally mild in nature. Although there is sufficient evidence in support of PRP in the treatment of OA, the long-term effects of PRP have not been reported. Further studies should focus on longer follow-up periods to identify the efficacy and safety of PRP in treating knee OA. There is also a need for standardization of PRP preparations in OA management.

Effect of allogeneic pure platelet-rich plasma, sodium cloxacillin, and their combination for the treatment of subclinical mastitis in crossbred cows.

Bovine subclinical mastitis (SCM) caused by Gram-positive bacteria is a major cause of economic loss in the dairy industry, exacerbated in situations where antimicrobial resistance is present. Pure platelet-rich plasma (P-PRP) may be a therapeutic alternative for SCM, when used alone or with antibiotics, such as sodium cloxacillin (SC). This study aimed 1) to evaluate the therapeutic efficacy of allogeneic P-PRP, SC, and their combination (P-PRP+SC) in cows with SCM caused by Staphylococcus aureus and by streptococci (Staphylococcus aureus and S. dysgalactiae); 2) to determine the concentrations of somatic cells (SCC), interleukin 1 beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and TGF-β1 in milk samples of the cows. 130 cows from 4 dairy herds completed the study, of which 40 cows were treated with P-PRP (10 mL), 28 cows with SC (5g), 36 with P-PRP+SC (10mL/5g), and 26 did not receive no treatment (negative control group, NCG). The overall bacteriological cure was observed in 10/40 (25%) cows in the P-PRP group, 9/28 (32.14%) animals in the SC group, 26/36 (72.22%) cows in the P-PRP+SC group, and 10/26 (38.46%) animals in the NCG. SCM caused by S. aureus (82/130, 63.08%), was cured in 6/24 (25%) cows treated with P-PRP, 7/24 (29.2%) cows treated with SC, 8/16 (50%) animals treated with P-PRP+SC, and in 8/18 (44.4%) cows in NCG. For SCM caused by the streptococci (48/130, 36.91%), the cure was achieved in 4/12 (33.3%) cows treated with P-PRP, 2/4 (50%) cows treated with SC, 18/20 (90%) cows treated with P-PRP+SC, and in 2/8 (25%) cows of the NCG. SCC was significantly (p < 0.001) affected by the treatment, herd, cure, bacteria group, and number of calvings factors. IL-1β milk concentrations were significantly (p < 0.001) influenced by treatment and farm factors, and the interaction between these factors. TNF-α milk concentrations were significantly (p < 0.001) influenced by time factor. TGF-β1 milk concentrations were significantly affected by the time and cure factors. The combination of P-PRP and SC showed the best therapeutic response (90%) against bovine SCM caused by streptococci. However, none of the treatments showed an effective therapeutic response against S. aureus.

The activity of monocyte-derived macrophages after stimulation with platelet-rich and platelet-poor concentrates. Study on an ovine model of insertion of a tibial implant coated with silicon-doped diamond-like carbon.

Macrophages are crucial immune cells that play a role in tissue repair and can exhibit pro- or anti-inflammatory behaviour based on environmental stimulation. Their functional phenotype can be affected by platelet-derived products as determined by those products' composition. When the inflammatory response caused by implantation is excessive, it can lead to rejection of the implant. Therefore, a thorough evaluation of implant haemocompatibility is necessary to minimise undesirable consequences. In an in vitro study, monocyte-derived macrophages (MDMs) were obtained from the whole blood of sheep after a silicon-doped diamond-like carbon-coated implant insertion. These MDMs were then exposed to autologous platelet-derived products for functional marker analysis. Platelet-poor plasma (PPP) and pure platelet-rich plasma (P-PRP) stimulation increased arginase-1 activity, while leukocyte-rich PRP stimulation produced a mixed response involving higher O2- (6.49 ± 2.43 nM vs non-stimulated 3.51 ± 1.23 nM, P-value < 0.05) and NO (3.28 ± 1.38 μM vs non-stimulated 2.55 ± 0.32μM, P-value < 0.05) generation. Using PPP and P-PRP stimulation in post-implantation procedures may contribute to the polarisation of macrophages towards the M2-like pro-resolving phenotype, thereby accelerating wound healing. This would also prevent implant degradation due to an excessive inflammatory process.

