Search Results (53)

Background: Although teriparatide is efficacious, its once-daily subcutaneous injections cause local adverse events, inconvenience, and higher cost, limiting long-term adherence. Therefore, this research aims to engineer a pH-responsive oral formulation of teriparatide for osteoporosis therapy. Methods: A layered silicate nanocomplex was obtained by spontaneous self-assembly of teriparatide (Teri) with 3-aminopropyl magnesium phyllosilicate (AC). The nanocomplex (AC-Teri) was then coated with a 1:1 blend of two polymethacrylic acid derivatives (Eudragit^® L100 and Eudragit^® S 100) to provide pH-triggered drug release along the gastrointestinal tract. Results: AC-Teri and the coated nanocomplex (EE/AC-Teri) displayed high encapsulation efficiency (>90%) with narrow size distributions. In a stepwise buffer transition system, EE/AC-Teri demonstrated pH-dependent release, with less than 25% drug liberated at pH 1.2, approximately 54% at pH 6.8, and 74% at pH 7.4 over 24 h. Particle size and ζ-potential of EE/AC-Teri shifted in parallel with dissolution of the outer polymer shell. EE/AC-Teri also protected the peptide against enzymatic degradation, preserving the secondary structure of encapsulated teriparatide in simulated intestinal fluids. Compared with free drug, EE/AC-Teri enhanced transcellular drug permeation 2.7-fold in Caco-2 cells. In dexamethasone-induced osteoporotic rats, oral EE/AC-Teri significantly stimulated bone formation while suppressing resorption; micro-CT and histology confirmed recovery of trabecular architecture. Conclusions: EE/AC-Teri represents a promising oral teriparatide formulation for the effective management of osteoporosis. Full article

Background/Objectives: Prussian blue (PB) is the agent of choice for internal cesium (Cs) decorporation, yet its performance is critically dependent on gastrointestinal (GI) pH and formulation attributes. Hypochlorhydria, common in patients treated with proton pump inhibitors, may alter the stagewise distribution of Cs binding during GI transit. This study aimed to compare the performance of different PB formulations in sequential in vitro models simulating normogastria and hypochlorhydria: normogastric regimen (NG) and hypochlorhydric regimen (HC). Methods: A static, enzyme-free sequential model was applied using compendial simulated fluids (SGFs pH 1.2 or acetate pH 4.0, SIF pH 6.8, and phosphate buffer pH 7.2). The formulations tested included PB active pharmaceutical ingredient (API) (bulk), compression blend, PB tablets 500 mg (PB tablets), and Radiogardase^®. For each stage, cesium bound (qs, mg/g PB), fractional contributions (fs), and total capture (qtotal) were quantified. Additional analyses included sensitivity to initial Cs concentration (C₀) and desorption in mineralized water. Results: Overall performance was primarily determined by formulation (p < 0.0001), with a significant formulation × regimen interaction. The compression blend and PB tablets exhibited the highest decorporation capacity, PB-API showed intermediate performance, and Radiogardase^® was clearly lower. Under HC, capture was concentrated in the gastric stage (44–47%), whereas in NG, it shifted toward intestinal stages. Desorption in the mineralized water was statistically significant but negligible compared with total capture, supporting the stability of cesium sequestration. Conclusions: Formulation and gastric acidity regimens not only determine the total cesium capture but also redistribute it across the GI tract. PB tablets represent an effective and accessible alternative to Radiogardase^®, maintaining robust decorporation capacity under clinically relevant pH conditions. Full article

Background/Objectives: Formulating recombinant therapeutic proteins is essential to ensure their safety, efficacy, and stability. A growing trend in biopharmaceutical development is the move toward buffer-free formulations, which aim to reduce immunogenicity, improve tolerability, and simplify production. This review explores technological advances, regulatory perspectives, and safety considerations related to this shift. Methods: A systematic documentary review was conducted using the PSALSAR framework. Scientific publications, patents, and regulatory documents (2020–2025) were retrieved from PubMed, Scopus, Web of Science, and regulatory databases (FDA, EMA). Inclusion criteria focused on recombinant proteins, buffer-free formulations, and regulatory alignment. Results: The findings reveal an increasing adoption of self-buffering strategies in high-concentration subcutaneous biologics. Technologies such as Fc-fusion, PASylation, and XTENylation enhance stability without conventional buffers. Regulatory bodies are progressively accepting minimalist formulations, provided safety and biosimilarity are demonstrated. However, intellectual property barriers limit formulation transparency. A synthesis of recent FDA and EMA approvals illustrates this formulation evolution. Conclusions: Buffer-free formulations offer a promising alternative for therapeutic protein development by improving patient experience and reducing formulation complexity. They align with biosimilar goals and regulatory trends, although long-term transparency and safety assessments remain critical for widespread adoption. Full article

