MEDICINA INTERNA: OPEN ACCESS (MI)

ICP Plasma Technology for Alternative Medicine and New Materials Fight Against Pathogenic Infection



Atsushi Takeda1, Akio Sanpei2*


1 Integrated Surface Interaction Inc, 330-0052, Japan


2 Department of Electronics, Kyoto Inst. Technology, 606-8585, Japan.


*Corresponding Author: Akio Sanpei, Department of Electronics, Kyoto Inst. Technology, Integrated Surface Interaction Inc, 330-0052, Japan, TEL: +81-80-5424-5041; FAX: 81-48-881-7098; E-mail:4megaicp@gmail.com


Citation: Atsushi Takeda, Akio Sanpei (2018) ICP Plasma Technology for Alternative Medicine and New Materials Fight Against Pathogenic Infection. Medcina Intern 2018 2: 127


Copyright: :© 2018 Akio Sanpei, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited


Received date: December 04, 2018; Accepted date: December 12, 2018; Published date: December 17, 2018.


Abstract

We developed brand new alternative medical technology due to ICP Plasma(RFP) Technology fight against various pathogenic infections which are caused by virus, resistant bacteria and protozoa. ICP Plasma synthesized Particles have been testing as Electroadsorptive particle against various proteins and inflammatory materials. We have obtained basic Bio Assay Result as for electroadsorption of typical proteins and chemical mediators. Depend upon these result, we detected very simple interaction rule. Further, we challenged to adsorb viruses and LPS of gram negative bacteria.


Envelope type H1N1Flu virus and Non enveloped Capsid type virus has been adsorbed by RFP particle but conventional particle does not adsorbed them. LPS has already adsorbed by RFP.


Therefore, almost pathogenic protein such as viral protein and LPS layer of Drug Resistant Bacteria will be adsorbed by RFP technology no matter what kinds of mutation occurs on pathogens. RFP particle dispersed emulsion will avoid pandemic and RFP particle’s aqueous dispersion will ameliorate the serious infection and Mortal Kidney Failure.


On the other hand,RFP damages bacteria’s LPS layer due to high plasma energy. So we can design the comprehensive Pandemic-Prevention System for global Airport no matter which kinds of infection risks.


Introduction

Plasma condition is an excited high energy atomic and/or ionic gas condition which are generating by high Frequency Waves such as Induction Coupled Plasma Device(ICP), Micro Waves(MWP). Although these plasma had been developed as low pressure plasma at the beginning but very useful atmospheric pressure glow plasma(APGP) has developed due to M.Kogoma (Sophia university, Tokyo) more than 20 years ago.


Further, high energy thermal plasma had been developed for various industrial application in 1980. Plasma gas energy is very high and energy degree is shown in “eV(electron Volt)”. The lowest plasma energy will be estimated to be 1ev. Large energy plasmas are estimating more than 100eV.


These high energy ICP has been used in the field of semiconductor industry and so on. In medical fields, dermatitis treatment by low temperature plasma has been developed in Germany for ameliorate the skin disease condition.


We developed ultra dispersible submicron spherical particles by using ICP Plasma device which have 4MHz high power supply in 1989-1996 in cooperation with Neturen Inc. Japan for developing ultraviolet rays(UVR) protection at the first time depend upon European cosmetic company and United States cosmetic company. They desired abnormal property against UVR’s radiation. They demanded perfect UVR screen particle between 254nm to 405nm(UVR), but there were no ideal material at the time. By our effort, such a property was invented in 1991. So one of Japanese cosmetic manufacture had started to compound this particle into their skincare formulation by 1 % only.


We also developed higher concentrated skincare emulsion against industrial dermatitis damages such as burn, static electricity, toxic particles etc. Our customers has been wearing our products onto their skin and they detected awesome effect beyond imagination as for allergy and atopy incurable diseases very quickly. We have been reported such a amazing result from our clients so many times.


