Note: Descriptions are shown in the official language in which they were submitted.
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GUM CONDITION ASSESSMENT
FIELD OF THE INVENTION
The invention is directed to methods of assessing a gum condition in a
individual.
BACKGROUND OF THE INVENTION
It is widely recognized by dental professionals that "gingival bleeding" or
"gum bleeding"
is a symptom of a potential gum condition. Dental professionals make this
assessment when
referring to bleeding that is induced by manipulating tissue with a
periodontal probe at the depth
of the gingival sulcus. There are many possible causes of gingival bleeding
including formation
and accumulation of plaque at the gum line. Some gum conditions, such as
gingivitis, can be
treated with proper brushing and flossing of teeth (i.e., good oral hygiene).
However, chronic
gingivitis can progress to periodontitis, which is an irreversible periodontal
infection
characterized by alveolar bone loss, and can eventually lead to tooth loss.
At least one report has over 70% of the Chinese population with varying
degrees of gum
disease. Furthermore, many individuals do not have access to dental
professionals. Even with
access to dental professions, there is a certain amount of subjectivity in
assessing, for example,
the relative status of a gum condition (e.g., mild, moderate, severe). One
reported approach is by
counting gum bleeding sites (induced with the periodontal probe). That is,
typical measures of
gum health or an improvement in a gum condition is by clinically observing a
reduction in the
number bleeding sites. However, the volume of bleeding is typically not taken
into account.
Although there may be a clinically evaluated reduction in the overall number
of bleeding sites,
the remaining (untreated) bleeding sites may still bleed at differing rates
due to pocket depth,
inflammation, and the like. There is a need to objectively quantifying the
amount blood to
provide a more objective measure of the relative status of a gum condition.
Furthermore,
objective measures such as quantifying blood volume, is more relevant and
meaningful to the
general public.
Accordingly, there is a need for a method to assess a gum condition of an
individual, and
to do so minimizing subjectivity and cost and while maximizing convenience. It
is an objective
to provide a measure this is more relevant and meaningful to the general
public. This is
particularly true in developing markets such as China.
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SUMMARY OF THE INVENTION
The present invention is based, in part, on the observation that hemoglobin
contained in
blood can be a proxy to assess a gum condition. In a nutshell, the more
gingival bleeding there is,
the more likely there is a gum condition (or the greater severity of the gum
condition). Indeed
hemoglobin that is contained in gingival bleeding can be quantified (and
correlated to clinical
assessments of gum conditions). The present invention is also based upon
incorporating tooth
brushing, a beneficial oral hygiene habit that can help in treating a gum
condition, into the
assessment method. Without wishing to be bound theory, such an approach
reduces barriers for
individuals to participate in the assessment, and re-enforces the need for
good oral hygiene such
as tooth brushing. In other words, the present invention incorporates a
relevant oral hygiene
activity into the gum condition assessment method.
The methods herein described can also provide a means to pre-screen
individuals that
many need further clinical evaluation.
The methods herein described can also provide a means to improve or assess
oral care
products or oral hygiene treatment regimens useful in the treatment of a gum
condition.
A first aspect of the invention provides a method of assessing a gum condition
of a
subject individual comprising the steps: (a) tooth brushing within an oral
cavity of the subject
individual; (b) post-brushing rinsing the brushed oral cavity with a post-
brushing rinse solution,
wherein the post-brushing rinse solution comprises a post-brushing rinse
solution volume from 1
ml to 50 ml, wherein the post-brushing rinsing is for a predetermined post-
brushing rinsing time
from 1 second to 1 minute; (c) collecting expectorated post-brushing rinse
solution from the
subject individual; (d) measuring hemoglobin quantity contained in the
collected post-brushing
rinse solution; and (e) comparing the measured hemoglobin quantity against a
known standard of
gum conditions corresponding to hemoglobin quantities to assesses the gum
condition of the
subject individual from the compared hemoglobin quantity.
A second aspect of the invention provides for a method of assessing a gum
condition of a
subject individual comprising the steps: (a) tooth brushing within an oral
cavity of the subject
individual; (b) post-brushing rinsing the brushed oral cavity with a post-
brushing rinse solution,
wherein the post-brushing rinse solution comprises a post-brushing rinse
solution volume from 1
ml to 50 ml, wherein the post-brushing rinsing is for a predetermined post-
brushing rinsing time
from 1 second to 1 minute; (c) collecting expectorated post-brushing rinse
solution from the
subject individual; and (d) visually comparing the collected post-brushing
rinse solution against a
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known standard of gum conditions corresponding to hemoglobin quantities to
assess the gum
condition of the subject individual.
