Beata Drzewiecka

Excessive inflammation following implant insertion may impair early healing and predispose to postoperative complications. Since neutrophils constitute the predominant leukocyte population during the initial inflammatory phase, and monocytes subsequently orchestrate tissue resolution and repair, understanding their direct response to biomaterials and bioactive compounds is essential. In the present study, we evaluated the effect of purified bioactive extract combined with biomaterial on neutrophil and monocyte activity in vitro, comparing their modulatory potential with that of non-purified extract. Porcine neutrophils and monocytes were cultured in contact with hydroxyapatite (HA) mixed with purified fractions obtained through sequential chromatographic isolation. Cellular responses were assessed by measuring enzyme release and reactive oxygen/nitrogen species (ROS/RNS) generation. Neutrophils exposed to HA alone displayed a pronounced oxidative response, confirming strong activation. Purified extracts, however, significantly reduced ROS/RNS production (NO: 2.71 ± 0.07 µM vs. HA-only group 5.72 ± 0.10 µM, p < 0.05), whereas non-purified extracts elicited only a partial reduction. Enzyme release followed the same trend, with purified fractions demonstrating a markedly higher inhibitory effect on neutrophil degranulation. Similarly, monocytes exposed to HA exhibited prolonged oxidative stress at 24 h and 48 h, which was markedly downregulated by the purified extract (HA + EX), demonstrating significant reductions in both nitric oxide (NO) and superoxide generation. These findings indicate that purification enhances the bioactivity and specificity of the tested extracts, improving their capacity to modulate excessive neutrophil and monocyte activation. To address this challenge, this study introduces a novel, advanced purification method designed to isolate and refine AMPNE-type particles from the crude extract. Furthermore, we investigated whether the purified particles retain their structural and functional integrity post-purification. By comparing the biological efficacy of the extract before and after purification, we evaluated whether the cleaning process maintains, enhances, or modifies their original bioactive properties, providing crucial insights into the viability of the proposed technology. Our study highlights the potential of purified bioactive molecules to more effectively regulate early inflammatory responses to biomaterials. These results provide a foundation for further research into targeted immunomodulatory strategies aimed at improving implant integration and reducing inflammatory complications.

Ozone (O₃) is a reactive oxidant increasingly applied in biomedical settings, yet its dose-dependent effects on innate immune cells, particularly those from non-human species, remain insufficiently defined. Within a One Health framework, this study examined how two clinically relevant O₃ exposure regimens (30 µg/mL and 90 µg/mL) affect porcine neutrophils and monocytes isolated from peripheral blood. Cell viability, reactive oxygen and nitrogen species (RONS) production, and the activity of key enzymes (myeloperoxidase, elastase, alkaline phosphatase, arginase) were assessed at 1 h and 24 h post-exposure. The lower dose induced mild functional activation without compromising viability, whereas the higher dose triggered pronounced oxidative stress, enhanced degranulation, and reduced neutrophil viability by more than 60%. Neutrophils exhibited a stronger and more dynamic response than monocytes, which retained viability and differentiation capacity at 30 µg/mL but showed impaired function at 90 µg/mL. These findings highlight the dual nature of O₃, where controlled exposure may support immunomodulation, while excessive dosing disrupts cell function. Defining safe and effective therapeutic windows remains critical for future applications.

Due to an aging population and the prevalence of illnesses associated with modern lifestyles, mushrooms, well known for their nutritional value and health-promoting properties, are becoming an increasingly important part of the diet. They are consumed in various forms, including food, nutraceuticals, and dietary supplements. A relatively new trend involves incorporating mushrooms or their components as additives and supplements to enhance the quality and functionality of traditional food products. The processing and preservation methods of fresh mushrooms can significantly impact the activity of resulting powders, extracts, or other functional forms used in food additives, supplements, and fortified foods. The functional benefits of mushrooms are frequently attributed to their antioxidant and anti-inflammatory properties. However, to date, the literature lacks comprehensive reviews that consolidate existing knowledge on mushroom-based food additives and products enriched with them. Therefore, this review aims to compile and methodically analyze the existing data in this field, identify existing knowledge gaps, and outline future perspectives for the development and application of such products. Special attention is given to food supplementation with microencapsulated additives, which represent a promising form of functional powders. All these aspects are evaluated in terms of their antioxidant and anti-inflammatory properties. Finally, future perspectives on improving the health benefits of food through mushroom-based additives are discussed.

