U.S. patent application number 14/167493 was filed with the patent office on 2014-05-29 for dental mineralization.
This patent application is currently assigned to The University of Melbourne. The applicant listed for this patent is The University of Melbourne. Invention is credited to Eric Charles REYNOLDS.
Application Number | 20140147512 14/167493 |
Document ID | / |
Family ID | 37498029 |
Filed Date | 2014-05-29 |
United States Patent
Application |
20140147512 |
Kind Code |
A1 |
REYNOLDS; Eric Charles |
May 29, 2014 |
DENTAL MINERALIZATION
Abstract
A method is provided for mineralizing a dental surface or
subsurface including contacting the dental surface with a protein
disrupting agent and stabilized amorphous calcium phosphate (ACP)
or amorphous calcium fluoride phosphate (ACFP).
Inventors: |
REYNOLDS; Eric Charles;
(Carlton, AU) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
The University of Melbourne |
Parkville |
|
AU |
|
|
Assignee: |
The University of Melbourne
Parkville
AU
|
Family ID: |
37498029 |
Appl. No.: |
14/167493 |
Filed: |
January 29, 2014 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
11916831 |
Dec 7, 2007 |
8673363 |
|
|
PCT/AU2006/000785 |
Jun 7, 2006 |
|
|
|
14167493 |
|
|
|
|
Current U.S.
Class: |
424/602 |
Current CPC
Class: |
A61K 8/24 20130101; A61K
8/22 20130101; A61K 8/20 20130101; A61P 1/02 20180101; A61K 33/42
20130101; A61P 43/00 20180101; A61K 8/66 20130101; A61Q 11/00
20130101; A61K 8/64 20130101; A61K 8/42 20130101; A61K 8/21
20130101; A61K 2800/70 20130101; A61K 8/19 20130101 |
Class at
Publication: |
424/602 |
International
Class: |
A61K 8/24 20060101
A61K008/24; A61Q 11/00 20060101 A61Q011/00; A61K 8/42 20060101
A61K008/42 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 7, 2005 |
AU |
2005902961 |
Claims
1. A method for mineralizing a dental enamel comprising
administering a protein disrupting agent to the enamel prior to
contacting the enamel with a phosphopeptide-stabilized amorphous
calcium phosphate (ACP) and/or amorphous calcium fluoride phosphate
(ACFP).
2. A method according to claim 1, wherein the protein disrupting
agent is selected from one or more of the group consisting of a
detergent, a chaotropic agent, a protease and a mixture of
proteases.
3. A method according to claim 2, wherein the chaotropic agent is
urea.
4. A method according to claim 2, wherein the protease or mixture
of proteases is selected from the group consisting of
endopeptidases, proteinases and exopeptidases.
5. A method according to claim 1, wherein the phosphopeptide is a
casein phosphopeptide.
6. A method according to claim 1, wherein the ACP or ACFP is in a
basic phase.
7. A method according to claim 1, wherein the dental enamel is in
an animal selected from the group consisting of humans, domestic
animals, companion animals and zoo animals.
8. A method for reducing a white spot lesion comprising
administering a protein disrupting agent to the enamel prior to
contacting the enamel with a phosphopeptide-stabilized amorphous
calcium phosphate (ACP) and/or amorphous calcium fluoride phosphate
(ACFP).
9. A method according to claim 8, wherein the white spot lesion is
caused by dental caries, dental erosion or fluorosis.
10. A method according to claim 8, wherein the protein disrupting
agent is selected from one or more of the group consisting of a
detergent, a chaotropic agent, a protease and a mixture of
proteases.
11. A method according to claim 10, wherein the chaotropic agent is
urea.
12. A method according to claim 10, wherein the protease or mixture
of proteases is selected from the group consisting of
endopeptidases, proteinases and exopeptidases.
13. A method according to claim 8, wherein the phosphopeptide is a
casein phosphopeptide.
14. A method according to claim 8, wherein the ACP or ACFP is in a
basic phase.
15. A kit for mineralizing a dental enamel including: (a) a first
composition comprising a protein disrupting agent; and (b) a second
composition comprising a phosphopeptide-stabilized amorphous
calcium phosphate (ACP) and/or amorphous calcium fluoride phosphate
(ACFP) complex in a pharmaceutically acceptable carrier, and
written instructions, wherein the written instructions direct the
user to administer the first composition prior to the second
composition.
16. A kit according to claim 15, wherein the protein disrupting
agent is selected from one or more of the group consisting of a
detergent, a chaotropic agent, a protease and a mixture of
proteases.