A third aspect of the invention provides for a method of assessing
effectiveness of a
product or regimen in the treatment of a gum condition comprising the steps:
(a) assessing an individual's first gum condition comprising the steps:
(i) tooth brushing within an oral cavity of the individual;
(ii) post-brushing rinsing the brushed oral cavity with a first post-brushing
rinse solution,
wherein the post-brushing rinse solution comprises a post-brushing rinse
solution volume
from 1 ml to 50 ml, wherein the post-brushing rinsing is for a predetermined
post-
brushing rinsing time, wherein the predetermined post-brushing rinsing time is
from 1
second to 1 minute;
(iii) collecting expectorated first post-brushing rinse solution from the
individual;
(iv) measuring hemoglobin quantity contained in the first post-brushing rinse
solution;
(v) comparing the first measured hemoglobin quantity against a known standard
of gum
conditions corresponding to hemoglobin quantities to assesses the first gum
condition of
the individual;
(b) subjecting the individual to the product or regimen;
(c) assessing the individual's second gum condition comprising the steps:
(i) tooth brushing within the oral cavity of the individual;
(ii) post-brushing rinsing the brushed oral cavity with a second post-brushing
rinse solution
having a post-brushing rinse solution volume, and for the predetermined post-
brushing
rinsing time;
(iii) collecting expectorated second post-brushing rinse solution from the
individual;
(iv) measuring hemoglobin quantity contained in the second post-brushing rinse
solution;
(v) comparing the second measured hemoglobin quantity against the known
standard of gum
conditions corresponding to hemoglobin quantities to assess the second gum
condition of
the individual; and
(d) comparing the first gum condition assessment to the second gum condition
assessment to
assess the effectiveness of the product or regimen in the treatment of the gum
condition in the
individual.
A fourth aspect of the invention provides for a computer-aided system of
assessing a gum
condition of a subject individual comprising: (a) a sampling section for
sampling expectorated
post-brushing rinsing solution of the subject individual; (b) a measuring
section in
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communication with the sampling section, wherein the measuring section is
configured to
measure hemoglobin quantity contained in the expectorated post-brushing
rinsing solution; and
(c) a computing section in communication with the measuring section, wherein
the computing
section is configured for receiving and statistically analyzing the measured
hemoglobin to the
gum condition.
These and other features, aspects, and advantages of the present invention
will become
evident to those skilled in the art from the detailed description which
follows.
DETAILED DESCRIPTION OF THE INVENTION
Instructions
An optional, but preferred, initial step is to provide instructions to the
individual
regarding the methods described herein. Instructions may be provided by a live
demonstration,
pre-recorded video, literature (e.g., text and pictures), web-based, or
combination thereof.
Preferably the instructions comprise a video since users typically find it
easier to model viewed
behavior as compared to reading about it. A pre-recorded video is more cost
effective than a
live demonstration by an instructor. Specific mention is made to instructing
the individual about
brushing; preferably having the individual tooth brushing their own oral
cavity. Alternatively, a
trained tester brushes the individual's oral cavity. Other oral hygiene
products and regimens
may be optionally included. The instructions may also provide the added
benefit of teaching or
reinforcing proper tooth brushing techniques (or other oral hygiene habits).
Pre-rinsing
An optional, but preferred step after optionally providing instructions to the
individual, is
pre-rinsing the oral cavity of the individual, before the brushing step
(described below), with a
pre-rinse solution. Preferably the pre-rinse solution comprises a
predetermined pre-rinse volume,
more preferably the predetermined pre-rinse volume is from 1 ml to 5 ml, yet
more preferably
from 1 ml to 2.5 ml. Without wishing to be bound by theory, having less than 1
ml will leave the
oral cavity too dry, even perhaps irritating the mouth. However, volumes
greater than about 2.5
ml may lead to undesirably variability in testing. Another advantage of the
pre-rinsing step is to
potentially remove contaminants from the oral cavity that could negatively
impact results.
Preferably the pre-rinsing step is for a predetermined pre-rinsing time, more
preferably
the predetermined pre-rinsing time is from 1 second to 1 minute, yet more
preferably from 2
seconds to 10 seconds. Preferably the pre-rinse solution comprises from 90% to
100%, more
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preferably from 95% to 100%, yet more preferably 98% to 100% water by weight
of the pre-
rinse solution. The pre-rinse solution may comprise minor ingredients such as
salt (to provide
an isotonic saline solution) or contain antibacterial agents and the like.
Preferably the pre-rinse
solution is simply water. Indeed simply using water is cost effective.
5
Immersing a Brush
An optional, but preferred step after optionally pre-rinsing the oral cavity
of the
individual, but before the brushing step (described below), comprises the step
of immersing a
brush in a brush immersing solution. Preferably the immersing solution
comprises from 90% to
100%, more preferably from 95% to 100%, yet more preferably 98% to 100% water
by weight of
the immersing solution. The brush immersing solution may comprise minor
ingredients such as
salt (to provide an isotonic saline solution) or contain antibacterial agents
and the like. More
preferably the immersing solution is simply water. Indeed simply using water
is cost effective.
