Fibroblasts Research Articles

In vitro anticancer efficacy of Calendula Officinalis extract-loaded chitosan nanoparticles against gastric and colon cancer cells

Objective This study assessed the anticancer activities of Calendula officinalis-loaded chitosan nanoparticles in gastric and colon cancer cells compared with fibroblast cells and examined the balance between ROS and antioxidants. Methods Considering this information, we synthesized Calendula officinalis-loaded chitosan nanoparticles (CO-CSNPs) via the ionic gelation method. Their characterizations were carried out with ZetaSizer, UV-Vis, FTIR and SEM devices including size, morphology and surface zeta potential analysis, loading capacity, encapsulation efficiency, in vitro drug release, and chemical interactions. The anticancer activities of CO, CSNPs, and CO-CSNPs were tested against AGS, Caco-2, and normal NIH-3T3 cells using an XTT assay. The anticancer effects were evaluated using DAPI staining, scratch assay, reactive oxygen species (ROS) detection and the CUPRAC method on cellular and non-cellular processes that promote anticancer mechanisms. Results Results showed that CO and CO-CNPs exhibited anticancer activity against AGS and Caco-2. Further, the formulation of CO with CSNPs enhanced the anticancer activity of CO while having no cytotoxicity on NIH-3T3. DAPI staining, scratch assay, ROS, and CUPRAC method confirmed the anticancer activity of CO and CO-CSNPs, which resulted in a reduction in the number of apoptotic cells, inhibited migration, triggered apoptotic pathway via ROS, and higher antioxidant activity. Conclusions The results of the study indicate that CO-CSNPs are a promising therapeutic formulation for gastric and colon cancer treatment. We consider that this study will lead to the investigation of molecular mechanisms of CO-CSNPs in cancer treatment and their investigation in clinical studies.

The Impact of Wharton's Jelly-derived Exosomes on the Production of Inflammatory Mediators from HIG-82 Synoviocytes.

Osteoarthritis (OA) is the most common joint disease worldwide. Routine treatment options are limited, and total knee replacement surgeries often come with complications. In recent years, the use of biologics, such as Wharton's jelly (Wj) derived from the umbilical cord (UC), has gained popularity. While mesenchymal stem cells (MSCs) derived from Wj show promise in restoring articular cartilage, they also have some limitations. Recent studies have indicated that exosomes isolated from acellular Wj may offer advantages under certain conditions. To investigate the anti-inflammatory properties of exosomes isolated from Wj in synoviocytes. Decellularization of Wj was performed using sterile umbilical cords obtained from patients. Next, the exosomes were isolated from Wj using ultracentrifugation. After characterizing the exosomes, they were co-cultured with inflammatory synovial fibroblast cells (HIG-82) for 24 hours. Then, the gene expression levels and protein contents of some important inflammatory mediators including metalloproteinase-13 (MMP-13), cyclooxygenase-2 (COX2) and inducible nitric oxide synthase (iNOS) were measured in the cells using real-time PCR and ELISA tests, respectively. The expression levels of MMP-13, COX-2, and iNOS genes were significantly reduced in the cultured cells treated with exosomes compared to untreated cells. Moreover, the content of MMP-13, COX-2, and iNOS proteins were significantly lower in the supernatant of the cultured cells compared to the control. Wj-derived exosomes exhibit notable anti-inflammatory properties, which can help mitigate inflammation in the synovial environment of joints. However, further research is required to fully understand their benefits and potential applications in treating osteoarthritis.

