Zeta potential refers to the electrical potential difference between the surface of a particle and the surrounding medium it is suspended in. It is a fundamental property of colloidal systems and is widely used to characterize the stability and behavior of particles in various applications such as pharmaceuticals, cosmetics, and water treatment[26]. Zeta potential (ZP) is a very important parameter in biological systems, particularly in the context of blood and other bodily fluids.
Zeta Potential in Biological Systems
Zeta potential is a measure of the surface charge on cells, proteins, and other biomolecules suspended in bodily fluids such as blood, lymph, and cerebrospinal fluid. Zeta potential plays a crucial role in maintaining the stability and proper functioning of biological systems by influencing the surface charge on cells, proteins, and other biomolecules suspended in bodily fluids[1][2].
Significance of Zeta Potential in Blood
Blood is a complex fluid composed of red blood cells, white blood cells, platelets, and various proteins suspended in a liquid called plasma. The zeta potential of blood components is essential for maintaining their stability and preventing unwanted agglomeration or adhesion.
1. Red Blood Cells: Red blood cells (RBCs) have a negative zeta potential, which helps them repel each other and maintain their discoid shape. A high negative zeta potential (around -15 mV) maintains the fluidity of blood by preventing RBC aggregation[6]. Changes in the zeta potential of red blood cells can lead to conditions such as sickle cell anemia and malaria[2].
2. Platelets: Platelets have a negative zeta potential, which prevents them from adhering to healthy blood vessel walls. However, in the event of injury, the zeta potential changes, allowing platelets to adhere and initiate the clotting process[2].
3. Proteins: Blood proteins, such as albumin and immunoglobulins, have a negative zeta potential, which helps maintain their stability and prevents unwanted interactions[2].
An ideal red blood cell (RBC) zeta potential is around -15 to -30 mV. This range represents a balance between having sufficient negative charge to maintain dispersion stability and avoid aggregation, while not being so highly negative that it causes excessive repulsion between RBCs[45].
Zeta Potential and Disease States
Alterations in the zeta potential of blood components can be indicative of various disease states:
1. Cardiovascular Diseases: Changes in the zeta potential of red blood cells, platelets, and proteins can be associated with cardiovascular diseases such as atherosclerosis and thrombosis. Hypertensive and cardiac patients have significantly reduced RBC zeta potential compared to healthy individuals. Microscopic imaging reveals increased abnormal RBC shapes in these patients[23].
2. Inflammatory Conditions: Inflammation can alter the zeta potential of blood components, leading to increased adhesion and agglomeration, which can exacerbate the inflammatory response.
3. Fertility issues: The zeta potential of sperm and ova can influence their ability to interact and fuse during fertilization. Altered zeta potential may contribute to fertility problems[11].
4. Cancer: Cancer cells often have a different zeta potential compared to healthy cells, which can be used as a diagnostic marker and a target for therapeutic interventions.
Why is Zetal Potential Little Known?
Zeta potential is not a well-known concept outside of certain scientific fields, despite its importance in understanding the behavior of colloidal systems like nanoparticles in the body. Zeta potential is a technical concept used primarily in materials science, chemistry, and pharmaceutical research. It is not a commonly discussed topic in medicine or biology courses. The term is rarely mentioned in popular science media or health news. The general public and even many healthcare professionals are not familiar with zeta potential.
There are a few key reasons for this:
Measuring zeta potential accurately in complex biological media is challenging. Factors like ionic strength, pH, and the presence of proteins can significantly alter the zeta potential[4]. Standardized protocols are still being developed. This makes it difficult to relate zeta potential to health effects.
Due to the difficulty of measuring it, there is limited clinical data demonstrating the diagnostic or prognostic value of zeta potential measurements. More research is needed to establish robust correlations with health outcomes[28][29]
Measuring Zeta Potential in Biological Systems
A high zeta potential (negative or positive) indicates that the particles will repel each other and there is dispersion stability. A low zeta potential means there is no force to prevent particles from coming together and aggregating[3].
Home measurement of zeta potential is simply not feasible with current technology and methods.
