Design of Carboxymethylcellulose/Poloxamer-Based Bioformulation Embedding Trichoderma afroharzianum for Agricultural Applications.

Microbial biological control agents are believed to be a potential alternative to classical fertilizers to increase the sustainability of agriculture. In this work, the formulation of Trichoderma afroharzianum T22 (T22) spores with carboxymethyl cellulose (CMC) and Pluronic F-127 (PF-127) solutions was investigated. Rheological and microscopical analysis were performed on T22-based systems at three different CMC/PF-127 concentrations, showing that polymer aggregates tend to surround T22 spores, without viscosity, and the viscoelastic properties of the formulations were affected. Contact angle measurements showed the ability of PF-127 to increase the wettability of the systems, and the effect of the formulations on the viability of the spores was evaluated. The viability of the spores was higher over 21 days in all the formulations, compared to the control in water, at 4 and 25 °C. Finally, the effectiveness of the formulations on sweet basil was estimated by greenhouse tests. The results revealed a beneficial effect of the CMC/PF-127 mixture, but none on the formulation with T22. The data show the potential of CMC/PF-127 mixtures for the future design of microorganism-based formulations.

Effect of Hyaluronic Acid and Mesenchymal Stem Cells Secretome Combination in Promoting Alveolar Regeneration.

Pharmacological therapies in lung diseases are nowadays useful in reducing the symptomatology of lung injury. However, they have not yet been translated to effective treatment options able to restore the lung tissue damage. Cell-therapy based on Mesenchymal Stem Cells (MSCs) is an attractive, as well as new therapeutic approach, although some limitations can be ascribed for therapeutic use, such as tumorigenicity and immune rejection. However, MSCs have the capacity to secrete multiple paracrine factors, namely secretome, capable of regulating endothelial and epithelial permeability, decrease inflammation, enhancing tissue repair, and inhibiting bacterial growth. Furthermore, Hyaluronic acid (HA) has been demonstrated to have particularly efficacy in promoting the differentiation of MSCs in Alveolar type II (ATII) cells. In this frame, the combination of HA and secretome to achieve the lung tissue regeneration has been investigated for the first time in this work. Overall results showed how the combination of HA (low and medium molecular weight HA) plus secretome could enhance MSCs differentiation in ATII cells (SPC marker expression of about 5 ng/mL) compared to the only HA or secretome solutions alone (SPC about 3 ng/mL, respectively). Likewise, cell viability and cell rate of migration were reported to be improved for HA and secretome blends, indicating an interesting potentiality of such systems for lung tissue repair. Moreover, an anti-inflammatory profile has been revealed when dealing with HA and secretome mixtures. Therefore, these promising results can allow important advance in the accomplishment of the future therapeutic approach in respiratory diseases, up to date still missing.

Preparation, Supramolecular Aggregation and Immunological Activity of the Bona Fide Vaccine Adjuvant Sulfavant S.

In aqueous conditions, amphiphilic bioactive molecules are able to form self-assembled colloidal structures modifying their biological activity. This behavior is generally neglected in preclinical studies, despite its impact on pharmacological development. In this regard, a significative example is represented by a new class of amphiphilic marine-inspired vaccine adjuvants, collectively named Sulfavants, based on the ß-sulfoquinovosyl-diacylglyceride skeleton. The family includes the lead product Sulfavant A (1) and two epimers, Sulfavant R (2) and Sulfavant S (3), differing only for the stereochemistry at C-2 of glycerol. The three compounds showed a significant difference in immunological potency, presumably correlated with change of the aggregates in water. Here, a new synthesis of diastereopure 3 was achieved, and the study of the immunomodulatory behavior of mixtures of 2/3 proved that the bizarre in vitro response to 1-3 effectively depends on the supramolecular aggregation states, likely affecting the bioavailability of agonists that can effectively interact with the cellular targets. The evidence obtained with the mixture of pure Sulfavant R (2) and Sulfavant S (3) proves, for the first time, that supramolecular organization of a mixture of active epimers in aqueous solution can bias evaluation of their biological and pharmacological potential.

