FROM THE IDEA TO THE PROTOTYPE
Plasma Life-Science Applications
R. Brandenburg, Th. von Woedtke, K.-D. Weltmann
Leibniz Institute for Plasma Science and Technology
Greifswald, Germany
With contributions from partner and colleagues in the network project:
2
H.M. Mott-Smith; Nature, 233, p. 219, (1971)
„ ... the discharge acted as a
sort of substratum carrying
particles of special kinds […]
This reminds him of the way
blood plasma carries around
red and white corpuscles and
germs. So he proposed to call
our ‚uniform discharge‘ a
‚plasma‘. Of course we all
agreed.“
First use of the term „plasma“: Langmuir (1928)
Langmuir‘s colleague
Harold M. Mott-Smith
remembers in 1971:
Credit:NicoleRager-Fuller,NSF
Irving Langmuir
(1881–1957)
at General Electric
Laboratory (1948)
Plasma for biomedical applications
Surface modification
Biocompatibility
Antifouling
Microfluidics
Cell culture systems
Plasma Medicine
“classical”: surgical applications
(coagulation or ablation of tissue)
new: wound and tissue treatment (“plasma
can heal”)
© INP
Erbe Elektromedizin
Bio-Decontamination/Sterilization
Inactivation of pathogens and microorganisms
on sensitive products (disinfection, sterilization)
as well as gases and liquids
Protection against biological warfare agents
© INP
4
Plasma life science
5
Implants
bone implants (joints, teeth)
tendons and ligaments
vascular grafts
stents
heart valves
Therapy devices
catheters
dialysis devices
pacemakers
Disposables
cell culture dishes
DNA chips
biosensors
high throughput screening systems
Vacuum Plasma Spraying of
titanium
DOT GmbH
DOT GmbH
Corrosion protection by PVD
Functionalized titer plates
Plasma
in use
Surface modification for biomedical applications
12
Plasma life science
13
Decontamination / Sterilization: some definitions
Decontamination:
Removal of dangerous contaminations
CBNR (Chemical, Biological, Radiological, Nuclear)
includes prions, pyrogens, viruses, chemical contaminations etc.
Antimicrobial Treatment (R. Koch, 1881):
Inactivation of microorganisms with the
aim to reduce/avoid infections
Disinfection:
to put dead or living material in a situation, that it is no longer able to
contaminate
Practical advise: reduction of infective micro organisms including viruses
Sterilization:
„Sterility is the absence of viable micro organisms.“ (Ph. Eur., USP)
Sterility Assurance Level SAL= 10-6: not more than one viable microorganism
in one million sterilized items of the final product
Robert Koch
(1843-1910)
14
Alternatives for heat sensitive products
Ionising radiation sterilisation (gamma radiation – radioisotopic source,
electron beam – electron accelerator): 25 kGy
Gas sterilisation (ethylene oxide)
LTSF - Low-temperature steam/formaldehyde
Chemical wet sterilisation (glutaraldehyde, peracetic acid, hydrogen
peroxide)
VHP - Vapor-phase hydrogen peroxide: hydrogen peroxide, peracetic
acid
UV irradiation (254 nm)
Sterilisation/decontamination procedures
Standard and reference procedures
Steam sterilisation (heating in an autoclave): 121°C, 2 bar, 15 min
Dry heat sterilisation : ≥ 160°C, ≥ 2 h
16
Specific antimicrobial effects of plasmas
Antimicrobial effect of plasmas
proven in many experiments with different plasma sources
but effects very specific
+ +
Plasma
parameters, components
Lethality of micro org.
e.g. UV sensitivity/resitivity
Preparation of samples
e.g. arrangement of micro org.