Preferably the step of immersing the brush is for a predetermined immersing
time,
preferably the predetermined immersing time is from 0.1 second to 5 seconds,
more preferably
from 0.5 seconds to 2 seconds, alternatively about 1 second.
Brushing
A step in the present invention is tooth brushing within an oral cavity of the
individual.
The individual may conduct the tooth brushing or a trained tester may conduct
the tooth brushing
to the individual.
Preferably the tooth brushing further comprises tooth brushing for a
predetermined
brushing time. Preferably the predetermined brushing time is from 30 seconds
to 5 minutes,
more preferably from 1 minute to 3 minutes, alternatively for about 2 minutes.
The tooth
brushing may by way of a manual toothbrush or an automated toothbrush (e.g.,
an electric
toothbrush, gum brush (e.g., Gum HealthTm)). For purposes of clarification,
the term "tooth
brushing" should be construed broadly to include those brushes designed to
brush gums. The
portion of the oral cavity that is brushed may include a tooth or gum, or
combination thereof. In
one non-limiting example, the interface between the tooth and gum is brushed.
Post-brushing Rinsing
A step in the present invention is post-brushing rinsing the oral cavity with
a post-
brushing rinse solution. Best results are typically obtained when the post-
brushing rinsing step is
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conducted immediately after the aforementioned tooth brushing step. Preferably
the post-
brushing rinse solution comprises a predetermined post-brushing rinse solution
volume.
Preferably the predetermined post-brushing rinse solution volume is from 1 ml
to 50 ml, more
preferably from 5 ml to 40 ml, yet more preferably from 5 ml to 15 ml,
alternatively about 10 ml.
Without wishing to be bound by theory, if the volume is less than 5 ml, it may
lead to
undesirably variability in testing. However, volumes greater than 50 ml may
provide for a
hemoglobin concentration that is too low to be accurately detected by some
methods (e.g., Ultra
Violet ¨ Visible Light Analysis described in further detail below). More than
one post-brushing
rinse can be employed.
Preferably the post-brushing rinsing step is for a predetermined post-brushing
rinsing
time. More preferably the predetermined post-brushing rinsing time is from 1
second to 1 minute,
yet more preferably from 5 seconds to 15 seconds, alternatively about 10
seconds. Preferably the
post-brushing rinse solution comprises from 90% to 100%, more preferably from
95% to 100%,
yet more preferably 98% to 100% water by weight of the pre-rinse solution. The
post-brushing
rinse solution may comprise minor ingredients such as salt (to provide an
isotonic saline solution)
or contain antibacterial agents and the like. Preferably the post-brushing
rinse solution is simply
water. Indeed simply using water is cost effective.
It is particularly preferred the step of collecting the saliva sample from the
brushed oral
cavity of the individual comprising collecting the post-brushing rinse
solution.
Collection
A step in the present invention is collecting expectorated post-brushing rinse
solution
from the individual. This can be way of a cup, test tube, vial, or the like.
Centrifugation
An optional, but preferred step after the aforementioned collection of
expectorated post-
brushing rinse solution is centrifuging the collected saliva sample before
measuring hemoglobin
quantity (described below). Preferably the centrifuging step is for a
predetermined centrifuging
time, more preferably the predetermined centrifuging time is from 1 minute to
20 minutes, yet
more preferably from 5 minutes to 15 minutes. Preferably the centrifuging step
is spun at a speed
as to reach at centrifugal force greater than 30 gravitational force (g-
force), preferably the
centrifugal force is between 30 g-force to 25,000 g-force, preferably from
8,000 g-force to
12,000 g-force, alternatively about 10,733 g-force. Without wishing to be
bound by theory,
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centrifuging the collected saliva sample removes solids or other ingredients
that may interfere
with measuring hemoglobin quantity.
Measuring Hemoglobin Quantity
A step in present invention is measuring hemoglobin quantity contained in the
collected
post-brushing rinse solution (or centrifuged post-brushing rinse solution).
Any known method
of measuring hemoglobin quantity may be used including, but not limited to,
colormetric,
spectrophotometric, chemical indicator, ELISA, colloidal gold techniques, and
other
immunological techniques, and the like.
Those tests that are simple, quick, and inexpensive are preferred. One
especially
preferred method of measuring hemoglobin quantity contained in the collected
saliva sample is
Ultra Violet ¨ Visible Light Analysis ("UVVLA"), more preferably wherein the
UVVLA is
conducted at a wavelength from 340 nm to 440 nm, preferably from 400 nm to 410
nm,
alternatively about 406 nm.
Comparing Against Known Standard
A step in present invention is comparing the measured hemoglobin quantity
against a
known standard of gum conditions corresponding to hemoglobin quantities. Such
standards are
commercially available. Of course the known standard will depend upon the
hemoglobin
quantity measuring test employed.
