In vitro biocompatibility analysis of protein-resistant amphiphilic polysulfobetaines as coatings for surgical implants in contact with complex body fluids.

The fouling resistance of zwitterionic coatings is conventionally explained by the strong hydrophilicity of such polymers. Here, the in vitro biocompatibility of a set of systematically varied amphiphilic, zwitterionic copolymers is investigated. Photocrosslinkable, amphiphilic copolymers containing hydrophilic sulfobetaine methacrylate (SPe) and butyl methacrylate (BMA) were systematically synthesized in different ratios (50:50, 70:30, and 90:10) with a fixed content of photo-crosslinker by free radical copolymerization. The copolymers were spin-coated onto substrates and subsequently photocured by UV irradiation. Pure pBMA and pSPe as well as the prepared amphiphilic copolymers showed BMA content-dependent wettability in the dry state, but overall hydrophilic properties a fortiori in aqueous conditions. All polysulfobetaine-containing copolymers showed high resistance against non-specific adsorption (NSA) of proteins, platelet adhesion, thrombocyte activation, and bacterial accumulation. In some cases, the amphiphilic coatings even outperformed the purely hydrophilic pSPe coatings.

Improvement of hepatic fibrosis after tenofovir disoproxil fumarate switching to tenofovir alafenamide for three years.

BACKGROUND: Both tenofovir alafenamide (TAF) and tenofovir disoproxil fumarate (TDF) are the first-line treatments for chronic hepatitis B (CHB). We have showed switching from TDF to TAF for 96 weeks resulted in further alanine aminotransferase (ALT) improvement, but data remain lacking on the long-term benefits of TDF switching to TAF on hepatic fibrosis. AIM: To assess the benefits of TDF switching to TAF for 3 years on ALT, aspartate aminotransferase (AST), and hepatic fibrosis improvement in patients with CHB. METHODS: A single center retrospective study on 53 patients with CHB who were initially treated with TDF, then switched to TAF to determine dynamic patterns of ALT, AST, AST to platelet ratio index (APRI), fibrosis-4 (FIB-4) scores, and shear wave elastography (SWE) reading improvement at switching week 144, and the associated factors. RESULTS: The mean age was 55 (28-80); 45.3%, males; 15.1%, clinical cirrhosis; mean baseline ALT, 24.8; AST, 25.7 U/L; APRI, 0.37; and FIB-4, 1.66. After 144 weeks TDF switching to TAF, mean ALT and AST were reduced to 19.7 and 21, respectively. From baseline to switching week 144, the rates of ALT and AST < 35 (male)/25 (female) and < 30 (male)/19 (female) were persistently increased; hepatic fibrosis was also improved by APRI < 0.5, from 79.2% to 96.2%; FIB-4 < 1.45, from 52.8% to 58.5%, respectively; mean APRI was reduced to 0.27; FIB-4, to 1.38; and mean SWE reading, from 7.05 to 6.30 kPa after a mean of 109 weeks switching. The renal function was stable and the frequency of patients with glomerular filtration rate > 60 mL/min was increased from 86.5% at baseline to 88.2% at switching week 144. CONCLUSION: Our data confirmed that switching from TDF to TAF for 3 years results in not only persistent ALT/AST improvement, but also hepatic fibrosis improvement by APRI, FIB-4 scores, as well as SWE reading, the important clinical benefits of long-term hepatitis B virus antiviral treatment with TAF.

Augmentation of Autologous Platelet-Rich Plasma in Hip Salvage in Stages I and II of Avascular Necrosis: Randomized Blinded Trial Comparing 3 Surgical Techniques in 82 Patients.