Synthesis, cytotoxic evaluation, and in silico studies of novel benzenesulfonamide-thiazolidinone derivatives against colorectal carcinoma

Thirteen new benzenesulfonamide derivatives containing 4-thiazolidinone were synthesized. Their cytotoxic properties were tested on colorectal carcinoma (HT-29, DLD-1, COLO-205) and normal colon fibroblast (CCD-986Sk) cell lines. Compounds 3l (IC50=114.62 µM), 3a (IC50=25.82 µM), 3k (IC50=30.99 µM), and 3i (IC50=62.94 µM) showed the highest cytotoxicity on HT-29, DLD-1, COLO-205, and CCD-986Sk cell lines, respectively. Compound 3e (IC50=1075.4 µM) exhibited the least cytotoxicity against CCD-986Sk. The apoptosis profile of 5-FU at 5 µM was similar to that of compound 3e at 30 µM. Molecular docking and MD simulations showed stable interactions for compounds 3i and 3e. Molecular docking and MD simulations revealed that compounds 3i and 3e exhibit stable interactions with key cancer-related protein targets, particularly TGFβ2. These interactions suggest their potential as therapeutic agents. The predicted ADME parameters further highlighted compounds 3e and 3i for their favorable lipophilicity, solubility, permeability, and oral absorption profiles, supporting their promise as candidates for future drug development.

Jellein-I-conjugated gold nanoparticles: Insights into the antibacterial, antibiofilm activities against MRSA, and anticancer properties

Antimicrobial peptides (AMPs) and their conjugation with nanoparticles hold promise for targeting Methicillin-resistant Staphylococcus aureus (MRSA). In this study, Jellein-I peptide, with a cysteine at the C-terminus, was conjugated to gold nanoparticles (GNPs) to enhance its antibacterial, antibiofilm, and anticancer activities. The GNPs were synthesized and then conjugated to Jellein-I, forming Jellein-I-Cys-GNPs. The nanoparticles were characterized using UV–visible spectroscopy, transmission electron microscopy (TEM), dynamic light scattering (DLS), and Fourier transform infrared (FTIR) spectroscopy. UV–visible spectroscopy, DLS, and TEM confirmed the formation of GNPs with an average size of 5.5 nm. The UV–visible spectroscopy and FTIR results indicated the presence of peptide functional groups on the surface of the GNPs. To assess antibacterial activity, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays were performed against MRSA strains. The conjugated Jellein-I exhibited lower MIC and MBC values (94 μM) compared to the free peptide (512 μM) against both clinical and standard MRSA strains. Additionally, GNP-conjugated Jellein-I demonstrated antibiofilm activity, inhibiting and removing biofilms while reducing metabolic activity by more than 82 %, 44 %, and 72 %, respectively. Cell viability studies on rat dermal fibroblast (RDF) cells showed over 80 % viability across all treatment concentrations. Furthermore, a clonogenic assay on the metastatic melanoma cell line (A375) demonstrated that GNP-conjugated Jellein-I had significantly better anticancer effects than the free peptide, even at a concentration as low as 9 μM. Our findings suggest that the conjugation of Jellein-I-Cys to GNPs could be a promising formulation for enhancing the antibacterial, antibiofilm, and anticancer activities of peptides. Further in vivo studies may establish this new nanoformulation as a potential therapeutic candidate for MRSA infections and cancer treatment.

The influence of blackcurrant extract–based cosmetic antimicrobial agent on skin cells and skin model membranes

The components of blackcurrant extract are known for their antimicrobial properties and the lipid membrane is the important target in terms of the activity of these compounds. In this work, a commercially available product that is PhytoCide Black Currant Powder (BCE), which can be used as a soothing, conditioning and antimicrobial agent in cosmetics, was investigated. The aim of this study was to explore its effect on skin cells and gain insight into the mechanism of its selectivity. The influence of this formulation on keratinocyte and fibroblast cells in in vitro tests was studied and the effect of BCE on model keratinocyte and fibroblast membranes was investigated. As model systems, the lipid monolayers of different compositions were used. The experiments involved the surface pressure/area measurement, penetration studies and Brewster angle microscopy experiments. The BCE formulation was shown to be non-toxic to the skin cells at the concentrations applied. Model membrane experiments confirmed that BCE components do not incorporate into keratinocyte and fibroblast membranes at membrane-related conditions. However, they alter morphology of the studied systems by acting in the region of polar head. Significant differences were found in the affinity of the BCE formulation for bacteria compared to skin lipids, consistent with the proven antibacterial effect of the preservative while lacking toxicity to skin cells. Thus, the membrane of the cell may be critical to the selectivity of the chemicals being tested.