MiBi procedures
e.g. detection limits, artefacts
17
Biologically active components of plasmas
(V)UV-radiation
damage of DNA
cell wall damage by photo-induced erosion
Global Effects
Temperature surface heating
Electric Fields electroporation
Radicals and Chemical products
cell wall damage by reactive etching (O, O2*)
DNA damage by oxidation
oxidation of proteins (O)
oxidation of lipid bilayers (fatty acids) (OH)
O3 cell respiration
natural signal stimulation (NO)
Charged particles (ions)
electrostatic pressure by accumulating charge
carriers on membrane charge lysis of cells
Plasma-cell interactions: various hypotheses
D. Dobrynin, G. Fridman, G. Friedman, A. Fridman, Physical and
biological mechnaisms of direct plasma interaction with living
tissue. New J. Phys 11 (2009) 115020
G. Fridman et al., Plasma Chem. Plasma Process. 26 (2006) 425
19Philip et al., IEEE Trans. Plasma Sci. 30 (2002) 1429
Low-pressure afterglow plasma, 915 MHz/2.45
GHz, 100-200 W, N2-O2 gas mixtures; spores
dried on a polystyrene surface
UV radiation
+
Erosion caused by reactive species
Low-pressure plasma
19
23R. Brandenburg, J. Ehlbeck, M. Stieber, Th. von Woedtke, J. Zeymer, O. Schlüter,
K.-D. Weltmann, Contrib. Plasma Phys. 47 (2007) 72-79
PE strip
2cm
Atmospheric pressure RF Argon Plasma jet
(27 MHz, 10…60 W)
Bacillus atrophaeus spores, dried on PE strips; Separation of plasma compounds by
quartz window or hot gas flow
Direct plasma
treatment
quartz
glass
plate
PE strip
UV only
PE
strip
Heated air flow
Q=20 slm
T=80-90°C
Heat only
Atmospheric-pressure plasma
24
R. Brandenburg, J. Ehlbeck, M. Stieber, Th. von Woedtke, J. Zeymer, O. Schlüter,
K.-D. Weltmann, Contrib. Plasma Phys. 47 (2007) 72-79
Treatment time (min)
0 1 2 3 4 5 6 7 8 9 10 11
CFU/strip
101
102
103
104
105
106
107
detection limit
plasma
UV only
Hot air only
(Median, error bars: Min./Max.; je n=5)
Atmospheric pressure RF Argon Plasma jet
(27 MHz, 10…60 W)
Bacillus atrophaeus spores, dried on PE strips
Separation of plasma compounds by quartz window or hot gas flow
Atmospheric-pressure plasma
28
Indirect surface DBD on air: influence of humidity
B. Atrophaeus spores dried on PE strips
M. Hähnel, Th. von Woedtke, K.-D. Weltmann, Plasma Process. Polym. 7 (2009) 244-249
Influence of humidity
29
Specific antimicrobial effects of plasmas
Antimicrobial effect of plasmas
proven in many experiments with different plasma sources
but effects very specific
+ +
Plasma
parameters, components
Lethality of micro org.
e.g. UV sensitivity/resitivity
Preparation of samples
e.g. arrangement of micro org.
MiBi procedures
e.g. detection limits, artefacts
30
Quantifiable effectivity of an antimicrobial process depends on nature and resistance
of the microorganisms used.
Use of test microorganisms with maximum resistance against the process to
be examined.
Bacterial spores vs. vegetative microorganisms
http://micro.magnet.fsu.ed
u/cells/procaryotes/images
/procaryote.jpg
# Resistance of microorganisms
Quantification of sterilisation effects
http://www.micro.cornell.
edu/cals/micro/research/l
abs/angert-
lab/images/endospore.jpg
Specific antimicrobial effects of plasmas
Different resistance against external factors:
heat, UV, chemicals, plasma components, …
High resistance against external factors:
Bioindicators for sterilization control
Size: 0.1-1 µm
32
Plasma treatment time (min)
0 2 4 6 8 10
cfu/sample
101
102
103
104
105
106
107
Bacillus atrophaeus spores
Escherichia coli
Staphylococcus aureus
detection
limit
Rf-driven Plasma Jet, 20 W, Argon 20 slm
Microorganism resistance
33
Specific antimicrobial effects of plasmas
Antimicrobial effect of plasmas
proven in many experiments with different plasma sources
but effects very specific
+ +
Plasma
parameters, components
Lethality of micro org.
e.g. UV sensitivity/resitivity
Preparation of samples
e.g. arrangement of micro org.
MiBi procedures
e.g. detection limits, artefacts
34
Bacterial load
Spray: 2 104 MO/cm2
Spot: 1.4 107 MO/cm2
... more or less single
spores on surface
... stacked spores
(several layers)
F. Rossi et al., Plasma Process. Polym. 3 (2006) 431; J. Wunderlich et al. ISPC 2003
Rossi et al.
Identical plasma treatment leads to
different lethal effects!
