The malarial parasite Plasmodium can acquire resistance to most mainstay antimalarial drugs, necessitating the development of new antiplasmodial agents with different modes of action. The innate defense protein, human platelet factor 4 (PF4), has a unique antiplasmodial action that involves selective entry into Plasmodium-infected red blood cells (RBC) and subsequent destruction of the parasite's digestive vacuole (DV). This activity is recapitulated in PF4-derived internalization peptides (PDIPs). Here, we characterized the actions of PDIP analogs and PF4 in live P. falciparum-infected human RBC to understand their kinetics, effects on cell and parasite viability, and molecular requirements for antiplasmodial activity. The entry and accumulation of PDIP, and peptide-induced DV destruction, were distinguishable as ordered and rapidly occurring events that were equivalent to PF4. Both host cell and parasite plasma membranes remained intact and undamaged following destruction of the DV, although modest changes in phosphatidylserine (PS) exposure on the surface of the host cells (indicative of changes to its phospholipid organization) and swelling (but not lysis) of the intracellular parasite were observed. PDIP retained its macrocyclic structure, and its activity depended on elevated levels of PS on the surface of infected versus uninfected cells. Neither the intramolecular disulfide bond of PDIP, nor the parasite's nutrient and ion transporter functions were required. These actions on the parasite DV were not detected for other antiplasmodial drugs and compounds. In conclusion, this study reveals the unique, rapid, and distinct antiplasmodial actions of PDIP, highlighting its potential for future antimalarial development.

Sepsis is a life-threatening condition that has challenged many clinicians over the years. The immune and hemostatic systems are the primary pillars of sepsis pathogenesis. Dysregulation of these intricate mechanisms significantly worsens the prognosis. Coagulopathy is a critical aspect of sepsis, with the degree of hemostatic impairment being a key determinant of poor outcomes. Although the concept of sepsis caused by bacteria has been well investigated, the fungal impact in the complexity of sepsis-related hemostatic derangement is not yet fully unraveled. In addition, sepsis occurs in patients across all age groups, with a particular concern for neonates, whose immature and vulnerable systems amplify the challenges. Notably, despite the high incidence of fungal septicemia in neonatal intensive care units (NICUs), along with its significant morbidity, mortality, and adverse neonatal outcomes, the impact of fungal sepsis on the neonatal hemostatic system-an essential determinant of prognosis-remains largely unexplored. The present review delves into the pathophysiologic mechanisms of sepsis-induced coagulopathy attributed to fungal infection, the mechanisms of fungal involvement in the hemostatic derangement, and attempts to contextualize this knowledge within the unique neonatal population. Finally, it aims to raise awareness of the critical need for a deep understanding of this hazardous condition to guide the development of optimal therapeutic strategies.

This study evaluated the impact of platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) on burn wound with dual-species biofilm. Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) were applied to infect the burn wound in rats to establish a dual-species biofilm model. After infection, the wound was treated with ionized silver (AG), PRF, and PRP. Silver scanning electron microscopy (SEM) was used to assess adhesion after infection. PRF and PRP reduced wound size from day 8 after burn injuries, while AG significantly promoted burn wound healing at day 12. New collagen was formed in the shortest time in PRF and PRP groups compared to AG and control groups. PRF and PRP greatly lowered the bacterial numbers in wounds with S. aureus and P. aeruginosa biofilm, whereas AG showed weak bacteriostatic effects. AG, PRF, and PRP treatments significantly reduced inflammatory mediators and induced VEGFA. However, AG treatment increased TNF-α. PRF and PRP accelerate wound healing in the presence of dual-species biofilm infection and show strong antibacterial activity against S. aureus and P. aeruginosa, indicating that PRF and PRP could be potential therapies for burn wounds with dual-species biofilm infection.

Diabetic foot ulcers (DFUs) are severe complications of diabetes, often leading to chronic wounds, amputations, and increased mortality risk. Platelet concentrates (PCs)-natural biomaterials utilized in regenerative medicine-have garnered attention for their capacity to enhance tissue repair and wound healing. This study reviews the preparation methods, biological mechanisms, and clinical efficacy of three generations of PCs: platelet-rich plasma (PRP), platelet-rich fibrin (PRF), and concentrated growth factors (CGF). Comparative analysis reveals that PRP, the first generation, provides abundant growth factors but relies on anticoagulants, which may hinder fibrin formation and tissue adhesion. PRF, as the second generation, eliminates anticoagulants, forming a fibrin matrix that sustains growth factor release and enhances cell migration. CGF, the latest advancement, employs refined centrifugation to achieve higher growth factor concentrations and a denser fibrin matrix, accelerating tissue regeneration. Case series results demonstrated superior wound healing outcomes with CGF, including faster epithelialization and reduced healing time compared to PRP and PRF. These findings underscore CGF's potential as the most effective PC for managing DFUs, supporting its broader clinical adoption in advanced wound care.

Platelet-rich plasma (PrP) is a blood derivative with positive roles in regenerative medicine, particularly in wound healing. Evidence has been reported for using peripheral blood-derived PrP in disease treatments, but umbilical cord blood (UCB)-derived PrP remains limited. Thus, we investigate the roles of UCB-derived PrP in cellular behaviours in vitro and in wound healing in vivo models.

We used 2D and 3D cell culture models to investigate the role of UCB-derived PrP gels in stimulating the attachment, proliferation, migration, and spheroid formation of umbilical cord-derived mesenchymal stem cell (UCMSC) and human dermal fibroblast (hFB). In addition, immunoassay and PCR were used to understand the enrichment of growth factors in UCB-derived PrP and the change of ECM genes in PrP-treated cells. Finally, a rat model was used to investigate the cutaneous wound healing process.

UCB-derived PrP gels were enriched with platelet-derived growth factor-BB (PDGF-BB) (3394.1 ± 2658.3 pg/mL), vascular endothelial growth factor-A (VEGF-A) (282.0 ± 53.0 pg/mL), hepatocyte growth factor (HFG) (762.7 ± 117.5 pg/mL), and fibroblast growth factor 2 (FGF-2) (17.734 ± 8 pg/mL). In addition, these UCB-derived PrP gels promoted cell attachment (> 154 % and > 117 % for UCMSCs and hFBs, respectively), proliferation (UCMSCs > 121 % and hFBs > 117 % at all time points), migration increased by 27 % and 26 % for UCMSCs and hFBs, and spheroid formation and fusion compared to the control. UCB-derived PrP gels also induced different expression of ECM genes, including COL1, COL3, HAS1, HAS2, HAS3, and ENL, in both UCMSCs and hFBs. Finally, this product from UCBs could enhance the wound healing process in excised skin rat models by reducing the wound area by 80 % compared to 27 % in controls after 14 days.

UCB-derived PrP gels facilitate cell behaviours in vitro, including cell adhesion, growth, and migration. In addition, in animal models, UCB-derived PrP reduced the wound healing time and enhanced the completion of skin tissues by increasing granulation tissue formation and reducing neutrophils at wound sites. These UCB-derived PrP gels will be used to support spheroid formation that will be used as biomaterials for 3D printing, engraftment, and wound healing treatment.