Effect of the rigid segment content on the properties of segmented polyurethanes conjugated with atorvastatin as chain extender.

Segmented polyurethanes were prepared with polycaprolactone diol as soft segment and various amounts of 4,4´-Methylenebis(cyclohexyl isocyanate) and atorvastatin, a statin used for lowering cholesterol, in order to obtain SPU with different content of rigid segments. Polyurethanes with 35% or 50% of rigid segment content were physicochemically characterized and their biocompatibility assessed with L929 fibroblasts. High concentrations of atorvastatin were incorporated by increasing the content of rigid segments as shown by FTIR, Raman, NMR, XPS and EDX. Thermal and mechanical characterization showed that polyurethanes containing atorvastatin and 35% of rigid segments were low modulus (13 MPa) semicrystalline polymers as they exhibited a glass transition temperature (Tg) at -38 °C, melting temperature (Tm) at 46 °C and crystallinity close to 35.9% as determined by DSC. In agreement with this, X-ray diffraction showed reflections at 21.3° and 23.6° for PCL without reflections for atorvastatin suggesting its presence in amorphous form with higher potential bioavailability. Low content of rigid segments led to highly degradable polymer in acidic, alkaline and oxidative media with an acceptable fibroblast cytotoxicity up to 7 days possibly due to low atorvastatin content.

Stimuli-responsive chitosan/poly (N-isopropylacrylamide) semi-interpenetrating polymer networks: effect of pH and temperature on their rheological and swelling properties.

The aim of this work was to synthesize semi-interpenetrating polymer networks (semi-IPNs) by free radical polymerization of N-isopropylacrylamide [poly (NIPAAm)], in the presence of chitosan (CHI), and to study the effect of pH and temperature changes on their rheological and swelling properties. The semi-IPNs are thermally stable up to about 400 °C and the presence of CHI increases the thermal degradation rate compared to bare poly (NIPAAm). The prepared systems presents a well-defined porosity and proved to be non-toxic, in vitro, on human embryonic skin fibroblast, thus offering appropriate support for cell proliferation. The semi-IPNs present, at physiological pH, swelling degrees well below those of the pure poly (NIPAAm). Differently, at acidic pH, the CHI macromolecules are protonated and become much more permeable to the diffusion of water giving a swelling degree that approaches that of bare poly (NIPAAm). The viscoelastic moduli of the semi-IPNs increase as a function of pH while the LCST remain unchanged. Moreover, the semi-IPNs viscoelastic moduli increase with the increase of CHI content and, in particular, the difference between the elastic modulus before and after the sol/gel transition is higher for the semi-IPN than for bare poly (NIPAAm) just at about physiological conditions.

Design of electrospayed non-spherical poly (L-lactide-co-glicolide) microdevices for sustained drug delivery.

Polymer chain entanglements in organic solvents can be considered a key parameter in the formation of non-spherical beads when electrospraying is employed. The shape of micro/nanometric drug delivery systems plays a major role since it can affect circulation, extravasation, distribution and in vivo clearance of the devices. In this frame, we investigated the influence of polymer processing parameters on the design of polylactic-co-glycolic acid non-spherical microdevices loaded with triamcinolone acetonide (TrA), a sparingly water soluble corticosteroid, prepared by electrospraying technique through a one-step process. In particular, we verified that the formation of non-spherical MDs is related to the presence of entanglements among polymer chains to select the optimal solution to be sprayed. The addition of TrA did not substantially affect the particle morphology in terms of size, size distribution and circularity at all the tested drug loadings. Furthermore, the drug could be released for a prolonged period, with controlled and reproducible kinetics for over 3 weeks. The mathematical modeling of release profiles highlighted that the release is mainly driven by degradation, at a higher extent in the case of low drug loading.

Collagen-Mesenchymal Stem Cell Microspheres Embedded in Hyaluronic Acid Solutions as Biphasic Stem Niche Delivery Systems for Pulmonary Differentiation.