Optimization of Platelet-Rich Plasma Fraction Selection for Surgical Treatment of Macular Holes

Objective: based on clinical and morpho — functional analysis of retinal restoration and assessment of the cellular composition of platelet-rich plasma, to determine its optimal fraction for surgical treatment of macular hole. Materials and methods. This study presents the results of 120 cases of surgical treatment of eyes with macular hole. The patients were divided into 2 groups. In the 1st group (60 eyes), the technology developed in our clinic was used without the use of postoperative tamponade of the vitreal cavity and the use of autologous platelet-rich plasma (PRP), in the 2nd — traditional technology for the treatment of macular hole s (60 eyes). The cellular composition of the PRP of 30 patients was analyzed. Results. 2 months after surgery, 119 patients had the restoration of all retinal layers in the macular hole area and positive dynamics of visual acuity was recorded in comparion with the preoperative period. After observation for 6 months after surgery, cases of recurrence of macular hole were not detected. Conclusion. The therapeutic value of BoTP lies in the tamponing and regenerative effect. According to the results of this study, for maculrupture surgery, it is rational to use plasma from a layer of pure platelet-rich plasma (P-PRP), which contains a sufficient number of platelets capable of having a positive therapeutic effect, and a minimum number of leukocytes capable of performing the functions of immune protion and repair of the retiand surrounding tissues.

Pure platelet-rich plasma promotes semaphorin-3A expression: a novel insight to ameliorate intervertebral disk degeneration in vitro

IntroductionIntervertebral disk degeneration (IVDD) can be effectively treated using platelet-rich plasma (PRP). While the exact process is fully understood, it is believed that using pure PRP (P-PRP) without leukocytes is a better option for preventing IVDD. Semaphorin-3A (Sema3A), an inhibitor of angiogenesis and innervation, is essential for preserving IVDD’s homeostasis. Whether PRP prevents IVDD by modifying Sema3A has yet to receive much research. This work aims to clarify how P-PRP affects Sema3A when IVDD develops in vitro.MethodsNucleus pulposus cells (NPCs) isolated from 8-week-old male Sprague-Dawley rats were exposed to 10 ng/ml IL-1β and then treated with P-PRP or leukocyte platelet-rich plasma (L-PRP) in vitro, followed by measuring cell proliferation, apoptosis and microstructures, inflammatory gene and Sema3A expression, as well as anabolic and catabolic protein expression by immunostaining, quantitative real-time polymerase chain reaction (qPCR), western blot, and enzyme-linked immunosorbent assay (ELISA).ResultsIn comparison with L-PRP, P-PRP had a higher concentration of growth factors but a lower concentration of inflammatory substances. P-PRP increased the proliferation of NPCs, while IL-1 relieved the amount of apoptosis due to its intervention. Anabolic genes, aggrecan, and collagen II had higher expression levels. MMP-3 and ADAMTS-4, two catabolic or inflammatory genes, showed lower expression levels. Sema3A activity was enhanced after P-PRP injection, whereas CD31 and NF200 expression levels were suppressed.ConclusionsP-PRP enhanced the performance of NPCs in IVDD by modifying the NF-κB signaling pathway and encouraging Sema3A expression, which may offer new therapy options for IVDD.The translational potential of this articleThe findings provide a new therapeutic target for the treatment of IVDD and show a novel light on the probable mechanism of PRP and the function of Sema3A in the progression of IVDD.

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.

Review of Dohan Eherenfest et al. (2009) on “Classification of platelet concentrates: From pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF)”

