NANOBUBBLES
It is my honor and great pleasure to introduce you to the world of nanobubbles!
For the last year it has been my privilege to learn from a team of extraordinary scientists who have educated me about the omissions in the biological and medical sciences, including physical chemistry, colloid and surface chemistry, and biochemistry.
The following summary is an attempt to introduce these concepts that remain largely unknown and is based on the seminal paper entitled “The endothelial surface layer-glycocalyx - Universal nano-infrastructure is fundamental to physiology, cell traffic and a complementary neural network” by Barry W. Ninham, Nikolai Bunkin, and Matthew Battye
This paper is the fourth in a series that attempts to apply well understood, yet ignored concepts, including self-assembly driven by molecular forces, to open up insights into central areas of physiology that have been a mystery.
The titles of those papers capture some of what their latest essay is about:
The two major omissions:
1. The effects of dissolved air
2. The unknown structure and function of the endothelial surface layer-glycocalyx complex that lines all cell membranes and tissues. It is massive in extent compared with the relatively tiny phospholipid membrane of cells which it partners. It is a complex, dynamic, organized nanostructured medium that both protects the cell and directs cell traffic.
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While the authors acknowledge that these investigations are speculative, they believe they offer support to the potential of this new approach in developing new understanding of many phenomena that have until now remained elusive. They accept also that extraordinary claims require extraordinary proof. As such they have done their best to provide multiple examples of how this proposed new paradigm might be useful in closing these gaps.
Key Concepts
1. Dissolved atmospheric gas is missing from theories of physical chemistry. It plays a central role. It is delivered from the lungs as nanobubbles.
These nanobubbles of oxygen and nitrogen circulate and are stable where bubble coalescence inhibition conditions exist, as they do in physiology, i.e. (137-142 mM NaCl)
Macro bubble coalescence is prevented by different solutes, including some but not all salts, amino acids and sugars. The phenomenon is not explained by classical (macroscopic ) theory. Hydrophobic interactions do not exist in gas free conditions. Depletion forces due to nanobubbles explain coalescence inhibition
Coalescence inhibition occurs above critical concentrations levels which vary for different solutes. There is a solute specific critical nanobubble concentration (CNC) for nanobubbles in different solutes. (Salt, glucose, amino acids)
Nanobubbles nucleate through spontaneous cavitation (e.g., when an enzyme and a substrate come together).
Cavitation produces highly reactive species and free radicals as a result of a co-operative harnessing of weak short range van der Waals molecular forces. This process provides the energy to drive enzymatic reactions
2. The Glycocalyx (GC) is a dynamic self-assembled nanostructure (a porous smart medium involving bicontinuous channels) which performs or supports many of the functions presently attributed to the cell membrane and various “binding sites”, ion pumps and “receptors”.
![]()
Credit: Hans Vink
The GC acts as chemical reactor, cell transport mediator, and even an extended neural network through its modulation of the rate of transport of specific ions. A hormone like insulin can change the size and connectivity of the bicontinuous channels of the porous GC medium (change in curvature induced by surface adsorption). The GC responds similarly to any changes in local environmental conditions – from salt ion levels, dissolved gas types and concentrations, to deuterium levels to the presence of drugs and/or hormones. These physical changes in structure alter function of the ESL-GC complex.
3. The Endothelial Surface Layer (ESL), an extension of the GC structure, acts as an exclusion zone to help protect the GC and cell membrane from everything from systolic sheer forces to viruses and bacteria, to red blood cells.
What is known is that endothelial surface layer (ESL) is very large in extent, stretching out from cell surfaces 0.5 –1.0 microns. It contains a very small amount of organic matter, but nonetheless it forms an exclusion zone. The exclusion zone repels red cells, T-cells, cancer cells, bacteria, and lipoprotein complexes. The glycocalyx is 20 times less in extent than the ESL. It is about 50 nm for T cells, 20nm for red cells. It is formed predominately by several sialic acid polymers that are heavily sulfated hydrocarbons: principally heparan sulfate and chondroitin sulfate. It also includes a 10% fraction of another polymer called sodium hyaluronate. This layer is analogous to the perfluorinated polymer membrane Nafion. It too has an exclusion zone, although orders of magnitude smaller, the ESL exclusion zone is 1 micron.
The structure of the ESL and its central role in protection against many diseases have been quantified by extensive pioneering work over the years by Vink’s School in the Netherlands.
The GC we will define as the interconnected matrix of nano-structured channels that run essentially along the cell membrane, whilst the we refer to the ESL as the balance of the extracellular matrix, i.e. remaining territory from these (relatively) horizontal channels, out through the regions containing the reed-like structure teased out from the GC, the gel-like foam and the entire exclusion zone out to the blood plasma region, as in the first figure above.