Our products which contains ICP particle by 2% in the emulsion had once saved the severest atopy dermatitis high school girl by 500grams of the emulsion without use of steroid and anti-histamine agents. We estimated the mechanism why such a rapid amelioration has been achieved. I established an unique hypothesis fight against allergen, inflammatory materials and the other microbes. ICP Plasma synthesized Ion doped Titanium Dioxide (type: MXCAI) has strong zeta potential(approx.-50mV), strongly positive charged on the surface, therefore negative molecular structure will be adsorbed on the surface of MXCAI. Stimulus COOH bonding is easily adsorbed.


As for “Active Amino Acid side chains such as NH2, ICP plasma synthesized spherical silica submicron particle (type: PFS4N) will adsorb them depend upon PFS4N’s surface negative charge. Furthermore, some patients who had been infected by bacteria and virus has been ameliorated by using the said particles. Thus, these result implied the possibility for reducing various pathogens due to the “Electroadsorption Theory” in biochemistry. Protein and/or LPS of gram negative bacteria and Peptidoglycan of gram positive bacteria.


In comparison with the said particles, plasma gas radiation against microorganisms has been testing by Japanese scientists. Because some of the microbe’s survival in space for one year in spite of the radiation. They have been testing the plasma collision effect against microbes by using Radio Frequency Plasma(RFP). According to chemical bonding energy for various bounding between C-O,H,N are distributing between 3.0-5 eV, and some of the organic bonding energy shows more than 6.0eV. Therefore, higher energy ion bombardment rapidly destroy or burn the surface layer of protein envelopes, capsid and bacteria surface layers.


In this paper, we present our Medical Plasma as alternative medicine for preventing infection and some examples. Details will be announced on the other papers because we have many experiment result as for multiple hybrid medical nanotechnology, bio assays, pharmaceutical analysis, animal test and in vitro tests. We show the essence of these result in this paper.


Experimental

Biomedical Particles Preparing due to ICP or MWP Plasmas

1) Ion Doped TiO2


By using 4MHz ICP Thermal Plasma in Commercial Plant since 1989, we produced Al-ion doped Anatase rich TiO2. Raw material is Metallic Titanium and Aluminum (3%) premixed fine grain particle, these metallic powder was supplied into thermal plasma tail on Ar gas and evaporated immediately at atomic condition. Metallic gases react with Oxygen in the plasma. TiO2 is instantly prepared and chilled rapidly so low temperature mineral assembly has occurred with specialized particle distribution which are much suitable to adsorb various pathogens not only virus but also trypanosoma.


2) Super high purity Silica


By using 5Nine Si, super high purity SiO2 was prepared by using above mentioned ICP Plasma for electroadsorption of inflammatory materials such as Histamine(H1). 5N-Si was fed into the said Oxygen/Ar plasma and instantly generated OH-free SiO2 with specified particle size distribution for avoid nano-risk against cell damage/death.


Also, we had produced high purity ultra fine silica particle depend upon MWP with organic silicon material’s evaporated gas and Ar/O2 thermal plasma. Further we have tested the production of PFS4N by using Silane(SiH4 gas). Figure 1-4.


Figure 1
Figure 1: ICP Plasma Device of Neturen Inc. Japan, 1991.
Figure 2
Figure 2: Biochemical Particle’s Preparation by ICP Plasma.
Figure 3
Figure 3: Medium Pressure He/Ar Plasma Discharge.
Figure 4
Figure 4: MWP Apparatus for PFS4N Silica (30-100nm in size).

Developed by Masuhiro Kogoma,Sophia Univ. Tokyo


3) Ion/Atom bombardment of the surface structure of Bacteria This experimental model was shown in Figure 5.


Figure 5
Figure 5: Schematic of the experimental setup (not to scale). A: 13.56-MHz RF source. B: piezoelectric vibrator. C and C’: powered and grounded electrode, respectively. D: illuminating laser (30 mW at 532 nm). E: microorganisms in plasma. F: aluminum foil. G: source of micro-organisms.

The treatment of the micro-organism sample was as follows: after dissolving the strains of micro-organisms with ultra pure water, we sterilized it by steaming it under pressure for 15 min at a temperature of 121 °C; it was confirmed that each micro-organism maintained its shape with staining; after centralization, we let the supernatant solution dry naturally; finally, the samples were broken up into smaller pieces with ultrasonic cleaning equipment, because the micro-organisms tend to aggregated as a consequence of the outlined treatment.


