1 Smith papyrus 2600 – 2200 BC grease-honey-lint as wound dressing Sumerian clay tablet c. 2000 BC various prescriptions Ebers papyrus c. 2000 BC 300 prescriptions Aristotle 350 BC sore eyes, wounds Dioscorides c. 1 AD sores, ulcers, throat & eye infections Dioscorides (c.50 AD) - an early herbalist Dioscorides wrote of honey being good for: • sunburn • spots on the face He also wrote that honey heals inflammation of the throat and tonsils Honey has been used for infected wounds throughout all ages – displaced by penicillin in 1940s It is being “rediscovered” now that the ‘antibiotic era’ is coming to an end
Transcript
1
Smith papyrus 2600 – 2200 BC grease-honey-lint as wound dressing
Sumerian clay tablet c. 2000 BC various prescriptions
Ebers papyrus c. 2000 BC 300 prescriptions
Aristotle 350 BC sore eyes, wounds
Dioscorides c. 1 AD sores, ulcers, throat & eye infections
Dioscorides (c.50 AD) - an early herbalist
Dioscorides wrote of honey being good for: • sunburn • spots on the face
He also wrote that honey heals inflammation
of the throat and tonsils
Honey has been used for infected
wounds throughout all ages –
displaced by penicillin in 1940s
It is being “rediscovered” now that the ‘antibiotic era’ is coming to an end
2
THE TENNESSEAN Apr. 28, 2012 MRSA is one of the more prevalent, potentially deadly, germs spread in gyms and locker rooms. MRSA (methicillin-resistant Staphylococcus aureus) is a type of Staph bacteria found on the skin and in the nose that is resistant to antibiotics. More than 90,000 Americans get potentially deadly MRSA infections every year and more than 18,000 die each year from this deadly disease.
Aristotle (350 BC), discussing differences in honeys, referred to pale honey being good as a salve for sore eyes and wounds
Aristotle (350 BC) Historia Animalium
Dioscorides (c.50 AD) stated that:
"Attick Hony is the best… That which is best… a pale yellow, not liquid…is good for all rotten & hollow ulcers.“
Gunther (1934) The Greek Herbal of Dioscorides
The potency of the anti-bacterial activity of honey can vary 100-fold
In present-day
folk medicine,
specific honeys
are known to be
the best to use
as medicines
NZ folk knowledge: manuka honey is best
The
pote
ncy
of th
e an
ti-ba
cter
ial
activ
ity o
f ho
ney
can
vary
100
-fold
3
All honey: glucose gluconic acid
+ + oxygen hydrogen peroxide
glucose oxidase
All honey: glucose gluconic acid
+ + oxygen hydrogen peroxide
glucose oxidase
Was there something else in manuka honey?
Tested honey solutions with
catalase added to destroy hydrogen
peroxide
Manuka honey is unique – it still has full antibacterial activity with catalase added
4
H2O2 H2O + O2
catalase
5
6
7
Honey does not produce hydrogen peroxide unless it is diluted
It continues to do so only for about 24 – 48 hours after it is diluted
8
A standard antiseptic, phenol, is used for comparison
Dormant protein-digesting enzymes in wound tissues are activated by oxidation by H2O2
The over-active enzymes digest the
matrix and tissue growth factors
The cytokines and cell growth factors produced by leukocytes stimulate growth of fibroblasts, epithelial cells and vascular cells (angiogenesis)
14
With matrix Matrix destroyed
The matrix in a healing wound is essential for cell attachment for
migration and multiplication
Attachment stimulates fibroblasts to
move along the strands of the matrix
It also stimulates them to multiply Epithelial cells migrate across the matrix
(using fibronectin-integrin contact)
Bacteria in wound
Inflammatory response
Exudation of serum
Good nutrition for bacteria
15
MIC values for species of bacteria for manuka honey with near median levels of antibacterial activity:
Staphylococcus aureus from infected wounds: 2%–3% (v/v) honey
Cooper R A et al. (1999) J. Royal Soc. Medicine 92: 283-285 Pseudomonas from infected wounds: 5.5%–8.7% (v/v) honey
Cooper, R. A.; Molan, P. C. (1999) J. Wound Care 8: 161-164 MRSA from infected wounds: 2.7–3.0% (v/v) honey VRE from hospital surfaces: 3.8–5.0% (v/v) honey
Cooper, Molan & Harding (2002) J. Applied Microbiology 93: 857-863
β-Haemolytic streptococci from wounds: 4.5%–9.7% (v/v) honey Cooper R A et al. (2000) First World Congress on Wound Healing: Melbourne
Coagulase-negative Staphylococci isolates: 3–3.5% (v/v) honey French, V.M. et al. (2004) J. Antimicrob. Chemo. 56: 228-231
5 isolates of Acinetobacter baumannii
MIC No. of isolates Manuka honey 6% 2
7% 1 8% 1
>8% 1 Pasture honey >7% 5
Unpublished work conducted with the Central Public Health Laboratory, Colindale, London
16
4 isolates of Stenotrophomonas maltophilia
MIC No. of isolates Manuka honey 3% 3
4% 1 Pasture honey <4% 1
5% 1 6% 2
Unpublished work conducted with the Central Public Health Laboratory, Colindale, London
George N (2004) Australian Infection Control Conference: Hobart