Anti-Inflammatory and Wound Healing Properties of Different Honey Varieties from Romania and Correlations to Their Composition.

The complex composition of honey plays a crucial role in wound healing, exhibiting varying effects at different stages of the healing process. This study investigated seven honey varieties sourced from different regions of Romania using in vitro experimental models developed in macrophage-like, fibroblast, and keratinocyte cell lines to explore the mechanisms by which honey promoted the healing process. This study assessed the impact of honey on inflammatory cytokine production in macrophage-like cells, cell proliferation and collagen synthesis in fibroblasts, and cell proliferation and migration in keratinocytes. Additionally, correlation analysis was conducted to examine the relationship between honey composition and its biological properties. Honey varieties presented both anti- and pro-inflammatory effects. Moreover, they displayed dose-dependent pro-proliferative effects, stimulating collagen synthesis and cell migration, thereby enhancing the re-epithelialization process. The Pearson coefficient analysis indicated a strong positive correlation between biological activities and phenolic content. Additionally, there was a medium positive correlation with the ascorbic acid content and a medium negative correlation with the glucose content in the different honey varieties. Romanian honey varieties rich in phenolics showed potential in modulating inflammation, proliferation, collagen synthesis, and cell migration, suggesting their suitability for further evaluation and development of innovative dressings for skin tissue regeneration.

Generation and characterization of giant panda induced pluripotent stem cells

The giant panda ( Ailuropoda melanoleuca ) stands as a flagship and umbrella species, symbolizing global biodiversity. While traditional assisted reproductive technology faces constraints in safeguarding the genetic diversity of giant pandas, induced pluripotent stem cells (iPSCs) known for their capacity to differentiate into diverse cells types, including germ cells, present a transformative potential for conservation of endangered animals. In this study, primary fibroblast cells were isolated from the giant panda, and giant panda iPSCs (GPiPSCs) were generated using a non-integrating episomal vector reprogramming method. Characterization of these GPiPSCs revealed their state of primed pluripotency and demonstrated their potential for differentiation. Furthermore, we innovatively formulated a species-specific chemically defined FACL medium and unraveled the intricate signaling pathway networks responsible for maintaining the pluripotency and fostering cell proliferation of GPiPSCs. This study provides key insights into rare species iPSCs, offering materials for panda characteristics research and laying the groundwork for in vitro giant panda gamete generation, potentially aiding endangered species conservation.

In Vitro Potential of Antioxidant Extracts from Gracilaria gracilis Cultivated in Integrated Multi-Trophic Aquaculture (IMTA) for Marine Biobased Sector

This study aimed to evaluate the antioxidant activity of bioactive compounds extracted from Gracilaria gracilis cultivated in an integrated multi-trophic aquaculture (IMTA) system by different extraction solvents and to investigate the potential capacity of the extracts in cellular systems against environmental pollutants. The global yields, total polyphenol contents, and antioxidant activity were assessed on G. gracilis by DPPH radical scavenging activity, comparing the antioxidant extraction efficiency of the different solvents (ethanol 80%, acetone 70%, N-hexane, and water). Ethanol extract, granted by the highest extractive yield and antioxidant capacity, was tested in vitro in the Sparus aurata fibroblast (SAF-1) cell line to evaluate its protective role against oxidative stress induced by the chemical flame retardant 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47). The results demonstrate that the cells pretreated with G. gracilis extract were protected against oxidative stress and had improved cell viability, cellular antioxidant defense system, and cell cycle control, as demonstrated by the gene expression on some biomarkers related to the cell cycle (p53) and oxidative stress (nrf2, sod, and cat). These results confirm that bioactive compounds obtained from seaweeds cultivated in IMTAs could contribute to producing high-value ingredients that are able to counteract environmental stressors, for the growth of the marine biobased industrial sector and the expansion of new value chains.