The zeta potential can be measured directly using techniques like electrophoretic light scattering[7][9]. Electrophoresis can quantify the zeta potential of RBCs and assess their surface charge and aggregation state[6][7][9].
Observing RBC stacking (rouleaux formation) under a microscope or measuring blood clotting time are not direct measurements of zeta potential. These are indirect indicators that may be influenced by zeta potential but are not quantitative measures of it[6]
Red Blood Cell Aging and Zeta Potential
As RBCs age in the human body over their typical 120-day lifespan, they undergo changes in their physical and biochemical properties. This includes a decrease in surface area, changes in membrane shear modulus, and loss of negatively charged sialic acid groups on the cell surface.
As RBCs age, their zeta potential becomes progressively less negative, ranging from around -30 mV in young cells to -11 mV in very old cells[11]. This is likely due to loss of negatively charged sialic acid groups on the membrane surface.
RBCs have a naturally negative surface charge due to sialic acid residues on their membrane glycoproteins and glycolipids. This gives them a zeta potential around -15 to -30 mV.
Particles with the same sign zeta potential (e.g. negative) repel each other and maintain dispersion stability
. RBCs with a zeta potential in the ideal range have enough negative charge to stay dispersed.
Red Blood Cell Turnover Rate
The human body contains approximately 20-30 trillion red blood cells (RBCs) at any given time, accounting for about 84% of the total cells in the body. Adult males typically have 5-6 million RBCs per microliter of blood, while females have 4-5 million[16].
RBCs are continuously produced in the bone marrow through a process called erythropoiesis, which takes about 7 days[16]. The body produces around 2 million new RBCs per second[17]. Once mature, RBCs circulate in the bloodstream for about 100-120 days before being removed and recycled by macrophages in the spleen and liver[16][17].
The turnover rate of RBCs is approximately 1% per day, meaning that the entire population of RBCs is replaced every 3-4 months[18]. This high turnover rate is necessary to maintain a constant supply of oxygen-carrying cells in the body.
Do Extracted RBCs have Different ZP?
Measured extracted RBCs may have a lower zeta potential than those in the body. Donating blood does not improve the RBC zeta potential of donated blood cells. The act of donating itself can modify donated RBCs surface properties in a way that lowers their zeta potential[22]. The causes include: Mechanical stress as RBCs are drawn from the vein, pass through tubing, and are centrifuged for separation. Oxidative stress from exposure to air and potential contamination. Changes in pH, temperature, and osmolarity as the blood is mixed with anticoagulants and stored. These stresses can induce: Conformational changes in membrane proteins, Redistribution of phospholipids, Oxidation of lipids and proteins and Vesiculation and loss of membrane surface area.
These modifications are very similar to the changes that occur when RBCs are infected by the malaria parasite Plasmodium falciparum. The parasite induces oxidative stress, alters membrane permeability, and modifies the display of host proteins on the RBC surface. Therefore, caution should be used when drawing conclusions about RBCs in the body based on analysis of RBCs taken for sampling. This is accounted for by appropriate controls and considering the potential impact of the donation process on RBC properties[27].
Does Donating A Pint of Blood Change RBC Population Age?
The average adult has about 2-3 liters of red blood cells (RBCs), which accounts for about 40% of their total blood volume. When donating a typical 470 mL unit of whole blood, approximately 200 mL of that is RBCs. This represents about 8% of the total RBC volume in an adult. Within 24-48 hours, the donor’s body will have fully restored the donated blood volume[24], but restoring the donated RBC volume takes 4-8 weeks after donation[25].
Donation and Replacement: When a donor gives a pint of blood, the body loses RBCs and needs to produce more to replace them. This process is stimulated by the release of erythropoietin from the kidneys, which signals the bone marrow to produce more RBCs[21].
Impact on RBC Age: The donated blood is typically composed of a mix of older and younger RBCs. The removal of these cells allows for the replacement with newer RBCs produced by the bone marrow. This process can lead to a shift towards a younger RBC population in the donor’s body[21].