4. Scale. Not only the scale of the structures we are discussing, but also the scale if you will of the forces between the ions that based on their concentrations can have significantly different effects on the curvature of GC components, resulting in fundamentally different ramifications for biological function.
Let’s compare sizes of what we are talking about and provide some context: (nm=nanometer)
So, the relative size of the lipid membrane to the thickness of the ESL (containing nanobubbles) is about 3 to 1000, about 300.
The relative size of the distance from earth of moon to that of the sun is 0.4 to 150, about 300 again.
All the pictures you see in the biomedical biochemistry books ignore size and give rise to absurd mis conceptualization.
So compared to the ESL-GC dimensions the cell lipid membrane is puny. It would be astonishing then if the “fuzzy layer” did not have a determining and dominating role in all things cellular. Unawareness of even the existence of the ESL-Glycocalyx complex has forced biologists /biochemists /biophysicists to adopt absurd, misleading models of the cell membrane that assign to it tasks impossible physically.
5. Forces. As mentioned, at critical physiological salt range (137-142 mM Na Cl) nanobubbles become stable, whilst at and above that same ionic strength, electrostatic forces between ions which provide the foundations of the theory that inform our intuition become unimportant. (pH, buffers, interfacial, tension activities, osmotic pressure). They are dominated rather by quantum mechanical dispersion forces that are highly specific
(Hofmeister effects).
Dissolved Air and Gases
Theories of the physical chemistry of solutions and colloid science do not include effects of dissolved air, oxygen, nitrogen and carbon dioxide. Nor do the medical text books and most research.
Degassing induces major qualitative changes. Some expamles of these are:
The way we utilize oxygen and how it enters red blood cells is not what we have been taught. Rather than the conventional assumption that oxygen “diffuses” through an amorphous undefined lung surfactant region to “bind” to hemoglobin, O2/N2 nanobubbles are taken up together.
In fact, inhaled air is compartmentalized and delivered to capillaries as nanobubbles of oxygen and nitrogen (20/80)%. In the lungs, nanobubble dimensions are typically of the order of 40 nm. They would not exist on inhalation without the self-organised templates provide by the lung surfactant. In circulation in capillaries, they are smaller.
Carbon dioxide is produced by metabolism. Molecular CO2 escapes through the cell membrane and forms nanobubbles by passage through the molecular frit that is the glycocalyx lining all cells and tissues.
![]()
A constantly replenished foam of CO2 nanobubbles is the main constituent of the endothelial surface layer. It extends up to a micron from the glycocalyx surface and forms a protective exclusion zone, protecting cells and tissues from red blood cells, T-cells, cancer cells, lipoproteins and pathogens, as well as the sheer forces resulting from rapid acceleration of such under systolic pressure. The CO2 nanobubbles are lethal to viruses and other pathogens. So are exhaust gases containing nitric oxide. NO is also produced from inhaled N2/O2 nanobubbles, an additional source to be recognized.
Nanobubbles
The nanobubbles are stable at physiological salt concentration, and circulate in the body as sources of O2 and NO. Further, stable CO2 nanobubbles formed by metabolism form the ubiquitous endothelial surface layer – the ESL.
Stability of the layer of nanobubbles is essential and determined by ion-pair concentrations, among other factors. Bubbles will not coalesce or fuse above a certain salt concentration, this happens to be the same concentration as our blood. This is specific to certain ion pairs. When bubbles fuse, they become larger forming macrobubbles and can cause embolism. Loss of coverage and integrity of this nanobubble layer causes a collapse of the ESL which will result in loss of defense against pathogens including cancer cells and T-cells. It will result in deposition of LDL plague deposition.
![]()
Credit: Hans Vink
It has not been recognized that nanobubbles are generalists and universal multitaskers in energy transfer.
Nanobubbles drive enzyme reactions
The O2/N2 nanobubbles access the cell interior from the blood stream through the ESL and GC [1]. These circulating nanobubbles are a major source of nitric oxide involved in driving many biochemical reactions like the tyrosine- noradrenaline- adrenaline story, the arginine paradox [1] and the mechanism of action of nitroglycerin, and, with CO2 nanobubbles the destruction of pathogens [2].
These concepts: nanobubbles existence, stability, dependence on solute, as energy sources, role of nitrogen, connection to enzymes are missing from the classical pantheon wherein an enzyme is activated by appending to a molecular name the post fix -ase. There is good reason for this. The entire venerable 200-year-old enabling disciplines of physical and colloid and surface chemistry omit dissolved atmospheric gas. This omission renders the theory and the intuition drawn from it at best misleading, and at worst useless. Once that is understood matters take on a different complexion and fall better into place.