36
Substrate material
Low-pressure glow-discharge oxygen RF plasma, 27.12 MHz, 300 W, continuous and pulse mode (5 s on,
25 s off) performance; forced-air cooled discharge vessel; B. subtilis spores dried on glass and Al-foil
Cvelbar et al., J. Phys. D: Appl. Phys. 39 (2006) 3487
Different substrate heating caused by surface
recombination of O-atoms
Combined effects of temperature and O-based
atom-by-atom etching of bacteria
37
Specific antimicrobial effects of plasmas
Antimicrobial effect of plasmas
proven in many experiments with different plasma sources
but effects very specific
+ +
Plasma
parameters, components
Lethality of micro org.
e.g. UV sensitivity/resitivity
Preparation of samples
e.g. arrangement of micro org.
MiBi procedures
e.g. detection limits, artefacts
38
Ph. Eur. [USP, JP]:
„Sterility is the absence of viable micro-organisms.“
Plasma “sterilisation“
What does it mean: sterility/sterilisation?
Sterility Assurance Level - SAL
10-6
denotes a probability
of not more than one viable micro-organism
in 1 × 106 sterilised items
Problem: Proof of sterility assurance
40
Exponential microorganism inactivation kinetics
Treatment intensity [x]
(time, dose, concentration, ...)
Numberofviablemicro-organisms[N]
0
200000
400000
600000
800000
1000000
Numberofviablemicro-organisms[N]
10-6
10-5
10-4
10-3
10-2
10-1
100
101
102
103
104
105
106
Treatment intensity [x]
(time, dose, concentration, ...)
Microbiological tests
general assumption!
41
Direct cell counting (spread-plate method)
Spread on culture medium
(agar) and incubation
Dilution series with
100 µl aliqout
Agitation in 10 ml 0.9 % NaCl
Treated test objects
(source: SÜßMUTH/EBERSPÄCHER/HAAG 1987)
Detection limit:
101-102 CFU/obj.
... applied if viable micro organisms present on all test objects
saline solution
42
Membrane filtration
Agitation in 10 ml 0.9 % NaCl
Membrane filtration
Filter on culture medium (agar)
and incubation
Detection limit:
1 cfu/obj.
... applied when less than a few hundred viable micro organisms on test object expected
Treated test objects
saline solution
43
Fraction negative method
Detection limit:
10-1 ... 10-2 cfu/obj.
... applied if test objects both with and without viable micro organisms expected
Incubation in nutrient solution
(broth)
turbidity of broth gives evidence
for microbial growth
m = - ln(n0/n)
m ... mean number of survivors per
test object
n0 ... number of test objects without
viable micro organisms
n ... number of identically treated
test objects n
Treated test objects
nutrient solution
44
0 2 4 6 8 10 12 14 16
Numberofviablemicroorganisms[N]
10-6
10-5
10-4
10-3
10-2
10-1
100
101
102
103
104
105
106
Extrapolation
Fraction negative
method
Direct cell counting
Treatment intensity [X]
Limits:
„Sterility is the absence of viable microorganisms.“
→ Sterility Assurance Level SAL= 10-6 (Ph. Eur.)
→ practical proof: RF ≥ 6 lg
Ideal:
linear inactivation
kinetics
↓
continuous effectivity
Real:
discontinuous
effectivity
Plasma-“Sterilisation“
45
Plasma treatment time (min)
0 2 4 6 8 10
cfu/sample
101
102
103
104
105
106
107
Bacillus atrophaeus spores
Escherichia coli
Staphylococcus aureus
detection
limit
Real inactivation kinetics
PE strip
2cm
Rf-driven Plasma Jet, 20 W, Argon 20 slm
48
Other non-thermal treatments
2% 3 % 4%
Glutaraldehyde treatment time [h]
0 1 2 3 4 5
meannumberofviablespores
pertestunit
10-2
10-1
100
101
102
103
104
105
106
107
2% 3 % 4%
UV irradiation time [s]
0 5 10 15 20 25 30
Numberof
colonyformingunits(cfu)perml
100
101
102
103
104
105
106
107
108
109
UV irradiation of
B. atrophaeus spore suspensuions
Direct cell counting
mean ± SD
n=10 per data point
Treatment of
B. atrophaeus spore strips
by glutaraldehyde solution