4) Basic Bio Assay for Proteins and some Inflammatory materials by MXCAI (Ion doped TiO2)/PFS4N (high purity Silica)


We have been collaborated with Japanese universities for carry out the bio assay examination as below.


1) Histamine adsorption: Prof.Dr.K.Maeyama, Ehime university, School of Medicine


2) Protein Adsorption: M.Mio, Ph.D,Prof.of Faculty of Pharmacology,Syujitsu University,


3) LPS Adsorption and other analysis: N.Hirasawa, Ph.D Prof. of Pharmaceutical faculty, Tohoku university.


4) Associated Analysis and animal test : H.Fukui, Medical doctor, Prof.of Pharmaceutical Science of Tokusima univ.


5)Consultation and advice: Medical Dr. M.Makino, National Sanatorium, Oku-Komyoen Hospital,


We selected some proteins for the adsorption test depend upon “Isoelectric point(IEP)” which is controlled by molecular structure.


Further, hydrophobic fluorescent material which has COOH at the tip of the molecule was selected.


To evaluate inflammation reducing effect, two steroid has tested as control sample.


5) Real adsorption of Fluvirus ( Type A Fluvirus)


This test has done at Tohoku University School of Medicine. Details has not translated into English but we prepared some TiO2 samples and examined the adsorption test by using “Living Active Type A Fluvirus.


(1) Particle concentration ratio: 10mg/mL


(2) N=1 as a pilot test


(3) 30min.incubation and 18,800g 4C 1hour centrifugal separation


(4) Final Result: Value is shown as TCID50 (unit by log.)


6) Human test for reducing Infection caused severe Inflammation


This test has done due to the requirement of the said patient who damaged small tick-bites because of no cure method.


We tried to reduce severe skin disease on foots, arms, back, etc. by using special compounds which are premixed with plasma synthesized particles (Al-Doped TiO2(MXCAI) or SiO2(PFS4N). These emulsion are formulated as Oil in Water type. We tested the emulsion which is not compounded with the said particle due to ulcer amelioration animal test.


As for ulcer amelioration, these emulsion has tested against burn by using animal.


Further, we examined a severe ulcer caused by Behchet’s Disease by using above mentioned emulsion and MXCAI.


Result

Biomedical Particles Preparing due to ICP or MWP Plasmas

1) Ion Doped TiO2, MXCAI


MXCAI particles are almost spherical particle and particle and mean particle size is approx.150nm(min.20nm,max.3000nm) Zeta potential at ph=6.0 is approx.-50mV (Control TiO2:nearly 0 mV) Figure 6,7.


Figure 6
Figure 6: TEM Picture for MXCAI Particle.
Figure 7
Figure 7: HREM Crystal Analysis for MXCAI.

Figure 7. Aluminum Ion doped MXCAI(TiO2) lattice structure is Perfectly new material, conventional TiO2 is poor property for biomedical application due to toxicity. Figure 8.


Figure 8
Figure 8: Aluminum ion doped MXCAI’s UVR screen profile (The definite material against Skin Cancer and DNA Protection).

PFS4N Plasma Synthesized ultra high purity spherical Silica

By using 5Nine Silicon powder, super high purity SiO2 was prepared by using above mentioned ICP Plasma for electroadsorption of inflammatory materials such as Histamine(H1). 5N-Si was fed into the said Oxygen/Ar plasma and instantly generated OH-free SiO2 with specified particle size distribution for avoid nano-risk against cell damage/death.


Figure 9 TEM Picture of PFS4N which has no impurities such as organic substances for dispersibility, The only material SiOH free surface and have negatively charged ultra dispersible particle. Toxins due to fatal bacteria is able to be adsorbed in colon.


Figure 9
Figure 9: TEM Picture of PFS4N which has no impurities such as organic substances for dispersibility, The only material SiOH free surface and have negatively charged ultra dispersible particle. Toxins due to fatal bacteria is able to be adsorbed in colon.

