Triangular silver nanoplates (TSNPs) exhibit unique optical and antimicrobial properties due to their shape, sharp edges, and vertices. In this study, TSNPs were incorporated into biopolymer blends (bacterial cellulose (BC) with polylactic acid (PLA), polycaprolactone (PCL), and polyhydroxybutyrate (PHB)). Antimicrobial activity of materials was tested against Escherichia coli ATCC 95922 and Staphylococcus aureus ATCC 25923 (10⁶ CFU/mL). After incubation (24 h at 37 °C, 100 rpm), optical density was measured at 630 nm. In order to assess biosensing applications, specifically fibronectin (Fn) behavior, TSNPs were protected with gold (AuTSNP) and analyzed via sucrose sensitivity test and monitored by localized surface plasmon resonance (LSPR). Additionally, AuTSNPs were coated with polyethylene glycol (PEGAuTSNP). Fibronectin functionalization of PEGAuTSNPs and pH-conformation was monitored (FnPEGAuTSNP). Eventually, adequate Fn and anti-Fn antibody concentrations were determined. BC/PHB/TSNPs showed antimicrobial activity against E. coli and S. aureus with 80 and 95% of growth inhibition, respectively. The sucrose sensitivity test indicated that the LSPR_λmax of the spectra is directly proportional to the sucrose concentration. LSPR_λmax of Fn-PEGAuTSNPs at pH 7 and pH 4 were measured at 633 and 643 nm, respectively. A total of 5 µg of Fn was determined to be adequate concentration, while 0.212 mg/mL of anti-Fn antibody indicatied system saturation.

Skin wound healing is coordinated by a delicate balance between proinflammatory and anti-inflammatory responses, which can be affected by opportunistic pathogens and metabolic or vascular diseases. Several antimicrobial peptides (AMPs) possess immunomodulatory properties, suggesting their potential to support skin wound healing. Here, we evaluated the proregenerative activity of three recently described AMPs (Clavanin A, Clavanin-MO, and Mastoparan-MO). Human primary dermal fibroblasts (hFibs) were used to determine peptide toxicity and their capacity to induce cell proliferation and migration. Furthermore, mRNA analysis was used to investigate the modulation of genes associated with skin regeneration. Subsequently, the regenerative potential of the peptides was further confirmed using an ex vivo organotypic model of human skin (hOSEC)-based lesion. Our results indicate that the three molecules evaluated in this study have regenerative potential at nontoxic doses (i.e., 200 μM for Clavanin-A and Clavanin-MO, and 6.25 μM for Mastoparan-MO). At these concentrations, all peptides promoted the proliferation and migration of hFibs during in vitro assays. Such processes were accompanied by gene expression signatures related to skin regenerative processes, including significantly higher KI67, HAS2 and CXCR4 mRNA levels induced by Clavanin A and Mastoparan-MO. Such findings translated into significantly accelerated wound healing promoted by both Clavanin A and Mastoparan-MO in hOSEC-based lesions. Overall, the data demonstrate the proregenerative properties of these peptides using human experimental skin models, with Mastoparan-MO and Clavanin A showing much greater potential for inducing wound healing compared to Clavanin-MO.

In skin lesions, the development of microbial infection affects the healing process, increasing morbidity and mortality rates in patients with severe burns, diabetic foot, and other types of skin injuries. Synoeca-MP is an antimicrobial peptide (AMP) that exhibits activity against several bacteria of clinical importance, but its cytotoxicity can represent a problem for its positioning as an effective antimicrobial compound. In contrast, the immunomodulatory peptide IDR-1018 presents low toxicity and a wide regenerative potential due to its ability to reduce apoptotic mRNA expression and promote skin cell proliferation. In the present study, we used human skin cells and a 3D skin equivalent models to analyze the potential of the IDR-1018 peptide to attenuate the cytotoxicity of synoeca-MP, as well as the influence of synoeca-MP/IDR-1018 combination on cell proliferation, regenerative processes, and wound repair. We found that the addition of IDR-1018 significantly improved the biological properties of synoeca-MP on skin cells without modifying its antibacterial activity against S. aureus. Likewise, in both melanocytes and keratinocytes, the treatment with synoeca-MP/IDR-1018 combination induces cell proliferation and migration, while in a 3D human skin equivalent model, it can accelerate wound reepithelization. Furthermore, treatment with this peptide combination generates an up-regulation in the expression of pro-regenerative genes in both monolayer cell cultures and in 3D skin equivalents. This data suggests that the synoeca-MP/IDR-1018 combination possesses a good profile of antimicrobial and pro-regenerative activity, opening the door to the development of new strategies for the treatment of skin lesions.