Fraction negative method
m = - ln (n0/n)
n=20 per data point
von Woedtke & Jülich, Pharmazie 56 (2001) 561
von Woedtke et al., Biosens. Bioelectron. 17 (2002) 373
von Woedtke et al., J. Hosp. Infect. 55 (2003) 204
49
Heat-based processes
• global excited energy state
• lethal changes of micro-organism structures resulting from a
constant and ubiquitous activity
⇒⇒⇒⇒ inactivation kinetics independent from number of
targets →→→→ extrapolation possible
Non-thermal processes
• discrete energy quanta or reactive molecules
• lethal changes of micro-organism structures resulting from „hits“ at
susceptible target structures
⇒⇒⇒⇒ inactivation kinetics strongly dependent from number of
targets →→→→ extrapolation problematic
Theory of antimicrobial activity
50
use of most resistant test micro-organisms
documentation of inactivation kinetics over ~ 5 log
first of all in the range of low contamination rates
(down to 10-2 if possible)
Resulting practical demands
for plasma „sterilization“
Proof of antimicrobial efficacy
on the highest experimentally accessible level
Plasma treatment time (min)
0 1 2 3 4 5 6 7 8 9 10 11
cfu/sample
101
102
103
104
105
106
107
Bacillus atrophaeus spores
51
Sterilisation and decontamination using low-pressure plasmas
52
Commercially available “plasma sterilization”:
Sterrad® Sterilization Systems
(Advanced Sterilization Products, Ethicon Inc., Johnson & Johnson)
Sterrad®
100S
1
2
3
4
5
Wrapped Samples
Trays
Plasma Region
Matching Network
Perforated Powered Electrode
6
7
8
9
To Vacuum Pump
Vacuum Chamber (grounded)
Pressure Gauge
H2O2 Vapour Injection Port
Lerouge et al., Plasmas and Polymers 6 (2001) 175; Moisan et al., Int. J. Pharm. 226 (2001) 1; http://www.aspjj.com
Sterilisation and decontamination using low-pressure plasmas
53
Commercially available “plasma sterilization”:
Sterrad® Sterilization Systems
(Advanced Sterilization Products, Ethicon Inc., Johnson & Johnson)
Sterrad®
100S
1
2
3
4
5
Wrapped Samples
Trays
Plasma Region
Matching Network
Perforated Powered Electrode
6
7
8
9
To Vacuum Pump
Vacuum Chamber (grounded)
Pressure Gauge
H2O2 Vapour Injection Port
Lerouge et al., Plasmas and Polymers 6 (2001) 175; Moisan et al., Int. J. Pharm. 226 (2001) 1; http://www.aspjj.com
inherent efficacy is above all due to hydrogen peroxide
plasma phase: residue detoxifying process
Krebs et al., Int J. Pharm. 160 (1998) 75
Sterilisation and decontamination using low-pressure plasmas
54
Commercially available atmosperic pressure plasma system:
TipChargerTM (Cerionx Inc., Rxton, PA, USA)
www.cerionx.com
low-temperature ('cold') atmospheric pressure plasma
(DBD) used to clean and sterilize pipet tips by removing
organic substances from treated surfaces.
Sterilisation and decontamination using atmospheric pressure
plasmas
55
additional features of atmospheric pressure plasma:
contracted plasmas
effects are limited locally
effects can be located exactly
Plasma-“Sterilisation“
56
INP
INP
300 mm
150 µm
gap
Chances of atmospheric pressure plasmas:
- customisation according to special product characteristics,
demands, geometries
Plasma-“Sterilisation“
INP
www.endoplas.de
5757
‘Sterilization’ of heat sensitive products
Medical devices (instruments, implants)
Pharmaceutical packagings
Pharmaceutical preparations
http://www.nanobionet.org
INP
INP
INP
INP
Vanguard AG INP
INP/dpa
INP
INP INP
INP
Plasma technology for biological decontamination
2004: Beginn der Verbundprojekts „PLASMOSE – Plasmagestützte
Oberflächenmodifizierung mittels modularer selektiver Plasmaquellen“
58
Endoscopes as medical instruments
control unit
light and video
connectors
distal end
tubular
device
4 mm
light guidecamera light guide
tubular device
Plasma inside closed packaging