This classic discusses the original publication of Dohan Eherenfest etal. on "Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF)", in which the authors propose four categories of platelet concentrates depending on their leucocyte and fibrin content (P-PRP, leucocyte- and platelet-rich plasma (L-PRP), pure platelet-rich fibrin (P-PRF), and L-PRF) to group a "jungle" of products in which the term platelet-rich plasma (PRP) was used indistinctly. They were able to identify common factors such as: (1) the use of anticoagulants and immediate centrifugation of the blood after its collection; (2) most preparation techniques allowed platelet concentrate preparation within an hour; (3) the centrifugation aimed to separate the blood in layers that would allow the extraction of specific fractions; and (4) the product was activated with thrombin or calcium chloride. The reviewed manuscript has been listed among the most cited PRP articles in regenerative medicine, with more than 800 citations, driving current scientific research and clinical practise by categorising L-PRP and P-PRP (now, leucocyte-poor PRP). The classification has also opened the door to understanding intrinsic biological mechanisms between platelets, leukocytes, fibrin, and growth factors, which will later be considered for studying the proliferation and differentiation of cells in different tissues affected by PRP. Since the initial classification of platelet concentrates, several other classification systems have been proposed and published in the current literature such as platelet, activation, white blood cell (PAW), Mishra, platelet, leucocyte, red blood cells, and activation (PLRA), dose of platelet, efficiency, purity, and activation (DEPA), method, activation, red blood cells, spin, platelets, image guidance, leukocytes, and light activation (MARSPILL), etc. These classifications have identified important aspects of PRP that affect the biological composition and, ultimately, the indications and outcomes. To date, there is still a lack of standardisation in sample preparation, cohort heterogeneity, and incomplete reporting of sample preparation utilised, leading to a lack of clarity and challenging researchers and clinicians.

Retrospective Analysis of Responders and Impaired Patients with Knee Osteoarthritis Treated with Two Consecutive Injections of Very Pure Platelet-Rich Plasma (PRP).

To assess the effectiveness of two consecutive intraarticular injections of PRP to treat knee osteoarthritis (KOA), discriminating between responders and impaired patients. This retrospective study included 73 consecutive patients who were referred for two intra-articular PRP injections (one week apart) for treating symptomatic moderate/severe KOA. Biological characterization of the PRP, including platelets, leukocytes and erythrocytes, was evaluated. Patient's subjective symptoms were recorded before the treatment and 1 year after the second injection using pain VAS and WOMAC scores. Responders were defined by an improvement of 10 points on WOMAC. At a 1-year follow up, we found 36 (49.3%) patients who fulfilled the criteria of responders, and 21 (28.8%) patients were impaired. A statistically and clinically significant global improvement of -29.2 ± 14.3 (p < 0.001) points in WOMAC score was observed 1 year after treatment in the responder group, with a higher response rate in patients with KL 2 (57.7%) compared to KL IV (28.6%). The percentage of patients with KL IV was higher in the impaired group (48.0%) compared to the responders (16.6%). As expected, the evaluation of the functionality of the knee in the impaired group indicates that it significantly worsened after one year from treatment (p = 0.027). However, the average pain score remained stable with no significant differences after 1 year (p = 0.843). No clinical complications or severe adverse events after the PRP injections were reported. The present study suggests that two intra-articular injections of 10 mL of very pure PRP provide pain and functional improvement in symptomatic KOA.

Исследование клеточного состава богатой тромбоцитами плазмы для оптимизации ее использования в хирургии макулярных разрывов. Часть 1

Currently, the use of platelet-rich plasma (PRP) is one of the successful areas of tissue engineering and cell therapy. In medicine PRP is widely used in ophthalmosurgery, in particular for macular hole (MH) closure. Purpose. To evaluate cellular composition of PRP in order to optimize its use in macular hole surgery. Material and methods. This study included platelet-rich plasma analysis of 20 patients (6 male and 14 female) who underwent surgical treatment of macular hole in Eye Microsurgery Center in the period from April till July 2021. Patients' age ranged from 57 to 82 (69,0±1.2) years. Ycellbio-Kit was used to obtain PRP. As a result of centrifugation 2 main blood fractions were obtained: plasma and erythrocyte layer. The study of the contents of the test tube was performed on 4 levels. The main attention was paid to the quantitative composition of platelets and leukocytes, which are the most important and significant in MH surgery. Results. The following platelet and leukocyte count in the general blood test and PRP were obtained as a result of the study. General blood test – platelet concentration was 140 to 285•10³/µL (215.85±9.23), P-PRP (pure platelet-rich plasma) – 699 to 5681•10³/µL (2600.4±329.9), L-PRP (plasma containing platelets and leukocytes) – 1995 to 9989•10³/µL (6555.8±506.7) (p&lt;0.001). General blood test – white blood cell concentration was 3.83 to 7.73•10³/µL (5.58±0.29), P-PRP was 0.02 to 7.08•10³/µL (1.49±0.58), L-PRP was 6.12–494.69•10³/µL (99.3±38.1) (p&lt;0.01). Conclusion. Platelet-rich plasma has regenerative activity and contractile properties in both fractions – in P-PRP and L-PRP, which simplifies the process of PRP extraction from the test tube and expands the possibilities for its use in MH surgery and other retinal pathologies associated with retinal tears. Given the lowest leukocyte concentration, the P-PRP fraction may be optimal for MH surgery. Key words: macular rupture, platelet-enriched plasma, platelets, leukocytes