Visually Comparing
A step in the present invention is visually comparing the collected post-
brushing rinse
solution against a known standard of gum conditions corresponding to
hemoglobin quantities.
When analytical materials or instruments are not available or less accurate or
precise results are
acceptable, the collected post-brushing rinse solution may simple be visually
compared, i.e., with
an unaided eye, to physical standards, pictures, photo, video or the like.
Communicating Gum Health
An optional, but preferred step in the methods of the present invention, is
communicating
the assessed gum health. This tool, much like the initial optional
instructions, can be provided
with a live consultation, pre-recorded video, literature (e.g., text and
pictures), web-based, or
combinations thereof. The communication is preferably tailored to the specific
gum health
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status assessed, most even more preferably contain information what can and
should be done by
the individual to remedy the individual's gum condition if an undesirable gum
condition is
assessed. Obtaining and showcasing an endorsement or certification from a well
recognized and
respected industry group may also be helpful in adoption of the methods
described herein.
Assessing Effectiveness of a Product or Regimen
Another aspect of the invention provides for a method of assessing
effectiveness of a
product or regimen in the treatment of a gum condition of an individual.
The method
comprising the steps: firstly assessing an individual's first gum condition
comprising the sub-
steps: (i) tooth brushing within an oral cavity of the individual; (ii) post-
brushing rinsing the
brushed oral cavity with a first post-brushing rinse solution for a
predetermined post-brushing
rinsing time, wherein the predetermined post-brushing rinsing time is from 1
second to 1 minute;
(iii) collecting expectorated first post-brushing rinse solution from the
individual; (iv) measuring
hemoglobin quantity contained in the first post-brushing rinse solution; and
(v) comparing the
first measured hemoglobin quantity against a known standard of gum conditions
corresponding
to hemoglobin quantities to assesses the first gum condition of the
individual.
A second step is subjecting the individual to the product or regimen. The
product or
regimen may comprise a toothbrush, toothpaste, mouth rinse, floss, teeth
whitening strips, or
combinations thereof. Indeed new ingredients or products may be designed or
tested.
A third step is assessing the individual's second gum condition (after the
individual is
subjected to the product or regimen) comprising the sub-steps: (i) tooth
brushing within the oral
cavity of the individual; (ii) post-brushing rinsing the brushed oral cavity
with a second post-
brushing rinse solution for the predetermined post-brushing rinsing time;
(iii) collecting
expectorated second post-brushing rinse solution from the individual; (iv)
measuring hemoglobin
quantity contained in the second post-brushing rinse solution; and (v)
comparing the second
measured hemoglobin quantity against the known standard of gum conditions
corresponding to
hemoglobin quantities to assess the second gum condition of the individual.
A fourth step is comparing the first gum condition assessment to the second
gum
condition assessment to assess the effectiveness of the product or regimen in
the treatment of the
gum condition in the individual.
Preferred aspects of the methods include those as previously described for the
earlier
aspects of the invention (that are not repeated here for the sake of brevity).
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Computer Aided System
Another aspect of the invention provides for a computer-aided system of
assessing a gum
condition of a subject individual comprising: (a) a sampling section for
sampling expectorated
post-brushing rinsing solution of the subject individual; (b) a measuring
section in
communication with the sampling section, wherein the measuring section is
configured to
measure hemoglobin quantity contained in the expectorated post-brushing
rinsing solution; (c) a
computing section in communication with the measuring section, wherein the
computing section
is configured for receiving and statistically analyzing the measured
hemoglobin to a gum
condition.
Preferably the computing section comprises: (i) an input module in
communication with
the measuring section, wherein the input module is for inputting input data,
wherein the input
data comprises demographic information of the subject individual or medical
information of the
subject individual; and (ii) a data processing module in communication with
the input module,
wherein the data processing module is configured for statistically analyzing
the input data to
correlate with a statistical significance to the gum condition.
Preferably the step of measuring hemoglobin quantity contained in the saliva
sample is
Ultra Violet ¨ Visible Light Analysis ("UVVLA") as described earlier.
EXAMPLES ¨ Clinical Study
The detection and quantification of gingival hemoglobin, using the inventive
method of
the present invention, in a clinical study is conducted.
Overall study design and plan is described
The study is a two-leg, 3 days clinical study. Around 40 qualified subjects
meeting all the
study entrance criteria are enrolled into the study. Gingivitis is measured by
hemoglobin sampled
at baseline and at day 3.
On the first day of baseline visit, subjects receive power brush sampling for
gingival
hemoglobin. Then all the subjects are given one tube of Crest Regular Caries
Protection
toothpaste and one Crest MeiLiLiangJie toothbrush and are asked to use in
their normal oral
hygiene habits for the duration of study. At the second day, subjects receive
manual brush
sampling for gingival hemoglobin. After sampling, the subjects are randomized
to two treatment
groups: Prophy and Non-prophy. The subjects who are assigned to Prophy group
receive dental
prophylaxis.