Most improtant take aways
Maintaing integrity of the nanobubble layer and thus the endothelial surface layer glycocalix complex is key for our health.
1. Exercise - Research consistently shows increasing exercise reduces not only the severity of many diseases, but particularly diseases of affluence, none more so than diabetes. VO2 max – the measure of a body’s ability to metabolise O2, is shown to be the key predictor of longevity Regular exercise has been calculated as reducing all-cause mortality risk by as much as 5-10x, a remarkable improvement, unmatched by any other intervention, whether behavioural/dietary, or medicinal/pharmaceutical. By contrast, it is even more positively effective than the leading cause of reduction (i.e. smoking) is negative. How can such a broad range of activities provide such a universal effect? When considered in the context of the effect of the ESL-GC complex, the answer becomes clearer. Increased intake of O2 necessarily increases intake of N2, (air being 4:1 N2/O2), providing ample opportunity for increased NO production to support enzymatic reactions. Increased metabolism producing a steady flow of CO2 throughout the body, especially via large primary muscles, but also the myriad of smaller skeletal and supporting muscles as well.
It is my honor and great pleasure to introduce you to the world of nanobubbles!
For the last year it has been my privilege to learn from a team of extraordinary scientists who have educated me about the omissions in the biological and medical sciences, including physical chemistry, colloid and surface chemistry, and biochemistry.
The following summary is an attempt to introduce these concepts that remain largely unknown and is based on the seminal paper entitled “The endothelial surface layer-glycocalyx - Universal nano-infrastructure is fundamental to physiology, cell traffic and a complementary neural network” by Barry W. Ninham, Nikolai Bunkin, and Matthew Battye
This paper is the fourth in a series that attempts to apply well understood, yet ignored concepts, including self-assembly driven by molecular forces, to open up insights into central areas of physiology that have been a mystery.
The titles of those papers capture some of what their latest essay is about:
-
Structure and function of the endothelial surface layer [1]
-
Pulmonary surfactant and COVID-19: A new synthesis [2]
-
Nafion: New and Old Insights into Structure and Function [3]
The two major omissions:
1. The effects of dissolved air
2. The unknown structure and function of the endothelial surface layer-glycocalyx complex that lines all cell membranes and tissues. It is massive in extent compared with the relatively tiny phospholipid membrane of cells which it partners. It is a complex, dynamic, organized nanostructured medium that both protects the cell and directs cell traffic.
While the authors acknowledge that these investigations are speculative, they believe they offer support to the potential of this new approach in developing new understanding of many phenomena that have until now remained elusive. They accept also that extraordinary claims require extraordinary proof. As such they have done their best to provide multiple examples of how this proposed new paradigm might be useful in closing these gaps.
Key Concepts
1. Dissolved atmospheric gas is missing from theories of physical chemistry. It plays a central role. It is delivered from the lungs as nanobubbles.
These nanobubbles of oxygen and nitrogen circulate and are stable where bubble coalescence inhibition conditions exist, as they do in physiology, i.e. (137-142 mM NaCl)
Macro bubble coalescence is prevented by different solutes, including some but not all salts, amino acids and sugars. The phenomenon is not explained by classical (macroscopic ) theory. Hydrophobic interactions do not exist in gas free conditions. Depletion forces due to nanobubbles explain coalescence inhibition
Coalescence inhibition occurs above critical concentrations levels which vary for different solutes. There is a solute specific critical nanobubble concentration (CNC) for nanobubbles in different solutes. (Salt, glucose, amino acids)
Nanobubbles nucleate through spontaneous cavitation (e.g., when an enzyme and a substrate come together).
Cavitation produces highly reactive species and free radicals as a result of a co-operative harnessing of weak short range van der Waals molecular forces. This process provides the energy to drive enzymatic reactions
2. The Glycocalyx (GC) is a dynamic self-assembled nanostructure (a porous smart medium involving bicontinuous channels) which performs or supports many of the functions presently attributed to the cell membrane and various “binding sites”, ion pumps and “receptors”.
Credit: Hans Vink
The GC acts as chemical reactor, cell transport mediator, and even an extended neural network through its modulation of the rate of transport of specific ions. A hormone like insulin can change the size and connectivity of the bicontinuous channels of the porous GC medium (change in curvature induced by surface adsorption). The GC responds similarly to any changes in local environmental conditions – from salt ion levels, dissolved gas types and concentrations, to deuterium levels to the presence of drugs and/or hormones. These physical changes in structure alter function of the ESL-GC complex.