Characterization of the Protein Corona of Three Chairside Hemoderivatives on Melt Electrowritten Polycaprolactone Scaffolds.

In tissue engineering, the relationship between a biomaterial surface and the host's immune response during wound healing is crucial for tissue regeneration. Despite hemoderivative functionalization of biomaterials becoming a common tissue-engineering strategy for enhanced regeneration, the characteristics of the protein-biomaterial interface have not been fully elucidated. This study characterized the interface formed by the adsorbed proteins from various hemoderivatives with pristine and calcium phosphate (CaP)-coated polycaprolactone (PCL) melt electrowritten scaffolds. PCL scaffolds were fabricated by using melt electrospinning writing (MEW). Three hemoderivatives (pure platelet-rich plasma (P-PRP), leucocyte platelet-rich plasma (L-PRP) and injectable platelet-rich fibrin (i-PRF)) and total blood PLASMA (control) were prepared from ovine blood. Hemoderivatives were characterized via SEM/EDX, cross-linking assay, weight loss, pH and protein quantification. The interface between PCL/CaP and hemoderivative was examined via FTIR, XPS and electrophoresis. i-PRF/PCL-CaP (1653 cm-1), PLASMA/PCL-CaP (1652 cm-1) and i-PRF/PCL (1651 cm-1) demonstrated a strong signal at the Amide I region. PLASMA and i-PRF presented similar N1s spectra, with most of the nitrogen involved in N-C=O bonds (≈400 eV). i-PRF resulted in higher adsorption of low molecular weight (LMW) proteins at 60 min, while PLASMA exhibited the lowest adsorption. L-PRP and P-PRP had a similar pattern of protein adsorption. The characteristics of biomaterial interfaces can be customized, thus creating a specific hemoderivative-defined layer on the PCL surface. i-PRF demonstrated a predominant adsorption of LMW proteins. Further investigation of hemoderivative functionalized biomaterials is required to identify the differential protein corona composition, and the resultant immune response and regenerative capacity.

Evaluation of Platelet-Rich Plasma by means of PRGF®-Endoret® protocol in leukemia cats: PDGF-BB and TGF-ß1 valuation.

Feline leukemia virus (FeLV) is a chronic disease that leads to the weakening of a cat's immune system. Platelet-rich plasma (PRP) offers therapeutic effects for multiple diseases, the use of PRP and growth factors (GFs) determination could be an alternative treatment to improve the quality of life in these patients. The objectives of this study were to determine and compare the concentration of platelets (PLTs), red blood cells (RBCs) and white blood cells (WBCs) between samples of whole blood (WB), PRP and platelet-poor plasma (PPP) fractions, and to evaluate the concentration of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor β1 (TGF-β1) in both fractions in FeLV cats using a PRGF®-Endoret® protocol previously standardized in this species. WB was collected from 11 asymptomatic FeLV-positive cats. PRP and PPP was obtained following PRGF®-Endoret® technology according to centrifugation at 265 g for 10 min. Cellular components, RBCs, WBCs, PLTs, and the PDGF-BB and TGF-β1 concentrations in PRP and PPP fractions were determined. PLT in the PRP fraction was statistically higher than WB and PPP fraction, with no statistical differences between WB and PPP. PLT concentration increased 1.4 times in PRP fraction compared to WB. Mean platelet volume (MPV) did not differ significantly between the WB, PRP, and PPP fractions. Compared to WB, the absolute numbers of RBCs and WBCs were decreased by 99% and more than 95% in the PRP and PPP fractions, respectively. TGF-ß1 concentrations increased in PRP vs. PPP, with no changes in PDGF-BB. Based on the degree of PLT enrichment and the absence of RBCs and WBCs, this blood product could be classified as a Pure Platelet-Rich Plasma (P-PRP). The presence of GFs in PRP and PPP samples suggests that the PRGF®-Endoret® methodology is suitable for obtaining PRP in FeLV cats, despite future studies are necessary to optimize the technique, standardize the results and assess clinical efficacy.