Cell therapy has the potential to become a feasible solution for several diseases, such as those related to the lungs and airways, considering the more beneficial intratracheal administration route. However, in lung diseases, an impaired pulmonary extracellular matrix (ECM) precludes injury resolution with a faulty engraftment of mesenchymal stem cells (MSCs) at the lung level. Furthermore, a shielding strategy to avoid cell damage as well as cell loss due to backflow through the injection path is required. Here, an approach to deliver cells encapsulated in a biomimetic stem niche is used, in which the interplay between cells and physiological lung ECM constituents, such as collagen and hyaluronic acid (HA), can occur. To this aim, a biphasic delivery system based on MSCs encapsulated in collagen microspheres (mCOLLs) without chemical modification and embedded in an injectable HA solution has been developed. Such biphasic delivery systems can both increase the mucoadhesive properties at the site of interest and improve cell viability and pulmonary differentiation. Rheological results showed a similar viscosity at high shear rates compared to the MSC suspension used in intratracheal administration. The size of the mCOLLs can be controlled, resulting in a lower value of 200 µm, suitable for delivery in alveolar sacs. Biological results showed that mCOLLs maintained good cell viability, and when they were suspended in lung medium implemented with low molecular weight HA, the differentiation ability of the MSCs was further enhanced compared to their differentiation ability in only lung medium. Overall, the results showed that this strategy has the potential to improve the delivery and viability of MSCs, along with their differentiation ability, in the pulmonary lineage.

The interplay between hyaluronic acid and stem cell secretome boosts pulmonary differentiation in 3D biomimetic microenvironments.

Mesenchymal stem cells (MCSs) secretome provide MSC-like therapeutic effects in preclinical models of lung injury, circumventing safety concerns with the use of live cells. Secretome consists of Extracellular Vesicles (EVs), including populations of nano- to micro-sized particles (exosomes and microvesicles) delimited by a phospholipidic bilayer. However, its poor stability and bioavailability severely limit its application. The role of Hyaluronic acid (HA) as potential carrier in biomedical applications has been widely demonstrated. Here, we investigated the interplay between HA and MSCs- secretome blends and their ability to exert a bioactive effect on pulmonary differentiation in a 3D microenvironment mimicking lung niche. To this aim, the physical-chemical properties of HA/Secre blends have been characterized at low, medium and high HA Molecular Weights (MWs), by means of SEM/TEM, DLS, confocal microscopy and FTIR. Collectively physical-chemical properties highlight the interplay between the HA and the EVs. In 3D matrices, HA/Secre blends showed to promote differentiation in pulmonary lineage, improved as the MW of the HA in the blends decreased. Finally, HA/Secre blends' ability to cross an artificial mucus has been demonstrated. Overall, this work provides new insights for the development of future devices for the therapy of respiratory diseases that are still unmet.

A Covalently Cross-Linked Hyaluronic Acid/Carboxymethyl Cellulose Composite Hydrogel as a Potential Filler for Soft Tissue Augmentation.

The use of fillers for soft tissue augmentation is an approach to restore the structure in surgically or traumatically created tissue voids. Hyaluronic acid (HA), is one of the main components of the extracellular matrix, and it is widely employed in the design of materials with features similar to human tissues. HA-based fillers already find extensive use in soft tissue applications, but are burdened with inherent drawbacks, such as poor thermal stability. A well-known strategy to improve the HA properties is to reticulate it with 1,4-Butanediol diglycidyl ether (BDDE). The aim of this work was to improve the design of HA hydrogels as fillers, by developing a crosslinking HA method with carboxymethyl cellulose (CMC) by means of BDDE. CMC is a water soluble cellulose ether, whose insertion into the hydrogel can lead to increased thermal stability. HA/CMC hydrogels at different ratios were prepared, and their rheological properties and thermal stability were investigated. The hydrogel with an HA/CMC ratio of 1/1 resulted in the highest values of viscoelastic moduli before and after thermal treatment. The morphology of the hydrogel was examined via SEM. Biocompatibility response, performed with the Alamar blue assay on fibroblast cells, showed a safety percentage of around 90% until 72 h.

