Inhibitory Effects of Lactoferrin on Growth and Biofilm Formation of
Porphyromonas gingivalis and Prevotella intermedia
Hiroyuki Wakabayashi,1* Koji Yamauchi,1 Tetsuo Kobayashi,2,3 Tomoko
Yaeshima,1 Keiji Iwatsuki,1 and Hiromasa Yoshie2
Food Science & Technology Institute, Morinaga Milk Industry Co., Ltd.,
Zama,1 Division of Periodontology, Niigata University Graduate School
of Medical and Dental Sciences, Niigata,2 General Dentistry and
Clinical Education Unit, Niigata University Medical and Dental
Hospital, Niigata, Japan3

Received 22 December 2008/ Returned for modification 3 February 2009/
Accepted 8 May 2009

Lactoferrin (LF) is an iron-binding antimicrobial protein present in
saliva and gingival crevicular fluids, and it is possibly associated
with host defense against oral pathogens, including periodontopathic
bacteria. In the present study, we evaluated the in vitro effects of
LF-related agents on the growth and biofilm formation of two
periodontopathic bacteria, Porphyromonas gingivalis and Prevotella
intermedia, which reside as biofilms in the subgingival plaque. The
planktonic growth of P. gingivalis and P. intermedia was suppressed
for up to 5 h by incubation with 130 µg/ml of human LF (hLF), iron-
free and iron-saturated bovine LF (apo-bLF and holo-bLF,
respectively), and 6 µg/ml of bLF-derived antimicrobial peptide
lactoferricin B (LFcin B); but those effects were weak after 8 h. The
biofilm formation of P. gingivalis and P. intermedia over 24 h was
effectively inhibited by lower concentrations (8 µg/ml) of various
iron-bound forms (the apo, native, and holo forms) of bLF and hLF but
not LFcin B. A preformed biofilm of P. gingivalis and P. intermedia
was also reduced by incubation with various iron-bound bLFs, hLF, and
LFcin B for 5 h. In an examination of the effectiveness of native bLF
when it was used in combination with four antibiotics, it was found
that treatment with ciprofloxacin, clarithromycin, and minocycline in
combination with native bLF for 24 h reduced the amount of a preformed
biofilm of P. gingivalis compared with the level of reduction achieved
with each agent alone. These results demonstrate the antibiofilm
activity of LF with lower iron dependency against P. gingivalis and P.
intermedia and the potential usefulness of LF for the prevention and
treatment of periodontal diseases and as adjunct therapy for
periodontal diseases.



--------------------------------------------------------------------------------
* Corresponding author. Mailing address: Food Science & Technology
Institute, Morinaga Milk Industry Co., Ltd., 5-1-83 Higashihara, Zama,
Kanagawa 228-8583, Japan. Phone: 81-46-252-3045. Fax: 81-46-252-3059.
E-mail:

Published ahead of print on 18 May 2009.



--------------------------------------------------------------------------------
Antimicrobial Agents and Chemotherapy, August 2009, p. 3308-3316, Vol.
53, No. 8
0066-4804/09/$08.00+0 doi:10.1128/AAC.01688-08
Copyright © 2009, American Society for Microbiology.

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Ho hum, one more time. Bacteria like all life must have iron to thrive.
If one remove one critial factor needed for life then it will not occur,
in this case iron.

Eskimos and others who ate a mostly meat diet were noted to have
exceptional dental health. It was when high carbohydrate bearing foods
were introduced into their diet from abroad did this quality dental
health decline.

Another nail in the coffin of the eating meat and it's iron is the cause
of *ALL* disease idea.

The iron of those nails is most surely killing it.

On Aug 7, 2:09*pm, ferr...@paris.com wrote:

> Bacteria like all life must have iron to thrive.

and water.. But must 'all life forms' have heme iron to thrive?? And
is there a shortage of iron to justify risks of HEME iron for humans?

Let's take a look...

> Eskimos

out of hominoidea ecological niche with pathetic life spans to
match...

> and others

(extirpated/feral primates)

> who ate a mostly meat

Humans cannot 'eat meat'. Enculturated humans may choose to consume
meat but it is not digested as fruit. Try to vary diet (exclude meat)
to notice the difference in digestion. 'Eating' connotes digestion.