Identification of antimicrobial bioactive peptides from the camel milk protein lactoferrin: Molecular docking, molecular dynamic simulation, and in vitro study.

This study explores the potential of antimicrobial peptides (AMPs) derived from camel milk lactoferrin as novel agents against antibiotic-resistant pathogens. Through in silico screening and molecular dynamics simulations, seven lactoferrin-derived peptides were identified as potential AMPs, with peptides P1 and P33 showing promising inhibitory effects against methicillin-resistant Staphylococcus aureus (MRSA). In vitro assays confirmed the efficacy of P33 against pathogenic strains, notably exhibiting activity against multi-drug-resistant strains. Moreover, the lactoferrin-derived peptide P33 demonstrated relatively low cytotoxicity on human fibroblast cells. These findings suggest that P33 holds significant potential as an antimicrobial agent, particularly against antibiotic-resistant bacteria, warranting further in vivo investigations for efficacy and safety. This study highlights the importance of exploring alternative sources of antimicrobial peptides, such as camel milk lactoferrin, in combating the growing threat of antibiotic resistance.

Silver(I) complexes with antifungal drug econazole: Structural characterization and antimicrobial activity study

Two new silver(I) complexes with the antifungal agent econazole (ecz) of the general formula [Ag(еcz)2]X, X = CF3SO3− (Ag1) and PF6− (Ag2), were synthesized and structurally characterized by spectroscopic (1H NMR, IR and UV–Vis) and electrochemical methods. The crystal structure of Ag2 was determined by single crystal X-ray diffraction analysis, confirming an ideal linear geometry of the silver(I) ion in this complex. In addition, by utilising Hirshfeld surface analysis (HSA), it was verified that the crystal structure of Ag2 is stabilized by H⋯H, H⋯Cl, C⋯H, and H⋯F interactions. Density functional theory (DFT) calculations provided additional evidence for formation of the complexes in the solid state and their stability in solution. The coordination of ecz to the silver(I) ion endowed this agent with antibacterial activity against Gram-positive (Pseudomonas aeruginosa and Escherichia coli) and Gram-negative (Staphylococcus aureus and Listeria monocytogenes) bacteria. The most significant antibacterial activity of Ag1 and Ag2 was observed against S. aureus with 56- and 73-fold improvement, respectively, compared to the parent drug. Moreover, a study of the bacterial antibiofilm activity revealed that these complexes were able to prevent approximately 90 % of biofilm formation at their low concentrations. Also, both complexes have shown more pronounced anti-Candida activity than econazole itself, being less toxic on the human normal fibroblast cell line MRC-5. The total amount of ergosterol was reduced in the presence of the subinhibitory concentrations of Ag1 and Ag2 complexes, which was also confirmed by a molecular docking study with two isomers of cytochrome P450 sterol 14α-demethylase, CYP51B and CYP130, as target receptors.

Increased susceptibility of human limbal aniridia fibroblasts to oxidative stress