Introduction: Avascular necrosis of the femoral head is common in routine orthopedic clinics. The challenge arises in managing early stages (I and II) without obvious radiological evidence. Authors explore this naïve research area by comparing surgical procedures in early AVN patients. Materials and Methods: A prospective multicentric study was performed from November 2020 to February 2023 on 82 patients treated with surgical decompression and adjuvants, concerning the defined inclusion and exclusion criteria. Radiopacity and intraosseous edema resolution and THA conversion rates were assessed. Hip pain VAS, groin/thigh pain, difficulty in sitting cross-legged incidence, pain-free walking distance, Harris hip scores, 30-s chair test, and complications were noted. Results: Among 82 patients, the mean age was 28.46 years. Male:female ratio of 3.9:1. 8.5% had bilateral affection and 48.78% had a positive family history. 93.90% presented with groin pain and difficulty in sitting cross-legged, restricted hip movements in 85.3%, and thigh pain in 54.87%. Harris hip scored worst in Group 3 followed by Group 2 and Group 1. 63.41% and 36.58% of patients had Grades 1 and 2 AVN, respectively. At 1 week post-operatively, 96.3% and 93.9% of patients were relieved from groin and thigh pain, respectively (p < 0.001); the trend being Group 3 > Group 2 > Group 1. Hip pain VAS followed a similar trend. At 4 weeks, Harris hip scores improved in Group 3 > Group 2 > Group 1. At 6 months, the trend was Group 2 > Group 3 > Group 1. Group 3 had better 30-s chair test results, pain-free walking distance, and longer cross-legged sitting time. Complication rate of 3.6%. 6.09% of patients underwent THA later. Sclerotic patch and marrow edema resolution early in Group 3, i.e., 46 and 31 days respectively, followed by Group 2 and Group 1. Conclusion: In Stages I and II AVN, biplanar core decompression (double) and intraosseous PRP injection is a promising salvage option; patients have better early hip scores (4 weeks), and early groin and thigh pain recovery. Patients treated early have better clinical and radiological recovery.

Exploring the Efficacy of Biologics in Knee Osteoarthritis: Ultrasound Evaluation of Cartilage Regeneration Effects.

Introduction: Biologics like growth factors, stem cells, and platelet-rich plasma show potential in stimulating cartilage regrowth and reducing inflammation. By synthesizing preclinical and clinical studies, this study offers insights into how these biologics work and their effectiveness in treating knee osteoarthritis. Methods and Materials: Twenty-four participants with knee osteoarthritis (Kellgren - Lawrence grade II or III) were enrolled after obtaining consent. They received three doses of 2 ml intraarticular platelet-rich plasma at 1 month intervals. The clinical assessment involved the oxford knee score (OKS) and visual analogue scale (VAS) for pain on Days 0, 90, and 180. Ultrasound measured femoral and trochlear cartilage thickness pre- (Day 0) and post-PRP (Day 90-180). Results: Before treatment, the average pain score was 7.2 (p = 1.03). On Day 90 post-PRP, it decreased to 5 (p = 0.81), and by Day 180, it further reduced to 4.5 (p = 0.97). The initial total OKS was 33.5 (p = 1.76), which increased to 36 (p = 1.71) on Day 90 and 38.5 (p = 1.89) on Day 180. The femoral and trochlear cartilage thickness also showed improvement from baseline (0.92) to Day 90 (0.96) and Day 180 (1.01), indicating significant cartilage healing post-PRP administration. Conclusion: Our study highlights the probability of PRP in treating knee OA, highlighting their ability to alleviate symptoms, enhance joint function, and promote articular cartilage regeneration.

Decoding the Decade: Exploring the Efficacy of Platelet-Rich Plasma (PRP) in Complex Wound Management - A Comprehensive Study.

Introduction: Autologous platelet-rich plasma (PRP) therapy has emerged as a promising regenerative treatment modality, offering potential improvements in healing outcomes through its rich content of growth factors and cytokines. We evaluated the effectiveness of PRP therapy in the management of complex wounds, using a decade-long retrospective analysis of treatments conducted at a tertiary care center from 2010 to 2020. The study introduces and assesses the efficacy of the Sandeep's Technique for Assisted Regeneration of Skin (STARS) in enhancing wound healing and quality of life for patients with complex wounds. Materials and methods: A prospective interventional study was conducted, involving two phases: the development and initial testing of PRP therapy (2010-2015) and the application and evaluation of the STARS protocol (2015-2020). The study included patients with complex wounds, utilizing autologous PRP prepared through a double spin centrifuge technique. Outcome measures included wound-healing rates, infection management, and complication rates, compared to conventional treatment methods. Results: The study treated 500 wounds in 432 patients with autologous PRP, noting significant improvements in wound-healing rates, 97.7% had infection control without antibiotics (even in MRSA cases), and all had a good pain control. Histopathological examinations confirmed collagen-rich healing with minimal scarring. The STARS protocol demonstrated the potential of PRP therapy in accelerating wound healing, reducing the need for additional surgical interventions, and enhancing patient outcomes. Conclusion: PRP therapy, particularly when administered following the STARS protocol, represents a safe, effective, and patient-friendly approach for the management of complex wounds. This study supports the integration of PRP therapy into regenerative care strategies, suggesting a shift toward more innovative and efficacious treatments in wound management.