Расширенный подход к первичной микрохирургической обработке обширных проникающих ранений склеры с одномоментной витрэктомией и герметизацией хориоретинального дефекта

Background: the current strategy for managing extensive zone 3 scleral lacerations implies delayed subtotal vitrectomy. A high rate of poor treatment outcomes requires novel approaches to surgical debridement of lacerations given anatomical and pathogenic specifics. Aim: to assess the treatment results of extensive zone 3 scleral laceration surgery by simultaneous vitrectomy and chorioretinal wound debridement. Patients and Methods: 14 patients (14 eyes) diagnosed with extensive penetrating scleral/corneoscleral laceration and polymorphic intraocular lesions associated with grade 4–5 visual impairment were enrolled. All patients underwent primary microsurgical debridement using a technique that involved external debridement and suturing of scleral laceration, single-step subtotal vitrectomy, and internal debridement of the chorioretinal wound. The chorioretinal wound was filled with pure platelet-rich plasma (P-PRP). Endotamponade was performed using silicone oil (n=9) or air-gas mixture (n=5). Results: after 6 months, visual acuity (VA) was estimated to be 0.02–0.5 in 12 patients and light perception in 2 patients. During the follow- up, no phthisis bulbi was reported. The progression of proliferative vitreoretinopathy (formation of intraretinal adhesions or subretinal bands involving healthy retina) was reported in 3 patients (21.4%). Chorioretinal scars were characterized by local nonaggressive proliferation, no prominence in the vitreous cavity, or proliferation of tissue beyond the scars. Conclusion: our approach to severe eye injury management has demonstrated positive outcomes by providing a stable anatomical result, significant improvement in early and late postoperative VA. KEYWORDS: open globe injury, primary microsurgical debridement, proliferative vitreoretinopathy, scleral lacerations, vitrectomy, platelet- rich plasma. FOR CITATION: Shamkin S.S., Subbotina S.N., Stepanyants A.B., Salov D.V. Advanced approach to primary microsurgical debridement of extensive scleral lacerations involving simultaneous vitrectomy and chorioretinal wound closure. Russian Journal of Clinical Ophthalmology. 2023;23(3):124–128 (in Russ.). DOI: 10.32364/2311-7729-2023-23-3-3.

Evaluation of the effect of injectionsof both platelet-rich plasma and hyaluronic acid in patients with early knee osteoarthritis via the concentration of interleukin-1β in serum

Background Early knee osteoarthritis, which causes the degeneration of articular cartilage and subchondral bone, is a disease that worsens with time and is a major contributor to physical disability. Several inflammatory cy- tokines released by chondrocytes work in concert to trigger the production of enzymes that break down cartilage. A pro-inflammatory cytokine called interleukin-1β (IL-1 β ) accelerates cartilage breakdown after trauma, re- duces matrix production, and results in chondrocyte death. Dendritic cells and macrophages are the main sources of IL-1β. On the other hand, os- teoblasts, periodontal ligament cells, and gingival fibroblasts have the abil- ity to release IL-1β. Treatment pure- platelet rich plasma (P-PRP) effec- tiveness of transforming growth factor, platelet-derived growth factor, type I insulin-like growth factor, and the vascular endothelial growth factor is thought to be related to their release. Because of their capacity to increase matrix formation, growth factors have been widely researched for OA and cartilage regeneration. Hyaluronic acid (HA) is a glycosaminoglycan poly- mer composed of N-acetyl glucosamine and D-glucuronic acid disaccharide molecules. Early embryonic development, inflammatory, wound repair, cell differentiation, and viscoelasticity are all affected by HA, and other biologi- cal processes. Materials and Methods Elabscience kits were used to assess the serum IL-1β level. The experimental investigation included 18 control groups, 10 HA injections, and 21 pure PRP injections. age ranged from 30 to 80. The study was excluded all individuals with advanced osteoarthritis in the knee, COVID-19, diabetes, and autoimmune diseases. the time frame running from November 2021 until June 2022. Other variables in our research were age, gender, family history, use of antihypertensive medications or medica- tions for other disorders, and body mass index (BMI). Results The findings of this study demonstrate elevated IL-1β levels in patients with early KOA before treatments injection compared with the healthy control. After treatments injection, the level of IL-1β was decreased compared with before injection. Conclusions IL-1β could be one of the prognostic signs of early KOA