The clean-label movement has markedly increased consumer demand for meat products free from synthetic additives, such as sodium nitrite, ascorbate, and phosphate. This review summarizes strategies to replace these additives with natural alternatives while preserving the functional and quality properties of traditionally cured meats. Nitrite replacement commonly employs nitrate-rich vegetables, alongside nitrate-reducing starter cultures or pre-converted nitrite powders for adequate nitric oxide production and meat pigment stabilization. Ascorbate substitutes include vitamin C-rich materials and polyphenol-based antioxidants from green tea and rosemary, supporting nitrite reduction and contributing to meat pigment and oxidative stability. To compensate for phosphate functions, natural substitutes such as hydrocolloids, dietary fibers, protein isolates, and calcium powders from eggshells or oyster shells have shown partial success in restoring water-holding capacity, pH buffering, and textural integrity. In addition, non-thermal processing technologies, such as high-pressure processing, ultrasound, and cold plasma are explored as complementary strategies to enhance the efficacy of natural ingredients and support industrial scalability. However, challenges persist regarding ingredient variability, dose-dependent effects, and consistency in functional performance. Future research should focus on synergistic ingredient combinations, formulation standardization, and scalable application in industrial production to ensure the production of high-quality clean-label meat products. Full article

L-lysine (L-Lys) is the first-limiting amino acid in swine nutrition, but free-form supplements exhibit poor intestinal absorption, reducing their bioavailability. This study aimed to enhance the gastric retention, controlled intestinal release, and systemic availability of L-Lys by optimizing solid lipid microcapsules (SLMs). SLMs were formulated using hydrogenated triglycerides (C16:0 or C18:1), free fatty acids, and varying emulsifier concentrations. Gastric retention and intestinal release were evaluated in vitro under simulated gastrointestinal conditions (a pepsin buffer at pH 5.0 for 2 h, followed by a pancreatin buffer at pH 6.5 for up to 8 h at 39 °C). SLMs with hydrogenated triglycerides showed significantly higher gastric retention (94–95%) than those with free fatty acids (48%). Specifically, C16:0 triglyceride-based SLMs achieved 74% intestinal release, which was enhanced to 90% with 1% emulsifier. This refined formulation was subsequently evaluated in vivo using weaned pigs (three groups; n = 4) fed a basal cornmeal diet. The treatments included a single oral administration of saline solution (placebo), free L-Lys (0.17 g/kg BW), or L-Lys SLMs (0.38 g/kg BW, equally providing L-Lys at 0.17 g/kg BW). The SLMs delayed the L-Lys plasma peak (T. max. 3–4 h vs. 1 h) and significantly increased the total L-Lys amount in the plasma over 24 h, demonstrating the enhanced relative bioavailability of encapsulated L-Lys. Full article

The formulation of biosimilar products critically determines their stability, safety, immunogenicity, and market accessibility. This article presents a novel integrative framework for biosimilar formulation that balances scientific, regulatory, and intellectual property dimensions, offering a holistic perspective rarely unified in the literature. It highlights the growing trend toward buffer-free, high-concentration systems that leverage protein self-buffering to improve patient comfort and formulation stability. The article also addresses regulatory flexibility from the FDA and EMA, which allows scientifically justified deviations from reference formulations to ensure pharmaceutical equivalence and minimize immunogenicity. A novelty of this article is its comprehensive analysis of how digital innovations, such as Quality-by-Design, Process-Analytical-Technology, and AI-based in silico simulations, are transforming formulation design and bioprocess optimization to reduce immunogenic risks and enhance bioequivalence. Two important key takeaways emerge: (1) strategic innovation in formulation, especially using buffer-free and high concentration systems, improve product stability and patient tolerability while complying with regulatory standards; and (2) intellectual property challenges, including patent thickets, strongly influence formulation decisions, making early legal-strategic alignment essential for market entry. The article confirms that practical recommendations for the selection of recombinant therapeutic protein formulations can effectively guide developers and regulators toward safer, more efficient, and commercially viable biosimilar products. Full article