> It was when high carbohydrate bearing foods
> were introduced

The second worst or very worst diet style? It's academic. Other
countries were also 'introduced' to processed/starchy food without the
same set backs.. Why? Inuit have also benefited from medical advances
too. Adopting SAD has produced disease risks for all countries
involved, not just Inuit, yet Inuit life expectancy remains relatively
low and certainly NOT a model in any aspect for natural/healthy diets
for H. sapiens.

Health Rep. 2008 Mar;19(1):7-19.
MAIN RESULTS: In 1991, life expectancy at birth in the Inuit-inhabited
areas was about 68 years, which was 10 years lower than for Canada
overall. From 1991 to 2001, life expectancy in the Inuit-inhabited
areas did not increase, although it rose by about two years for Canada
as a whole. As a result, the gap widened to more than 12 years.

So why are humans to consume diets of high iron now?

> into their diet from abroad did this quality dental
> health decline.

Maybe humans can brush their teeth with heme iron toothpaste. Inuit
diet has been watered down in some respects, as has any traditional
diet supplemented w/junk food, the fact remains there is no evidence
Inuit are healthier overall now. And with what traditional food Inuit
do subsist on, it appears they are unable to keep pace w/the rest of
the world in terms of overall health despite increased health care
access for Inuit that comes w/the modern diet changes. Canada has far
outpaced Inuit in terms of life expectancy and disease avoidance.

Overall health status and life expectancy of Inuit did NOT decline
with introduction of processed/Westernized diet while access to modern
medical advances came in its train. So Inuit had access to med care as
well as junk food JUST LIKE THE REST OF THE MODERN WORLD THAT
INCREASED LIFE EXPECTANCY, INSTEAD. What was the confounding variable?
Perhaps animal fat as it limits/is not life promoting (only survival
food). Cows can consume heme iron too. So Inuit have cleaner teeth but
die sooner?? You can keep teeth clean and consume cardboard and die
quickly. How do you separate excess iron from meat?

> Another nail in the coffin of the eating meat and it's iron is the cause
> of *ALL* disease idea.

But why does it have to be 'heme iron' as opposed to plant iron? Why
the disease risks? Where did Ironjustice advocate 'high carbohydrate
bearing foods' without sufficient plant iron and vitamin C as opposed
to the problematic heme iron??

Can J Public Health. 2000 Nov-Dec;91(6):407-10. Anemia and iron status
in Inuit infants from northern Quebec.The corresponding prevalence of
microcytic anemia was 0.0%, 4.3% and 21.3%. At 2, 6 and 12 months,
iron-deficiency anemia was present in 1.3% (1/79), 24.4% (21/86) and
26.3% (25/95) of infants, respectively. This study shows iron-
deficiency anemia to be a problem in Inuit infants as young as 6
months old.

So what good comes from a heme iron diet?

> The iron of those nails is most surely killing it.

"The paradox of anemia with high meat intake: a review of the
multifactorial etiology of anemia in the Inuit of North America.

Anemia is a serious concern among indigenous populations in North
America, and it appears to be widespread among the Inuit despite
abundant intakes of heme iron. It is therefore hypothesized that
anemia for the Inuit involves other dietary factors not usually
associated with animal foods, such as low intakes of vitamin A and/or
folate, riboflavin, and vitamin C.
Nutr Rev. 2008 May;66(5):256-71.


Yes, Ironjustice may be fixated on the iron issue but what are you
trying to prove by defending a risky heme iron diet? It is more absurd
and popular to be oblivious to the excess iron consumed in animals
with numerous/varied disease risks than insufficient iron risk of a
high vitamin C diet without the meat/varied disease risks.

“There is a remarkably close association between the amount of iron
stored in our tissues and the risk of death from cancer, heart
disease, or from all causes.. The shape of this curve, representing
the iron burden, is amazingly similar to the curves representing the
rate of death in general, and the rate of death from cancer. There is
no other relationship in biology that I know of that has this peculiar
shape” Iron's Dangers Ray Peat 2006.