Recovery Time: It takes several weeks for the body to fully replace the lost RBCs. Male donors typically need to wait 12 weeks and female donors 16 weeks before donating again to ensure that their RBC levels are fully restored[21].
The ZP of newly produced RBCs after donation is likely higher than the average ZP of the remaining RBC collection, as they are younger. Therefore, the net effect of blood donation on the overall ZP of the RBC collection in the body is expected to be a slight increase, as the newly produced RBCs with higher ZP replace the lost RBCs. However, the magnitude of this effect is expected to be small. Also, this increase is expected to be temporary as the new RBCs age and their ZP decreases over time.
More research is needed to quantify these effects.
How Rapidly Does RBC Zeta Potential Change?
Red blood cell (RBC) zeta potential is highly dynamic and can change rapidly after a blood draw:
– RBC zeta potential exhibits circadian rhythms, with values becoming progressively less negative over the course of the day[11]. This suggests RBCs may compensate for changes in blood plasma composition to maintain a constant level of electrostatic repulsion.
– Immediately after resuspending RBCs in a low ionic strength solution, zeta potential shows a rapid peak before reaching a new steady-state value over approximately 20 minutes[12]. Interestingly, zeta potential exhibits a 5-15 minute delay before beginning equilibration compared to membrane potential.
– Storage of RBC units leads to a reduction in zeta potential without a commensurate loss of sialic acid, possibly due to leakage of cytosolic K+ causing membrane depolarization[12]. The zeta potential of RBCs decreases by around 42% over the course of storage, from -14.5 mV on day 1 to around -8.5 mV by day 29[34].
Significant changes can occur within minutes to hours after a blood draw. Measuring zeta potential at a single timepoint may not be representative of conditions in the body (in vivo).
Demonstrated EMF Impacts on Zeta Potential
Electric fields can significantly impact RBC morphology, water permeability, and electrophoretic properties.
The application of an alternating current (AC) electromagnetic field (EMF) can significantly impact the zeta potential of calcium carbonate (CaCO3) particles in aqueous solutions[36][4][5]. The zeta potential, which is a measure of the surface charge of particles, changes as a function of the frequency of the applied EMF[1][4]. When an EMF with a frequency between 1-5 kHz is applied, the zeta potential of CaCO3 particles becomes positive[36]. However, when the frequency ranges from 6-10 kHz, the zeta potential changes from positive to negative[36][4]. This change in sign indicates a shift in the electrostatic repulsion between the CaCO3 particles and the surfaces they interact with, such as vessel walls or pipes[36].
The application of an AC electric field of 50 Hz to inclined electrodes causes RBCs to move in the vertical direction, i.e. in the direction of the gradient of the electric field intensity, due to dielectrophoresis. In contrast, under a DC electric field, RBCs move horizontally in the direction of the electric field due to electrophoresis[44]. The velocity is linearly proportional to the field strength.
The magnitude of the zeta potential also depends on the EMF frequency. As the frequency increases, the zeta potential initially rises, reaches a maximum, and then decreases[4][36]. At a specific frequency, the zeta potential reaches a zero value, where the particles have no net surface charge[36][4].
These changes in zeta potential can have significant implications for various applications, such as scale formation in geothermal systems[36]. By applying an EMF with an appropriate frequency, it is possible to either accelerate or inhibit scale deposition, depending on the desired outcome[36].
In addition to CaCO3, EMFs can also affect the zeta potential of other materials, such as iron oxide nanoparticles and viral particles[2][3]. The changes in zeta potential can influence the interactions between these particles and biological systems, such as cells[38][39].
Some key effects of AC electric fields on RBCs include:
- Changes in cell diameter: There is a small but statistically significant increase in RBC diameter upon exposure to AC electric current[35][37]. This effect is positively correlated with the duration of exposure.
- Increased echinocyte formation: The percentage of echinocytes (spiny RBCs) increases in a duration-dependent manner with AC electric current exposure. This is attributed to modifications in the RBC membrane structure[35].