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At the first day of day 3 visit: the subjects receive power tooth brush
sampling for
gingival hemoglobin. At the second day, the subjects receive manual tooth
brush sampling.
Specific procedures are listed below (Table 1).
Tablel. Study Schedule by Procedure Type and Visit
BASELINE
DAY 3 VISIT
PROCEDURE: VISIT
Day 1 Day 2 Day 1 Day 2
INFORMED CONSENT X
MEDICAL HISTORY X
DEMOGRAPHICS X
INCLUSION/EXCLUSION X
CONTINUANCE CRITERIA X X X
HEMOGLOBIN SAMPLING
X X X X
METHOD
DENTAL PROPHYLAXIS X
ADVERSE EVENT
X X X
RECORDING
SUBJECT ACCOUNTABILITY X
5
Baseline Visit: At the first day, subjects are asked to read and sign an
informed consent
and receive a signed copy for their records. Personal medical history
information is obtained and
reviewed and retained as site source documentation. Demographic information
and inclusion
criteria are obtained and documented on the appropriate case report form
(CRF). Then subjects
10 receive power brush sampling for gingival hemoglobin. Then all the
subjects are given one tube
of CREST Regular Caries Protection toothpaste (Lot # 40671864AA) and one
CREST
MeiLiLianghe toothbrush (Lot #0M003). Subjects are asked to use the assigned
product during
the course of the study and are reminded to refrain from smoking and any oral
hygiene after
1 lpm the night before and morning of their next visit and refrain from
consuming anything by
mouth (except a small amount of water) at least 4 hours prior to their next
visit. General
comments, if applicable, are recorded in the appropriate CRF.
At the second day, continuance criteria is assessed and documented on the
appropriate
CRF. Subjects receive manual brush sampling. Then the subjects are randomized
to two
treatment groups based on Ultra Violet ¨ Visible Light Analysis ("UVVLA")
score of power
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brush, age, gender and smoking status: prophylaxis and non prophylaxis. The
subjects who are
assigned to Prophy group receive dental prophylaxis. Subjects are asked to use
the assigned
product during the course of the study and are reminded to refrain from
smoking and any oral
hygiene after 1 lpm the night before and morning of their next visit and
refrain from consuming
anything by mouth (except a small amount of water) at least 4 hours prior to
their next visit.
General comments, if applicable, are recorded in the appropriate CRF.
Day 3 visit:
At the first day, continuance criteria is assessed and documented on the
appropriate CRF.
Then the subjects receive power brush sampling. Subjects are asked to use the
assigned product
during the course of the study and reminded to refrain from smoking and any
oral hygiene after
1 lpm the night before and morning of their next visit and refrain from
consuming anything by
mouth (except a small amount of water) at least 4 hours prior to their next
visit. General
comments, if applicable, are recorded in the appropriate CRF.
At the second day, continuance criteria is assessed and documented on the
appropriate
CRF. Subjects receive manual brush sampling. After completion of all
procedures, final subject
accountability is conducted, and the subjects are terminated from the study
after receiving
compensation.
Inclusion Criteria:
In order to be included in the study, each subject must:
(i) be at least 18 years of age; (ii) provide written informed consent prior
to participation
and be given a signed copy of the informed consent form; (iii) possess a
minimum of 12 natural
anterior teeth; (iv) have gingivitis but without periodontitis; (v) be in good
general health as
determined by the Investigator/designee based on a review of the medical
history/update for
participation in the study; and (vi) a normal healthy adult.
Subjects reporting for their visit refrain from: performing any oral hygiene
prior to their
appointment; and consuming anything by mouth (except a small amount of water)
at least 4
hours prior to their next visit. Subjects agree to receive dental prophylaxis
required by study.
Subjects self-report if there is visible bleeding when brushing in past 2
weeks. No periodontal
disease and other oral diseases except tartar.
Exclusion Criteria:
Subjects are excluded from study participation where there is evidence of any
one the
following: (i) periodontal disease beyond gingivitis (as noted above in
inclusion criteria); (ii)
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teeth that are carious, fully-crowned, extensively restored and have heavy
dental calculus; (iii)
teeth with unresolved carious lesions; (iv) any condition which requires
antibiotic premedication
for the administration of a dental prophylaxis; (v) self-reported pregnancy or
intent to become
pregnant during the course of the study and nursing females; (vi) atypical
discoloration or
pigmentation in the gingival tissue; (vii) fixed facial orthodontic
appliances; (viii) use of
antibiotics any time during the study; (ix) any diseases or conditions that
could be expected to
interfere with the subject safely completing the study; and (x) inability to
undergo imaging or
other study procedures.