What It Takes To Be a Platelet: Evolving Concepts in Platelet Production.

Platelets are among the most abundant cells within the circulation. Given that the platelet lifespan is 7 to 10 days in humans, a constant production of around 100 billion platelets per day is required. Platelet production from precursor cells called megakaryocytes is one of the most enigmatic processes in human biology. Although it has been studied for over a century, there is still controversy about the exact mechanisms leading to platelet release into circulation. The formation of proplatelet extensions from megakaryocytes into bone marrow sinusoids is the best-described mechanism explaining the origin of blood platelets. However, using powerful imaging techniques, several emerging studies have recently raised challenging questions in the field, suggesting that small platelet-sized structures called buds might also contribute to the circulating platelet pool. How and whether these structures differ from microvesicles or membrane blebs, which have previously been described to be released from megakaryocytes, is still a matter of discussion. In this review, we will summarize what the past and present have revealed about platelet production and whether mature blood platelets might emerge via different mechanisms.

Flow Cytometry and Platelets.

Clinical assessment of platelet activation by flow cytometry is useful in the characterization and diagnosis of platelet-specific disorders and as a measure of risk for thrombosis or bleeding. Platelets circulate in a resting, "unactivated" state, but when activated they undergo alterations in surface glycoprotein function and/or expression level, exposure of granule membrane proteins, and exposure of procoagulant phospholipids. Flow cytometry provides the means to detect these changes and, unlike other platelet tests, is appropriate for measuring platelet function in samples from patients with low platelet counts. The present review will focus on flow cytometric tests for platelet activation markers.

Autolougous platelet concentrates in esthetic medicine.

This narrative review summarizes current knowledge on the use of autologous platelet concentrates (APCs) in esthetic medicine, with the goal of providing clinicians with reliable information for clinical practice. APCs contain platelets that release various growth factors with potential applications in facial and dermatologic treatments. This review examines several facial esthetic applications of APCs, including acne scarring, skin rejuvenation, melasma, vitiligo, stretchmarks, peri-orbital rejuvenation, peri-oral rejuvenation, hair regeneration and the volumizing effects of APC gels. A systematic review of literature databases (PubMed/MEDLINE) was conducted up to October 2023 to identify randomized controlled trials (RCTs) in the English language on APCs for facial rejuvenation and dermatology. A total of 96 articles were selected including those on platelet rich plasma (PRP), plasma-rich in growth factors (PRGF), and platelet-rich fibrin (PRF). Clinical recommendations gained from the reviews are provided. In summary, the use of APCs in facial esthetics is a promising yet relatively recent treatment approach. Overall, the majority of studies have focused on the use of PRP with positive outcomes. Only few studies have compared PRP versus PRF with all demonstrating superior outcomes using PRF. The existing studies have limitations including small sample sizes and lack of standardized assessment criteria. Future research should utilize well-designed RCTs, incorporating appropriate controls, such as split-face comparisons, and standardized protocols for APC usage, including optimal number of sessions, interval between sessions, and objective improvement scores. Nevertheless, the most recent formulations of platelet concentrates offer clinicians an ability to improve various clinical parameters and esthetic concerns.

Platelet-Rich Plasma Treatment for the Lumbar Spine: A Review and Discussion of Existing Gaps.