- Adapted Dielectric Barrier Discharge (DBE)
Plasma ignited inside of the packaging
- Prooven by FTIR-Gasanalyses within packaging
65
Chances and perspectives:
1. Antimicrobial effectivity of atmospheric pressure plasmas against a
broad spectrum of microorganisms was proved in many cases
3. Several possibilities to integrate atmospheric pressure plasma sources
as part of special product and/or process-related master plans of
(micro-) biological decontamination, above all for manufaacturing and
(re-)processing of medical devices
4. Plasma treatment of liquids expandable:
microbiological decontamination
Investigation of biological plasma effects
2. Ubiquitous applicable sterilization procedures based on atmospheric
pressure plasma are hardly to realize
5. Huge potential in the field of in vivo antiseptics → plasma medicine
Sterilisation and decontamination using atmospheric pressure
plasmas
Plasma life science
„Recent demonstrations of plasma technology in
the treatment of living cells, tissues, and organs
are creating a new field at the intersection of
plasma science and technology with biology and
medicine – Plasma Medicine.“
M. Laroussi & A. Fridman, Editorial
Plasma Process. Polym. 5 (2008) 501
Plasma medicine
67
68
‘Violet Rays’ or ‘Violet Wands’ & the Zeileis method
www.electrotherapymuseum.com
Antique 'quack' medical devices
claimed to be useful in electrotherapy
The Zeileis method (1912)
– HV treatment
1 H. BURGER: The Doctor, the Quack, and
the Appetite of the Public for Magic in
Medicine,Proceedings of the Royal Society
of Medicine, 1.11.1933 [1]
2 Zeileis-Institut, Valentin-Zeileis-Straße 33,
A-4713 Gallspac
„Conventional“ surgical plasma applications
Zenker M, GMS Krankenhaushyg Interdiszip 3 (2008) Doc15;
Raiser & Zenker, J. Phys. D: Appl. Phys. 39 (2006) 3520
Argon Plasma Coagulation (APC)
ERBE Elektromedizin GmbH,Tübingen, Germany
Coblation® (cold ablation)
ArthroCare Corp., Synnyvale, CA,USA
Stalder KR, Woloszko J, Contrib Plasma Phys 47 (2007) 64;
www.arthrocareent.com/
Plasma medicine: step forward
lethal plasma effects
(antimicrobial; surgical)
Selectivity of plasma treatment
Plasma can heal!
+
non-lethal plasma effects
Chronic wounds
4.5 to 5 million people in Germany suffer from chronic wounds
i.e. ca. 5% of all stationary patients in hospitals and rehab hospitals
loss quality of life for patients
health economic problem: > 5 Mrd. € per year in Germany
increasing problem because of ageing population
e.g. pressure ulcers, venous insufficiency ulcers, diabetic/neuropathic ulcers
Actual Focus: Wound Healing
Wound antiseptics
Chronic wounds (no healing within 8 weeks)
Frequent cause: wound infections
Staphylococcus aureus, Staphylococcus epidermidis
Streptococcus pyrogenes, MRSA
Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa
anaerobians
expenditure of time,
adverse effects
!
Conventional therapy: chemical antiseptics
74
Plasma Medicine: vision
Superficial cleaning and antiseptics
+
Stimulation of tissue regeneration in deeper layers
Integrated concept of plasma-based wound treatment
Plasma-cell interactions
PLASMA CELLSGAS PHASE LIQUID PHASE
76
78
Experimental Plasma Medicine
Plasma sources Biological effects
development
adaptation
diagnostics
optimization, control,
monitoring
experimental applications
Therapeutic Applications
physiological liquids
cells:
- microorganisms
- mammalian cells
cell and tissue cultures:
- not contaminated/infected
- contaminated/infected
isolated tissues/organs
organisms:
- animal experiments
- clinical investigations
in vitro
in vivo
79
The bans for new methods in medicine …
Non-thermal Atmospheric Pressure
Plasma Sources
INP
Basic requirements for therapeutic applications
Plasmas which ...
