Vasco D. B. Bonifácio

Supercritical carbon dioxide (scCO2) offers a sustainable medium for molecularly imprinted polymer (MIP) synthesis, yet rational monomer selection under supercritical conditions remains largely unexplored. Here, molecular dynamics (MD) simulations were used as a pre–polymerization screening tool to investigate template–monomer interactions for L–leucine (LEU) imprinting in scCO2. A CO2 force field was adapted for relevant temperature and pressure conditions, and relative binding free energies together with cluster population analysis were used to compare candidate systems. A beryllium–curcumin complex (BeCU) combined with 2–vinylpyridine was identified as a favorable system at 60 °C and 300 bar. Guided by these computational trends, BeCU–based MIPs were synthesized in scCO2 and experimentally evaluated. The optimized LEU/BeCU (1:1) system exhibited an imprinting factor of 11.6 in static binding assays. The materials were subsequently integrated into heat–transfer method (HTM) thermal sensors using two different surface–functionalization strategies: particle immobilization and direct scCO2–assisted grafting onto plasma–activated aluminum substrates. Both sensing platforms enabled selective thermal detection of LEU over L–isoleucine in the 0.23–0.92 mM range, with limits of detection of 0.03 mM for the particle immobilization method and 0.06 mM for the direct scCO2–assisted grafting method. This work demonstrates that MD–assisted screening can guide scCO2–based MIP synthesis and supports metal–coordinated imprinting as a viable strategy for thermal biosensing platforms and promotes a scalable and sustainable solution for selective amino acid detection.

Metabolic reprogramming in cancer cells induces profound alterations in membrane lipid organization, affecting phospholipid distribution, surface charge, and overall lipid composition. These modifications contribute to key oncogenic processes, including tumor progression, immune evasion, and resistance to therapy. Importantly, they also give rise to distinct biophysical features that may serve as selective targets for therapeutic intervention. Among these, the externalization of anionic phospholipids and the overexpression of negatively charged glycoconjugates render cancer cell membranes particularly susceptible to cationic membrane-active agents. Cationic anticancer peptides (ACPs) and their synthetic mimics (SMACPs) have emerged as promising candidates for selectively disrupting malignant membranes; however, limitations related to stability, bioavailability, and toxicity have restricted their clinical utility. To address these challenges, nanotherapeutic platforms—particularly dendrimers—have emerged as promising tools to enhance peptide delivery, enable precise targeting, and support multifunctional therapeutic strategies. Dendrimers, including the most recent class of polyurea (PURE) dendrimers, feature modular and biodegradable architectures that enable concurrent drug delivery, gene silencing, and imaging functionalities. This chapter examines how membrane lipid remodeling contributes to cancer pathophysiology and highlights recent advances in dendrimer-based nanocarriers as precision tools for exploiting membrane-associated vulnerabilities in oncology.

Lung cancer is a heterogeneous disease and the leading cause of cancer-related deaths worldwide. The aggressiveness of non-small-cell lung cancer (NSCLC) is often associated with oncogenic activation of the mitogen-activated protein kinase (MAPK) pathway. Among the most prevalent mutations in NSCLC, epidermal growth factor receptor 1 (EGFR) and Kirsten rat sarcoma (KRAS) alterations play a key role in cancer progression and patient survival. In this study, we investigated the influence of EGFR and KRAS mutational status on the response to polycationic core–shell dendrimers, using NSCLC cell lines harboring different EGFR and KRAS profiles. Cells were treated with PURE _G4 -OEI ₄₈ and PURE _G4 -OCEI ₂₄ dendrimers, which showed heterogeneous responses. In PC-9 ^EGFREx19Del cells, PURE _G4 -OEI ₄₈ induced elevated p-ERK/ERK ratios, indicating activation of the ERK-MAPK pathway and resistance. Notably, the EGFR ^Ex19Del mutation contributed to this resistance. To overcome this effect, a synergistic strategy combining PURE _G4 -OEI ₄₈ and PURE _G4 -OCEI ₂₄ with the tyrosine kinase inhibitor (TKI) gefitinib was explored. This combination sensitized PC-9 cells, reducing ERK activation and enhancing cell death. Ex vivo chick chorioallantoic membrane (CAM) assays confirmed that EGFR ^Ex19Del drives resistance to PURE _G4 -OEI ₄₈ , whereas co-treatment with gefitinib improves efficacy. The use of complex dendrimer systems allows precise modulation of membrane-targeted interactions and intracellular signaling, providing mechanistic insights into overcoming EGFR ^Ex19Del -driven resistance. Overall, these findings highlight the translational potential of integrating membrane-targeted nanotherapeutics with EGFR-directed therapies to improve outcomes in NSCLC patients.

Breast carcinoma (BC) is the most common malignancy in women, with triple‐negative breast cancer (TNBC) making up 10–20% of cases. TNBC has limited targeted therapies and poor survival due to late diagnosis and metastasis. Dendrimers are precise nanostructures with a three‐dimensional globular architecture designed to target the negatively charged membranes of cancer cells. This study evaluated the anticancer potential of two novel core–shell polycationic polyurea (PURE) dendrimers, PURE _G4 ‐OEI ₄₈ and PURE _G4 ‐OCEI ₂₄ , targeting BC cell membranes. Both dendrimers selectively interacted with TNBC cells, inducing apoptosis, necroptosis, and ferroptosis. In vivo , they reduced tumor volume in HCC1806 xenografts, with PURE _G4 ‐OEI ₄₈ showing no toxicity, while PURE _G4 ‐OCEI ₂₄ induced mild hepatic toxicity. These results suggest PURE dendrimers are promising TNBC treatments, with further modifications needed to enhance efficacy and reduce toxicity.

The development of environmentally friendly materials for forensic applications is a growing area of interest. Traditional forensic methods often rely on resource-intensive processes and hazardous materials, and thus a demand for sustainable efficient materials without compromising performance is needed. Fluorescent, regular and magnetic powders were prepared through ball-milling-assisted coating using biodegradable matrices such as silica, chitosan, and tri-sodium citrate. The effect of the magnetic core on the optical properties, along with the influence of matrix type on the photophysical and magnetic properties of the powders, was assessed. The results suggest that the polymeric matrix effectively prevented fluorescence quenching, although a reduction in fluorescence intensity was observed when comparing magnetic and non-magnetic powders. For core–shell structures based on chitosan and tri-sodium citrate, the reduction in fluorescence caused by the absorption of the magnetic core (Fe₃O₄) was less pronounced. Additionally, these structures exhibited better magnetic properties when compared with a silica-based core–shell. All fluorescent powders proved highly efficient in fingerprint detection on various surfaces, yielding similar results to commercially available powders. The produced powders are not only safe and cost-effective but also environmentally friendly, making them an alternative to the current commercial powders used in forensic applications.

The anticancer activity of two polycationic core–shell PURE biodendrimers is demonstrated. PUREG4-OEI48 induces apoptosis through mitochondrial disruption, whereas more hydrophobic PUREG4-OCEI24, shows cytotoxic and antimigratory effects.

To meet clinical demands, industry must enhance extracellular vesicle (EV) yields. Transitioning to 3D culture systems is a primary strategy, while emerging approaches explore stimuli or alternative cell sources to further boost EV secretion.