Bone-Regenerative Ability of Platelet-Rich Plasma Following Sinus Augmentation with Anorganic Bovine Bone: A Systematic Review with Meta-Analysis.

Background: The objective of this systematic review is to assess the effect of the adjuvant use of platelet-rich plasma (PRP) and its type on new bone formation by anorganic bovine bone during maxillary sinus floor augmentation procedure. Methods: PubMed, Cochrane Central Register of Controlled Trials, and Ovid databases were searched for relevant studies published up to 16 September 2021. Randomized clinical trials (RCTs) and non-randomized controlled clinical trials (CCTs) that reported data on the new bone formation (measured by histomorphometric analysis) were considered. Risk of bias and quality assessment of included studies were evaluated following the Cochrane Handbook for Systematic Reviews of Interventions and the Risk Of Bias In Non-randomised Studies of Interventions (ROBINS-I) tool. Strength of evidence was assessed following the approach of the Agency for Healthcare Research and Quality (AHRQ) through its evidence-based practice center (AHRQ EPC). The meta-analysis was based on the primary outcome of newly formed bone, for which the standard mean difference was calculated. Results: After the application of eligibility criteria, six clinical trials (three RCTs and three CCTs) covering 85 maxillary sinus floor elevation procedures were included. The pooled new bone formation value for PRP was 1.67 (95% CI: −0.15 to 3.49; I2: 86%), indicating the absence of significant effect. Plasma rich in growth factors (PRGF) was the pure PRP tested in five of the included studies. When sub-group (type of PRP) meta-analysis was performed, significantly higher new bone formation was observed in the PRGF group [2.85 (95% CI: 0.07 to 5.64; I2: 88%)] in comparison to the control group. Conclusions: A beneficial effect on new bone formation after maxillary sinus floor elevation can be obtained when anorganic bovine bone is mixed with PRGF.

Оцінка ефективності різних режимів подвійного центрифугування для отримання чистої збагаченої тромбоцитами плазми

The purpose - to determine the optimal parameters of double centrifugation in the preparation of pure platelet-rich plasma with a maximum reduction of other blood cellular elements. Materials and methods. The study included 30 apparently healthy individuals aged 18 to 60 years who underwent venous blood sampling. The first centrifugation of all blood samples was performed in the 160 g × 10 min mode. The second centrifugation was performed in 12 different modes: 190 g × 10 min, 190 g × 15 min, 220 g × 10 min, 220 g × 15 min, 250 g × 10 min, 250 g × 15 min, 290 g × 10 min, 290 g × 15 min, 320 g × 10 min, 320 g × 15 min, 360 g × 10 min, 360 g × 15 min. The number of erythrocytes, leukocytes, platelets (in whole blood, after the first and second centrifugation), platelet capture efficiency, platelet enrichment factor, erythrocytes and leukocytes reduction efficiency, concentration of major growth factors in plasma after the second centrifugation were determined. Results. In whole blood, the concentration of erythrocytes was 4.455±0.301 × 1012/l, leukocytes - 6.485±0.501 × 109/l, platelets - 281.5±22.7 × 109/l. After the first centrifugation, the volume of the obtained plasma containing platelets was 4.51±0.25 ml, the number of platelets was 482.1±47.4 × 109/l, with platelet reduction rates of 14.39±1.1%, their preservation - 85.6±1.1% and enrichment factor - 1.71±0.1; the number of erythrocytes was 0.104±0.006 × 1012/l, leukocytes - 0.205±0.025 × 109/l, and their reduction efficiency - 98.83±0.09% and 98.42±0.14%, respectively. After the second centrifugation in different modes, the 250 g × 15 min mode turned out to be the most effective, which made it possible to achieve a platelet enrichment factor of about 4.93 with a concentration of platelets in the final product of 1386.7±116.5 × 109/l, their capture efficiency of 82.1±1.1% and erythrocytes and leukocytes reduction efficiency of 98.86±0.08% and 98.45±0.14%, respectively. Conclusions. In case of double-spin centrifugation, the sequential combination of 160 g × 10 min and 250 g × 15 min regimens is most effective for the preparation of pure platelet-rich plasma. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of all participating institutions. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