3. The Endothelial Surface Layer (ESL), an extension of the GC structure, acts as an exclusion zone to help protect the GC and cell membrane from everything from systolic sheer forces to viruses and bacteria, to red blood cells.
What is known is that endothelial surface layer (ESL) is very large in extent, stretching out from cell surfaces 0.5 –1.0 microns. It contains a very small amount of organic matter, but nonetheless it forms an exclusion zone. The exclusion zone repels red cells, T-cells, cancer cells, bacteria, and lipoprotein complexes. The glycocalyx is 20 times less in extent than the ESL. It is about 50 nm for T cells, 20nm for red cells. It is formed predominately by several sialic acid polymers that are heavily sulfated hydrocarbons: principally heparan sulfate and chondroitin sulfate. It also includes a 10% fraction of another polymer called sodium hyaluronate. This layer is analogous to the perfluorinated polymer membrane Nafion. It too has an exclusion zone, although orders of magnitude smaller, the ESL exclusion zone is 1 micron.
The structure of the ESL and its central role in protection against many diseases have been quantified by extensive pioneering work over the years by Vink’s School in the Netherlands.
The GC we will define as the interconnected matrix of nano-structured channels that run essentially along the cell membrane, whilst the we refer to the ESL as the balance of the extracellular matrix, i.e. remaining territory from these (relatively) horizontal channels, out through the regions containing the reed-like structure teased out from the GC, the gel-like foam and the entire exclusion zone out to the blood plasma region, as in the first figure above.
4. Scale. Not only the scale of the structures we are discussing, but also the scale if you will of the forces between the ions that based on their concentrations can have significantly different effects on the curvature of GC components, resulting in fundamentally different ramifications for biological function.
Let’s compare sizes of what we are talking about and provide some context: (nm=nanometer)
-
The cell lipid membrane is 3 nm thick. 3x 10-9 (billionths)
of a meter.
-
Nanobubbles of O2/N2/ CO2 approx. 2-30 nm
-
Thickness of glycocalyx helical tubes say 10-20 nm
-
Thickness of glycocalyx 50 nm for a T cell; 20 nm Red
blood cell
-
Thickness of ESL 1,000 nm
-
Thickness of red blood cell 8,000 nm
-
Thickness of gram-positive bacterial wall 80 nm
-
Size of bacteria1000 - 10,000 nm
-
[Distance of earth to sun 150 x 10+9 meters]
-
[Distance of moon to earth 0.4 x 10+9]
-
[Diameter of earth 13 x 10+6 meters]
So, the relative size of the lipid membrane to the thickness of the ESL (containing nanobubbles) is about 3 to 1000, about 300.
The relative size of the distance from earth of moon to that of the sun is 0.4 to 150, about 300 again.
All the pictures you see in the biomedical biochemistry books ignore size and give rise to absurd mis conceptualization.
So compared to the ESL-GC dimensions the cell lipid membrane is puny. It would be astonishing then if the “fuzzy layer” did not have a determining and dominating role in all things cellular. Unawareness of even the existence of the ESL-Glycocalyx complex has forced biologists /biochemists /biophysicists to adopt absurd, misleading models of the cell membrane that assign to it tasks impossible physically.
5. Forces. As mentioned, at critical physiological salt range (137-142 mM Na Cl) nanobubbles become stable, whilst at and above that same ionic strength, electrostatic forces between ions which provide the foundations of the theory that inform our intuition become unimportant. (pH, buffers, interfacial, tension activities, osmotic pressure). They are dominated rather by quantum mechanical dispersion forces that are highly specific
(Hofmeister effects).
Dissolved Air and Gases
Theories of the physical chemistry of solutions and colloid science do not include effects of dissolved air, oxygen, nitrogen and carbon dioxide. Nor do the medical text books and most research.
Degassing induces major qualitative changes. Some expamles of these are:
- Emulsions become stable.
-
“Hydrophobic” interactions disappear.
-
Polymerization, enzyme-substrate and other reactions cease.
-
Measurements of pH, pKas, activities, buffers, and molecular forces all change.
-
Cavitation stops.
The way we utilize oxygen and how it enters red blood cells is not what we have been taught. Rather than the conventional assumption that oxygen “diffuses” through an amorphous undefined lung surfactant region to “bind” to hemoglobin, O2/N2 nanobubbles are taken up together.
In fact, inhaled air is compartmentalized and delivered to capillaries as nanobubbles of oxygen and nitrogen (20/80)%. In the lungs, nanobubble dimensions are typically of the order of 40 nm. They would not exist on inhalation without the self-organised templates provide by the lung surfactant. In circulation in capillaries, they are smaller.