- Altered water permeability: AC electric current exposure is strongly and positively correlated with increased water permeability across the RBC membrane. This leads to decreased tonicity and further contributes to echinocyte formation[35].
- Ultrastructural changes: Scanning electron microscopy reveals various ultrastructural changes in the RBC membrane upon AC electric current exposure, including the formation of spiny projections and membrane blebbing[35].
Cell Membrane Permeability Induction by High Frequency EMF
Exposure to high-frequency EMFs, such as 18 GHz, can induce increased membrane permeability in RBCs, allowing the uptake of nanoparticles or other substances without compromising cell viability. This effect is believed to be caused by mechanical disturbance of the RBC membrane due to the EMF exposure.
Based on the search results, both Starlink and Dish Network utilize frequencies around 18 GHz for their satellite internet and television services, respectively.
Starlink uses the Ku band, which ranges from 12-18 GHz, for its satellite internet services. Specifically, Starlink is authorized to use the frequency ranges of 17.8-18.6 GHz and 18.8-19.3 GHz for space-to-Earth transmissions.
While the search results do not explicitly mention Dish Network’s frequencies, it is well-known that Dish Network uses the Ku band, including frequencies around 18 GHz, for its satellite television services. The Ku band, which ranges from 12-18 GHz, is commonly used for satellite television broadcasting and communication services.
Therefore, depending on the power levels reaching consumers, there may be a scientificially and medically valid concern that both Starlink and Dish Network which operate in the Ku band are causing a variety of health issues. If a consumer had both services, would this not be twice the cause for concern?
Regulatory bodies such as the FCC are supposed to impose limits on the maximum permissible exposure levels for radiofrequency (RF) emissions to ensure public safety. Both Starlink and Dish Network would need to comply with these regulations for their services to be approved for residential use.
Studies: 18GHz EMF Changes Membrane Permeability
Exposure to high-frequency electromagnetic fields (EMFs), such as 18 GHz, can indeed induce increased membrane permeability in red blood cells (RBCs), facilitating the uptake of nanoparticles or other substances without compromising cell viability. This phenomenon is believed to be caused by mechanical disturbances of the RBC membrane due to the EMF exposure.
Several studies have demonstrated this effect:
1. A study published in *Scientific Reports* showed that exposing RBCs to an 18 GHz EMF led to the uptake of nanospheres with high efficiency (96% for 23.5 nm nanospheres and 46% for 46.3 nm nanospheres). The study confirmed that the nanospheres were translocated through the membrane into the cytosol, likely due to the EMF-induced rotation of water dipoles causing membrane disturbance and enhanced membrane trafficking through a quasi-exocytosis process. The membrane permeability was transient and restored after approximately 10 minutes[46].
2. Another study investigated the effect of 18 GHz EMF on Gram-positive bacteria and found that the exposure induced permeability in the bacterial membranes, allowing the uptake of silica nanospheres. The cells remained permeable for at least nine minutes after exposure, and up to 84% of the cells remained viable. The study suggested that the mechanical stimulation of cellular membranes from high-frequency vibrations contributed to the increased permeability[47].
3. Research on mammalian cell lines, such as pheochromocytoma cells, also indicated that 18 GHz EMF exposure triggered a transient increase in membrane permeability. This effect was attributed to the interaction of the EMF with polar molecules like water, causing oscillations and potentially inducing local elastic tension on the membrane, leading to increased ion mobility and conductivity across the cell membrane[48].
4. A study published in *BioRisk* confirmed that exposure of RBCs to 18 GHz EMF resulted in cell membrane permeabilization and efficient uptake of nanospheres. The study emphasized that this effect was not due to bulk heating but rather to the mechanical disturbance of the membrane caused by the EMF[49].
These studies collectively support the hypothesis that high-frequency EMF exposure can induce increased membrane permeability in RBCs through mechanical disturbances, facilitating nanoparticle uptake without compromising cell viability.
Can 18 GHz EMF Cause Tinnitus?