... operate at atmospheric pressure
treatment of living objects
manageability (operation at open atmospheres)
... operate stable
reproducibility of treatment results
reliability
... are well characterized
quantitative knowledge on plasma parameters and
“macroscopic” characteristics
process monitoring
influence by operation parameters
81
INP
MPE
INP
TU/e
MPE
GREMI
Lough-
borough
ODU
Atmospheric pressure plasmas for biomedicine
Plasma
Nozzle
Isolation
Gas
High voltage
supply
Electrode
Plasma
Object (Tissue)
Stray capacity
Drexel
Cinogy
Surface Discharge
Volume Barrier Discharge
Plasma jet
K.-D. Weltmann et al., Pure Appl. Chem., Vol. 82, No. 6, (2010)
From kINPen 09 to kINPen MED
Simple and safe use under consideration of the excisting and known basic plasma
properties
Requirements according to DIN EN 60601-1
PLL-Electronic for optimal Plasmaignition
Gas-flow-control, automatic shut down
Timercontrol to secure a maximum dosage
Burst-Mode: lower irritation by current and temperature to the surface, lower
radiation output
Distance holder for reproducable treatment
83
90
Adapted device for petri dishes
closed confinement
variable distance DBD to surface/suspension
well defined and stable discharge and
treatment parameters possible
60mm
95 mm
Flexible discharge arrangement
94
Experimental Plasma Medicine
Plasma sources Biological effects
development
adaptation
diagnostics
optimization, control,
monitoring
experimental applications
Therapeutic Applications
physiological liquids
cells:
- microorganisms
- mammalian cells
cell and tissue cultures:
- not contaminated/infected
- contaminated/infected
isolated tissues/organs
organisms:
- animal experiments
- clinical investigations
in vitro
in vivo
95
In-vitro characterization of plasma-cell interactions
- General estimation of biological effectivity
- Estimation of therapeutic options
- Estimation of possible risk factors
- Insights into reaction mechanisms
97
Neutral red uptake (NRU) assay 48 and 72 h after plasma treatment
(uptake of the dye into lysosomes of vital cells → dyed cells = living cells)
Influence of plasma treatment on HaCaT- RPMI Influence of plasma treatment on HaCaT- IMDM
Cells in suspension →→→→ cytotoxicity
Keratinocytes (HaCaT) in suspension, different media; kINPen (Ar 3.8 slm); seeding after plasma
treatment
Cytotoxic effects dependent on plasma treatment time and on cell culture
medium
Vitality(%)
Vitality(%)
RPMI – Rosewell Park Memorial Institute,
IMDM – Iscoves Modivied Dulbeccos Media (more glucose, buffer, pyruvate)
Source: University Düsseldorf
Scratch assay: mechanical scratching of a 2D keratinocyte (HaCaT); detection of timedependent
scratch-width reduction
t24
t10t0
Adherent 2D cell cultures →→→→ scratch assay
≈≈≈≈ 3 mm
Modified scratch assay
K. Wende, K. Landsberg, U. Lindequist, K.-D.. Weltmann, Th. von Woedtke, IEEE Trans. Plasma Sci. 38 (2010) 2479 - 2485
Vessel with adherent cells
(w/ or w/o scratch)
Agarose layer (< 1mm)
Cell culture medium
Cell culture medium
Low melting agarose
HaCaT monolayer
agarose with scratch
HaCaT cell line with
scratch
petri dish
microorganism
Adherent 2D cell cultures →→→→ scratch assay
2D keratinocytes (HaCaT) culture (RPMI), co-cultivated with S. epidermidis;
40 s APPJ (Ar gas)
Modified scratch assay: protection of cells by agarose overlay
0
0,02
0,04
0,06
0,08
0,1
0,12
0 10 20 30 40 50
time [h]
scratchwidthreduction[cm]
infected, plasma
infected
Demonstration of selective
antiseptic plasma treatment
K. Wende, K. Landsberg, U. Lindequist, K.-D.. Weltmann, Th. von Woedtke, IEEE Trans. Plasma Sci. 38 (2010) 2479 - 2485
103
Key results Plasma Vitro
Stimulation of cell regeneration by
atmospheric pressure plasma treatment
No substantial structural changes of
intracellular proteins (charge influenced)