Platelets, traditionally known for their role in hemostasis, have emerged as key players in immune response and inflammation. Sepsis, a life-threatening condition characterized by systemic inflammation, often presents with thrombocytopenia, which at times, can be significant. Platelets contribute to the inflammatory response by interacting with leukocytes, endothelial cells, and the innate immune system. However, excessive platelet activation and consumption can lead to thrombocytopenia and exacerbate the severity of sepsis. Understanding the multifaceted roles of platelets in sepsis is crucial for developing effective therapeutic strategies. Targeting platelet-mediated inflammatory responses and promoting platelet production may offer potential avenues for improving outcomes in septic patients with thrombocytopenia. Future research should focus on elucidating the mechanisms underlying platelet dysfunction in sepsis and exploring novel therapeutic approaches to optimize platelet function and mitigate inflammation. This review explores the intricate relationship between platelets, inflammation, and thrombosis in the context of sepsis.

The survival of an organism relies on its ability to repair the damage caused by trauma, toxic agents, and inflammation. This process involving cell proliferation and differentiation is driven by several growth factors and is critically dependent on the organization of the extracellular matrix. Since autologous platelet concentrates (APCs) are fibrin matrices in which cells, growth factors, and cytokines are trapped and delivered over time, they are able to influence that response at different levels. The present review thoroughly describes the molecular components present in one of these APCs, leukocyte- and platelet-rich fibrin (L-PRF), and summarizes the level of evidence regarding the influence of L-PRF on anti-inflammatory reactions, analgesia, hemostasis, antimicrobial capacity, and its biological mechanisms on bone/soft tissue regeneration.

Platelets are anucleate blood cells traditionally associated with hemostasis but now increasingly recognized for their multifaceted roles in immunity, inflammation, and tissue repair. Advances in platelet proteomics, employing high-throughput techniques such as mass spectrometry, have significantly enhanced our understanding of platelet biology and its clinical implications in transfusion medicine. Platelet proteomics offers a retrospective view of physiological and pathological changes over the platelet's 7-10-day lifespan, making it a unique tool for studying cumulative biological events. Recent applications include the identification of biomarkers for cardiovascular, infectious, autoimmune diseases and cancer. In neurodegeneration and aging, platelets have been explored for their shared molecular pathways with neurons, with findings implicating Tau, amyloid-beta, and alpha-synuclein as potential biomarkers. Proteomics is also emerging as an important factor in the development of evidence-based, tailor-made platelet-derived therapies. While promising, platelet proteomics requires further standardization and computational advances to support transitioning from research to routine clinical practice.

Platelet lysate (PL) demonstrates antimicrobial and anti-inflammatory properties offering potential for treatment of bacterial pneumonia in horses. It remains unknown whether nebulization is suitable for PL administration in horses. This pilot study characterized particle size and flow rate of pooled equine PL (single preparation) nebulized using an equine-specific nebulizer (Flexivent®).

Protein composition and antimicrobial activity were compared before and after nebulization. Protein composition was evaluated according to growth factor, antimicrobial peptide and cytokine concentrations and proteomic analysis. To evaluate antimicrobial activity, bacterial growth inhibition [maximum growth (μmax); carrying capacity (K)] were determined for E. coli, Streptococcus equi subsp zooepidemicus and Rhodococcus equi (WT and MDR) using pre- and post-nebulized PL concentrations of 50%.

Flow rate and median particle size were 0.8 ml/min and 4.991 μm with 52% of particles ≤ 5 μm. Differences in PL protein composition were detected with nebulization. For E. coli and S. zooepidemicus, nebulization did not alter effect of PL on growth parameters. PL treatments decreased K for S. zooepidemicus (p = 0.009) compared to BHI. For R. equi K was increased post- vs. pre-nebulization (WT and MDR) and μmax increased pre- vs, post-nebulization (MDR). PL treatments increased K and μmax for MDR R. equi and μmax for WT R. equi compared to BHI (p ≤ 0.05).

Nebulization of PL in vitro is technically feasible. The results of this study support further investigation to better characterize the effect of nebulization on PL and its suitability for nebulization in horses.

Atherosclerosis is a chronic, multifactorial inflammatory disorder of large and medium-size arteries, which is the leading cause of cardiovascular mortality and morbidity worldwide. Although platelets in cardiovascular disease have mainly been studied for their crucial role in the thrombotic event triggered by atherosclerotic plaque rupture, over the last two decades it has become clear that platelets participate also in the development of atherosclerosis, owing to their ability to interact with the damaged arterial wall and with leukocytes. Platelets participate in all phases of atherogenesis, from the initial functional damage to endothelial cells to plaque unstabilization. Platelets deposit at atherosclerosis predilection sites before the appearance of manifest lesions to the endothelium and contribute to induce endothelial dysfunction, thus supporting leukocyte adhesion to the vessel wall. In particular, platelets release matrix metalloproteinases, which interact with protease-activated receptor 1 on endothelial cells triggering adhesion molecule expression. Moreover, P-selectin and glycoprotein Ibα expressed on the surface of vessel wall-adhering platelets bind PSGL-1 and β2 integrins on leukocytes, favoring their arrest and transendothelial migration. Platelet-leukocyte interactions promote the formation of radical oxygen species which are strongly involved in the lipid peroxidation associated with atherosclerosis. Platelets themselves actively migrate through the endothelium toward the plaque core where they release chemokines that modify the microenvironment by modulating the function of other inflammatory cells, such as macrophages. While current antiplatelet agents seem unable to prevent the contribution of platelets to atherogenesis, the inhibition of platelet secretion, of the release of MMPs, and of some specific pathways of platelet adhesion to the vessel wall may represent promising future strategies for the prevention of atheroprogression.

Platelets are considered one of the most important reservoirs not only of growth factors, but also of neurotrophic factors that could contribute to the repair of vascular lesions and the prevention of neurological deterioration. Among these factors, Brain-Derived Neurotrophic Factor (BDNF) - a protein belonging to the neurotrophin family - is widely expressed both in the hippocampus and in platelets. Platelets constitute an important reservoir of BDNF; however, little is known about the factors modulating its release into the circulation and whether anti-platelet drugs affect this secretion. In this review, we have discussed the link between BDNF and platelets and their role in neurological disorders.