While 18 GHz EMF has been shown to transiently increase membrane permeability in some cell types, there is no direct evidence linking this effect to tinnitus. Increased membrane permeability can cause significant damage to cochlear hair cells through several mechanisms:
1. Disruption of ion homeostasis: Increased membrane permeability allows abnormal ion fluxes across the cell membrane, particularly calcium ions. This can lead to a rapid increase in intracellular calcium levels, which is toxic to hair cells[51][52].
2. Mitochondrial dysfunction: The influx of calcium can cause loss of mitochondrial membrane potential, leading to decreased energy production and increased oxidative stress[52].
3. Activation of cell death pathways: Elevated intracellular calcium and disrupted ion balance can trigger apoptotic and necrotic cell death pathways in hair cells[52].
4. Entry of toxic substances: Increased membrane permeability allows potentially harmful substances from the extracellular environment to enter the hair cells, causing further damage[51].
5. Release of cellular contents: Membrane leakage can result in the release of cellular contents, including enzymes and other molecules, which may impact surrounding cells and tissues[51].
6. Impaired mechanoelectrical transduction: Damage to the hair cell membrane can disrupt the delicate structures necessary for converting sound waves into electrical signals, compromising hearing function[53].
7. Oxidative stress: The influx of ions and disruption of cellular processes can lead to increased production of reactive oxygen species (ROS), causing oxidative damage to cellular components[52][54].
8. Inflammation: Membrane damage can trigger inflammatory responses, potentially exacerbating hair cell injury[54].
Notably, studies have shown that membrane permeability increases rapidly following exposure to intense noise, preceding other signs of cellular damage such as nuclear condensation[51]. This suggests that membrane disruption is an early event in noise-induced hair cell damage and may be a direct consequence of mechanical stress rather than a secondary effect of apoptosis[51].
Understanding these mechanisms is crucial for developing potential therapeutic interventions to prevent or mitigate hair cell damage in conditions such as noise-induced hearing loss and other forms of sensorineural hearing loss.
Health Correlates with Increased Echinocytes (spiny RBCs)
The percentage of echinocytes (spiny RBCs) increases in a duration-dependent manner with AC electric current exposure[35][37][43]. A study exposed red blood cells to AC electric current for 0, 0.5, 3, and 6 hours. The researchers found a strong and statistically significant correlation between electric current exposure duration and the percentage of echinocytes. The overall Pearson’s correlation coefficient was 0.8886 (P<0.0001), and it increased to 0.9556 (P<0.0001) when excluding the control samples. The study also found statistically significant differences in the percentage of echinocytes between the control samples and the 0.5, 3, and 6 hour exposure samples (P<0.0001). More echinocytes were found in the samples exposed to electric current compared to the controls[35].
Echinocytes have an increased membrane stiffness compared to normal discocytes[42]. The altered mechanical properties of echinocytes may impair their ability to deform and flow through small capillaries.
Transformation of red blood cells from the normal biconcave disc shape to echinocytes is associated with increased membrane stiffness.
Several studies have found that red blood cell shape changes, including echinocyte formation, are characteristic of certain neurodegenerative disorders like Parkinson’s disease, Alzheimer’s disease, and amyotrophic lateral sclerosis (ALS)[41][42]. This suggests a potential link between RBC morphology and neurodegeneration. This shape change occurs in some neurodegenerative diseases like Parkinson’s, ALS and Alzheimer’s, but the clinical significance is unclear.
How to Increase Zeta Potential (General)
If the zeta potential (ZP) is low, it indicates that the particles in the suspension are unstable and may aggregate, coagulate, or flocculate. This true both outside and inside the body. To address this issue, several strategies can be employed:
1. Adjust pH: Zeta potential is highly influenced by pH. By adjusting the pH of the suspension, the surface charge of the particles can be altered, which can improve stability. For example, if the particles have a negative charge, increasing the pH (making a solution more alkaline and less acidic) can enhance the negative charge, leading to greater stability[30][31].