BACKGROUND: Platelet-rich plasma (PRP) is obtained by centrifuging autologous whole blood to extract a layer concentrated with platelets, growth factors found in platelet granules, and cytokines. These components work together to promote and facilitate the healing process at sites of injury. An increasing number of clinical studies are assessing the efficacy of PRP as a treatment for lower back pain. OBJECTIVES: Lumbar back pain is a significant cause of years lived with disability. This paper conducts a thorough review of clinical studies on intradiscal, facet-joint, epidural, and mixed-target PRP interventions in the lumbar spine. Furthermore, gaps in the current literature regarding lumbar spinal PRP injections are identified to help guide future clinical trials. STUDY DESIGN: Literature review. METHODS: An initial search was conducted using Ovid MEDLINE, focusing on PRP injections in the spine. Boolean operators were used to combine MeSH terms and key words such as "spine," "lumbar spine," "thoracic spine," "cervical spine," "intervertebral disc," "platelet-rich plasma," and "inject." The search revealed an absence of papers about PRP injections into the cervical and thoracic spine, so the review was written with a specific focus on the lumbar spine. For the purposes of this paper, the selected manuscripts were separated into categories of intradiscal, facet-joint, epidural, and mixed-target PRP injections. RESULTS: A multitude of case reports, case series, prospective clinical studies, and randomized controlled trials have yielded results supporting the use of intradiscal, facet-joint, and epidural PRP injections in the lumbar spine. However, a handful of papers suggest that PRP lacks efficacy in improving lumbar back pain and function. With the relative dearth of literature assessing the effects of spinal PRP injections, additional double-blinded randomized trials are needed. Important findings from available studies include the observation of PRP's increased efficacy over time, the correlation of the number of targeted injection sites with the efficacy of PRP injections, and the correlation of platelet count with PRP injections' efficacy. LIMITATIONS: There exists wide variability in PRP preparation protocols and in the methods of assessing PRP's therapeutic benefits between each study that evaluates PRP's effects in the lumbar spine. CONCLUSIONS: All clinical studies evaluating PRP as a form of treatment for the lumbar spine should include full transparency and details about the methods used for PRP preparation and injection. Future double-blinded randomized trials can fill in existing gaps by assessing the effects of platelet concentration and dose on the extent of clinical improvement as well as by establishing an expected timeline for clinical improvement after PRP injections. Cross-study standardization of which pain scoring systems to utilize for study evaluation would increase comparability among different papers.

Does one size really fit all? The case for personalized antiplatelet therapy in interventional cardiology.

Cardiovascular disease is the leading cause of death worldwide. Dual antiplatelet therapy (DAPT), with aspirin plus a P2Y12 inhibitor, is currently recommended as a default for patients after acute coronary syndrome (ACS) and following percutaneous coronary intervention (PCI). However, controversies arise over the role of aspirin, the optimal duration of DAPT after drug-eluting stent (DES) implantation, the choice of P2Y12 inhibitor and the variability in individual responses to antiplatelet agents. Recent data indicate that monotherapy with a P2Y12 inhibitor may have adequate anti-ischemic effects with lower bleeding risk. Additionally, discrepancies in DAPT duration recommendations and the optimal P2Y12 inhibitor, provides more uncertainty. We ask the question "does one size really fits all?" or should a more personalized strategy should be implemented.

Diseases affecting the heart and blood circulation are the leading cause of death worldwide. Treatment with drugs that prevents platelets from clumping (called antiplatelets) like aspirin plus another drug group (called P2Y12 inhibitors) like clopidogrel, ticagrelor and prasugrel, is currently recommended as a default for patients after heart attack and/or in whom coronary stents are inserted. However, it is very well documented that the response of any individual to these drugs is highly variable, and that the patients who don't respond as well to them are at increased risk of having clot events in their coronary arteries. On the other hand, people who respond to the drugs very sensitively have a higher bleeding risk. Despite these observations, there is no attempt to test the response of individuals patients to their antiplatelet drugs in routine practice. This review article looks in detail and whether the currently used strategy of "One size fits all" should be changed, given that there may well now be the chance to perform routine testing on everyone, and personalize their treatment accordingly.

Clinical values of preoperative red blood cell distribution width and platelet parameters in patients with papillary thyroid carcinoma.

The prevalence of thyroid carcinoma is increasing, and papillary thyroid carcinoma (PTC) is the most frequent subtype. More and more attention is being concentrated on the association between inflammation indicators and malignant tumors. The aim of the present study was to analyze whether the preoperative red blood cell distribution width (RDW) and platelet parameters, including mean platelet volume (MPV) and platelet distribution width (PDW), can be applied to distinguish between patients with PTC or papillary thyroid microcarcinoma (PTMC) and healthy controls, and to explore the associations with clinicopathological characteristics. The study retrospectively compared the RDW, MPV and PDW values of 780 patients with PTC or PTMC against a healthy control group. Receiver operating characteristic (ROC) curves were conducted to determine diagnostic accuracy. Furthermore, the clinicopathological features of the patients with PTC or PTMC were compared between higher and lower platelet parameter groups based on the RDW, MPV and PDW values. Significantly higher preoperative RDW, MPV and PDW values were found in patients with PTC or PTMC compared with those of the healthy group. ROC curve analysis showed that the area under the curve (AUC) plus 95% confidence interval (95% CI) values of RDW, MPV and PDW were 0.808 (0.780-0.835), 0.771 (0.743-0.799) and 0.711 (0.681-0.742), respectively. When RDW and MPV were combined together, the AUC (95% CI) value was enhanced to 0.858 (0.835-0.881) for the patients with PTC. For the patients with PTMC, RDW, MPV and PDW had AUC (95% CI) values of 0.812 (0.783-0.840), 0.779 (0.749-0.808) and 0.718 (0.685-0.751), respectively. When RDW and MPV were combined together, the AUC (95% CI) value was enhanced to 0.858 (0.835-0.881). A higher RDW was significantly associated with being female, deeper tumor infiltration, and normal FT3 and FT4 levels. A higher PDW was significantly associated with elevated thyrotropin receptor antibody levels. In conclusion, as convenient and available inflammation indicators, RDW, PDW and MPV have diagnostic ability and can distinguish between patients with PTC or PTMC and healthy controls. In addition, the combined application of RDW and MPV can improve the diagnostic power. The values of RDW and MPV were associated with clinicopathological characteristics. To the best of our knowledge, this is the first study to prove the usefulness of preoperative RDW combined with MPV in diagnosing patients with PTC or PTMC.