Macrophages and osteocytes are important regulators of inflammation, osteogenesis and osteoclastogenesis. However, their interactions under adverse conditions, such as biomaterial-associated infection (BAI) are not fully understood. We aimed to elucidate how factors released from macrophages modulate osteocyte responses in an in vitro indirect 3D co-culture model. Human monocyte-derived macrophages were cultured on etched titanium disks and activated with either IL-4 cytokine (anti-inflammatory M2 phenotype) or Staphylococcus aureus secreted virulence factors to simulate BAI (pro-inflammatory M1 phenotype). Primary osteocytes in collagen gels were then stimulated with conditioned media (CM) from these macrophages. The osteocyte response was analyzed by gene expression, protein secretion, and immunostaining. M1 phenotype macrophages were confirmed by IL-1β and TNF-α secretion, and M2 macrophages by ARG-1 and MRC-1.Osteocytes receiving M1 CM revealed bone inhibitory effects, denoted by reduced secretion of bone formation osteocalcin (BGLAP) and increased secretion of the bone inhibitory sclerostin (SOST). These osteocytes also downregulated the pro-mineralization gene PHEX and upregulated the anti-mineralization gene MEPE. Additionally, exhibited pro-osteoclastic potential by upregulating pro-osteoclastic gene RANKL expression. Nonetheless, M1-stimulated osteocytes expressed a higher level of the potent pro-osteogenic factor BMP-2 in parallel with the downregulation of the bone inhibitor genes DKK1 and SOST, suggesting a compensatory feedback mechanisms. Conversely, M2-stimulated osteocytes mainly upregulated anti-osteoclastic gene OPG expression, suggesting an anti-catabolic effect. Altogether, our findings demonstrate a strong communication between M1 macrophages and osteocytes under M1 (BAI)-simulated conditions, suggesting that the BAI adverse effects on osteoblastic and osteoclastic processes in vitro are partly mediated via this communication. STATEMENT OF SIGNIFICANCE: Biomaterial-associated infections are major challenges and the underlying mechanisms in the cellular interactions are missing, especially among the major cells from the inflammatory side (macrophages as the key cell in bacterial clearance) and the regenerative side (osteocyte as main regulator of bone). We evaluated the effect of macrophage polarization driven by the stimulation with bacterial virulence factors on the osteocyte function using an indirect co-culture model, hence mimicking the scenario of a biomaterial-associated infection. The results suggest that at least part of the adverse effects of biomaterial associated infection on osteoblastic and osteoclastic processes in vitro are mediated via macrophage-to-osteocyte communication.

Equine joint disease including septic arthritis (SA) and osteoarthritis (OA) is a critical cause of equine lameness. Platelet-rich plasma (PRP) is one of the most popular regenerative therapies to treat equine OA, even SA, but the evidence in support of the treatment is conflicting.

The aim of the study was to systematically review the current evidence on PRP products used for SA and OA, as well as the efficacy of PRP products as treatment for OA on the basis of a meta-analysis of the available literature.

Systematic review and meta-analysis.

A systematic search of relevant databases (PubMed, Web of Science, Scopus) was performed to identify studies published from 2013 to 2023, in accordance with the PRISMA guidelines. Randomised controlled trials, non-randomised trials and controlled laboratory studies that used at least one type of PRP products were included. Dichotomous outcomes were presented using odds ratios (ORs) and 95% confidence intervals (95% CIs).

A total of 21 publications were identified in the systematic review and 5 of them in the meta-analysis. These publications involved various types of PRP products and reported different outcomes. Although most of the studies were associated with a high risk of bias, the overall estimated effect was consistent with a significant improvement in the PRP products treatment group compared with the control group (OR: 15.32; 95% CI: 3.00-78.15; p < 0.05). There was a significant improvement in clinical performance outcomes between the groups (OR: 36.64; 95% CI: 3.69-364.30; p < 0.05).

PRP products as intra-articular treatment are likely efficacious for treatment of equine OA and have potential for treating SA. These conclusions might be affected by the limited number of randomised controlled studies and high variability of different types of PRP products. To better evaluate the efficacy of PRP, a widely recognised classification system and the utilisation of randomised, blinded, equivalency or non-inferiority trials are required.

There is a growing interest in using extracellular vesicles (EVs) for therapeutic applications. EVs are composed of cytoplasmic proteins and nucleic acids and an external lipid bilayer containing transmembrane proteins on their surfaces. EVs can alter the state of the target cells by interacting with the receptor ligand of the target cell or by being internalised by the target cell. Blood cells are the primary source of EVs, and 1 μL of plasma contains approximately 1.5 × 107 EVs. Owing to their easy acquisition and the avoidance of cell amplification in vitro, using blood cells as a source of therapeutic EVs has promising clinical application prospects. This review summarises the characteristics and biological functions of EVs derived from different blood cell types (platelets, erythrocytes, and leukocytes) and analyses the prospects and challenges of using them for clinical therapeutic applications. In summary, blood cell-derived EVs can regulate different cell types such as immune cells (macrophages, T cells, and dendritic cells), stem cells, and somatic cells, and play a role in intercellular communication, immune regulation, and cell proliferation. Overall, blood cell-derived EVs have the potential for use in vascular diseases, inflammatory diseases, degenerative diseases, and injuries. To promote the clinical translation of blood cell-derived EVs, researchers need to perform further studies on EVs in terms of scalable and reproducible isolation technology, quality control, safety, stability and storage, regulatory issues, cost-effectiveness, and long-term efficacy.

Chronic rhinosinusitis (CRS) is a prevalent chronic disease observed on a global scale. The utilization of endoscopic sinus surgery (ESS) has gained significant recognition as an effective intervention for individuals with CRS and nasal polyps who have not responded to conventional treatments. The need (or not) for revision surgery frequently relies on the promotion of optimal wound healing. The impact of platelet-rich plasma (PRP) on tissue healing has been extensively examined in various surgical fields.

The present prospective study involved 30 patients suffering with nasal polyposis who underwent endoscopic sinus surgery. 15 patients were assigned to the PRP group, and 15 patients to the control group. The clinical follow-up of the patients took place at specific intervals, at weeks 1, 2, 3, 4, 8, and 12 after the surgical procedure. The evaluator identified the existence of adhesions, crusting, bleeding, granulation and infection using a visual analogue scale score. The patients also completed the SNOT 22 questionnaire prior to surgery and at each postoperative visit.

The present study observed a lower incidence of adhesion, infection, hemorrhage and granulation in the PRP group. Furthermore, a statistically significant difference was detected between the groups.

Based on the findings of the present investigation, it seems that platelet-rich plasma (PRP) is beneficial on wound healing during the early stages following the surgical procedure. The technique is characterized by its limited invasiveness, which contributes to its low risk profile and the achievement of clinically good outcomes.

A simple method to generate antibacterial peptides by alkaline hydrolysis of hen egg whites is reported. The method reproducibly generates short peptides with molecular weight of less than 14.4 kDa that exhibit low to no cytotoxicity on RAW 264.7 macrophage cells, but do inhibit the bacterial growth of Cutibacterium acnes (C. acnes), Staphylococcus aureus (S. aureus) and antibiotic-resistant S. aureus (MRSA), while also reducing nitric oxide production from heat-killed C. acnes-treated RAW 264.7 cells. Peptidomics revealed at least thirty peptides within the complex mixture, of which eight were evaluated individually. Three peptides (PK8, EE9 and RP8) were potent anti-inflammation and antibacterial agents, but notably the complex egg white hydrolysate (EWH) was more effective than the individual peptides. Electron microscopy suggests the antibacterial mechanism of both the hydrolysate and the selected peptides is through disruption of the cell membrane of C. acnes. These findings suggest that EWH and EWH-derived peptides are promising candidates for infection and inflammation treatment, particularly in managing acne and combating antibiotic-resistant bacteria like MRSA.