Objectives: Thermo-gelling hydrophilic polymers like PLGA–PEG–PLGA are known as injectable sustained-release depots for biologics, but they face challenges due to the occurrence of severe burst release. This study aimed to develop a strategy to avoid the initial burst release by pre-encapsulating proteins in polysaccharide microparticles through an aqueous–aqueous emulsion mechanism, thereby enhancing therapeutic retention and linear release kinetics. Methods: Five model proteins (G-CSF, GM-CSF, IGF-1, FVIII, BSA) were encapsulated in dextran microparticles, using an organic solvent-free aqueous–aqueous emulsion method. These particles were dispersed in a 23% (w/w) PLGA–PEG–PLGA solution and injected into a 37 °C release buffer to form a gel depot. The in vitro release profiles were quantified using ELISA and MicroBCA assays over 9–42 days. The bioactivity of the proteins was validated using cell proliferation assays (NFS-60, TF-1, MCF-7) and chromogenic kits. The in vivo pharmacokinetics of the FVIII-loaded formulations were evaluated in Sprague–Dawley rats (n = 5/group) over 28 days. Results: Protein-loaded dextran particles retained their structural integrity within the hydrogel and exhibited minimal burst release (≤5% within 30 min vs. >25% for free proteins). Sustained near-linear release profiles were observed for all the proteins, with complete release by day 9 (G-CSF, GM-CSF, BSA) or day 42 (FVIII). Rats administered with the thermal gel with FVIII–dextran particles showed a significantly lower peak plasma concentration (C_max: 88.25 ± 30.21 vs. 132.63 ± 66.67 ng/mL) and prolonged therapeutic coverage (>18 days vs. 15 days) compared to those administered with the thermal gel with the FVIII solution. The bioactivity of the released proteins remained at ≥90% of the native forms. Conclusions: Pre-encapsulation in dextran microparticles effectively mitigates burst release from thermosensitive hydrogels, while preserving protein functionality. Full article

Background: Graphene Oxide (GO) has shown great potential in biomedical applications for cancer therapeutics. The biosafety and stability issues of GO in biological media have been addressed by functionalization with polyethylene glycol (PEG). Methods: In this work, carboxylated, nanosized GO (nCGO) was evaluated as a potential carrier of paclitaxel (PCT). The effect of PEG characteristics on particle size and surface charge, colloidal stability, drug, and release, and the hemolytic potential of nCGO, was investigated. Optimum PEG-nCGO/PCT formulations based on the above properties were evaluated for their anticancer activity (cytotoxicity and apoptosis induction) in the A549 lung cancer cell line. Results: An increase in the length of linear PEG chains and the use of branched (4-arm) instead of linear PEG resulted in a decrease in hydrodynamic diameter and an increase in ζ potential of the pegylated nCGO particles. Pegylated nCGO exhibited high colloidal stability in phosphate-buffered saline and in cell culture media and low hemolytic effect, even at a relatively high concentration of 1 mg/mL. The molecular weight of PEG and branching adversely affected PCT loading. An increased rate of PCT release at an acidic pH of 6.0 compared to the physiological pH of 7.4 was observed with all types of pegylated nCGO/PCT. Pegylated nCGO exhibited lower cytotoxicity and apoptotic activity than non-pegylated nCGO. Cellular uptake of pegylated nCGO increased with incubation time with cells leading to increased cytotoxicity of PEG-nCGO/PCT with incubation time, which became higher than that of free PCT at 24 and 48 h of incubation. Conclusions: The increased biocompatibility of the pegylated nCGO and the enhanced anticancer activity of PEG-nCGO/PCT compared to free PCT are desirable properties with regard to the potential clinical application of PEG-nCGO/PCT as an anticancer nanomedicine. Full article