There is no shortage of protein, air, fat, water, bacteria or iron.
There is an abundance of preventable/acquired diseases of affluence.

With health,
Chris

>> Bacteria like all life must have iron to thrive.

>and water.. But must 'all life forms' have heme iron to thrive?? And is
>there a shortage of iron to justify risks of HEME iron for humans?"

And several other factors for life also, for the purposes of the study
posted, dental health, iron is iron. Your question irrelevant.

>Let's take a look..."

>out of hominoidea ecological niche with pathetic life spans to

m>atch...

Irrelevant to the question at hand, dental health and iron intake.
Eskimos traditionally had excellent dental health and high iron intake,
which contridicts our rusty friend's compulsive conclusion, end of
story. I see the direction you want to go, not interested, that line
more religion then science.

Your eskimo assertions are difficult to support. What little quality
data about traditional diet and health shows a typical short life as was
the case for most pre-agricultural cultures. Death from dangers other
then health were the principal reason. The little eskimo data shows
some people living into 70's and 80's if they survived the dangers of
the environment.

A Canadian health survey found that the more the use of the traditional
diet the more healthy the individual.

On Aug 7, 7:20Â*am, ironjustice <teamtan...@hotmail.com> wrote:
Inhibitory Effects of Lactoferrin on Growth and Biofilm Formation of
Porphyromonas gingivalis and Prevotella intermedia <<


The effect of lactoferrin on oral bacterial attachment
Oral Microbiology and Immunology
S. Y. Arslan 1 , K. P. Leung 2 , C. D. Wu 1
1 Department of Pediatric Dentistry, College of Dentistry,
University of Illinois at Chicago, Chicago, IL, USA ,
2 US Army Dental and Trauma Research Detachment, Great Lakes, IL, USA
Correspondence to Christine D. Wu, Department of Pediatric Dentistry,
College of Dentistry, MC850,
University of Illinois at Chicago, 801 S. Paulina Street,
Room 469J, Chicago, IL 60612-7212, USA
Tel.: + 1 312 355 1990;
fax: + 1 312 996 1981;
e-mail:
Volume 24 Issue 5, Pages 411 - 416
Published Online: 17 Aug 2009

Abstract

Introduction: Lactoferrin (Lf), an iron-binding salivary glycoprotein,
plays an important role in human innate defense against local mucosal
infection. We hypothesized that Lf interferes with initial oral
bacterial attachment to surfaces by iron sequestration, so inhibiting
subsequent biofilm formation. The objective was to investigate the
effect of Lf on the early stages of single-species and multi-species
oral biofilm development.

Methods: Streptococcus gordonii, Streptococcus mutans, Fusobacterium
nucleatum and Porphyromonas gingivalis were used in this study. Glass
disks of a two-track flow cell coated with flowing artificial saliva
(0.3 ml/min) with and without Lf (100 μg/ml) were used for studying
bacterial attachment (3 h, 37°C). Attachment was also examined by
incubating single or multiple species of test bacteria (107 colony-
forming units/ml) with Lf-coated (20–100 μg/ml) and uncoated glass
slides. The effects of β-lactoglobulin, 2,2'-dipyridyl (25–100μg/ml),
an iron chelator, and FeCl3 on attachment were also examined.

Results: Lf inhibited the initial attachment of S. gordonii (50.3%, P
< 0.05) but not that of F. nucleatum and P. gingivalis. However, the
attachment of a dual-species biofilm containing S. gordonii (i.e. S.
gordonii/F. nucleatum or S. gordonii/P. gingivalis) was significantly
reduced (48.7% or 62.1%, respectively, P < 0.05) in the presence of
Lf. β-Lactoglobulin did not affect the attachment of S. gordonii. In
the presence of 100 μm 2,2'-dipyridyl, attachment of S. gordonii was
reduced by 53.87%. No reduction in attachment was noted in S. gordonii
pretreated with Lf (100 μg/ml) and FeCl3 (20–200 μm).

Conclusion: Lf suppresses initial attachment of S. gordonii and S.
gordonii coaggregates by iron sequestration. This may lead to
subsequent inhibition of oral biofilm development.