Periodontitis is a dysbiosis-driven inflammatory disease affecting the tooth-supporting tissues, characterized by their progressive resorption, which can ultimately lead to tooth loss. A step-wise therapeutic approach is employed for periodontitis. After an initial behavioral and non-surgical phase, intra-bony or furcation defects may be amenable to regenerative procedures. This review discusses the regenerative technologies employed for periodontal regeneration, highlighting the current limitations and future research areas. The search, performed on the MEDLINE database, has identified the available biomaterials, including biologicals (autologous platelet concentrates, hydrogels), bone grafts (pure or putty), and membranes. Biologicals and bone grafts have been critically analyzed in terms of composition, mechanism of action, and clinical applications. Although a certain degree of periodontal regeneration is predictable in intra-bony and class II furcation defects, complete defect closure is hardly achieved. Moreover, treating class III furcation defects remains challenging. The key properties required for functional regeneration are discussed, and none of the commercially available biomaterials possess all the ideal characteristics. Therefore, research is needed to promote the advancement of more effective and targeted regenerative therapies for periodontitis. Lastly, improving the design and reporting of clinical studies is suggested by strictly adhering to the Consolidated Standards of Reporting Trials (CONSORT) 2010 statement.

The fungus Candida albicans can cause mucosal infections including oropharyngeal candidiasis (OPC) in immunocompromised patients. In humans, an increased risk of fungal infections correlates with thrombocytopenia. However, our understanding of platelets and megakaryocytes (Mks) in mucosal fungal infections is almost entirely unknown. When megakaryocyte- and platelet-depleted mice were infected with OPC, the tongue showed higher fungal burden, due to decreased neutrophil accumulation. Protection depended on a distinct population of oral-resident Mks. Interleukin-17, important in antifungal immunity, was required since mice lacking the IL-17 receptor had decreased circulating platelets and their oral Mks did not expand during OPC. The secretion of the peptide toxin candidalysin activated human Mks to release platelets with antifungal capacity. Infection with a candidalysin-deficient strain resulted in decreased expansion of tongue Mks during OPC. This is the first time that a distinct megakaryocyte population was identified in the oral mucosa which is critical for immunity against fungal infection.

This study aims to evaluate the clinical effects of different blood derivatives on wound healing using network meta-analysis. PubMed, Embase, OVID, Web of Science, SCOPUS and Cochrane Central were searched to obtain studies about blood derivatives on wound healing until October 2023. R 4.2.0 and Stata 15.0 softwares were used for data analysis. Forty-four studies comprising 5164 patients were included. The results of network meta-analysis showed that the healing area from high to low was GF + ORCCB, ORCCB, GF, PRF, Unnas paste dressing, APG, PRP injection, PRP, PRP + thrombin gel, PPP, HPL, CT. The healing time from low to high was PRP + thrombin gel, GF, PRP, PC + K, PC, APG, PRF, CT, Silver sulfadiazine ointment. The number of patients cured from high to low was APG, PRP injection, PRP, Aurix, PRF, Leucopatch, HPL, Antimicrobial Ointment Dressing, CT, 60 μg/cm2 repifermin, 120 μg/cm2 repifermin, AFG, PPP. The order of analgesic effect from high to low was AFG, Aminogam gel, PRF, PRP, Oxidised oil, APG, GF, CT. The order of the number of wound infection cases from low to high is APG, 20 μg/cm2 repifermin, 60 μg/cm2 repifermin, PRP, LeucoPatch, CT, PPP, Antiseptic ointment dressing. Healing area: GF + ORCCB had the best effect; Healing time: PRP + thrombin gel took the shortest time. The number of cured patients and the reduction of wound infection: APG has the best effect. Analgesic effect: AFG has the best effect. More studies with large sample sizes are needed to confirm the above findings.

Thrombocytopenia, characterized by a decrease in platelet count, is a multifaceted clinical manifestation that can arise from various underlying causes. This review delves into the intriguing nexus between viruses and thrombocytopenia, shedding light on intricate pathophysiological mechanisms and highlighting the pivotal role of platelets in viral infections. The review further navigates the landscape of thrombocytopenia in relation to specific viruses, and sheds light on the diverse mechanisms through which hepatitis C virus (HCV), measles virus, parvovirus B19, and other viral agents contribute to platelet depletion. As we gain deeper insights into these interactions, we move closer to elucidating potential therapeutic avenues and preventive strategies for managing thrombocytopenia in the context of viral infections.

The determination of lipopolysaccharide (endotoxin) in serum or plasma samples using Limulus amebocyte lysate (LAL)-based assays is currently not sufficiently reliable in clinical diagnostics due to numerous interfering factors that strongly reduce the recovery of LPS in clinical samples. The specific plasma components responsible for the endotoxin neutralizing capacity of human blood remain to be identified. There are indications that certain endotoxin-neutralizing proteins or peptides, which are part of the host defense peptides/proteins of the innate immune system may be responsible for this effect. Based on our finding that several antimicrobial peptides can be neutralized by the polyanion heparin, we developed a heparin-containing diluent for serum and plasma samples, which enables reliable quantification of LPS measurement in clinical samples using the LAL assay. In a preclinical study involving 40 donors, this improved protocol yielded an over eightfold increase in LPS recovery in serum samples, as compared to the standard protocol. This modified protocol of sample pretreatment could make LPS measurement a valuable tool in medical diagnostics.

Effective wound treatment has become one of the most important challenges for healthcare as it continues to be one of the leading causes of death worldwide. Therefore, wound care technologies significantly evolved in order to provide a holistic approach based on various designs of functional wound dressings. Among them, hydrogels have been widely used for wound treatment due to their biocompatibility and similarity to the extracellular matrix. The hydrogel formula offers the control of an optimal wound moisture level due to its ability to absorb excess fluid from the wound or release moisture as needed. Additionally, hydrogels can be successfully integrated with a plethora of biologically active components (e.g., nanoparticles, pharmaceuticals, natural extracts, peptides), thus enhancing the performance of resulting composite hydrogels in wound healing applications. In this review, the-state-of-the-art discoveries related to stimuli-responsive hydrogel-based dressings have been summarized, taking into account their antimicrobial, anti-inflammatory, antioxidant, and hemostatic properties, as well as other effects (e.g., re-epithelialization, vascularization, and restoration of the tissue) resulting from their use.

The skin is the largest organ in the human body and serves as a barrier for protective, immune, and sensory functions. Continuous and permanent exposure to the external environment results in different levels of skin and extracellular matrix damage. During skin wound healing, the use of good dressings and addition of growth factors to the wound site can effectively modulate the rate of wound healing. A dressing containing bioactive substances can absorb wound exudates and reduce adhesion between the wound and dressing, whereas growth factors, cytokines, and signaling factors can promote cell motility and proliferation.

We prepared a functional wound dressing by combining platelet-rich plasma (PRP) and zwitterionic hydrogels. Functional wound dressings are rich in various naturally occurring growth factors that can effectively promote the healing process in various types of tissues and absorb wound exudates to reduce adhesion between wounds and dressings. Furthermore, PRP-incorporated zwitterionic hydrogels have been used to repair full-thickness wounds in Sprague-Dawley rats with diabetes (DM SD).

Fibroblasts and keratinocytes were cultured with PRP, zwitterionic hydrogels, and PRP-incorporated zwitterionic hydrogels to assess cell proliferation and specific gene expression. Furthermore, PRP-incorporated zwitterionic hydrogels were used to repair full-thickness skin defects in DM SD rats.