2. Lower Electrolyte Concentration: In general, adding more electrolytes increases the ionic strength of a solution. Increasing the ionic strength can reduce the zeta potential, while decreasing it can increase the zeta potential. High ionic strength can lead to particle aggregation[30][31]. To lower elevated electrolyte levels in the blood, the most effective approach is to stay well-hydrated by drinking plenty of water. Proper hydration throughout the day helps dilute the concentration of electrolytes in the blood and promotes their excretion through urine.
3. Optimize Temperature: The zeta potential of some solutions is strongly dependent on temperature, while others show almost no temperature dependence[32]. In some cases, adjusting the temperature can improve stability by altering the surface charge or the ionic strength of the suspension[4]. The magnitude of the zeta potential typically decreases with increasing temperature[33]. RBCs at 96.8°F (36°C) will have a higher zeta potential compared to RBCs at 98.6°F (37°C), but the exact extent of the difference is was not found in this search.
By implementing these strategies, the stability of the particles can be improved, and the zeta potential can be optimized for the specific application.
How to Increase Zeta Potential (of Blood Cells, In Vivo)
Increasing the zeta potential of blood cells in vivo (in the body) can be achieved through several methods, each targeting different aspects of the cell membrane and surrounding environment. Here are some key strategies::
1. Grounding (Earthing): Grounding or earthing, which involves direct physical contact with the Earth, has been shown to increase the zeta potential of RBCs. This method involves connecting the body to the Earth using conductive patches, which can increase the surface charge on RBCs. This increase in zeta potential reduces blood viscosity and RBC aggregation, thereby improving blood flow and reducing cardiovascular risk factors[58]. Research does indicate that grounding or earthing (direct skin contact with the earth’s surface) can increase the zeta potential of red blood cells, which reduces blood viscosity and clumping.
- A study showed that 2 hours of grounding increased the zeta potential in all subjects by an average of 2.70 and significantly reduced red blood cell aggregation[61].
- Another study found that just one hour of grounding during yoga exercises significantly reduced post-exercise blood viscosity compared to a control group[63].
- The effects of grounding on zeta potential and blood viscosity appear to occur relatively quickly, with one study noting changes after only 45 minutes of earthing[62].
While these studies demonstrate positive effects on zeta potential and blood viscosity from grounding, they do not specify an exact daily duration that would be universally effective. The time required may vary depending on individual factors and the specific grounding method used. It’s important to note that the research suggests even short periods of grounding can produce measurable effects on blood properties. However, for optimal benefits, regular and sustained contact with the earth may be recommended, though an exact daily duration is not specified in these results.
2. Re-sialylation of Aged Red Blood Cells: One effective method involves the re-sialylation of aged red blood cells (RBCs). Sialic acid (SA) is a crucial component of the RBC membrane that contributes to its negative charge and, consequently, its zeta potential. Aging reduces the SA content on RBC membranes, leading to a decrease in zeta potential. By incubating aged RBCs with SA in the presence of cytidine triphosphate (CTP) and α-2,3-sialyltransferase, the SA content can be restored, effectively increasing the zeta potential to levels comparable to young RBCs[57][60]. While the re-sialylation itself is performed in vitro, outside of the body, the treated RBCs could potentially be reintroduced into the circulation for in vivo effects. A study mentioned that after re-sialylation, the lifespan of these treated RBCs in circulation was extended twofold.
3. Use of Dextrans: Medically, dextrans are used as plasma volume expanders, antithrombotics, and in some drug delivery systems. Dextrans, particularly those with higher molecular weights, can also increase the zeta potential of RBCs when used in saline solutions. Dextrans enhance the electrophoretic mobility of RBCs, thereby increasing their zeta potential. This method is particularly effective in saline environments and can help in reducing RBC aggregation[49]. Dextrans are not intended for consumption as food. They are primarily used for medical and research purposes. Dextrans are not digestible by humans and are not meant to be eaten. They are typically administered intravenously for medical purposes.
In summary, increasing the zeta potential of blood cells in vivo can be achieved through re-sialylation, grounding, the use of dextrans, and optimizing measurement conditions. Each method targets different mechanisms to enhance the electrochemical properties of RBC membranes, thereby improving their overall function and reducing aggregation.
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