Pomegranate extract-loaded sphingosomes for the treatment of cancer: Phytochemical investigations, formulation, and antitumor activity evaluation.

AIM: Formulation of Pomegranate Extracts (PE)-loaded sphingosomes as an antitumor therapy for the intravenous and passive targeted delivery to various tumor types, especially that of the breast, colon, and uterus; to increase the therapeutic activity and decrease the adverse effects profile. METHODS: The pericarp and seeds' juice of Punica granatum were each extracted using D.W. and ethanol. Phytochemical investigation of all extracts was carried out including total phenolics, flavonoids, and anthocyanins contents, the antioxidant activity, as well as HPLC analysis of phenolics and flavonoids. The antitumor potential of all extracts was also tested utilizing three cell lines: MCF-7, HeLa, and HCT116. The candidate extract was chosen for the formulation phase and was entrapped into the sphingosomes using the thin-film hydration method and employing three different PE: lipids weight ratios. The synthesized formulations were characterized for their size, morphological features, zeta potential, entrapment efficiency, and in vitro drug release and kinetics modeling studies. The optimized formula was further analyzed by FTIR spectroscopy and electron microscopy. The antitumor activity of F2 was also investigated using the same cancer cell lines compared to the plant extract. RESULTS: The highest phenolics, flavonoids, and anthocyanins contents were observed in the ethanolic pericarps extract (EPE), followed by the ethanolic seeds extract (ESE). Consequently, EPE showed a higher antitumor activity hence it was selected for the formulation phase. PE-loaded sphingosomes formula (F2) was selected for having the highest EE% (71.64%), and a sustained release profile with the highest in vitro release (42.5±9.44%). By employing the DDSolver, the Weibull model was found the most suitable to describe the PE release kinetics compared to other models. The release mechanism was found to follow Fickian diffusion. Simulated pharmacokinetic parameters have portrayed F2 as the candidate formula, with the highest AUC (536.095) and slowest MDT (0.642 h). In addition, F2 exhibited a significant (p>0.05) stronger and prolonged anticancer effect against MCF-7, HeLa, and HCT116 cell lines at all concentrations tested compared to the free extract. CONCLUSION: The results proved that sphingosomes are an effective delivery system, improving pharmacological efficacy and reducing serious side effects of anticancer medications and natural products.

Correction: Pomegranate extract-loaded sphingosomes for the treatment of cancer: Phytochemical investigations, formulation, and antitumor activity evaluation.

[This corrects the article DOI: 10.1371/journal.pone.0293115.].

Bioengineered Abiotic Nanomaterials Through Cell Membrane-Camouflaging: Advancements and Challenges in Lung Cancer.

Lung cancer remains a major global health concern with high mortality rates and poor prognosis. Bridging the gap between the chemical and cellular understanding of cell-decorated biomimetic nanocomposites and their clinical translation is crucial for developing effective therapies. Nanocomposites show promise in targeted drug delivery and diagnostics, but their clinical application is hindered by biocompatibility and clearance issues. To overcome these challenges, biomimetic approaches utilizing cell membrane-coated nanomaterials emerge. By camouflaging nanomaterials with cell membranes, the biointerfaces are enhanced, and the inherent properties of the donor cell membranes are acquired. This review provides an overview of recent advancements on cell membrane-coated nanocomposites for lung cancer diagnosis and treatment. It discusses fabrication techniques, biomedical applications, challenges, and future prospects. The incorporation of cell membranes into nanocomposites holds potential for improved lung cancer therapy, but further development and refinement are needed for precise tumor targeting. Addressing the identified challenges will pave the way for clinical translation of these biomimetic nanoplatforms and advance lung cancer diagnosis and treatment.

Fermentation-Derived Albumin-Based Hydrogels for Tissue Adhesion Applications.