The swelling ratio of hydrogel, hydrogel + PRP1000 (108 platelets/mL), and hydrogel + PRP1000 (109 platelets/mL) groups were similar (~07.71% ± 1.396%, 700.17% ± 1.901%, 687.48% ± 4.661%, respectively) at 144 hours. The tensile strength and Young modulus of the hydrogel and hydrogel + PRP10000 groups were not significantly different. High concentrations of PRP (approximately 108 and 109 platelets/mL) effectively promoted the proliferation of fibroblasts and keratinocytes. The zwitterionic hydrogels were not cytotoxic to any cell type. High PRP concentration-incorporated zwitterionic hydrogels increased the rate of cell proliferation and significantly increased the expression of characteristic genes such as collagen, fibronectin, involucrin, and keratin. Subsequently, zwitterionic hydrogels with high PRP concentrations were used to repair full-thickness skin defects in DM SD rats, and a wound healing rate of more than 90% was recorded on day 12.

PRP contains high concentrations of growth factors that promote cell viability, enhance specific gene expression, and have a high medical value in cell therapy. Zwitterionic hydrogels have a 3-dimensional interconnected microporous structure and can resist cell adhesion without causing cytotoxicity. Platelet-rich plasma-incorporated zwitterionic hydrogels further enhance the cellular properties and provide an effective therapeutic option for wound healing.

This study aims to examine the effect of Leukocyte-Rich Platelet-Rich Plasma (LR-PRP) on bacterial translocation in an experimental peritonitis model in rats. Secondary peritonitis occurs due to the loss of integrity in the mucosal barrier of the gastrointestinal system, resulting from contamination of the peritoneal cavity by microorganisms. LR-PRP has been shown to have positive anti-infectious, immunomodulatory, and angiogenetic effects.

Twenty-seven Wistar-Albino rats were divided into three groups: Sham, Control, and Experimental. Laparotomy was performed on the rats under anesthesia, and the cecum was isolated. No procedure was performed on the Sham group. The cecums of the rats in the Control and Experimental groups were punctured twice within 5 minutes using an 18-gauge needle. The blood product of each rat in the Experimental group was prepared for autologous use as LR-PRP and administered intraperitoneally. The abdomens of rats in all groups were closed after 8 minutes. After 8 hours, the rats were sacrificed, and tissue and blood samples were collected. Inflammatory parameters (TNF-α, IL-1, and IL-6) and blood cultures were analyzed from the blood samples. Cultures were also performed on liver, spleen, and mesenteric lymph node tissue samples.

Liver tissue culture growth was not detected in rats in the sham group. It was detected in 6 rats in the control group, and in 1 rat in the experimental group. Mesenteric lymph node tissue culture growth was detected in 2 rats in the sham group, in 7 in the control group and in 1 in the experimental group. Blood culture growth was not detected in rats in the sham group, but detected in 8 rats in the control group, and 3 in the experimental group. In terms of liver tissue culture, mesenteric lymph node tissue culture, and blood culture; a significant relationship was statistically observed between the control and experimental groups (p=0.049, p=0.008, p=0.015, respectively). It was infered that a statistically significant relationship in the mean TNF-alpha, IL-1, IL-6 values was not seen between the control and experimental groups (p=0.999, p=0.999, p=0.590, respectively).

LR-PRP's ability to suppress bacterial translocation was statistically significant in liver tissue culture, mesenteric lymph node tissue culture, and blood culture when comparing the Control and Experimental groups. LR-PRP was found to be effective in preventing bacterial translocation without suppressing inflammation and exhibited antimicrobial properties as supported by the literature.

Teleost fish possess a diversity of type Ⅰ interferons (IFNs) repertoire, which play a crucial role in antiviral and antimicrobial immune responses. In our previous study, IFNe1-3 and IFNb were identified and cloned from Chinese sturgeon (Acipenser sinensis), an acipenseriform fish. However, the absence of Chinese sturgeon genome data has left the question of whether there are other type Ⅰ IFN members in this species unresolved. In this study, we have identified and characterized a novel IFN, IFNf in Chinese sturgeon (AsIFNf). Bioinformatics analysis revealed that the AsIFNf contains a unique disulfide bond (2 cysteines) located in the second exon and fifth exon region, distinguishing it from other reported teleost type I IFNs. Meanwhile, qPCR results showed that AsIFNf mRNA was detectable in all examined tissues and up-regulated in the spleen or kidney in response to poly I: C, Citrobacter freundii, and Spring Viremia of Carp Virus (SVCV), but not by LPS. Furthermore, compared to recombinant AsIFNe2 protein (rAsIFNe2), rAsIFNf exhibited a stronger protective effect on Chinese sturgeon fin cells against SVCV and also induced higher expression of antiviral genes Mx and viperin. Importantly, AsIFNf displayed characteristics similar to antimicrobial peptides (AMPs) with a positive charge and demonstrated a broad spectrum of antimicrobial activity in vitro. These findings provide a theoretical foundation for understanding the primitive structure and function of interferon, as well as deepening our comprehension of the innate immune system and disease defense in the endangered Chinese sturgeon.

Platelet-rich plasma (PRP), derived from the centrifugation and subsequent separation of whole blood, results in an unusually high concentration of platelets. A newer form of platelet concentrate, platelet-rich fibrin (PRF), has also been developed. There has been significant research into the therapeutic effects of PRP, particularly in enhancing wound healing and preventing infections in surgical wounds. This scoping review aims to thoroughly evaluate preclinical and clinical evidence regarding the antimicrobial effects of PRP and PRF. In conducting this review, 612 records were examined, and 36 articles were selected for inclusion. The studies reviewed include preclinical research, such as in-vitro and in-vivo studies, and clinical trials involving human participants. The current clinical evidence suggests a notable trend towards the antimicrobial capabilities of PRP and PRF, underscoring their potential benefits in treating wounds. The application of PRP and PRF in wound management shows encouraging outcomes, but further investigation is needed to optimize their use as antimicrobial agents. Additional research, particularly randomized controlled trials, is essential to substantiate their antimicrobial effectiveness in specific diseases and types of wounds, considering their potential impact on clinical results.

Increasing antimicrobial resistance in veterinary practice has driven the investigation of novel therapeutic strategies including regenerative and biologic therapies to treat bacterial infection. Integration of biological approaches such as platelet lysate and mesenchymal stromal cell (MSC) therapy may represent adjunctive treatment strategies for bacterial infections that minimize systemic side effects and local tissue toxicity associated with traditional antibiotics and that are not subject to antibiotic resistance. In this review, we will discuss mechanisms by which biological therapies exert antimicrobial effects, as well as potential applications and challenges in clinical implementation in equine practice.

To examine the use of platelet-rich plasma (PRP) for treatment of pilonidal disease (PD) and thus provide a reference for clinical application.

A systematic review of PubMed and the Cochrane Library was performed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses. We considered all studies that reported the use of PRP for treatment of PD. Extracted data included the first author's name, year of publication, study type, number of included patients, inclusion and exclusion criteria, interventions, anesthesia, application of PRP (source, preparation, dose, and operation), antibiotics, follow-up time, therapeutic outcomes, and adverse events.

In total, eight randomized controlled trials and one prospective cohort study involving 809 patients were included. PRP reduced pain, accelerated healing, and reduced adverse events. The application of combined minimally invasive surgery achieved better results. However, overfilling of the wound with PRP in minimally invasive surgeries was shown to potentially increase the risk of adverse events.

PRP can be used as an adjuvant treatment in PD surgery to improve the therapeutic effect and reduce adverse events. The optimal combination of PRP and various factors is an important direction of future research.INPLASY registration number: INPLASY2023100070.