Aniridia-associated keratopathy originates from a haploinsufficiency of the transcription factor PAX6 (PAX6+/−). In the corneal epithelium of PAX6+/− mice, a significant increase in oxidized proteins was observed, accompanied by impaired compensation for elevated oxidative stress (OS). The extent to which limbal fibroblast cells (LFCs) are affected by an increased susceptibility to OS in cases of congenital aniridia (AN) has not been determined, yet. Our aim was to examine the impact of OS on antioxidant enzyme expression in normal and AN-LFCs. Following isolation and culture of primary LFCs (n = 8) and AN-LFCs (n = 8), cells were treated with cobalt chloride for 48 h to chemically induce hypoxic conditions and OS. Subsequently, HIF-1α/-2α, PHD1/2, Nrf2, CAT, SOD1, PRDX6, and GPX1 gene expression was examined by qPCR. SOD1, PRDX6, and GPX1 protein levels were assessed from the cell lysate by Western blot. The induction of hypoxia led to reduced HIF-1α gene expression in both fibroblast groups (p≤0.008), while the decrease in PHD1 was limited to AN-LFCs (p = 0.0007). On the other hand, under hypoxic conditions, PHD2 showed higher mRNA expression in AN-LFCs compared to normal LFCs (p = 0.013). As a result of OS, the mRNA levels of Nrf2 (p<0.0001) and the antioxidant enzymes CAT (p = 0.005), SOD1 (p = 0.005), GPX1 (p = 0.002) decreased in AN-LFCs. This was accompanied by an increased protein expression of SOD1 (p = 0.019) and PRDX6 (p=0.0009). In the normal LFC group, the induced extent of OS had no impact on the gene (p≥0.151) and protein expression (p ≥ 0.629) of antioxidant enzymes, except for the GPX1 mRNA level (p = 0.027). AN-LFCs exhibit higher susceptibility to OS than normal LFCs. Therefore, in AN-LFCs, there are sustained alterations in gene and protein expression of antioxidative enzymes even after 48 h of CoCl2 treatment.

Design, synthesis, biological evaluation and molecular docking study of thiadiazole-isatin hybrid analogues as potential anti-diabetic and anti-bacterial agents

We have synthesized thiadiazole-isatin hybrid analogues (1–20), characterized through different spectroscopic techniques such as 1HNMR, 13CNMR, HREI-MS, and evaluated against α-glucosidase and α-amylase enzymes. All analogues showed potent inhibitory potentials, having an IC50 values ranged from 22.08 ± 0.09 to 54.03 ± 0.07 μM (against α-amylase) and 19.03 ± 0.04 to 50.03 ± 0.02 μM (α-glucosidase) when compared with the standard drug acarbose (IC50 = 22.07 ± 0.02 μM for α-amylase and IC50 = 18.01 ± 0.06 μM for α-glucosidase respectively). Among the series, analogues 13 (IC50 = 19.03 ± 0.04 and 22.08 ± 0.09 μM), 12 (IC50 = 21.03 ± 0.07 and 24.03 ± 0.07 μM), and 5 (IC50 = 22.02 ± 0.03 and 26.02 ± 0.04 μM) were identified as the most active inhibitors. The structure–activity relationship was carried out, mainly associated with the nature, number, position, and electron-donating and electron-withdrawing effects of the substituent (s) on the phenyl ring. With the help of molecular docking, the binding interaction of the most active analogues within the active site of enzymes was confirmed. Almost twelve compounds were also screened for antibacterial potential, and few were found to be effective against Bacillus subtilis. All compounds were verified for cytotoxicity against the 3 T3 mouse fibroblast cell line and detected non-toxic.

Synthesis and Characterization of Multifunctional Chitosan-Silver Nanoparticles: An In-Vitro Approach for Biomedical Applications.

Antibiotics are successful in promoting health quality by preventing various infectious diseases and minimizing mortality and morbidity all over the world. However, the indiscriminate use of antibiotics has led to the emergence of multi-drug-resistant bacteria, which pose a serious threat to health care sector. Therefore, it is necessary to develop novel antimicrobial agents with versatile characteristics, such as antibacterial activity, low toxicity, wound healing potency, and antioxidant property. In this context, silver chitosan nanoparticles were synthesized in the present study, and their physical characterization revealed that the size of synthesized chitosan-silver nanoparticles was 14-25 nm, with positive surface charge. The functional groups and crystalline nature of the nanoparticles were confirmed by FT-IR and XRD analysis. Further, the silver chitosan nanoparticles showed antibacterial activity against two important clinical pathogens, S. aureus and E. coli. The MTT assay carried out in the present study showed that the synthesized nanoparticles are non-toxic to host cells. A scratch assay on fibroblast cells (L292) demonstrated that the silver chitosan nanoparticles showed promising wound healing activity. A fluorescent DCFH-DA staining assay revealed anantioxidant property of the synthesized nanoparticles. Overall, the study emphasizes the versatile nature of synthesized chitosan-silver nanoparticles, suggesting their great compatibility for biomedical applications.