Continuance Criteria:
Subjects may be excluded from the study or the analysis due to any one of the
following:
(i) use of antibiotics any time during the study; (ii) pregnancy or
development an intercurrent
medical condition which puts the subject at increased risk or invalidates the
results of the study;
(iii) use of any oral care products other than assigned study products; (iv)
subject reporting for
their visit having performed any oral hygiene prior to their appointment; (v)
subjects reporting
for their visit having consumed anything by mouth (except a small amount of
water) at least 4
hours prior to their next visit; and (vi) UVVLA score >0.2.
Oral Care Product(s):
No investigational product is used in this study. Subjects used CREST Regular
Caries
Protection toothpaste and CREST MeiLingLinaghe toothbrush in their normal
habits during
the study (per the Lot numbers previously described).
Method of Assigning Subjects to Treatment Sequences:
To ensure sampling method randomization, subjects are randomized to two
methods
according to their visit order. To ensure treatment randomization, at the
baseline visit, subjects
are stratified based on the power brush UVVLA scores, age, gender, and smoking
status within
each cohort. Subjects are randomly assigned to two cohorts (Prophylaxis/Non-
prophylaxis).
Subjects who reside in the same household are assigned to the same group.
Primary Efficacy Variable(s):
RGB Digital Imaging for sample picture taking: This system consists of a S2
Pro CCD
high resolution digital camera manufactured by Fuji or a similar camera. It is
equipped with a
Nikon Micro Nikkor lens with a linear polarizer to permit cross-polarized
light or similar optical
system. Dedo lights, mounted to an imaging rig, provides lighting.
Prior to daily use, the system is calibrated to assure proper operation.
Additionally, a
color standard is centered and imaged every hour, and then is removed prior to
imaging samples.
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For each examination period, the room is dark. The sample tube is placed on
the chin rest, and a
white balance is placed behind the sample tube. Operator follows standard
capture image
process and capture sample imaging.
Gingival hemoglobin method sampling procedure sampling and storage: Sampling
method "1" by manual brush brushing and sample method "2" by Power brush
brushing, both
follow the same procedure listed as follows: (1) Watch the video of brush
instruction; (2) Take
an empty tube; (3) Take 2m1 water to rinse the whole mouth for 5 seconds; (4)
Open the
toothbrush from its packaging and put the whole brush head into 30m1 water for
1 second; (5)
Brush whole teeth thoroughly for 2 minutes by oneself, expectorate in the
tube; (6) After
brushing, immediately rinse with 10m1 water for 10 seconds, expectorate in the
tube; (7) Label
the sample; (8) Centrifuge the sample at 10,733 g-force for 10 minutes; (9)
Take lml into a 2m1
tube, for UVVLA and sample picture taking immediately.
Ultraviolet¨visible spectroscopy method (UVVLA): is used to find the specific
absorbance of wavelength for hemoglobin, which is used for samples
measurement. The
absorbance of a sample is proportional to the concentration of Hemoglobin.
Determination of Sample Size: The sample size is based on logistical
considerations and
not power implications. Approximate 40 qualified subjects are enrolled into
the study.
Hypothesis:
The following hypothesis is tested separately each sample method (method 1 and
2) of
UVVLA at Day 3:
Null: The mean change from baseline score is equal to 0.
Alternative: The mean change from baseline score is not equal to 0.
The following hypothesis are tested separately between groups (Prophy/Non-
prophy) for
each sample method of UVVLA at Day 3:
Null: The mean score for the Prophy group is equal to the mean score of the
Non-
prophy group.
Alternative: The mean score for the Prophy group is not equal to the mean
score of the
Non-prophy group.
The following hypothesis is tested of end point at Day 3:
Null: The mean change from baseline score is equal to 0.
Alternative: The mean change from baseline score is not equal to 0.
Statistical and Analytical Plans:
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Change from baseline is investigated using paired t-tests. Analysis of
covariance is used
to compare groups (Prophy/Non-prophy) with respect to each efficacy variable:
UVVLA. A
separate statistical model is used to test group difference at each visit. The
baseline value of each
respective endpoint is used as a covariate in each model. Statistical
comparisons are two- sided
with a 0.05 significance level.
Overall results of the study are provided
Statistical Methods: The minimum detection level for UVVLA is 20. If the score
is less
than 20, the level is changed to 10 as a final score. Log 10 transformations
are applied to
UVVLA variables to better meet the normal distribution hypothesis. The change
from baseline is
investigated using paired t-tests. Analysis of covariance is used to compare
groups (Prophy/Non-
prophy) with the efficacy variable of UVVLA. A separate statistical model is
used to test group
difference at each visit. The baseline value of UVVLA endpoint is used as a
covariate in each
model. Statistical comparisons are two-sided with a 0.05 significance level.
Study Population: Referring to Table 1 below, the study population consists of
forty-two
(42) subjects enrolled at the first visit and forty-two (42) subjects
completing the study. Subjects'
age ranged from 20 to 53 years with a mean of 33.74 and standard deviation of
7.75 years.