Freeze-drying protocols and methods of maintaining the in-vitro biological activity of horse platelet lysate.

Platelet lysate, derived from platelets, are valuable biological products rich in bioactive molecules. Their use promotes tissue healing and modulates inflammation. However, maintaining the stability and bioactivity of platelet lysate is challenging since they degrade rapidly at room temperature. This study focused on the possibility to confer enhanced stability to freeze-dried equine platelet lysate as an alternative to platelet-rich plasma (PRP). Platelet lysate (PL) was derived from PRP and freeze-dried either as such or using various adjuvants. Primary cell cultures of porcine Vascular Wall-Mesenchymal Stem Cells were treated with different PL formulations, and cell viability was assessed using an MTT assay. Overall, the addition of PL significantly improved cell viability as compared to controls without growth factor supplementation or with foetal bovine serum. Notably, the freeze-drying process maintained the effectiveness of the PL for at least a week. Furthermore, the study revealed that varying the horse as the source of PL could yield varying effects on cell viability. Detailed freeze-drying protocols were established, including freezing, primary drying and secondary drying phases, and the type of adjuvant. This study demonstrated the potential of freeze-dried equine PL as a viable alternative to PRP and highlighted the importance of precise freeze-drying protocols and adjuvants for standardization. Equine PL showed promise for medical treatment in horses, offering advantages such as extended shelf life, ease of handling, and reduced transportation costs, with the potential for broadened therapeutic usage.

Biomedical application of TiO2NPs can cause arterial thrombotic risks through triggering procoagulant activity, activation and aggregation of platelets.

BACKGROUND: Titanium dioxide nanoparticles (TiO2NPs) are widely used in medical application. However, the relevant health risk has not been completely assessed, the potential of inducing arterial thrombosis (AT) in particular. METHODS: Alterations in platelet function and susceptibility to arterial thrombosis induced by TiO2NPs were examined using peripheral blood samples from healthy adult males and an in vivo mouse model, respectively. RESULTS: Here, using human platelets (hPLTs) freshly isolated from health volunteers, we demonstrated TiO2NP treatment triggered the procoagulant activity of hPLTs through phosphatidylserine exposure and microvesicles generation. In addition, TiO2NP treatment increased the levels of glycoprotein IIb/IIIa and P-selectin leading to aggregation and activation of hPLTs, which were exacerbated by providing physiology-mimicking conditions, including introduction of thrombin, collagen, and high shear stress. Interestingly, intracellular calcium levels in hPLTs were increased upon TiO2NP treatment, which were crucial in TiO2NP-induced hPLT procoagulant activity, activation and aggregation. Moreover, using mice in vivo models, we further confirmed that TiO2NP treatment a reduction in mouse platelet (mPLT) counts, disrupted blood flow, and exacerbated carotid arterial thrombosis with enhanced deposition of mPLT. CONCLUSIONS: Together, our study provides evidence for an ignored health risk caused by TiO2NPs, specifically TiO2NP treatment augments procoagulant activity, activation and aggregation of PLTs via calcium-dependent mechanism and thus increases the risk of AT.

The Prognostic Value of Preoperative Inflammatory Markers for Pathological Grading of Glioma Patients.