17. A method according to claim 16, wherein the chaotropic agent is
urea.
18. A method according to claim 16, wherein the protease or mixture
of proteases is selected from the group consisting of
endopeptidases, proteinases and exopeptidases.
19. A kit according to claim 15, wherein the phosphopeptide is a
casein phosphopeptide.
20. A kit according to claim 15, wherein the ACP or ACFP is in a
basic phase.
Description
[0001] This application is a Continuation of U.S. Ser. No.
11/916,831, filed 7 Dec. 2007, which is a National Stage
Application of PCT/AU2006/000785, filed 7 Jun. 2006, which claims
benefit of Serial No. 2005902961, filed 7 Jun. 2005 in Australia
and which applications are incorporated herein by reference. To the
extent appropriate, a claim of priority is made to each of the
above disclosed applications.
[0002] The present invention relates to a method of mineralizing a
dental surface, in particular tooth enamel. Methods of mineralizing
hypomineralized lesions (including subsurface lesions) in the tooth
enamel caused by dental caries, dental erosion and fluorosis are
also provided.
BACKGROUND
[0003] Common causes of hypomineralized lesions are caries and
fluorosis.
[0004] Dental caries is initiated by the demineralization of hard
tissue of the teeth usually by organic acids produced from
fermentation of dietary sugar by dental plaque odontopathogenic
bacteria. Dental caries is still a major public health problem.
Further, restored tooth surfaces can be susceptible to further
dental caries around the margins of the restoration. Even though
the prevalence of dental caries has decreased through the use of
fluoride in most developed countries, the disease remains a major
public health problem. Dental erosion or corrosion is the loss of
tooth mineral by dietary or regurgitated acids. Dental
hypersensitivity is due to exposed dentinal tubules through loss of
the protective mineralized layer, cementum. Dental calculus is the
unwanted accretion of calcium phosphate minerals on the tooth
surface. All these conditions, dental caries, dental erosion,
dental hypersensitivity and dental calculus are therefore
imbalances in the level of calcium phosphates.
[0005] Enamel fluorosis (mottling) has been recognized for nearly a
century, however, the aetiological role of fluoride was not
identified until 1942 (Black and McKay, 1916). The characteristic
appearance of fluorosis may be differentiated from other enamel
disturbances (Fejerskov et al., 1991). The clinical features of
fluorotic lesions of enamel (FLE) represent a continuum ranging
from fine opaque lines following the perikymata, to chalky, white
enamel (Fejerskov et al., 1990; Giambro et al., 1995). The presence
of a comparatively highly mineralized enamel outer surface and a
hypomineralized subsurface in the fluorotic lesion simulates the
incipient enamel "white spot" carious lesion (Fejerskov et al.,
1990). With increasing severity, both the depth of enamel involved
in the lesion and the degree of hypomineralization increases
(Fejerskov et al., 1990, Giambro at al., 1995). The development of
fluorosis is highly dependent on the dose, duration and timing of
fluoride exposure (Fejerskov et al., 1990, Fejerskov et al., 1996;
Aoba and Fejerskov, 2002) and is believed to be related to elevated
serum fluoride concentrations. Chalky "white spot" lesions may also
form on developing teeth in children such as after treatment with
antibiotics or fever. Such lesions indicate areas of
hypomineralization of the tooth enamel.
[0006] Depending on lesion severity, fluorosis has been managed
clinically by restorative replacement or micro-abrasion of the
outer enamel (Den Besten and Thariani, 1992; Fejerskov at al.,
1996). These treatments are unsatisfactory because they involve
restorations or removal of tooth tissue. What is desired is a
treatment that will mineralize the hypomineralized enamel to
produce a natural appearance and structure.
[0007] Specific complexes of casein phosphopeptides and amorphous
calcium phosphate ("CPP-ACP", available commercially as
Recaldent.TM.) have been shown to remineralize enamel subsurface
lesions in vitro and in situ (Reynolds, 1998; Shen at al., 2001;
Reynolds et al., 2003).
[0008] WO 98/40406 in the name of The University of Melbourne (the
contents of which are herein incorporated fully by reference)
describes casein phosphopeptide-amorphous calcium phosphate
complexes (CPP-ACP) and CPP-stabilised amorphous calcium fluoride
phosphate complexes (CPP-ACFP) which have been produced at alkaline
pH. Such complexes have been shown to prevent enamel
demineralization and promote remineralization of enamel subsurface
lesions in animal and human in situ caries models (Reynolds,
1998).
[0009] The CPP which are active in forming the complexes do so
whether or not they are part of a full-length casein protein.