Figure 10 FTIR data for PFS4N, Ultra high adsorption effect for H2O and CO2 gas, SiOH freeness and new SiO2 has invented due to IR’s Absorption position. Figure 11.


Figure 10
Figure 10: FTIR data for PFS4N, Ultra high adsorption effect for H2O and CO2 gas, SiOH freeness and new SiO2 has invented due to IR’s Absorption position.
Figure 11
Figure 11: PFS4N(SiH4) FTIR data shows the SiOH absorption.

Plasma treatment of Bacteria

By using 2 types of bacteria, plasma treatment has done due to the following model. Figure 12-15.


Figure 12
Figure 12: Schematic model for RF plasma treatment for leviating bacteria.
Figure 13
Figure 13: bacillus micro-organisms.
Figure 14
Figure 14: SEM image of gram-negative bacteria (Klebsiella pneumoniae) Before experiment.
Figure 15
Figure 15: SEM image of Surface LPS layer of Klebsiella pneumoniae After 1 hour exposed.

Typical Bio Assay for P-roteins and Organic material

These tests has been done by the financial assist of Japanese Government and detail has already uploading on JSP’s web site in Japanese language, Editor M.Mio et al. [1]


Here, we will explain two analysis result, further information should be on request. Bio assay was done by pharmaceutical group by using Al doped TiO2 and high purity SiO2. These data have been reported on 18th International Symposium Plasma Chemistry, Kyoto(ISPC18) and we will report further new data about adsorption property between organic fluorescent substance which has COOH function group and particles.


4-1) Basic IEP showing Protein’s Electroadsorption result for Cytochrome C(CYCS), Heme protein having NH, NHR in molecule and IEP is basic. Figure 16,17.


Figure 16
Figure 16: Adsorption of CYCS test By PFS4N (-) charged.
Figure 17
Figure 17: Result for the same by using MXCAI (+) charged.

Protein adsorption in buffer solution


In the case of OVA, the UV absorption spectra of OVA at 0.05-10 mg/ml were not affected by silica(PFS4N) nano-particles. From these observations, we concluded that OVA do not bind to silica nano-particles at the pH and ionic conditions used in the present study. On the other hand, when BSA was incubated with silica nano-particles, 0.05 mg/ml of BSA was almost completely adsorbed by 10 mg/ml of PFS4N nano-particles and the UV absorption around 280 nm of BSA almost completely disappeared (Figure 18). When the concentration of BSA was raised to 0.25 mg/ml and 0.5 mg/ml, UV absorption around 280 nm of BSA was diminished by incubation with PFS4N nano-particles. Cytochrome C(CYCS), the solution of which is red, has a broad spectrum in the ultraviolet area [2]. When 0.25 mg/ml of CYCS was incubated with 10mg/ml of PFS4N nano-particles, CYCS was almost completely adsorbed by CYCS, as seen in the absorption spectrum (Figure 19). In the same way, when 0.5 mg/ml of CYCS was incubated with 10 mg/ml of PFS4N nano-particles, CYCS protein was almost completely precipitated with PFS4N nano-particle (approx.200nm) (Figure 16). In the spectra, a residual peak (about 220nm) was assigned as that of the buffer material (MES). nano-particles. Although we tried to use much higher concentrations of cytochrome c, the UV absorption became too high to determine the changes in the absorption spectra. In the case of histamine, weak adsorption in phosphate buffer (pH 6.5) was observed (data not shown).


Figure 18
Figure 18: absorption spectra of BSA(0.25mg/ml) with PSF4N Silica nano-particles(10mg/ml) in buffer water.
Figure 19

By the similar Bio Assay examination, we have detected the relation between Plasma Synthesized Particles and Proteins as follows. These relation are depending upon IEP of each materials. But these relation will not be applied to Conventional Inorganic Particles(CIP) and Non-Effective Plasma Method(NEP) by using low frequency power generator such as 500KHz. Table 1.