Introduction: The independent diagnostic value of inflammatory markers neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) and the diagnostic efficacy of NLR, derived neutrophil to lymphocyte ratio (dNLR), PLR, and lymphocyte-to-monocyte ratio (LMR) in glioma cases remain unclear. We investigated the correlation of preoperative peripheral blood inflammatory markers with pathological grade, Ki-67 Proliferation Index, and IDH-1 gene phenotype in patients with glioma, focusing on tumor grade and prognosis. Methods: We retrospectively analyzed the clinical, pathological, and laboratory data of 334 patients with glioma with varying grades and 345 with World Health Organization (WHO I) meningioma who underwent initial surgery at the Affiliated Hospital of Jining Medical University from December 2019 to December 2021. The diagnostic value of peripheral blood inflammatory markers for glioma was investigated. Results: The proportion of men smoking and drinking was significantly higher in the glioma group than in the meningioma group (P < .05); in contrast, the age and body mass index (Kg/m2) were significantly lower in the glioma group (P = .01). Significant differences were noted in the pathological grade (WHO II, III, and IV), Ki-67 Proliferation Index, and peripheral blood inflammatory markers such as lymphocyte median, NLR, dNLR, and PLR between the groups (P < .05). No significant correlation existed between peripheral blood inflammatory factors and IDH-1 gene mutation status or tumor location in patients with glioma (P > .05). LMR, NLR, dNLR, and PLR, varied significantly among different glioma types (P < .05). White blood cell (WBC) count, neutrophil, NLR, and dNLR correlated positively with glioma risk. Further, WBC, neutrophil, NLR, dNLR, and LMR had a high diagnostic efficiency. Conclusion: Peripheral blood inflammatory markers, serving as noninvasive biomarkers, offer high sensitivity and specificity for diagnosing glioma, differentiating it from meningioma, diagnosing GBM, and distinguishing GBM from low-grade glioma. These markers may be implemented as routine screening tools.

Worse fibro-inflammatory activity on diagnostic liver biopsy adversely impacts biochemical remission in autoimmune hepatitis.

BACKGROUND: Autoimmune hepatitis (AIH) patients can present with advanced fibrosis at diagnosis or may progress to the same if biochemical remission on treatment is not achieved. METHODS: We conducted a single-center retrospective analysis of 34 pediatrics and 39 adult AIH patients. Three pathologists, blinded to clinical information, reviewed the diagnostic liver biopsy (DLB) slides of AIH patients. We evaluated the impact of clinical, laboratory, and histopathologic parameters on outcomes including biochemical remission (BR). RESULTS: Incidence of advanced (Ludwig stage 3 or 4) fibrosis on DLB was 45.2 %. AIH patients with advanced fibrosis had higher median Ishak score (p < 0.001) and higher IgG level (p = 0.01) at diagnosis. The incidence of BR at 6-month (31.2% vs. 88.6 %, p = 0.001) and 1-year (68.8% vs. 88.6 %, p = 0.04) post-diagnosis was significantly lower in AIH patients with advanced fibrosis. Although not statistically significant, a higher proportion of AIH patients with advanced fibrosis were on high dose of steroids (58% vs. 37.9 %, p = 0.1) at 1 year post diagnosis. Higher serum IgG level at diagnosis was associated with lower odds of achieving BR at 6-month (p = 0.004) and 1-year (p = 0.03) post-diagnosis in multivariate analysis. Pediatric age at diagnosis (p = 0.02) was associated with higher steroid dose at 1-year post-diagnosis in univariate analysis. CONCLUSIONS: Findings of advanced fibrosis on DLB of AIH patients was accompanied by more pronounced necro-inflammatory activity and higher serum IgG level, which translated to lower rates of BR and higher exposure to steroids during the first year after diagnosis.

Qualitative and quantitative status of cytochrome P450s after the administration of a liposomal platelet substitute in rat liver.