Medullins A-H, Sesquiterpenes from Perenniporia medulla-panis, and Their Cellular Signaling Mechanism in HDF Cells.

A chemical investigation of an ethyl acetate-soluble layer in the culture broth of Perenniporia medulla-panis resulted in the isolation of eight novel sesquiterpenes conjugated Gly (1), l-Val (2), l-Ala (3), l-Tyr (4), l-Thr (5), l-Ile (6), l-Leu (7), and l-Phe (8). Elucidation of their structures was performed through comprehensive spectroscopic analysis. The absolute configuration of the sesquiterpene skeleton was ascertained using modified Mosher's methods. The configurations of the amino acid units in compounds 2-8 were identified through acid hydrolysis followed by LC-MS analysis employing Marfey's method. Compounds 1-3 and 5-8 showed significant regulating effect on MAP kinase activity (p-ERK and p-JNK) in human diploid fibroblast (HDF) cells.

Anti-Inflammatory Activity and Wound Healing Ability of Coconut Oil Mouthwash on Gingival Fibroblast Cell In Vitro

Coconut oil-pulling therapy is used for maintaining oral health. The procedure has benefits for the prevention of oral disease, including dental caries, oral malodor, bleeding gums. However, virgin coconut oil (VCO) has unsatisfying oily taste. Therefore, coconut oil mouthwash (CoMW) was recently developed. This in vitro study aimed to evaluate the anti-inflammatory activity of coconut oil mouthwash consisting of 60% v/v virgin coconut oil (VCO), 30% v/v propylene glycol (PG), and 10% v/v distilled water on human gingival fibroblast (HGF) cells compared with the activity on murine macrophage (Raw 264.7) cells. The cytotoxicity of CoMW, 0.12% chlorhexidine gluconate (CHX), VCO, and PG was assessed. IC50 concentration of CoMW, CHX, and PG were 1:8 (v/v), 1:32 (v/v), and 1:16 (v/v), respectively. All tested concentrations of VCO had no impact on cell viability. Their anti-inflammatory effects of each IC50 concentration were further studied. Notably, the IC50 concentration of CHX also significantly inhibited nitric oxide production in lipopolysaccharide (LPS)- activated Raw 264.7 cells. Moreover, the IC50 concentrations of CoMW, VCO, PG, and CHX could suppress the interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2) gene expressions in LPS-activated HGF cells, while also enhancing the cell migration of HGF cells as likely to the effect observed with the IC50 concentration of CHX. Wound healing ability of CoMW was also demonstrated after testing with a scratch assay. These findings indicated promising potential for coconut oil mouthwash as an effective agent in reducing inflammation and facilitating wound healing.

Impact of kV-cone beam computed tomography dose on DNA repair mechanisms: A pilot study

PurposeIn head and neck squamous cell carcinoma (HNSCC), early complications of the radiotherapy (RT) are observed from the beginning of the treatment to a few months after its end. During external radiotherapy treatment, several patient-dependent parameters can cause a modification of the dose distribution compared to the planned distribution due to variation in patient positioning, anatomy, or intra-fractional movements for example. To verify these parameters during treatment sessions, one of the most commonly used solutions is the cone-beam computed tomography (CBCT). Nowadays, the use of CBCT may constitutes a significant part of the total dose at the end of treatment (up to 10 cGy per session) and more often the volume irradiated by imaging is larger than the one irradiated by the treatment, leading to unintentional irradiation of nearby organs.In this study, we asked whether the imaging low dose added to a following fraction dose (2Gy) may affect the biological response in terms of DNA repair. Material and methodsUsing an IVInomad dosimeter and scintillating fiber probes specially designed for this exploratory study, we exposed fibroblasts cells from head and neck cancer (HNC) patients to a CBCT dose followed by a radiotherapy fraction dose. DNA double strand breaks and DNA repair were assessed by immunofluorescence using the biomarkers gamma H2AX (γH2AX) and pATM. ResultsThe median dose of CBCT was measured between 17 to 21 mGy per session. The kinetics of both biomarkers were found to be strongly dependent on the individual factor in radiosensitive patients. For HNC patients, a prior CBCT dose applied few minutes before the 2Gy dose may have a sublinear effect on the DNA repair mechanisms and potentially on observed health tissue toxicity. ConclusionThe preliminary results obtained highlight the importance of individual and tissue factors for recognizing and repairing DSB during a treatment by radiotherapy using CBCT.