Currently, most of the clinically available surgical glues and sealants lack elasticity, good adhesion and biocompatibility properties. Hydrogels as tissue adhesives have received extensive attention for their tissue-mimicking features. Here, a novel surgical glue hydrogel based on a fermentation-derived human albumin (rAlb) and biocompatible crosslinker for tissue-sealant applications has been developed. In order to reduce the risks of viral transmission diseases and an immune response, Animal-Free Recombinant Human Albumin from the saccharomyces yeast strain was used. A more biocompatible crosslinking agent, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), was used and compared with glutaraldehyde (GA). The design of crosslinked albumin-based adhesive gels was optimized by varying the albumin concentration, the mass ratio between albumin and the crosslinking agent as well as the crosslinker type. Tissue sealants were characterized in terms of mechanical (tensile and shear), adhesive and in vitro biocompatibility properties. The results indicated that the mechanical and adhesive properties improved as the albumin concentration increased and the mass ratio between albumin and crosslinker decreased. Moreover, the EDC-crosslinked albumin gels have better biocompatibility properties than GA-crosslinked glues.

Effect of Composition of Lung Biomimetic Niche on the Mesenchymal Stem Cell Differentiation toward Alveolar Type II Pneumocytes.

Pulmonary niche dynamically orchestrates the signals, such as proliferation or differentiation of mesenchymal stem cells (MSCs), which allows inducing tissue repair. Lung niche includes extracellular matrix (ECM), comprising hyaluronic acid (HA) and collagen (COLL), and several types of MSCs. Impaired ECM, in lung pathologies, makes the promising therapies based on MSCs ineffective, as it results in a reduced attachment and homing of MSCs, precluding their differentiation and viability. To overcome this problem, in this study a pulmonary biomimetic niche based on HA and COLL hydrogel is developed, with the specific aim to elucidate the role of COLL and HA/COLL semi-interpenetrating polymer networks (SIPNs) in directing the differentiation of MSCs into Alveolar Type II (ATII) cells. The effect of low (L), medium (M), and high (H) molecular weight (MW) HA is investigated, both like structural component of the SIPNs hydrogel and like trophic factor in cell culture media solution. HA in the culture media significantly improves surfactant protein (SP)-C expression (≈2 ng mL-1 ), without showing difference in the MW tested, compared to control only (≈1 ng mL-1 ). Furthermore, LMWHA/COLL hydrogel promotes the SPC expression (approximately two times) compared to COLL, MMWHA/COLL, and HMWHA/COLL hydrogels.

Design of functional nanoparticles by microfluidic platforms as advanced drug delivery systems for cancer therapy.

Nanoparticle systems are functional carriers that can be used in the cancer therapy field for the delivery of a variety of hydrophobic and/or hydrophilic drugs. Recently, the advent of microfluidic platforms represents an advanced approach to the development of new nanoparticle-based drug delivery systems. Particularly, microfluidics can simplify the design of new nanoparticle-based systems with tunable physicochemical properties such as size, size distribution and morphology, ensuring high batch-to-batch reproducibility and consequently, an enhanced therapeutic effect in vitro and in vivo. In this perspective, we present accurate state-of-the-art microfluidic platforms focusing on the fabrication of polymer-based, lipid-based, lipid/polymer-based, inorganic-based and metal-based nanoparticles for biomedical applications.

Emerging treatment approaches for triple-negative breast cancer.

Approximately, 15% of global breast cancer cases are diagnosed as triple-negative breast cancer (TNBC), identified as the most aggressive subtype due to the simultaneous absence of estrogen receptor, progesterone receptor, and HER2. This characteristic renders TNBC highly aggressive and challenging to treat, as it excludes the use of effective drugs such as hormone therapy and anti-HER2 agents. In this review, we explore standard therapies and recent emerging approaches for TNBC, including PARP inhibitors, immune checkpoint inhibitors, PI3K/AKT pathway inhibitors, and cytotoxin-conjugated antibodies. The mechanism of action of these drugs and their utilization in clinical practice is explained in a pragmatic and prospective manner, contextualized within the current landscape of standard therapies for this pathology. These advancements present a promising frontier for tailored interventions with the potential to significantly improve outcomes for TNBC patients. Interestingly, while TNBC poses a complex challenge, it also serves as a paradigm and an opportunity for translational research and innovative therapies in the field of oncology.