Perianal abscess is a common disease of the anus and intestine. Surgery is an important treatment option for perianal abscess. However, some patients have a long healing time, poor healing effect after surgery, or even pseudo-healing. Platelet-rich plasma (PRP) is rich in platelets that can release a large number of factors when activated and promote wound healing. Moreover, there are few reports on the use of PRP for wounds that are difficult to heal after perianal abscess surgery.

The patient had reported a complaint of perianal swelling and discomfort associated with anal pain, which was considered a perianal abscess. Ceftriaxone, fumigation, and sitz bath were administered after mixed hemorrhoid and perianal abscess surgeries were performed; however, the wound remained unhealed for more than 3 months, and there was a fistula under the skin.

Perianal color ultrasonography revealed perianal abscess.

Autologous PRP treatment was performed 5 times for each patient.

The postoperative wound healed within 15 days after 5 times PRP treatments.

PRP is a novel treatment option for pseudo-healing.

Enteric fistula (EF), a serious complication after abdominal surgery, refers to unnatural communication between the gastrointestinal tract and the skin or other hollow organs. It is associated with infection, massive fluid/electrolyte loss, and malnutrition, resulting in an unhealed course. Despite advances in surgical techniques, wound care, infection control, and nutritional support, EF remains associated with considerable morbidity and mortality. Autologous platelet-rich plasma (PRP) containing elevated platelet concentrations has been proposed to promote healing in many tissues. However, the mechanism of action of PRP in EF treatment remains unclear owing to its complicated clinical manifestations. In this review, we summarized the clinical approaches, outlined the principal cytokines involved in the healing effects, and discussed the advantages of PRP for EF therapy. In addition, we defined the mechanism of autologous PRP in EF management, which is essential for further developing EF therapies.

Coronavirus disease 2019 (COVID-19), which was caused by the coronavirus - severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was globally responsible for remarkable morbidity and mortality. Several highly effective vaccines for COVID-19 were developed and disseminated worldwide within an unprecedented timescale. Rare but dangerous clotting and thrombocytopenia events, and subsequent coagulation abnormalities, have been reported after massive vaccination against SARS-CoV-2. Soon after their global rollout, reports of a morbid clinical syndrome following vaccination with adenovirus-DNA-based vaccines appeared. In the spring of 2021, reports of a novel, rare and morbid clinical syndrome, with clinically devastating and fatal complication after vaccination with adenovirus-based coronavirus vaccines (Janssen/Johnson & Johnson and Astra-Zeneca vaccines) led to a brief suspension of their use by several countries. Those complications were associated with unusual cerebral and splanchnic venous thrombosis, and circulating autoantibodies directed against anti-platelet factor 4 (PF4), a protein secreted from platelets, leading to the designation: Vaccine-Induced Immune Thrombotic Thrombocytopenia (VITT). The reported VITT incidence remains very low and does not affect the overall benefit of immunization, however, if left untreated, VITT can be debilitating or even fatal. VITT resembled specific adverse drugs' reactions that also involved the production of autoantibodies and subsequent abnormal platelet activation through platelet FcγRIIa. These unusual but well-documented drug reactions were heparin-induced thrombocytopenia (HIT), streptokinase- (SK), and anisoylated plasminogen-streptokinase activator complex- (APSAC) associated with platelet-activating antibodies. There was considerable overlapping of clinical features between VITT, COVID-19 and these adverse drugs' reactions. We review the phenomenon of VITT against the backdrop of shared and common mechanisms that underlie HIT-, SK-, and APSAC-platelet FcγRIIa-dependent platelet activation. An understanding of VITT's pathogenesis may be achieved by comparing and contrasting VITT-, HIT-, SK- and APSAC-induced platelet activation mechanisms, their respective physiopathology and similarities. Discussing these conditions in parallel provides insight into complex immunological disorders and diseases associated with abnormal hemostasis and thrombosis in particular.

Platelet factor 4 (PF4, CXCL4), the most abundant α-granule platelet-specific chemokine, forms tetramers with an equatorial ring of high positive charge that bind to a wide range of polyanions, after which it changes conformation to expose antigenic epitopes. Antibodies directed against PF4 not only help to clear infection but can also lead to the development of thrombotic disorders such as heparin-induced thrombocytopenia (HIT) and vaccine-induced thrombocytopenia and thrombosis (VITT). This review will outline the different mechanisms through which PF4 engagement with polyanions combats infection but also contributes to the pathogenesis of inflammatory and thrombotic disease states.

Recent work has shown that PF4 binding to microbial polyanions may improve outcomes in infection by enhancing leukocyte-bacterial binding, tethering pathogens to neutrophil extracellular traps (NETs), decreasing the thrombotic potential of NET DNA, and modulating viral infectivity. However, PF4 binding to nucleic acids may enhance their recognition by innate immune receptors, leading to autoinflammation. Lastly, while HIT is induced by platelet activating antibodies that bind to PF4/polyanion complexes, VITT, which occurs in a small subset of patients treated with COVID-19 adenovirus vector vaccines, is characterized by prothrombotic antibodies that bind to PF4 alone.

Investigating the complex interplay of PF4 and polyanions may provide insights relevant to the treatment of infectious disease while also improving our understanding of the pathogenesis of thrombotic disorders driven by anti-PF4/polyanion and anti-PF4 antibodies.

The complement receptor CR3, also known as integrin Mac-1 (CD11b/CD18), is one of the major phagocytic receptors on the surface of neutrophils and macrophages. We previously demonstrated that in its protein ligands, Mac-1 binds sequences enriched in basic and hydrophobic residues and strongly disfavors negatively charged sequences. The avoidance by Mac-1 of negatively charged surfaces suggests that the bacterial wall and bacterial capsule possessing net negative electrostatic charge may repel Mac-1 and that the cationic Mac-1 ligands can overcome this evasion by acting as opsonins. Indeed, we previously showed that opsonization of Gram-negative Escherichia coli with several cationic peptides, including PF4 (Platelet Factor 4), strongly augmented phagocytosis by macrophages. Here, we investigated the effect of recombinant PF4 (rPF4) on phagocytosis of Gram-positive Staphylococcus aureus in vitro and examined its impact in a mouse model of S. aureus peritonitis. Characterization of the interaction of rPF4 with nonencapsulated and encapsulated S. aureus showed that rPF4 localizes on the bacterial surface, thus making it available for Mac-1. Furthermore, rPF4 did not have direct bactericidal and bacteriostatic activity and was not toxic to host cells. rPF4 enhanced phagocytosis of S. aureus bioparticles by various primary and cultured Mac-1-expressing leukocytes by several folds. It also increased phagocytosis of live nonencapsulated and encapsulated bacteria. Notably, the augmentation of phagocytosis by rPF4 did not compromise the intracellular killing of S. aureus by macrophages. Using a murine S. aureus peritonitis model, we showed that treatment of infected mice with rPF4 caused a significant increase in the clearance of antibiotic-susceptible S. aureus and its methicillin-resistant (MRSA) variant and markedly improved survival. These findings indicate that rPF4 binding to the bacterial surface circumvents its antiphagocytic properties, improving host defense against antibiotic-susceptible and antibiotic-resistant bacteria.