Table 1: Electroadsorption Behavior of PFS4N and MXCAI 4-2, Adsorption test for hydrophobic fluorescent material. Table Absorption behavior of SiO2 and TiO2 nano-powders


Substances Classification (charge) Iso electric point/pH Results in SiO2 Results in TiO2
OVA Egg protein (-) 4.5 no excellent
BSA Cow blood serum (+) 5.3 good -
Cyiochrome c Hema protein (+) 10.5 excellent no
SiO2 Inorganic (-) 1.8-2.5 - -
TiO<>2 Inorganic (+) 6.0 - -

• To demonstrate such mechanism, we measure the proteins absorption on the TiO2 nano-powder that is produced in ICP plasma. Because TiO2 powder has positive surface charge that is reverse charging with silica powder.


4-2) Adsorption test for hydrophobic fluorescent material


Adsorption result are shown in Figure 20 and in this test, PFS4N and MXCAI also adsorbed BODIPY 500/510. COOH was adsorbed by MXCAI (Ion doped TiO2) and hydrophobic group was adsorbed by PFS4N.


Figure 20
Figure 20: Florescent emission for PFS4N and MXCAI.

High purity ultra dispersible Plasma Synthesized Silica nano-particle which has adsorbed the said florescent material shows fluorescent emission by UVR radiation, Although MXCAI shows extinction of florescent emission due to static removing [3].


These result suggest us the significant importance for the electroadsorption of various pathogens such as virus, bacteria and protozoans, especially as for Glycoprotein of viral envelope and protozoan such as Trypanosomas.


4-3) Real adsorption test against active virus


Real adsorption of Fluvirus (Type A Fluvirus) This test has done at Tohoku University School of Medicine. Details has not translated into English but we prepared some TiO2 samples and examined the adsorption test by using “Living Active Type A Fluvirus.


(5) Particle concentration ratio: 10mg/mL


(6) N=1 as a pilot test


(7) 30min.inculcation and 18,800g 4C 1hour centrifugal


8) Final Result: Value is shown as TCID50(unit by log.)


1) TiO2 particle size: 4groups have been obtained such as 20-100nm, 100-300nm, 300-600nm, -2micron meter. Mean particle size is approx.150nm.


2) SiO2 particle size: ditto. Table 2.


Table 2: viral adsorption result for plasma synthesized particles Analyzed result: shown in Logarithm.


Influenza A Virus RNA Envelope (+) Entevovirus RNA Envelope (-)
Control 4.1 Control 4.1
PFS4N Not Detective PFS4N 3.8
MXCAI Not Detective MXCAI Not Detective

5 Human test for reducing Infection caused severe Inflammation


5-1) Test for Infection caused abnormal inflammation


This test has done due to the requirement of the said patient who damaged small tick-bites. There is no remedy method. We tried to reduce severe skin disease on foots, arms, back, etc. by using special compounds which are premixed with plasma synthesized particles (Al-Doped TiO2(RCP), SiO2(FC), RCP and FC must not be mixed. Finally we detected the best item fight against the severest inflammation and Anaphylaxis shock [4].


PFS4N 4-4% compounded oil in water emulsion(FC) was the best to reduce swollen legs and arms, inflammation, itch and pain. Plasma synthesized particles extracted lymph fluids through Aquaporin(AQP) so much and ameliorated the symptom.


Histamine H1 has been reduced by electroadsorption due to PFS4N particle and abnormal inflammatory materials which binds with NH,NH2 has removed. Figure 21.


Figure 21
Figure 21: A result for the amelioration against the severest inflammation and swelling caused by tick bites, 69year old woman.

5-2) Typical insect bite management to reduce pathogens.


This test has done by mosquito’s bite to the skin. Recently some serious infections has been spreading in the whole world. Vaccination and vaccine itself has not been yet completed [5]. Therefore, we have to protect by ourselves against small insects.


This test has done with PFS4N and/or MXCAI 3% compounded emulsion. Result is shown in Figure22. Lymph fluid were able to be excluded very quickly and itch is removed within 2-30min.