Ultrasmall Solid-Lipid Nanoparticles via the Polysorbate Sorbitan Phase-Inversion Temperature Technique: A Promising Vehicle for Antioxidant Delivery into the Skin.

Solid lipid nanoparticles promote skin hydration via stratum corneum occlusion, which prevents water loss by evaporation, and via the reinforcement of the skin's lipid-film barrier, which occurs through the adhesion of the nanoparticles to the stratum corneum. The efficacy of both phenomena correlates with lower nanoparticle size and the increased skin permeation of loaded compounds. The so-called Polysorbate Sorbitan Phase-Inversion Temperature method has, therefore, been optimized in this experimental work, in order to engineer ultrasmall solid-lipid nanoparticles that were then loaded with α-tocopherol, as the anti-age ingredient for cosmetic application. Ultrasmall solid-lipid nanoparticles have been proven to be able to favor the skin absorption of loaded compounds via the aforementioned mechanisms.

A Hyaluronic Acid-Based Formulation with Simultaneous Local Drug Delivery and Antioxidant Ability for Active Viscosupplementation.

Hyaluronic acid (HA) and its derivatives are widely used for intra-articular injection to augment compromised viscoelastic properties of damaged synovial fluid. Combining HA-based devices with anti-inflammatory drugs or bioactive principles in order to provide an additional benefit to the viscosupplementation is emerging as a new promising approach to improve the clinical outcome. Here, we aim to design a novel active viscosupplementation agent that can load and release hydrophobic drugs and at the same time possessing antioxidant properties. Optimized ternary systems named HCV based on HA, (2-hydroxypropyl)-ß-cyclodextrin (CD), and vitamin E (VE), without being engaged in formal chemical bonding with each other, showed the best viscoelastic and lubrication properties along with antioxidant capabilities, able to solubilize and release DF. The physical-chemical characterization suggested that the HCV system displayed rheological synergism and higher thermal stability because of the presence of VE and its antioxidant activity, and the loading of hydrophobic drugs was improved by the presence of CD and VE. Cell morphology and viability tests on L929 cells exhibited high biocompatibility of the HCV system with higher level expression of anti-inflammatory interleukin-10.

Hyaluronan-coated nanoparticles for active tumor targeting: Influence of polysaccharide molecular weight on cell uptake.

Here we aimed to correlate different molecular weights of hyaluronic acid (HA), 200, 800 and 1437 kDa, used to decorate poly(lactic-co-glycolic acid) (PLGA)-based nanoparticles (NPs), to their cell uptakes. NP internalization kinetics in CD44-overexpressing breast carcinoma cells were quantified, using healthy fibroblast cells as reference. Actually, NP uptake and selectivity by tumor cells were maximized for NPs HA 800 kDa, while being minimum for NPs HA1400 kDa. This unexpected result could be explained considering that the interaction between NPs and tumor cells is dictated by rearrangement and conformation of that segment of HA chain that actually protrudes from the NPs. Overall, results obtained in this work point at how HA molecular weight, is pivotal project parameter in NP formulation to promote active targeting in the CD44 overexpressing cancer cells.

Advances in Hyaluronic-Acid-Based (Nano)Devices for Cancer Therapy.

Cancer is the main cause of fatality all over the world with a considerable growth rate. Many biologically active nanoplatforms are exploited for tumor treatment. Of nanodevices, hyaluronic acid (HA)-based systems have shown to be promising candidates for cancer therapy due to their high biocompatibility and cell internalization. Herein, surface functionalization of different nanoparticles (NPs), e.g., organic- and inorganic-based NPs, is highlighted. Subsequently, HA-based nanostructures and their applications in cancer therapy are presented.