Thymosin beta-4 (Tβ4) is a ubiquitous protein with multiple and diverse intracellular and extracellular functions in vertebrates, which play fundamental roles in innate immune against pathogens and wound healing. In this study, the full-length cDNA of Tβ4 was cloned from Penaeus monodon (designated as PmTβ4), using the technology of rapid amplification of cDNA ends (RACE). The cDNA of PmTβ4 was 1361 bp with an open reading frame (ORF) of 501 bp, which encoding a polypeptide of 166 amino acid. The Quantitative Real-time PCR (qRT-PCR) analysis results showed that PmTβ4 was ubiquitously expressed in all the tested shrimp tissues, with the highest expression level was detected in the hemolymph, while the lowest expression level in the muscle. The expression level of PmTβ4 was significantly up-regulated in hepatopancreas after challenged by Vibrio parahaemolyticus, Vibrio harveyi and Staphylococcus aureus. In vitro antimicrobial test showed that the recombinant protein of PmTβ4 (rPmTβ4) had broad-spectrum of antimicrobial activity, which could inhibit both the growth of gram-negative bacteria and gram-positive bacteria, including Vibrio vulnificus, V. parahaemolyticus, Streptococcus agalactiae, S. aureus and Aeromonas hydrophila. Moreover, rPmTβ4 had a certain binding ability to different bacteria, and this binding ability exhibits a strong dose-dependent effect. In vivo, PmTβ4 could facilitate external bacterial clearance in shrimp, and have beneficial to shrimp survival post V. parahaemolyticus infection. Furthermore, wound-healing assay was carried out to study the role of PmTβ4 in the process of wound healing. The results showed that the PmTβ4 expression was significantly up-regulated by injury treatment, and exerted positive effects to promote wound healing. In addition, PmTβ4 can significantly increase the expression level of superoxide dismutase (SOD) and Catalase (CAT) after injury treatment in shrimp, which would involve in scavenging reactive oxygen species (ROS) caused by the wound. In conclusion, these results indicated that PmTβ4 may play important roles in antibacterial immunity and wound healing in Penaeus monodon.

The SARS-CoV-2 virus, causing the devastating COVID-19 pandemic, has been reported to affect platelets and cause increased thrombotic events, hinting at the possible bidirectional interactions between platelets and the virus. In this review, we discuss the potential mechanisms underlying the increased thrombotic events as well as altered platelet count and activity in COVID-19. Inspired by existing knowledge on platelet-pathogen interactions, we propose several potential antiviral strategies that platelets might undertake to combat SARS-CoV-2, including their abilities to internalize the virus, release bioactive molecules to interfere with viral infection, and modulate the functions of immune cells. Moreover, we discuss current and potential platelet-targeted therapeutic strategies in controlling COVID-19, including antiplatelet drugs, anticoagulants, and inflammation-targeting treatments. These strategies have shown promise in clinical settings to alleviate the severity of thrombo-inflammatory complications and reduce the mortality rate among COVID-19 patients. In conclusion, an in-depth understanding of platelet-SARS-CoV-2 interactions may uncover novel mechanisms underlying severe COVID-19 complications and could provide new therapeutic avenues for managing this disease.

Antimicrobial peptides (AMPs) are essential elements of thehost defense system. Characterized by heterogenous structures and broad-spectrumaction, they are promising candidates for combating multidrug resistance. Thecombined use of AMPs with other antimicrobial agents provides a new arsenal ofdrugs with synergistic action, thereby overcoming the drawback of monotherapiesduring infections. AMPs kill microbes via pore formation, thus inhibitingintracellular functions. This mechanism of action by AMPs is an advantage overantibiotics as it hinders the development of drug resistance. The synergisticeffect of AMPs will allow the repurposing of conventional antimicrobials andenhance their clinical outcomes, reduce toxicity, and, most significantly,prevent the development of resistance. In this review, various synergies ofAMPs with antimicrobials and miscellaneous agents are discussed. The effect ofstructural diversity and chemical modification on AMP properties is firstaddressed and then different combinations that can lead to synergistic action,whether this combination is between AMPs and antimicrobials, or AMPs andmiscellaneous compounds, are attended. This review can serve as guidance whenredesigning and repurposing the use of AMPs in combination with other antimicrobialagents for enhanced clinical outcomes.

Platelets, the smallest cells in human blood, known for their role in primary hemostasis, are also able to interact with pathogens and play a crucial role in the immune response. In severe coronavirus disease 2019 (COVID-19) cases, platelets become overactivated, resulting in the release of granules, exacerbating inflammation and contributing to the cytokine storm. This study aims to further elucidate the role of platelets in COVID-19 progression and to identify predictive biomarkers for disease outcomes. A comparative proteome analysis of highly purified platelets from critically diseased COVID-19 patients with different outcomes (survivors and non-survivors) and age- and sex-matched controls was performed. Platelets from critically diseased COVID-19 patients exhibited significant changes in the levels of proteins associated with protein folding. In addition, a number of proteins with isomerase activity were found to be more highly abundant in patient samples, apparently exerting an influence on platelet activity via the non-genomic properties of the glucocorticoid receptor (GR) and the nuclear factor κ-light-chain-enhancer of activated B cells (NFκB). Moreover, carbonic anhydrase 1 (CA-1) was found to be a candidate biomarker in platelets, showing a significant increase in COVID-19 patients.

The complement receptor CR3, also known as integrin Mac-1 (CD11b/CD18), is one of the major phagocytic receptors on the surface of neutrophils and macrophages. We previously demonstrated that in its protein ligands, Mac-1 binds sequences enriched in basic and hydrophobic residues and strongly disfavors negatively charged sequences. The avoidance by Mac-1 of negatively charged surfaces suggests that the bacterial wall and bacterial capsule possessing net negative electrostatic charge may repel Mac-1 and that the cationic Mac-1 ligands can overcome this evasion by acting as opsonins. Indeed, we previously showed that opsonization of Gram-negative Escherichia coli with several cationic peptides, including PF4 (Platelet Factor 4), strongly augmented phagocytosis by macrophages. Here, we investigated the effect of recombinant PF4 (rPF4) on phagocytosis of Gram-positive Staphylococcus aureus in vitro and examined its impact in a mouse model of S. aureus peritonitis. Characterization of the interaction of rPF4 with nonencapsulated and encapsulated S. aureus showed that rPF4 localizes on the bacterial surface, thus making it available for Mac-1. Furthermore, rPF4 did not have direct bactericidal and bacteriostatic activity and was not toxic to host cells. rPF4 enhanced phagocytosis of S. aureus bioparticles by various primary and cultured Mac-1-expressing leukocytes by several folds. It also increased phagocytosis of live nonencapsulated and encapsulated bacteria. Notably, the augmentation of phagocytosis by rPF4 did not compromise the intracellular killing of S. aureus by macrophages. Using a murine S. aureus peritonitis model, we showed that treatment of infected mice with rPF4 caused a significant increase in the clearance of antibiotic-susceptible S. aureus and its methicillin-resistant (MRSA) variant and markedly improved survival. These findings indicate that rPF4 binding to the bacterial surface circumvents its antiphagocytic properties, improving host defense against antibiotic-susceptible and antibiotic-resistant bacteria.