Thirty-five (83%) of the subjects are female.
Table 1: Demographics Summary; All Subjects Treated
Demographic/
Statistic or Non-Prophy Prophy Overall
Category (n=21) (n=21) (n=42) P-value
Age (Years)
Mean (SD) 33.19 (7.33) 34.29 (8.28) 33.74 (7.75) 0.6525'
Min.-Max. 24 - 46 20 - 53 20 - 53
Sex
Femaleb 18 (86%) 17 (81%) 35 (83%) 1.0000d
Maleb 3 (14%) 4 (19%) 7 (17%)
Smoking
Nb 20 (95%) 20 (95%) 40 (95%) 1.0000d
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Table 1: Demographics Summary; All Subjects Treated
Demographic/
Statistic or Non-Prophy Prophy Overall
Category (n=21) (n=21) (n=42) P-value
yb
1 (5%) 1 (5%) 2 (5%)
a Two-sided Analysis of Variance for (ANOVA) p-value for the treatment
comparison.
b The number (percent) of subjects in each category.
c Two-sided chi-square p-value for the treatment comparison.
d Two-sided p-value for the treatment comparison from a Kruskal
Wallis test.
Efficacy Data
Manual brush: Referring to Table 2, at Baseline visit, Non-Prophy group and
Prophy
group starts from a Mean (SD) value of 1.93 (0.56) and 2.05 (0.49),
respectively. At Day 3 visit,
5 for Non-Prophy group and Prophy group, the Mean (SD) value is 1.80 (0.57)
and 1.61 (0.51),
respectively.
Referring to Table 3, the Non-Prophy group exhibits a difference from baseline
relative to
Prophy group with change from baseline means of 0.13 (0.36) (P=0.1162) and
0.44 (0.48)
(P=0.0005). The Prophy group exhibits significantly lower (P=0.0325) mean by
15.0% relative
10 to Non-Prophy group.
Power brush: Referring to Table 2, at Baseline visit, Non-Prophy group and
Prophy
group starts from a Mean (SD) value of 2.29 (0.50) and 2.31 (0.37),
respectively. At Day 3 visit,
for Non-Prophy group and Prophy group, the Mean (SD) value is 2.07 (0.55) and
1.78 (0.51),
respectively.
Table 2: Hemoglobin --- Day 03
Treatment N Mean (SD) Median Min.-Max.
Manual--Baseline (overall treatment difference p-value: 0.4631)
Non-Prophy 21 1.93 (0.56) 2.01 1.00 to 2.79
Prophy 21 2.05 (0.49) 2.06 1.00 to 3.03
Overall 42 1.99 (0.52) 2.02 1.00 to 3.03
Manual--Day 03
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Non-Prophy 21 1.80(0.57) 1.84 1.00
to 2.83
Prophy 21 1.61(0.51) 1.65 1.00
to 2.64
Power--Baseline (overall treatment difference p-value: 0.8927)
Non-Prophy 21 2.29 (0.50) 2.21 1.65
to 3.19
Prophy 21 2.31(0.37) 2.25 1.68
to 3.01
Overall 42 2.30 (0.43) 2.25 1.65
to 3.19
Power--Day 03
Non-Prophy 21 2.07 (0.55) 2.21 1.00
to 3.26
Prophy 21 1.78(0.51) 1.71 1.00
to 3.10
Referring to Table 3, the Non-Prophy group exhibited difference from baseline
relative to
Prophy group with changed from baseline means of 0.21 (0.49) (P=0.0606) and
0.53 (0.45)
(P<0.0001). Prophy group exhibited significantly lower (P=0.0341) mean by
14.7% relative to
Non-Prophy group.
Table 3: Change from Baseline; Hemoglobin --- Day 03
Treatment Mean (SD) P-valuea Median Min.-Max.
Manual--Day 03
Non-Prophy -0.13 (0.36) 0.1162 -0.11 -0.96 to
0.47
Prophy -0.44 (0.48) 0.0005 -0.39 -1.47 to
0.58
Power--Day 03
Non-Prophy -0.21 (0.49) 0.0606 -0.10 -1.21 to
0.56
Prophy -0.53 (0.45) <0.0001 -0.58 -1.39 to
0.22
a Tests mean change from baseline versus 0 (2-sided paired-
difference t-test)
Lastly, Table 4 below summarizes the efficacy results of the study. For the
Powerbrush,
the Prophy group exhibited significantly lower (P=0.0341) mean by 14.7%
relative to Non-
Prophy group. For the Manual brush, the Prophy group exhibited significantly
lower (P=0.0325)
mean by 15.0% relative to Non-Prophy group. This method is validated in Prophy
vs. Non-
prophy study (which is the best accepted standard in oral care industry and
accordingly is a
positive control).