For centuries, humans have used mushrooms as both food and pro-health supplements. Mushrooms, especially those related to the functions of the human immune system, are rich in dietary fiber, minerals, essential amino acids, and various bioactive compounds and have significant health-promoting properties. Immunoregulatory compounds in mushrooms include lectins, terpenes, terpenoids, polysaccharides, and fungal immunomodulatory proteins (FIPs). The distribution of these compounds varies from one species of mushroom to another, and their immunomodulatory activities depend on the core structures and chemical modifications in the composition of the fractions. In this review, we describe active compounds from medical mushrooms. We summarize potential mechanisms for their in vitro and in vivo activities and detail approaches used in developing and applying bioactive compounds from mushrooms. Finally, we discuss applications of fungal peptides and highlight areas that require improvement before the widespread use of those compounds as therapeutic agents and explore the status of clinical studies on the immunomodulatory activities of mushrooms and their products, as well as the prospect of clinical application of AMPs as ‘drug-like’ compounds with great potential for treatment of non-healing chronic wounds and multiresistant infections.

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

Implant insertion can evoke excessive inflammation which disrupts the healing process and potentially leads to complications such as implant rejection. Neutrophils and macrophages play a vital role in the early inflammatory phase of tissue repair, necessitating the study of cellular responses in host–implant interactions. In order to deepen the knowledge about these interactions, the response of neutrophils and macrophages to contact with selected biomaterials was examined in vitro on the basis of secretory response as well as reactive oxygen species/reactive nitrogen species (ROS/RNS) generation. Porcine neutrophils exposed to hydroxyapatite (HA) released more enzymes and generated higher levels of ROS/RNS compared to the control group. The addition of AMPNE diminished these responses. Although the results from porcine cells can provide valuable preliminary data, further validation using human cells or clinical studies would be necessary to fully extrapolate the findings to human medicine. Our study revealed that human neutrophils after contact of with HA increased the production of nitric oxide (NO) (10.00 ± 0.08 vs. control group 3.0 ± 0.11 µM, p < 0.05), while HAP or FAP did not elicit a significant response. Human macrophages cultured with HA produced more superoxide and NO, while HAP or FAP had a minimal effect, and curdlan reduced ROS/RNS generation. The addition of AMPNE to cultures with all biomaterials, except curdlan, reduced neutrophil activity, regardless of the peptides’ origin. These results highlight the potential of antimicrobial peptides in modulating excessive biomaterial/host cell reactions involving neutrophils and macrophages, enhancing our understanding of immune reactions, and suggesting that AMPNE could regulate leukocyte response during implantation.

Osteoarthritis (OA) is a chronic, progressive, multifactorial disease resulting in a progressive loss of articular cartilage structure and function that is most common in middle-aged and older patients. OA is involved in the loss of extracellular matrix and cartilage as well as cell number decreases within the matrix, especially in the further stages of the disease. The immune system plays a pivotal role in the pathomechanism of this condition. Both humoral and cellular mediators contribute to cartilage destruction, abnormal bone remodeling, synovitis, and joint effusion. The increasing prevalence of this disease has led to a growing interest in using animal models as the primary way to broaden the knowledge of the pathogenesis of OA and possible therapies at each stage of disease development. This review aims to describe the signs, pathogenesis, and classification of OA as well as discuss the advantages and disadvantages of some animal models. The currently used treatment methods include mesenchymal stem cells, exosomes, gene therapies, and blood-derived products. In addition, exogenous growth factors, platelet-rich plasma (PRP), platelet lysate, and autologous conditioned serum (ACS) are discussed with the application of tissue engineering techniques and biomaterials.