Polysorbates are the predominant surfactants used to stabilize protein formulations. Unfortunately, polysorbates can undergo hydrolytic degradation, which releases fatty acids that can accumulate to form visible particles. The detection and quantitation of these fatty acid degradation products are critical for assessing the extent of polysorbate degradation and the associated risks of particle formation. We previously developed a user-friendly mass spectrometric method called Fatty Acids by Mass Spectrometry (FAMS) to quantify the free fatty acids. The FAMS method was validated according to ICH Q2 (R1) guidelines and is suitable for a wide range of products, buffers and protein concentrations. The end-to-end workflow can be automated from sample preparation to data analysis. To broaden method accessibility, the QDa detector selected for fatty acid measurement does not require specific mass spectrometry experience. We provide here a detailed procedure for both manual and automated sample preparation for high-throughput analysis. In addition, we highlight in this protocol the critical operational details, procedural watchouts and troubleshooting tips to support the successful execution of this method in another laboratory. Full article

Breast cancer is one of the leading causes of death in the female population because of the resistance of cancer cells to many anticancer drugs used. Curcumin has cytotoxic activities against breast cancer cells, although it has limited use due to its poor bioavailability and rapid metabolic elimination. The synthesis of metal complexes of curcumin and curcuminoids is a relevant topic in the search for more active and selective derivatives of these molecular scaffolds. However, solubility and bioavailability are concomitant disadvantages of these types of molecules. To overcome such drawbacks, the preparation of inclusion complexes offers a chemical and pharmacologically safe option for improving the aqueous solubility of organic molecules. Herein, we describe the preparation of the inclusion complex of dimethoxycurcumin magnesium complex (DiMeOC-Mg, (4)) with beta-cyclodextrin (DiMeOC-Mg-BCD, (5)) in the stoichiometric relationship 1:1. This new inclusion complex’s solubility in aqueous media phosphate buffer saline (PBS) was improved by a factor of 6x over the free metal complex (4). Furthermore, 5 affects cell metabolic rate, cell morphology, cell migration, induced apoptosis, and downregulation of the matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9), interleukin-6 (IL-6), and signal transducer and activator of transcription-3 (STAT3) expression levels on MD Anderson metastasis breast-231 cancer (MDA-MB-231) cell lines. Results of an antitumor assay in an in ovo model showed up to 30% inhibition of tumor growth for breast cancer (MDA-MB-231) when using (5) (0.650 mg/kg dose) and 17.29% inhibition with the free homoleptic metal complex (1.5 mg/kg dose, (4)). While the formulation of inclusion complexes from metal complexes of curcuminoids demonstrates its usefulness in improving the solubility and bioavailability of these metallodrugs, the new compound (5) exhibits excellent potential for use as a therapeutic agent in the battle against breast cancer. Full article

In the present study we report the development of an advantageous optical sensor for the speciation of Fe(III)/Fe(II). The sensor is based on the selective reaction of Fe(III) with a Desferal (Deferoxamine) reagent at pH = 2, while both Fe(III) and Fe(II) react with the reagent at pH = 5 using an acetate/glycine buffer. In this way, frequently used extra oxidation (H₂O₂) or reduction (ascorbic acid or hydroxylamine) steps are avoided. Both species can be determined in the range of 25 to 150 μM using a 96-well plate platform and the instrument-free detection of the colored complex with an overhead book scanner. The LOD is 4 μM, and an additional advantage is that a single calibration curve can be utilized for quantitation. The applicability of the sensor was demonstrated by analyzing commercially available pharmaceutical formulations for quality control purposes. Full article

Nanomedicine has garnered significant attention due to the advantages it offers in the treatment of cancer-related disorders, some of the deadliest diseases affecting human lives. Conventional medication formulations often encounter issues of instability or insolubility in biological environments, resulting in low bioavailability. Nanocarriers play a crucial role in transporting and safeguarding drugs at specific sites of action, enabling gradual release under particular conditions. This study focuses on methotrexate (MTx) and cytarabine (Cyt), essential antitumoral drugs, loaded into PEGylated squalene micellar structures to enhance therapeutic effectiveness and minimize drawbacks. The micelles were prepared using ultrasound-assisted methods in both water and phosphate buffer saline solutions. Evaluation of drug-loaded micelles encompassed parameters such as particle size, colloidal stability, surface charge, morphology, encapsulation efficiency, drug loading capacity, and in vitro release profiles under simulated physiological and tumoral conditions. In vitro cell inhibition studies conducted on MCF-7 and HeLa cell lines demonstrated higher antitumoral activity for the drug-encapsulated micelles compared to free drugs. The encapsulation effectively addressed the burst effect, providing sustained release for at least 48 h while enhancing the drug’s protection under physiological conditions. Full article