Alandejani, T. et al. (2009). Effectiveness of honey on Staphylococcus aureus and Pseudomonas aeruginosa biofilms. Otolaryngology–Head and Neck Surgery 141(1): 114-118
Maddocks, S. E. et al. (2012). Manuka honey inhibits the
development of Streptococcus pyogenes biofilms and causes reduced expression of two fibronectin binding proteins. Microbiology 158: 781-790
Merckoll, P. et al. (2009). Bacteria, biofilm and honey: A study of
the effects of honey on 'planktonic' and biofilm-embedded chronic wound bacteria. Scandinavian Journal of Infectious Diseases 41(5): 341-347
Okhiria, O. A. et al. (2009). Honey modulates biofilms of
Pseudomonas aeruginosa in a time and dose dependent manner. Journal of ApiProduct and ApiMedical Science 1(1): 6-10
Honey has other advantages for wound
treatment besides antibacterial activity
• moist healing environment
• autolytic debriding action
• anti-inflammatory action
• stimulates cell growth
Wounds heal 50% faster if a moist environment is maintained
Winter, GD & Scales JT 1963 Nature 197: 91-92
17
So why are wounds often kept dry? Bacteria thrive in moist conditions
A moist environment can also cause maceration of skin around the wound
Slough harbours bacteria ��inflammatory response
Debridement is needed to remove harboured bacteria
Slough harbours bacteria ��inflammatory response
Streptokinase is expensive
Maggots are problematical
Moist autolysis �� bacterial growth
Published evidence is:
• honey works faster than autolytic debridement
• honey compares well with surgical debridement
• honey stops slough forming on fresh wounds
Molan P. C. (2009) ‘Debridement of wounds with honey’ Journal of Wound Technology 5: 12-7
3 days after using honey dressings
19
Proposed mechanism of action:
• Slough is attached by a fibrin clot which is digested by a protease
• Is there activation of plasmin, which digests only fibrin, not matrix?
Conclusions from the study: • A mechanism for the debriding action
of honey has been found
• This activity varies in potency between different batches of honey
• The debriding activity of honey dressings can now be standardised
Anti-inflammatory effect of honey on wounds • observed clinically (reported in many papers):
visibly reduces inflammation reduces oedema, exudation and pain
• observed histologically: fewer neutrophils present in: biopsy samples from burns 5 animal studies with no infection present
Reviewed in: Molan, P. C. (2002)
Ostomy/Wound Management 48: 28
20
6 RCTs have been published on using honey on burns, demonstrating its effectiveness in reducing inflammation
21
Chapter 2 Characterising THP-1
55
It can be seen that all three treatments gave a significant increase in
phagocytosis with the treatments giving significantly different degrees of
activation of phagocytosis from each other. The effect of PMA was
approximately twice that of Vit-D3 and of LPS was a further three times greater.
Figure 2.12. Time lapse photographs under a fluorescent microscope of an LPS-activated macrophage (100 ng/ml LPS for 1 day) phagocytising a FITC-labelled latex particle. The whole event took approximately 25 seconds. As the cell moves, other particles already phagocytosed become visible (400 x magnification).
Time-lapse photography of phago-cytosis of fluorescent latex beads
The effect of manuka honey on phagocytosis with zymosan (yeast cell wall fragments)
The effect of 0.25% various honeys on phagocytosis (The error bars show the standard deviation)
!
The anti-inflammatory activity was not correlated with the rating of antibacterial activity of the honeys
The inflammatory component of honey
has been identified as Apalbumin-1
The
effe
ct o
f 0.2
5% v
ario
us h
oney
s on
pha
gocy
tosi
s (T
he e
rror
bar
s sh
ow th
e st
anda
rd d
evia
tion)
!
The
anti-
infla
mm
ator
y ac
tivity
was
not
cor
rela
ted
with
the
ratin
g of
ant
ibac
teria
l act
ivity
of t
he h
oney
s
22
Apalbumin-1 is a bee protein –
so why is manuka honey more active?
and why some batches more than others?
The special antibacterial activity of
manuka honey is due to MGO
MGO reacts with Apalbumin-1 to make
it more potently anti-inflammatory
H2O2 ��������������������
� ������������������ ������������
H2O2
H2O2
H2O2
H2O2
������������������������������ ������������
Fibroblasts are stimulated by hydrogen peroxide
23
Sutures removed 10 days post-op.
2 weeks post-op.)
4 weeks post-op.
12 weeks post-op.
Several studies with experimentally inflicted wounds have demonstrated that honey hastens the healing process.
Case studies have shown ‘kick-starting’ of healing in recalcitrant wounds
24
Studies with leukocytes in culture have shown that honey stimulates the production of cytokines
31 days of honey
dressings
From C. Goldberg MD, UCSD From C. Goldberg MD, UCSD