In the process of the drug development, studies on the cytochrome P450 (CYP) profiles after its administration provided fundamental information regarding drug interactions with concomitantly administered drugs. Here, we evaluated the influence of the administration of H12-(ADP)-liposomes, a platelet substitute, on the mRNA and protein expression, and metabolic activity of CYPs, with focus on the CYP1A2, CYP2C11 and CYP3A2, in rat liver.At 24 h after administering saline or H12-(ADP)-liposomes (10 mg of lipids/kg), a quantitative RT-PCR and western blot analysis revealed that the mRNA and proteins expression of all of the target hepatic CYP isoforms were not different between the saline and H12-(ADP)-liposome groups. Furthermore, an ex vivo CYP metabolic activity assay showed that hepatic CYP metabolic activities in the H12-(ADP)-liposome group were comparable to the corresponding saline group. On the other hand, the area under the blood concentration-time curve for substitutes for CYP1A2 and CYP2C11 was higher in H12-(ADP)-liposome group than in saline group, but the degree of elevations was negligible levels.At a minimum, based on these results, we conclude that H12-(ADP)-liposomes have no quantitative and qualitative effect on the hepatic CYP isoforms, indicating that the drug interactions of H12-(ADP)-liposomes with CYP-metabolizing drugs would be negligible.

Comprehensive analysis of L-PRF exudate components and their impact on whole blood platelets.

OBJECTIVE: This study assessed the cellular composition and effects of leukocyte-platelet-rich fibrin (L-PRF) exudate on whole blood platelets from healthy volunteers. Key objectives included evaluating leukocyte subpopulations, platelet activation markers, platelet-leukocyte interactions and quantifying inflammatory cytokines within the L-PRF exudate. MATERIALS AND METHODS: L-PRF was obtained from 20 healthy donors. Flow cytometry methodologies were used to assess intracellular calcium kinetics and activated GPIIbIIIa, and P-selectin expression. Leukocyte subpopulations and platelet-leukocyte interactions were characterized using monoclonal antibodies. Inflammatory cytokines (IL-8, IL-1ß, IL-6, IL-10, TNF, IL-12p70) within L-PRF exudate were quantified using a cytometric bead array. RESULTS: The expression of activated GPIIbIIIa, and P-selectin exhibited a significant increase (p < 0.001) when L-PRF exudate was added to platelets of whole blood. Regarding intracellular Ca2+ mobilization, the L-PRF exudate elicited significant responses (p < 0.001). L-PRF exudate contained different leukocytes populations, being TCD4 + the most representative of T cells. It was possible to stablish a profile of cytokines produced by the L-PRF exudate, with human IL-8 cytokine exhibiting the highest average (16.90 pg/mL). CONCLUSIONS: Despite the study limitations, the research yielded important insights: 1- L-PRF exudate can stimulate platelet activation, essential in healing, tissue inflammation and remodeling. 2-The presence of leukocyte subpopulations within L-PRF exudate reflexes its complexity and potential to enhance immune responses. 3-The analysis of inflammatory cytokines within L-PRF exudate revealed its immunomodulatory potential. These findings are valuable evidences for understanding the potential role of L-PRF exudate in regenerative dentistry and medicine, offering innovative therapeutic strategies. CLINICAL RELEVANCE: This research highlights crucial aspects that could significantly influence the clinical use of L-PRF exudate in the oral cavity. The findings support the application of L-PRF exudate in both surgical and regenerative dentistry, facilitating the development of innovative therapeutic strategies to enhance patient outcomes.

Pharmacological inhibition of Septins with Forchlorfenuron attenuates thrombus formation in experimental thrombotic mice models with modulating multiple signaling pathways in platelets.

INTRODUCTION: The Septin family of cytoskeletal proteins is abundant in platelets. When these proteins are functionally blocked using the compound forchlorfenuron (FCF), it hampers the normal activation processes of purified human platelets. OBJECTIVES: To evaluate the in vivo effects of FCF on physiological haemostasis and pathological thrombosis in mice and to investigate possible molecular mechanisms. METHODS: The impact of FCF on haemorrhage risk in the brain, liver, and tail of mice was investigated. Using several experimental models, thrombus development in the lung, mesenteric arteries, and postcava was studied. Functional assays were performed on mice and human platelets, both with and without FCF pretreatment. These tests included aggregation, granule release, ROS production, integrin αIIbß3 activation, cytoskeletal remodeling imaging, and clot retraction. RESULTS: Neither oral nor intravenous administration of FCF showed any apparent impairment of haemostasis in the tissues studied, but only later administration resulted in a significant reduction in thrombus formation in different mice vessel types. FCF generally inhibited agonist-induced platelet aggregation, degranulation, ROS burst, morphological expansion on the fibrinogen matrix with completely disordered dynamic organizations of the cytoskeleton for septin, tubulin and actin. In addition, FCF was found to antagonise agonist-induced dephosphorylation of VASP (Ser239) and PI3K/AKT and ERK1/2 phosphorylation. CONCLUSION: FCF showed preferences in attenuating pathological thrombus formation, apart from physiological haemostasis, with possible mechanisms to prevent cytoskeletal remodelling and signal transduction of AKT, ERK1/2 and VASP signalling pathways, suggesting that Septin may serve as a promising target for the prevention and treatment of thrombotic diseases.