Climate change results in exceptional environmental conditions and drives the migration of pathogens to which local plants are not adapted. Biotic stress disrupts plants’ metabolism, fitness, and performance, ultimately impacting their productivity. It is therefore necessary to develop strategies for improving plant resistance by promoting stress responsiveness and resilience in an environmentally friendly and sustainable way. The aim of this study was to investigate whether priming tobacco plants with a formulation containing silicon-stabilised hybrid lipid nanoparticles functionalised with quercetin (referred to as GS3 phyto-courier) can protect against biotic stress triggered by Agrobacterium tumefaciens leaf infiltration. Tobacco leaves were primed via infiltration or spraying with the GS3 phyto-courier, as well as with a buffer (B) and free quercetin (Q) solution serving as controls prior to the biotic stress. Leaves were then sampled four days after bacterial infiltration for gene expression analysis and microscopy. The investigated genes increased in expression after stress, both in leaves treated with the phyto-courier and control solutions. A trend towards lower values was observed in the presence of the GS3 phyto-courier for genes encoding chitinases and pathogenesis-related proteins. Agroinfiltrated leaves sprayed with GS3 confirmed the significant lower expression of the pathogenesis-related gene PR-1a and showed higher expression of peroxidase and serine threonine kinase. Microscopy revealed swelling of the chloroplasts in the parenchyma of stressed leaves treated with B; however, GS3 preserved the chloroplasts’ mean area under stress. Furthermore, the UV spectrum of free Q solution and of quercetin freshly extracted from GS3 revealed a different spectral signature with higher values of maximum absorbance (A_max) of the flavonoid in the latter, suggesting that the silicon-stabilised hybrid lipid nanoparticles protect quercetin against oxidative degradation. Full article

In this study, novel materials have been obtained via a dual covalent and ionic crosslinking strategies, leading to the formation of a fully interpenetrated polymeric network with remarkable mechanical performances as drug delivery platforms for dermal patches. The polymeric network was obtained by the free-radical photopolymerization of N-vinylpyrrolidone using tri(ethylene glycol) divinyl ether as crosslinker in the presence of sodium alginate (1%, weight%). The ionic crosslinking was achieved by the addition of Zn²⁺, ions which were coordinated by the alginate chains. Bentonite nanoclay was incorporated in hydrogel formulations to capitalize on its mechanical reinforcement and adsorptive capacity. TiO₂ and ZnO nanoparticles were also included in two of the samples to evaluate their influence on the morphology, mechanical properties and/or the antimicrobial activity of the hydrogels. The double-crosslinked nanocomposite hydrogels presented a good tensile resistance (1.5 MPa at 70% strain) and compression resistance (12.5 MPa at a strain of 70%). Nafcillin was loaded into nanocomposite hydrogel films with a loading efficiency of up to 30%. The drug release characteristics were evaluated, and the profile was fitted by mathematical models that describe the physical processes taking place during the drug transfer from the polymer to a PBS (phosphate-buffered saline) solution. Depending on the design of the polymeric network and the nanofillers included, it was demonstrated that the nafcillin loaded into the nanocomposite hydrogel films ensured a high to moderate activity against S. aureus and S. pyogenes and no activity against E. coli. Furthermore, it was demonstrated that the presence of zinc ions in these polymeric matrices can be correlated with the inactivation of E. coli. Full article

In recent years, oncolytic viruses (OVs) have emerged as an effective means of treating cancer. OVs have multiple oncotherapeutic functions including specifically infecting and lysing tumor cells, initiating immune cell death, attacking and destroying tumor angiogenesis and triggering a broad bystander effect. Oncolytic viruses have been used in clinical trials and clinical treatment as drugs for cancer therapy, and as a result, oncolytic viruses are required to have long-term storage stability for clinical use. In the clinical application of oncolytic viruses, formulation design plays a decisive role in the stability of the virus. Therefore, this paper reviews the degradation factors and their degradation mechanisms (pH, thermal stress, freeze–thaw damage, surface adsorption, oxidation, etc.) faced by oncolytic viruses during storage, and it discusses how to rationally add excipients for the degradation mechanisms to achieve the purpose of maintaining the long-term stability of oncolytic viral activity. Finally, the formulation strategies for the long-term formulation stability of oncolytic viruses are discussed in terms of buffers, permeation agents, cryoprotectants, surfactants, free radical scavengers, and bulking agent based on virus degradation mechanisms. Full article