Dose dependent, but reversible influences on
cell DNA
Dose dependent emergence/generation of
ROS in cells
Important influence of cell culture medium on
in vitro results
106C. Bender et al., Plasma Process. Polym. 7 (2010) 318–326
HET-CAM: Hens‘s Egg Test – Chorioallantoic Membrane
• Standard test for eye and skin irritating potential of chemicals
• Fertilized eggs, incubated for 10 d
Living tissue – irritation potential
Key results Plasma Cure
Proof of tissue compatibility of plasma
treatment in antiseptically effective doses
Inactivation of bacteria without irritation of the
surrounding cells/tissue
Antiseptic plasma effect on human skin
comparable to conventional antiseptics
No influence on skin barrier function as well as
on the antioxidative potential of the skin
C. Bender et al., Plasma Process.
Polym. 7 (2010) 318–326
A. Hammann et al., Skin Pharmacol. Physiol. 23 (2010) 328-332
109
Key results Plasma Derm
Antimicrobial effect on bacteria, fungi and
parasites shown
Safety of application proven e.g. no skin irritation,
no influence on skin barrier function, no
desiccation, no change of skin pH-value, no other
visible side effects
Successful treatment of athlet‘s foot (Tinea pedis)
Successful single treatment of inflammatory skin
diseases e.g. psoriasis, acne, lichen ruber
G. Daeschlein et al., Plasma Process. Polym. 7 (2010) 224-230
G. Daeschlein et al, IEEE Trans. Plasma Sci. 38 (2010) 2969-2973
G. Daeschlein et al., IEEE Trans. Plasma Sci. 39 (2011) 815-821
110
Key results Plasma Dent
Successful treatment of teeth root channel in vitro
Inactivation of biofilms on teeth and on dental implants
Improved cell adhesion on implant surfaces
Successful decontamination and coating of dental prosthesis
Th. Kocher et al., Greifswald University
I. Koban et al., GMS Krankenhaushyg Interdiszip. 2009;4(2):Doc09.
I. Koban et al., New J. Phys. 12 (2010) 073039
I. Koban et al., Plasma Process. Polym, submitted
Highlight Plasma Dent (example)
Endodontics – reduction of E. feacalis
0
1
2
3
4
5
6
7
8
9
lg(KbE/ml)
1 NaCl- Spül. 2 Gaskontr. 3 Chx- Spül. 4 KINPen 08
Abtötung von E.faecalis im Wurzelkanalmodell
Error Bars: 95% Confidence Interval
Chlorhexidine (standard): 2 log
(after 5 min. treatment)
APPJ (kINPen): 3…4 log
(after 1 min. treatment)
Root canal model
Th. Kocher et al., Greifswald University
Enterococcus faecalis inactivation
(Error bars: 95% confidence interval)
Numberofbacteria(lgcfu/ml)
NaCl
rinsing
Ar gas
flow
Chlorhexidine
rinsing
APPJ
treatment
control
Persisting 4 years Plasma + Polihexanide
Example
German shepherd dog with chronic wound
‘Harras’, 9 years
After 6.5 weeks After 11 weeks
C. Bender et al.
Univ. Greifswald
Plasma + Polihexanide
Example
Labrador with chronic wound
‚Astor, 12 years
2010-12-08 2011-01-10
C. Bender et al.
Univ. Greifswald
Practical needs
For the patient
no fear
no pain
no delay
For the doctor
simple („one doctor – one knob“)
robust (mm not µm)
small (ambulant treatment)
Outlook – „Plasma goes into hospital“
In vitro tests
susceptibility
toxicity
selectivity
Clinical tests
susceptibility
compatibility
long-term effects
Clinical needs
Indications (examples)
chronic inflammatory diseases
acute infections
auto immune diseases
effectivity
safety
TISSUE TOLERABLE PLASMA - TTP
Summary: chances & risks
Chances of Tissue Tolerable Plasma:
1. Prevention and treatment of diseases (chronic wounds, skin and
mucosal infectious diseases, localized tumors, keloid formation,
promotion of angiogenesis, tissue ablation, hemostasis)
2. Inhibition of biofilm formation by surface treatment and by direct action
on biofilms
3. Promotion of incorporation of implants into viable tissue by changing the
surface (hydrophobicity, plasma steered application of antimicrobial
active layers with drug delivery function)
4. Promotion of improved penetration of topically applied drugs with
therapeutic outcome
5. Improved cleaning performance in the treatment process of medical
devices by surface modification.
Risks of plasma medicine:
• A few physical, more technological and administrative
• Generation of false expectations (ahead of time)
• Basic scientific understanding is lacking or inadequate
• Costs for implementation into practice (approval of medical devices,
health insurance companies, …)
The field is new,
with huge chances,
with significant research potential,
is broad enough to promote co-operations
is an opportunity for plasma technology.
Summary: statement
www.campus-plasmamed.de
Contact
INP
Leibniz Institute for Plasma Science and Technology
Adresse: Felix-Hausdorff-Str. 2, 17489 Greifswald
Telefon: +49 - 3834 - 554 300, Fax: +49 - 3834 - 554 301
E-Mail: welcome@inp-greifswald.de, Web: www.inp-greifswald.de
This study was realized within the joint research project
“Campus PlasmaMed” supported by the German Federal
Ministry of Education and Research (grants no. 13N9779 and 13N11188).
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