Kelvin probe force microscopy (KPFM) is a highly advanced technique offering notable surface sensitivity and high lateral resolution, ranging from micrometres to the sub-nanometre scale. This scanning probe technique effectively detects local electrical surface potential (ESP), influenced charge distribution, and work function differences, making it essential for studying biological and biochemical processes, from single molecules to complex cellular structures. By enabling nanometre-resolution analysis under simulated conditions, KPFM provides crucial insights into the physicochemical evolution, functionality, and structural organization of biomolecular systems. Recent advancements have significantly expanded KPFM's capabilities, revealing ESP characteristics in diverse biological entities, including single proteins, DNA strands, lipid films, fibrils, and complex neuronal structures. The technique also facilitates the study of biomolecular nanolayers on advanced nanomaterials like gold nanoparticles and carbon nanotubes, enhancing its role in bio-nanotechnology. Such versatility highlights KPFM's transformative potential in elucidating biomolecular interactions at unprecedented resolutions. This review critically analyses recent advancements, addresses ongoing challenges in measuring ESP in biological samples, and highlights emerging strategies to improve resolution and sensitivity. Additionally, KPFM's implications in diagnostics, biosensing, tissue engineering, therapeutics, drug screening, and Alzheimer's research are explored, establishing it as a powerful tool at the intersection of nanotechnology and biomedical innovation.

The biodegradation of therapeutic magnetic-oxide nanoparticles (MONPs) in the human body raises concerns about their lifespan, functionality, and health risks. Interactions between apoferritin proteins and MONPs in the spleen, liver, and inflammatory macrophages significantly accelerate nanoparticle degradation, releasing metal ions taken up by apoferritin. This can alter the protein’s biological structure and properties, potentially causing health hazards. This study examines changes in apoferritin’s shape, electrical surface potential (ESP), and protein-core composition after incubation with cobalt-ferrite (CoFe₂O₄) oxide nanoparticles. Using atomic force microscopy (AFM) and scanning Kelvin probe force microscopy (SKPFM), we observed changes in the topography and ESP distribution in apoferritin nanofilms over time. After 48 h, the characteristic apoferritin hole (∼1.35 nm) vanished, and the protein’s height increased from ∼3.5 to ∼7.5 nm due to hole filling. This resulted in a significant ESP increase on the filled-apoferritin surface, attributed to the formation of a heterogeneous chemical composition and crystal structure (γ-Fe₂O₃, Fe₃O₄, CoO, CoOOH, FeOOH, and Co₃O₄). These changes enhance electrostatic interactions and surface charge between the protein and the AFM tip. This approach aids in predicting and improving the MONP lifespan while reducing their toxicity and preventing apoferritin deformation and dysfunction.

The last two decades have witnessed the emergence of micro- and nanoswimmers (MNSs). Researchers have invested significant efforts in engineering motile micro- and nanodevices to address current limitations in minimally invasive medicine. MNSs can move through complex fluid media by using chemical fuels or external energy sources such as magnetic fields, ultrasound, or light. Despite significant advancements in their locomotion and functionalities, the gradual deterioration of MNSs in human physiological media is often overlooked. Corrosion and biodegradation caused by chemical reactions with surrounding medium and the activity of biological agents can significantly affect their chemical stability and functional properties during their lifetime performance. It is therefore essential to understand the degradation mechanisms and factors that influence them to design ideal biomedical MNSs that are affordable, highly efficient, and sufficiently resistant to degradation (at least during their service time). This review summarizes recent studies that delve into the physicochemical characteristics and complex environmental factors affecting the corrosion and biodegradation of MNSs, with a focus on metal-based devices. Additionally, different strategies are discussed to enhance and/or optimize their stability. Conversely, controlled degradation of non-toxic MNSs can be highly advantageous for numerous biomedical applications, allowing for less invasive, safer, and more efficient treatments.

Functional oxide nanoparticles are extensively utilized in the last decades for biomedical purposes due to their unique functional properties. Nevertheless, their biodegradation mechanism by biological species, particularly by proteins at oxide/protein interfaces, still remains limited. Here, a systematic approaches is provided to investigate electrochemical behavior, electronic properties, and biodegradation mechanism of cobalt ferrite (CFO) and cobalt ferrite-bismuth ferrite (CFO-BFO) core-shell nanoparticles in apoferritin-containing media. Scanning Kelvin probe force microscopy results indicate that the presence of a thin shell (≈5 nm) of BFO on CFO causes a significant increase in surface potential. The potentiodynamic polarization measurements in different solutions showed higher anodic current densities for both samples when decreasing pH and increasing apoferritin concentration. Notably, CFO-BFO exhibits lower anodic current densities than CFO due to a slightly higher flat band potential and lower donor density distribution on CFO-BFO than on CFO, which results in lower electrochemical activity. Long-term monitoring reveals that biodegradation of both nanoparticles is accelerated by high apoferritin concentrations and acidic media, resulting in the decrease of electrochemical potentials and impedance values, and enhancement of metal ion release. Thus, this systematic biodegradation study can help to predict the lifespan and toxicity of these functional nanoparticles in biological environments.