KEYWORDS
iron • iron chelator • lactoferrin • oral bacterialattachment • oral
biofilm
Arslan SY, Leung KP, Wu CD. The effect of lactoferrin on oral
bacterial attachment.
Oral Microbiol Immunol 2009: 24: 411–416.© 2009 John Wiley & Sons A/S.


Accepted for publication April 28, 2009

DIGITAL OBJECT IDENTIFIER (DOI)
10.1111/j.1399-302X.2009.00537.x About DOI
Copyright © 2009 John Wiley & Sons A/S


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> Hiroyuki Wakabayashi,1* Koji Yamauchi,1 Tetsuo Kobayashi,2,3 Tomoko
> Yaeshima,1 Keiji Iwatsuki,1 and Hiromasa Yoshie2
> Food Science & Technology Institute, Morinaga Milk Industry Co., Ltd.,
> Zama,1 Division of Periodontology, Niigata University Graduate School
> of Medical and Dental Sciences, Niigata,2 General Dentistry and
> Clinical Education Unit, Niigata University Medical and Dental
> Hospital, Niigata, Japan3
>
> Received 22 December 2008/ Returned for modification 3 February 2009/
> Accepted 8 May 2009
>
> Lactoferrin (LF) is an iron-binding antimicrobial protein present in
> saliva and gingival crevicular fluids, and it is possibly associated
> with host defense against oral pathogens, including periodontopathic
> bacteria. In the present study, we evaluated the in vitro effects of
> LF-related agents on the growth and biofilm formation of two
> periodontopathic bacteria, Porphyromonas gingivalis and Prevotella
> intermedia, which reside as biofilms in the subgingival plaque. The
> planktonic growth of P. gingivalis and P. intermedia was suppressed
> for up to 5 h by incubation with 130 µg/ml of human LF (hLF), iron-
> free and iron-saturated bovine LF (apo-bLF and holo-bLF,
> respectively), and 6 µg/ml of bLF-derived antimicrobial peptide
> lactoferricin B (LFcin B); but those effects were weak after 8 h. The
> biofilm formation of P. gingivalis and P. intermedia over 24 h was
> effectively inhibited by lower concentrations (8 µg/ml) of various
> iron-bound forms (the apo, native, and holo forms) of bLF and hLF but
> not LFcin B. A preformed biofilm of P. gingivalis and P. intermedia
> was also reduced by incubation with various iron-bound bLFs, hLF, and
> LFcin B for 5 h. In an examination of the effectiveness of native bLF
> when it was used in combination with four antibiotics, it was found
> that treatment with ciprofloxacin, clarithromycin, and minocycline in
> combination with native bLF for 24 h reduced the amount of a preformed
> biofilm of P. gingivalis compared with the level of reduction achieved
> with each agent alone. These results demonstrate the antibiofilm
> activity of LF with lower iron dependency against P. gingivalis and P.
> intermedia and the potential usefulness of LF for the prevention and
> treatment of periodontal diseases and as adjunct therapy for
> periodontal diseases.
>
> ---------------------------------------------------------------------------Â*-----
> * Corresponding author. Mailing address: Food Science & Technology
> Institute, Morinaga Milk Industry Co., Ltd., 5-1-83 Higashihara, Zama,
> Kanagawa 228-8583, Japan. Phone: 81-46-252-3045. Fax: 81-46-252-3059.
> E-mail: h_wak...@morinagamilk.co.jp
>
> Â*Published ahead of print on 18 May 2009.
>
> ---------------------------------------------------------------------------Â*-----
> Antimicrobial Agents and Chemotherapy, August 2009, p. 3308-3316, Vol.
> 53, No. 8
> 0066-4804/09/$08.00+0 Â* Â* doi:10.1128/AAC.01688-08
> Copyright © 2009, American Society for Microbiology.
>
> Who loves ya.
> Tom
>
> Jesus Was A Vegetarian!http://tinyurl.com/2r2nkh
>
> Man Is A Herbivore!http://tinyurl.com/a3cc3
>
> DEAD PEOPLE WALKINGhttp://tinyurl.com/zk9fk