Carbon dioxide is produced by metabolism. Molecular CO2 escapes through the cell membrane and forms nanobubbles by passage through the molecular frit that is the glycocalyx lining all cells and tissues.
A constantly replenished foam of CO2 nanobubbles is the main constituent of the endothelial surface layer. It extends up to a micron from the glycocalyx surface and forms a protective exclusion zone, protecting cells and tissues from red blood cells, T-cells, cancer cells, lipoproteins and pathogens, as well as the sheer forces resulting from rapid acceleration of such under systolic pressure. The CO2 nanobubbles are lethal to viruses and other pathogens. So are exhaust gases containing nitric oxide. NO is also produced from inhaled N2/O2 nanobubbles, an additional source to be recognized.
Nanobubbles
The nanobubbles are stable at physiological salt concentration, and circulate in the body as sources of O2 and NO. Further, stable CO2 nanobubbles formed by metabolism form the ubiquitous endothelial surface layer – the ESL.
Stability of the layer of nanobubbles is essential and determined by ion-pair concentrations, among other factors. Bubbles will not coalesce or fuse above a certain salt concentration, this happens to be the same concentration as our blood. This is specific to certain ion pairs. When bubbles fuse, they become larger forming macrobubbles and can cause embolism. Loss of coverage and integrity of this nanobubble layer causes a collapse of the ESL which will result in loss of defense against pathogens including cancer cells and T-cells. It will result in deposition of LDL plague deposition.
Credit: Hans Vink
It has not been recognized that nanobubbles are generalists and universal multitaskers in energy transfer.
Nanobubbles drive enzyme reactions
The O2/N2 nanobubbles access the cell interior from the blood stream through the ESL and GC [1]. These circulating nanobubbles are a major source of nitric oxide involved in driving many biochemical reactions like the tyrosine- noradrenaline- adrenaline story, the arginine paradox [1] and the mechanism of action of nitroglycerin, and, with CO2 nanobubbles the destruction of pathogens [2].
These concepts: nanobubbles existence, stability, dependence on solute, as energy sources, role of nitrogen, connection to enzymes are missing from the classical pantheon wherein an enzyme is activated by appending to a molecular name the post fix -ase. There is good reason for this. The entire venerable 200-year-old enabling disciplines of physical and colloid and surface chemistry omit dissolved atmospheric gas. This omission renders the theory and the intuition drawn from it at best misleading, and at worst useless. Once that is understood matters take on a different complexion and fall better into place.
Most improtant take aways
Maintaing integrity of the nanobubble layer and thus the endothelial surface layer glycocalix complex is key for our health.
1. Exercise - Research consistently shows increasing exercise reduces not only the severity of many diseases, but particularly diseases of affluence, none more so than diabetes. VO2 max – the measure of a body’s ability to metabolise O2, is shown to be the key predictor of longevity Regular exercise has been calculated as reducing all-cause mortality risk by as much as 5-10x, a remarkable improvement, unmatched by any other intervention, whether behavioural/dietary, or medicinal/pharmaceutical. By contrast, it is even more positively effective than the leading cause of reduction (i.e. smoking) is negative. How can such a broad range of activities provide such a universal effect? When considered in the context of the effect of the ESL-GC complex, the answer becomes clearer. Increased intake of O2 necessarily increases intake of N2, (air being 4:1 N2/O2), providing ample opportunity for increased NO production to support enzymatic reactions. Increased metabolism producing a steady flow of CO2 throughout the body, especially via large primary muscles, but also the myriad of smaller skeletal and supporting muscles as well.
Here is a series of videos by Barry Ninham and Matt Battye explaining the principles:
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1. Reines BP, Ninham BW. Structure and function of the endothelial surface layer: unraveling the nanoarchitecture of biological surfaces. Q Rev Biophys. 2019 Nov 27;52:e13. doi: 10.1017/S0033583519000118. PMID: 31771669.
2. Ninham B, Reines B, Battye M, Thomas P. Pulmonary surfactant and COVID-19: A new synthesis. QRB Discov. 2022 Apr 22;3:e6. doi: 10.1017/qrd.2022.1. PMID: 37564950; PMCID: PMC10411325.
3. Ninham BW, Battye MJ, Bolotskova PN, Gerasimov RY, Kozlov VA, Bunkin NF. Nafion: New and Old Insights into Structure and Function. Polymers (Basel). 2023 May 7;15(9):221 doi: 10.3390/polym15092214. 4.PMID: 37177360; PMCID: PMC10181149.