Figure 22

Discussion

Fatal Infection and various damages against internal organs are spreading into the world. Although so many morphological analysis of various pathogens such as viruses, bacteria and trypanosoma have been disclosed but there is no systematic textbooks as for the surface details. So, we had to seek out the real protein structure of them. For example, many viral structure are divided into two groups. As for bacteria, there are 2 types roughly. Almost pathogenic surface is formed by special protein group, but biochemically we decided the tip molecule bonding to COOH (Negatively charged). The most typical structure of this type is shown in Fluvirus (Envelope type).


According to highly charged surface condition of Plasma Synthesized Biochemical Particle, the said proteins which has COOH group at the tip of viral envelope layer is easily adsorbed by “Positively charged Ion Doped TiO2” as biochemical particle. This inference has been obtained due to prior bio assay examinations as for protein-adsorption. In case of bacteria, gram negative bacteria has LPS surface and LPS charged to be negative, and gram positive bacteria charged positive because of lack of LPS* layer. Trypanosoma is covered with Glycoprotein envelope which is similar to viral envelope protein. Therefore we can adsorb “Almost Fatal Pathogens(FP)” no matter what type, species they are. This “Electroadsorption” is a basic biochemistry theory. As for inflammation removing, we have already proved the said particle’s efficiency and safety.


So many fatal viruses causes severe dangerous fatal inflammation, so our Thermal Plasma Technology may conquer the world. We would like to propose that new prevention system and a ideal canister which attached with ICP or RF Plasma device to transport the dangerous patient at airport or aircraft. Plasma energy will damages pathogenic proteins and RNA/DNA also.


Further, pathogens particles will be adsorbed on the surface of MXCAI immediately. Further, Drug Resistant Bacteria will be adsorbed by MXCAI or PFS4N no matter what mutation generated onto the surface layer. The most important things is the possibility of binding and detoxification of endotoxin of bacteria such as cholera toxin. Viral infection causes rapid kidney failure like bacteria, but MXAI will reduce them. MXCAI has removed Indole(C8H7N) and/or Skatole(C9H9N,C8H6NCH3) perfectly.


We have to take great care of ICP or RF Plasmas which have high frequency power supply because low frequency plasma (less than 1MHz) had never obtained the effective crystals yet. We suggest following important schematic morel fight against viral pandemic. Figure 23.


Figure 23
Figure 23: Electroadsorption model for Envelope (+) (-) virus and viral proteins.

Almost viruses have COOH tips on the surface layer (partly mutated and NHx binding), so MXCAI-type special TiO2 can adsorb them however these virus are capsid type or mutated type.


Conclusion

For reducing the various infection risk in the whole world, we have been developing very unique materials and infection and inflammation preventive method due to medical nano technology based on “Electroadsorption between biomedical particle and pathogens. Although ICP system could developed amazing result against viral infection and infection causing serious inflammation, but huge amount production by cheaper cost are desired by medical doctors. So we have to develop a new system for producing the said materials as soon as we can. Therefore, we have started with some possible method for saving tremendous people in Africa and developing countries. Further, very dangerous “Droplet Infection” due to intense cough due to infected patients will be ameliorated by reducing Leukotoriene (LT) due to Plasma Synthesized Silica(4% compounded emulsion).


References

  1. Kogoma M, Takeda A (20011) Generation and Application of Atmospheric Pressure Plasma. Nova Science Pub Inc. ISBN-10:1612097170
  2. Takeda A (2017) Medical Evidence for Biochemical Plasma Particle against Pathogenic Infection and Infection caused Inflammation’s Reducing Human Experiment. 18th Workshop of Fine Particle Plasmas, textbook
  3. Mio M, Takeda A, et al. (2010) A research for Inflammation reducing due to the absorptive material against inflammatory materials. New Technology Presentation Meetings, 29thJan.2010, Japan Science and Technology Agency. textbook 11-16
  4. Takeda A, Thin Solid Films (2003) Structure of metal doped TiO2 particles produced by RF plasma, 435: 211-214.
  5. Sanpei A, Hayashi Y (2018) IEEE TRANSACTIONS ON PLASMA SCIENCE, Levitation of Microorganisms in the Sheath of an RF Plasma,718-722, VOL.46, NO.4 .