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Table 4: Efficacy
Results
Analysis of Covariance: Adjusting for Baseline Score
Hemoglobin -- Day03
P-Value"
% Reduction
from Non-
Treatment N Adj. Mean (SE) Prophy' Prophy
Manual (error variance=0.161, baseline mean=1.99)
Non-Prophy 21 1.844 (0.088) 0.0325
Prophy 21 1.568(0.088) 15.0%
Power (error variance=0.204, baseline mean=2.30)
Non-Prophy 21 2.081 (0.099) 0.0341
Prophy 21 1.774 (0.099) 14.7%
a Percent Change versus Control = 100 x (( Control - Experimental )/ Control).
b
2-sided p-value comparing treatments using analysis of covariance.
Sample pictures of the clinical study are evaluated by visual perception
study. The details
are provided below:
Study Design
9 healthy female subjects aged 18-35 years old in Beijing as a naive grader
joined the
study.
The variable of deeper color is analyzed. The range of the score comparing pre
and post
treatment images is -4 to 4. The Prophy group treatment is dental prophylaxis;
and the Non-
Prophy group treatment, is the normal oral hygiene habit group.
The average score of all graders are calculated for each set of images (pre &
post images).
A simple t-test method is applied to test the score to assess if there is a
significant change after
treatment.
Treatment is considered statistically significantly better after applying
treatment if the p-
value is less than or equal to 0.05 (2-sided) and the average is negative.
Results
A lower score means less redness after day 3.
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Manual brush: In the Non-prophy group, the average mean is -0.85; in the
Prophy group,
the average mean is -1.51; the average means of both groups are statistically
significantly lower
than the hypothesis average mean 0 with P<0.039 and 0.001, respectively.
Comparing the 2
groups, no statistically significant difference is observed with P=0.225;
Power brush: In the Non-prophy group, the average mean is -0.531; in Prophy
group, the
average mean is -2.05; the average mean of Prophy group is statistically
significantly lower than
the hypothesis average mean 0 with P<0.0001, the average mean of Non-prophy
group does not
show statistically significant lower than the hypothesis average mean 0.
Comparing the 2 groups,
statistically significant difference is observed with P=0.018.
Table 5
T-Test: Mean = 0
Sampling Confidence
Method Treatment Mean (SD) P-Valuea Interval
Non- -0.85 (-1.644, -
Manual .
0039
Prophy (1.752) 0.049)
-1.51 (-
2.313, -
Manual Prophy .
0001
(1.757) 0.714)
-0.53
Power Non-phy 6)
0.320 (-1.611,0.552)
Pro (2.37
-2.05 (-
2.740, -
Power Prophy .
<00001
(1.520) 1.356)
a T-Test P-Value against the hypothesis that the average score is
Zero.
Table 6
T-Test: Comparing Treatment Mean
Confidence
Sampling Method Treatment Mean (SD) Diff P-Value Interval
Non- -0.85
Manual Prophy (1.752) 0.6667 0.2254a (-0.428,
1.761)a
-1.51
Manual Prophy
(1.757)
Non- -0.53
Power Prophy (2.376) 1.5185 0.0180a (0.275, 2.763)a
-2.05
Power Prophy
(1.520)
a T-Test in the Pooled method for Equal Variance between groups.
b
T-Test in the Scatterthwaite method for Unequal Variance between groups.
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Conclusion
Power brush: Prophy group shows significantly better improvement after
treatment and it
exhibits significantly better change (P=0.018) relative to Non-prophy group.
Manual brush: Both Prophy group and Non-prophy group show the significantly
better
improvement after treatment, but Prophy group is not significantly better
(P=0.225) than the
Non-prophy group.
This sampling method by power brush is validated in Prophy vs. Non-prophy
study
(which is best accepted standard in oral care industry and accordingly is a
positive control).
The dimensions and values disclosed herein are not to be understood as being
strictly
limited to the exact numerical values recited. Instead, unless otherwise
specified, each such
dimension is intended to mean both the recited value and a functionally
equivalent range
surrounding that value. For example, a dimension disclosed as "40 mm" is
intended to mean
"about 40 mm."
Every document cited herein, including any cross referenced or related patent
or
application and any patent application or patent to which this application
claims priority or
benefit thereof, is hereby incorporated herein by reference in its entirety
unless expressly
excluded or otherwise limited. The citation of any document is not an
admission that it is prior
art with respect to any invention disclosed or claimed herein or that it
alone, or in any
combination with any other reference or references, teaches, suggests or
discloses any such
invention. Further, to the extent that any meaning or definition of a term in
this document
conflicts with any meaning or definition of the same term in a document
incorporated by
reference, the meaning or definition assigned to that term in this document
shall govern.
While particular embodiments of the present invention have been illustrated
and
described, it would be obvious to those skilled in the art that various other
changes and
modifications can be made without departing from the spirit and scope of the
invention. It is
therefore intended to cover in the appended claims all such changes and
modifications that are
within the scope of this invention.