The aim of the present study was to compare the barrier function during bacterial invasion and wound healing properties of 3 commonly used membranes including horizontal platelet-rich fibrin (H-PRF) against two commercially available resorbable collagen membranes.

H-PRF membranes were prepared by collecting venous blood from 3 healthy volunteers using a 700 g for 8-min centrifugation protocol followed by compression into membranes. To evaluate their barrier function, 3 groups (H-PRF membrane, collagen membrane A (Bio-Gide, Geistlich), collagen membrane B (Megreen, Shanxi Ruisheng Biotechnology Co) were placed between an inner chamber and outer chamber and inoculated with S. aureus. At 2 h, 24 h, and 48 h post-inoculation, cultures from the inner and outer chambers were assessed for bacterial CFUs. Then, scanning electron microscope (SEM) was utilized to visualized the morphological destruction by bacteria of the inner and outer surfaces of the membranes. To assess the wound healing properties of each membrane, leachates from each group were applied to human gingival fibroblasts (HGF) and a scratch assay was performed at 24 h and 48 h.

S. aureus showed a minimal bacterial attachment or invasion rate through either collagen membranes at 2 h post-inoculation, yet over time demonstrated rapid degradation, especially on the rougher surface. While PRF demonstrated higher number of CFUs after 2 h, no significant penetration/degradation of the H-PRF membranes was observed at 24 h and 48 h in the H-PRF group. Both collagen membranes demonstrated significant morphological changes 48 h post-bacterial innoculation, while minimal obvious morphological changes were observed in the H-PRF group. The wound healing assay also demonstrated significantly better wound closure rates in the H-PRF group.

H-PRF membranes exhibited better barrier function towards S. aureus over 2 days of innoculation and better wound healing ability when compared to two commercially available collagen membranes.

This study provides further evidence for the application of H-PRF membranes during guided bone regeneration by minimizing bacterial invasion. Furthermore, H-PRF membranes have significantly better ability to promote wound healing.

Angiogenesis is the formation of new blood vessel from existing vessels and is a critical first step in tissue repair following chronic disturbances in healing and degenerative tissues. Chronic pathoanatomic tissues are characterized by a high number of inflammatory cells; an overexpression of inflammatory mediators; such as tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1); the presence of mast cells, T cells, reactive oxygen species, and matrix metalloproteinases; and a decreased angiogenic capacity. Multiple studies have demonstrated that autologous orthobiological cellular preparations (e.g., platelet-rich plasma (PRP)) improve tissue repair and regenerate tissues. There are many PRP devices on the market. Unfortunately, they differ greatly in platelet numbers, cellular composition, and bioformulation. PRP is a platelet concentrate consisting of a high concentration of platelets, with or without certain leukocytes, platelet-derived growth factors (PGFs), cytokines, molecules, and signaling cells. Several PRP products have immunomodulatory capacities that can influence resident cells in a diseased microenvironment, inducing tissue repair or regeneration. Generally, PRP is a blood-derived product, regardless of its platelet number and bioformulation, and the literature indicates both positive and negative patient treatment outcomes. Strangely, the literature does not designate specific PRP preparation qualifications that can potentially contribute to tissue repair. Moreover, the literature scarcely addresses the impact of platelets and leukocytes in PRP on (neo)angiogenesis, other than a general one-size-fits-all statement that "PRP has angiogenic capabilities". Here, we review the cellular composition of all PRP constituents, including leukocytes, and describe the importance of platelet dosing and bioformulation strategies in orthobiological applications to initiate angiogenic pathways that re-establish microvasculature networks, facilitating the supply of oxygen and nutrients to impaired tissues.

Platelets play a crucial role in hemostasis and the immune response, mainly by recognizing signals associated with vascular damage. However, it has recently been discovered that the antimicrobial peptide LL-37 activates platelets in functions related to thrombus formation and inflammation. Therefore, this work aims to evaluate the effect of LL-37 on the activation of antimicrobial functions of human platelets. Our results show that platelets treated with LL-37 increase the surface expression of receptors (Toll-like receptors (TLRs) 2 and -4, CD32, CD206, Dectin-1, CD35, LOX-1, CD41, CD62P, and αIIbβ3 integrins) for the recognition of microorganisms, and molecules related to antigen presentation to T lymphocytes (CD80, CD86, and HLA-ABC) secrete the antimicrobial molecules: bactericidal/permeability-increasing protein (BPI), azurocidin, human neutrophil peptide (HNP) -1, and myeloperoxidase. They also translate azurocidin, and have enhanced binding to Escherichia coli, Staphylococcus aureus, and Candida albicans. Furthermore, the supernatant of LL-37-treated platelets can inhibit E. coli growth, or platelets can employ their LL-37 to inhibit microbial growth. In conclusion, these findings demonstrate that LL-37 participates in the antimicrobial function of human platelets.

Exploring innovative techniques and treatments to improve spinal fusion procedures is a global challenge. Here, we provide a scientific opinion on the ability of a vertebral bone marrow (vBM) clot to provide a local combined delivery system not only of stem cells, signaling biomolecules and anti-inflammatory factors but also of molecules and proteins endowed with antimicrobial properties. This opinion is based on the evaluation of the intrinsic basic properties of the vBM, that contains mesenchymal stem cells (MSCs), and on the coagulation process that led to the conversion of fibrinogen into fibrin fibers that enmesh cells, plasma but above all platelets, to form the clot. We emphasize that vBM clot, being a powerful source of MSCs and platelets, would allow the release of antimicrobial proteins and molecules, mainly cathelicidin LL- 37, hepcidin, kinocidins and cationic host defense peptides, that are per se gifted with direct and/or indirect antimicrobial effects. We additionally highlight that further studies are needed to deepen this knowledge and to propose vBM clot as multifunctional bioscaffold able to target all the main key challenges for spinal fusion surgery.

In recent decades, the potential of PRF has been extensively studied. The number of studies about PRF has increased three times since the year 2012, but the full spectrum of its fundamental properties, such as antimicrobial and anti-inflammatory activity, is not clearly described. In oral and maxillofacial surgery, PRF is described in alveolar ridge preservation, orthognathic surgery, cleft lip and palate surgery, maxillary sinus augmentation, and dental implant placement as demonstrating favorable results and its clinical advantages. The structural complexity, inhomogeneous nature, and clotting ability of PRF make its antimicrobial effect evaluation complicated. Nevertheless, most of the used antimicrobial testing methods are based on antibacterial agent diffusion ability in culture media. Because the oral and maxillofacial region is the most frequent area of PRF application, its antimicrobial activity evaluation also prevails in the oral microbiome. PRF's biological potential is highly dependent on the specific preparation protocol and methodology used; it should be carefully prepared and kept under proper conditions to keep cellular content alive. PRF's influence on living cells demonstrates a stimulating effect on bone regeneration, and an angiogenetic effect, and it provides anti-inflammatory activity. According to analyzed studies, PRF demonstrated success in oral and maxillofacial surgery in various methods of application. Antibacterial and anti-inflammatory properties were proven by antibacterial activity against different bacterial species, sustained growth factor, sustained release, and cell activity on the material application. Accurately and correctly prepared PRF can ensure antibacterial and anti-inflammatory properties, and it can be a beneficial clinical tool in oral and maxillofacial surgery.