Ingestion of the Non-Nutritive Sweetener Erythritol, but Not Glucose, Enhances Platelet Reactivity and Thrombosis Potential in Healthy Volunteers.

BACKGROUND: Although artificial and non-nutritive sweeteners are widely used and generally recognized as safe by the US and European Union regulatory agencies, there have been no clinical trials to assess either long-term cardiovascular disease risks or short-term cardiovascular disease-relevant phenotypes. Recent studies report that fasting plasma levels of erythritol, a commonly used sweetener, are clinically associated with heightened incident cardiovascular disease risks and enhance thrombosis potential in vitro and in animal models. Effects of dietary erythritol on thrombosis phenotypes in humans have not been examined. METHODS: Using a prospective interventional study design, we tested the impact of erythritol or glucose consumption on multiple indices of stimulus-dependent platelet responsiveness in healthy volunteers (n=10 per group). Erythritol plasma levels were quantified with liquid chromatography tandem mass spectrometry. Platelet function at baseline and following erythritol or glucose ingestion was assessed via both aggregometry and analysis of granule markers released. RESULTS: Dietary erythritol (30 g), but not glucose (30 g), lead to a >1000-fold increase in erythritol plasma concentration (6480 [5930-7300] versus 3.75 [3.35-3.87] µmol/L; P<0.0001) and exhibited acute enhancement of stimulus-dependent aggregation responses in all subjects, agonists, and doses examined. Erythritol ingestion also enhanced stimulus-dependent release of the platelet dense granule marker serotonin (P<0.0001 for TRAP6 [thrombin activator peptide 6] and P=0.004 for ADP) and the platelet α-granule marker CXCL4 (C-X-C motif ligand-4; P<0.0001 for TRAP6 and P=0.06 for ADP). In contrast, glucose ingestion triggered no significant increases in stimulus-dependent release of either serotonin or CXCL4. CONCLUSIONS: Ingestion of a typical quantity of the non-nutritive sweetener erythritol, but not glucose, enhances platelet reactivity in healthy volunteers, raising concerns that erythritol consumption may enhance thrombosis potential. Combined with recent large-scale clinical observational studies and mechanistic cell-based and animal model studies, the present findings suggest that discussion of whether erythritol should be reevaluated as a food additive with the Generally Recognized as Safe designation is warranted. REGISTRATION: URL: https://www.clinicaltrials.gov; Unique identifier: NCT04731363.

Prospective Outcomes After Serial Platelet-Rich Plasma (PRP) Injection in Vocal Fold Scar and Sulcus.

OBJECTIVE: Vocal fold scar and sulcus pose significant treatment challenges with no current optimal treatment. Platelet-rich plasma (PRP), an autologous concentration of growth factors, holds promise for regenerating the superficial lamina propria. This study aims to evaluate the potential benefits of serial PRP injections on mucosal wave restoration and vocal function. METHODS: In a prospective clinical trial across two institutions, patients with vocal fold scar underwent four serial PRP injections, one month apart. Blinded independent laryngologists and expert listeners used pretreatment and one-month post-fourth injection videostroboscopy and CAPE-V assessments to evaluate mucosal wave and voice quality changes, respectively. Additionally, patient reported outcome measures (PROMs) were evaluated. RESULTS: In the study, 15 patients received 55 PRP injections without adverse effects. Eight patients (53.3%) had mild, three patients (20%) had moderate, and four patients (26.7%) had severe scar. There was an average reduction of 8.7 points in post-treatment VHI-10 scores (p = 0.007). The raters observed an improvement in post-treatment voice in 73.4% of cases, and CAPE-V scores showed a reduction of 18.8 points on average (p = 0.036). The videostroboscopic VALI ratings showed an improvement in mucosal wave rating from 2.0 to 4.0. On average, the raters perceived the post-PRP exams to be better in 56.7% of cases. CONCLUSIONS: PRP has been validated as a safe autologous option for treatment of vocal fold scar. While results for mucosal wave and voice quality varied, there was a consistent improvement in PROMs. LEVEL OF EVIDENCE: Level 3: Prospective cohort study, with blinded analysis Laryngoscope, 2024.