L-Arginine-Loaded Oxidized Isabgol/Chitosan-Based Biomimetic Composite Scaffold Accelerates Collagen Synthesis, Vascularization, and Re-epithelialization during Wound Healing in Diabetic Rats.

Impaired wound healing in diabetic wounds is common due to infection, inflammation, less collagen synthesis, and vascularization. Diabetic wound healing in patients is still a challenge and needs an ideal wound dressing to treat and manage diabetic wounds. Herein, an efficacious wound dressing biomaterial was fabricated by cross-linking oxidized isabgol (Oisab) and chitosan (Cs) via trisodium trimetaphosphate and Schiff base bonds. l-Arginine (l-Arg) was incorporated as a bioactive substance in the Oisab + Cs scaffold to promote cell adhesion, cell proliferation, collagen synthesis, and vascularization. The fabricated scaffolds showed microporous networks in the scanning electron microscopy analysis. The scaffold also possessed excellent hemocompatibility. In vitro studies using fibroblasts (L929 and human dermal fibroblast cells) confirmed the cytocompatibility of these scaffolds. The results of the in vivo chicken chorioallantoic membrane assay confirmed the proangiogenic activity of the Oisab + Cs + l-Arg scaffolds. The wound-healing potential of these scaffolds was studied in streptozotocin-induced diabetic rats. This in vivo study showed that the period of epithelialization in the Oisab + Cs + l-Arg scaffold-treated wounds was 21.67 ± 1.6 days, which was significantly faster than the control (30.33 ± 2.5 days). Histological and immunohistochemical studies showed that the Oisab + Cs + l-Arg scaffolds significantly accelerated the rate of wound contraction by reducing inflammation, improving collagen synthesis, and promoting neovascularization. These findings suggest that the Oisab + Cs + l-Arg scaffolds could be beneficial in treating diabetic wounds in clinical applications.

Evaluation of cell adhesion and inflammatory response of chitin and chitosan nanofibers patterned by inkjet printing

AbstractChitin and chitosan nanofibers (ChNF and CtsNF) are promising biomaterials due to their biocompatibility, biodegradability, and non‐toxicity. This study investigates the cell adhesion properties and inflammatory responses of CtsNF, ChNF, and their mixtures when patterned on cellophane films using inkjet printing technology, keeping in mind their potential applications as cell culture scaffolds. The viscosities of 0.1 wt% aqueous dispersions of CtsNF, ChNF, and their mixtures were confirmed to be suitable for inkjet printing. Microstructures with varying thicknesses were fabricated by adjusting the printing parameters. Mouse fibroblast cells (L929) and mouse macrophages (RAW264.7) were used to evaluate cell adhesion and inflammatory responses. The results demonstrated that CtsNF microstructures exhibited excellent cell adhesion even for those as thin as ~140 nm and low inflammatory potential. This finding provides valuable insights into the development of advanced biomaterials for medical applications and could be instrumental in optimizing dosage settings for wound healing treatments as well.

The Remarkable and Selective In Vitro Cytotoxicity of Synthesized Bola-Amphiphilic Nanovesicles on Etoposide-Sensitive and -Resistant Neuroblastoma Cells.