Unravelling Alveolar Bone Regeneration Ability of Platelet-Rich Plasma: A Systematic Review with Meta-Analysis.

Background: This systematic review aims to evaluate the efficacy of the available platelet-rich plasma (PRP) products and composition to regenerate alveolar bone after tooth extraction. Methods: PubMed, Cochrane Central Register of Controlled Trials, and EBSCO databases were searched up to 2 July 2021. Only randomized clinical trials using leukocyte-rich plasma (L-PRP) or pure-platelet rich plasma (P-PRP) for bone regeneration in alveolar ridge preservation were selected. The following outcomes were considered: (1) new bone formation (primary outcome) and (2) bone density (secondary outcome). A meta-analysis for PRP, P-PRP, and L-PRP using a fixed effect model was performed with Review Manager 5.4 software. Overall evidence was qualified using GRADE. Results: Six randomized clinical trials from 2639 unique articles initially identified met the inclusion criteria. The meta-analysis showed a significant effect of the P-PRP on the outcome of new bone formation (SMD, 1.44; 95% CI, 0.84 to 2.03) for P-PRP treatment. No information was retrieved for L-PRP. A statistically significant difference was also observed in the P-PRP group for bone density outcome (SMD, 1.24; 95% CI, 0.81 to 1.68). The L-PRP treated sockets also showed higher bone density (SMD, 0.88; 95% CI, 0.31 to 1.45) in comparison to control sockets. The quality of evidence was moderate for both outcomes in the P-PRP group and low for the L-PRP group. Conclusions: Despite the limitations of the included studies, our data suggest that P-PRP, in comparison to unassisted healing, can improve alveolar bone regenerative potential. However, more high-quality clinical studies are needed.

Temporal Release and Denature of Several Mediators in Pure Platelet-Rich Plasma and Temperature-Induced Platelet Lysates Derived from a Similar Bovine Platelet Concentrate.

There is scarce information about bovine platelet-rich plasma/platelet-rich gel (PRP/PRG) and related hemocomponents (HCs), such as platelet lysates (PLs), including growth factor (GF) and cytokine concentrations, and how the stability of these biomolecules could be affected by time and temperature. This study aimed to evaluate the release and stability of transforming growth factor beta 1 (TGF-β1), interleukin 4 (IL-4), and tumor necrosis factor alpha (TNF-α) contained in bovine pure PRP (P-PRP) and temperature-induced PL (TIPL) coming from a similar platelet concentrate (PC) at 4 and 37°C at 3 and 96 h. Platelet concentrates (PCs) presented a 1.7-fold concentration of platelets (PLTs) with negligible counts of white blood cells (WBCs) when compared to the counts for these cells in whole blood. TGF-β1 concentrations were significantly lowest in plasma followed by TIPL, chemical-induced PL (CIPL), and P-PRP. IL-4 and TNF-α concentrations did not differ between HCs. TGF-β1 concentrations were negatively affected in P-PRPs stored at 4°C at 3 and 96 h, whereas those from P-PRP maintained at 37°C presented similar concentrations to TIPL stored at both temperatures over time. IL-4 and TNF-α concentrations were not affected by time or temperature in any of the HCs evaluated. Pure PRGs released additional quantities of GF and cytokines over time when compared with HCs stored over 96 h at 4 and 37°C. The method, either chemical or physical, used for platelet activation or damage produces a different GF and cytokine release pattern, which makes to each evaluated HCs different despite they come from a similar bovine PC. P-PRP activated with calcium gluconate and maintained at 37°C, which polymerizes in P-PRG, showed the best GF and cytokine release/denature profile compared with the rest of the HCs evaluated.