In the cellular environment, high noise levels, such as fluctuations in biochemical reactions, protein variability, molecular diffusion, cell-to-cell contact, and pH, can both mediate and interfere with cellular functions. In this work, gold edge-coated triangular silver nanoparticles (AuTSNP) were validated as a promising new tool to indicate protein conformational transitions in cultured cells and to monitor essential protein activity in the presence of an optimized bone biomimetic chitosan-based scaffold whose rational design mimics the ECM as a natural scaffold. A chitosan-based scaffold formulation with hydroxyapatite (CS/HAp) was selected due to its promising features for orthopedic applications, including combined high mechanical strength biocompatibility and biodegradability. Functionalized AuTSNP-based tests with the model ECM protein, fibronectin (Fn), illustrate that the protein interactions can be clearly sensed over time through the local surface plasmon resonance (LSPR) technique. This demonstrates that AuTNSP are a powerful tool to detect protein conformational activity in the presence of biomimetic bone tissue regeneration scaffolds within a cellular environment that comprises a diversity of molecular cues.

Extrachromosomal circular DNAs (eccDNAs) are heterogeneous circular DNA molecules derived from genomic DNA, and believed to be involved in intercellular communication and in natural biological processes. Extracellular vesicles (EVs) are membrane-bound particles released from all cells, and have been shown to contain various classes of nucleic acids. EVs can play a role in intercellular communication and may be used as biomarkers. This constitutes the first study to demonstrate that EVs derived from healthy human dermal fibroblasts carry eccDNA. eccDNA from EVs and their corresponding donor cells were isolated and sequenced on the Oxford Nanopore MinIon platform, followed by the identification of potential eccDNAs through four different bioinformatic pipelines, namely ecc_Finder, cyrcular-calling, CReSIL, and Flec. Our main findings demonstrate that EVs derived from human dermal fibroblasts carry eccDNA; there is variability in the number of eccDNAs identified in the same sample through different pipelines; and there is variability in the identified eccDNAs across biological replicates. Additionally, eccDNAs characterized in this research had (a) sequences as small as 306 base pairs and as large as 28,958 base pairs across all samples, (b) uneven chromosomal distribution, and (c) an average of 49.7% of the identified eccDNAs harboring gene fragments. Future implications for this novel research include using this framework method to elucidate factors and conditions that may influence the skin aging process and related biogenesis in human dermal cells.

Inflammation contributes to the onset and exacerbation of numerous age-related diseases, often manifesting as a chronic condition during aging. Given that cellular senescence fosters local and systemic inflammation, senotherapeutic interventions could potentially aid in managing or even reducing inflammation. Here, we investigated the immunomodulatory effects of the senotherapeutic Peptide 14 (Pep 14) in human peripheral blood mononuclear cells (PBMCs), monocytes, and macrophages. We found that, despite failing to significantly influence T cell activation and proliferation, the peptide promoted a Th2/Treg gene expression and cytokine signature in PBMCs, characterized by increased expression of the transcription factors GATA3 and FOXP3, as well as the cytokines IL-4 and IL-10. These observations were partially confirmed through ELISA, in which we observed increased IL-10 release by resting and PHA-stimulated PBMCs. In monocytes from the U-937 cell line, Pep 14 induced apoptosis in lipopolysaccharide (LPS)-stimulated cells and upregulated IL-10 expression. Furthermore, Pep 14 prevented LPS-induced activation and promoted an M2-like polarization in U-937-derived macrophages, evidenced by decreased expression of M1 markers and increased expression of M2 markers. We also showed that the conditioned media from Pep 14-treated macrophages enhanced fibroblast migration, indicative of a functional M2 phenotype. Taken together, our findings suggest that Pep 14 modulates immune cell function towards an anti-inflammatory and regenerative phenotype, highlighting its potential as a therapeutic intervention to alleviate immunosenescence-associated dysregulation.