Neuroblastoma (NB) is a solid tumor occurring in infancy and childhood. Its high-risk form has currently a survival rate <50%, despite aggressive treatments. This worrying scenario is worsened by drug-induced secondary tumorigenesis and the emergency of drug resistance, calling for the urgent development of new extra-genomic treatments. Triphenyl phosphonium salts (TPPs) are mitochondria-targeting compounds that exert anticancer effects, impair mitochondria functions, and damage DNA at the same time. Despite several biochemical applications, TPP-based bola-amphiphiles self-assembling nanoparticles (NPs) in water have never been tested as antitumor agents. Here, with the aim of developing new antitumor devices to also counteract resistant forms of HR-NB, the anticancer effects of a TPP-based bola-amphiphile molecule have been investigated in vitro for the first time. To this end, we considered the previously synthesized and characterized sterically hindered quaternary phosphonium salt (BPPB). It embodies both the characteristics of mitochondria-targeting compounds and those of bola-amphiphiles. The anticancer effects of BPPB were assessed against HTLA-230 human stage-IV NB cells and their counterpart, which is resistant to etoposide (ETO), doxorubicin (DOX), and many other therapeutics (HTLA-ER). Very low IC50 values of 0.2 µM on HTLA-230 and 1.1 µM on HTLA-ER (538-fold lower than that of ETO) were already determined after 24 h of treatment. The very low cell viability observed after 24 h did not significantly differ from that observed for the longest exposure timing. The putative future inclusion of BPPB in a chemotherapeutic cocktail for HR-NB was assessed by investigating in vitro its cytotoxic effects against mammalian cell lines. These included monkey kidney cells (Cos-7, IC50 = 4.9 µM), human hepatic cells (HepG2, IC50 = 9.6 µM), a lung-derived fibroblast cell line (MRC-5, IC50 = 2.8 µM), and red blood cells (RBCs, IC50 = 14.9 µM). Appreciable to very high selectivity indexes (SIs) have been determined after 24 h treatments (SIs = 2.5-74.6), which provided evidence that both NB cell populations were already fully exterminated. These in vitro results pave the way for future investigations of BPPB on animal models and upon confirmation for the possible development of BPPB as a novel therapeutic to treat MDR HR-NB cells.

Pneumolysin-responsive liposomal platform for selective treatment of Streptococcus pneumoniae.

The bacterium Streptococcus pneumoniae has become a leading cause of meningitis, sepsis, and bacterial pneumonia worldwide, with increased prevalence of antibiotic-resistant serotypes serving to exacerbate the issue. The main factor responsible for colonization and immune response escape in pneumococcal infections is the secreted molecule pneumolysin, which is a subset within a family of related toxins that form transmembrane pores in biological membranes through cholesterol recognition and binding. The conserved activity and structure of pneumolysin between all observed S. pneumoniae serotypes, along with its requirement for pathogenicity, has made this molecule an attractive target for vaccination, diagnostic, and sequestration platforms, but not yet as a facilitative agent for therapeutic treatment. Consequently, the present work aimed to examine the impact of liposomal cholesterol content for pneumolysin-induced release of the encapsulated antimicrobial peptide nisin. It was determined that a cholesterol content above 45mol% was necessary to facilitate interactions with both purified pneumolysin toxin and S. pneumoniae culture, demonstrated through enhanced nisin release and a reduction in hemolytic rates upon exposure of the toxin with cholesterol-rich vesicles. Antibacterial testing highlighted the ability of the developed platform to elicit a potent and specific bactericidal response in vitro against cultured S. pneumoniae when compared to a control strain, Staphylococcus epidermidis. It further improved viability of a fibroblast cell line upon S. pneumoniae challenge, outperforming free nisin via the synergistic impact of simultaneous bacterial clearance and pneumolysin neutralization. These findings collectively indicate that cholesterol-rich liposomes hold promise as a selective treatment platform against pneumococcal infections.