Effect of cell-free and platelet plasma on burn wound healing in rats

The presented study analyzes the healing effect of pure cell-free plasma and plateletrich plasma. The study was conducted on Wistar rats. Three groups of rats were formed for the experiment: a control group and two experimental ones (No. 1, No. 2). Burn wounds were inflicted on animals under sevoflurane anesthesia in the interscapular region with a copper-plated disc with a diameter of 20.0 mm heated in the flame of a gas burner to a temperature of 300.0 OC by applying it for 2.0 - 3.0 seconds. In the control group, treatment of burn wounds was not carried out, but surface application of aerosol 10% lidocaine was used to reduce the pain syndrome. For rats of experimental groups No. 1 and No. 2, treatment of burn wounds with plasma was started on the 5thday after injury. Rats of experimental group No. 1 were treated by injecting cell-free plasma under and around the burn. Rats of experimental group No. 2 were treated by injecting platelet-rich plasma under and around the burn. Monitoring and evaluation of wound healing was carried out by measuring their area. Photo fixation and photo documentation were carried out at the time of the burn wound and then on the fifth day after the injury (the first injection of plasma samples), on the 14th day after the injury, on the 23rd day after the injury and on the 30thday after the injury. The result of the study showed that both cell-free plasma and platelet-rich plasma accelerate the healing of a burn wound in comparison with the control. But platelet plasma, in comparison with cell free plasma, has a stronger therapeutic effect, significantly accelerating the contraction of the wound edges in the very first days after administration.

Intramammary treatment using allogeneic pure platelet-rich plasma in cows with subclinical mastitis caused by Gram-positive bacteria

The aims of the study were (1) to compare the cure risk of intramammary treatment of pure platelet rich plasma (P-PRP) or cefquinome sulfate (CS) in cows with subclinical mastitis (SCM) caused by Gram-positive bacteria, evaluated via somatic cell count (SCC) and the microbiological analysis of milk; (2) to compare the inflammatory/anti-inflammatory response of mammary gland to both treatments through the analyses of interleukins (IL), interferon gamma (IFN-γ), and tumour necrosis factor alpha (TNF-α) in milk. A non-inferiority randomized clinical trial was conducted. The null hypothesis was that cure risk in the experimental group (P-PRP) was inferior to the reference group (CS). A total of 103 cows were selected according to SCC and presence of Gram-positive bacteria, 49 cows were treated with CS and 54 cows were treated with P-PRP after determination of its cellular and molecular quality control. Cure was assessed by milk analyses at day 21 and 22 after treatment. Cows that remained with SCM were retreated at day 26, and cure assessed at day 47 and 48. Overall, bacteriological cure was observed in 16 cows (30%) of the P-PRP group, and 35 cows (71%) in CS group. Staphylococcus aureus cure risk was higher in CS group, but inconclusive for Streptococcus spp. The mean SCC increased in relation to time only in the P-PRP group. A direct relation between time and treatment for IL-1, IL-2, and IL-6 was observed, while no differences were observed for IL-4. Furthermore, IL-1 and IL-2 increased in cows treated twice in both groups. IL-8, IFN-γ, and TNF-α showed a significant interaction between time and treatment. IFN-γ concentration was lower in the P-PRP group compared to the CS on days 0 and 22. Leukocyte counts were lower in P-PRP when compared to whole blood. TGF-β1 and PF4 concentrations were higher in platelet lysates in comparison to P-PRGS and plasma. Moreover, PDGF-BB concentration was significantly higher in platelet lysates in comparison to plasma. Results obtained in this study demonstrate that SCM treated with PRP showed a lower rate of bacteriologic cure when compared to animals treated with CS.