Examples of active (CPP) that can be isolated after tryptic
digestion of full length casein have been specified in U.S. Pat.
No. 5,015,628 and include peptides Bos .alpha..sub.s1-casein X-5P
(f59-79) [1], Bos .beta.-casein X-4P (f1-25) [2], Bos
.alpha..sub.s2-casein X-4P (f46-70) [3] and Bos
.alpha..sub.s2-casein X-4P (f1-21) [4] as follows:
TABLE-US-00001 [1]
Gln.sup.59-Met-Glu-Ala-Glu-Ser(P)-Ile-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-Il-
e-Val-Pro-Asn- Ser(P)-Val-Glu-Gln-Lys.sup.79 .alpha..sub.s1(59-79)
[2]
Arg.sup.1-Glu-Leu-Glu-Glu-Leu-Asn-Val-Pro-Gly-Glu-Ile-Val-Glu-Ser(P)-L-
eu-Ser(P)- Ser(P)-Ser(P)-Glu-Glu-Ser-Ile-Thr-Arg.sup.25
.beta.(1-25) [3]
Asn.sup.46-Ala-Asn-Glu-Glu-Glu-Tyr-Ser-Ile-Gly-Ser(P)-Ser(P)-Ser(P)-Gl-
u-Glu-Ser(P)- Ala-Glu-Val-Ala-Thr-Glu-Glu-Val-Lys.sup.70
.alpha..sub.s2(46-70) [4]
Lys.sup.1-Asn-Thr-Met-Glu-His-Val-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-Ser-Ile-
-Ile-Ser(P)- Gln-Glu-Thr-Tyr-Lys.sup.21 .alpha..sub.s2(1-21)
[0010] The access of mineralizing ions to the tooth enamel in many
cases can be limited by the layer of salivary proteins that forms
over the surface of the enamel, termed the pellicle. The proteins
of the pellicle can also accumulate in sub-surface enamel lesions,
thereby inhibiting the mineralization of these lesions. Such
accumulations of proteins can discolour over time, leaving
unsightly patches on the tooth. Accordingly, there is a need to
remove these proteins to remove discolouration and avoid
limitations of access to the enamel by remineralizing ions. To
overcome these and other limitations of known treatments, research
to this end has been conducted.
SUMMARY OF THE INVENTION
[0011] In one aspect, the present invention provides a method of
mineralizing a dental surface or sub-surface including contacting
the dental surface with a protein disrupting agent, and contacting
the dental surface with stabilized amorphous calcium phosphate
(ACP) or amorphous calcium fluoride phosphate (ACFP). The dental
surface is preferably dental enamel. In one embodiment the dental
surface is a lesion in the enamel, such as a lesion caused by
caries, dental erosion or fluorosis.
[0012] Mineralization of dental surfaces can be significantly
enhanced by the disruption of pellicle proteins from the dental
surface prior to the application of a remineralizing material, such
as stabilised ACP and/or ACFP. In particular, it has been found
that the mineralization of enamel by stabilized soluble forms of
ACP (CPP-ACP) and ACFP (CPP-ACFP) is enhanced by pre-treatment of
the enamel surface with a protein disrupting agent such as alkaline
bleach.
[0013] Preferably the ACP and/or ACFP is phosphopeptide
(PP)-stabilized. Preferably, the phosphopeptide (as defined below)
is a casein phosphopeptide.
[0014] In a preferred embodiment the ACP and/or ACFP is in the form
of a casein phosphopeptide stabilized ACP and/or ACFP complex.
[0015] Preferably, the phase of the ACP is predominantly a basic
phase, wherein the ACP comprises predominantly the species
Ca.sup.2+, PO.sub.4.sup.3- and OH.sup.-. The basic phase of ACP may
have the general formula
[Ca.sub.3(PO.sub.4).sub.2].sub.x[Ca.sub.2(PO.sub.4)(OH)] where
x.gtoreq.1. Preferably x=1-5. More preferably, x=1. Preferably the
two components of the formula are present in equal proportions.
Accordingly, in one embodiment, the basic phase of ACP has the
formula Ca.sub.3(PO.sub.4).sub.2Ca.sub.2(PO.sub.4)(OH).
[0016] Preferably, the phase of the ACFP is predominantly a basic
phase, wherein the ACFP comprises predominantly the species
Ca.sup.2+, PO.sub.4.sup.3- and F.sup.-. The basic phase of ACFP may
have the general formula
[Ca.sub.3(PO.sub.4).sub.2].sub.x[Ca.sub.2(PO.sub.4)F].sub.y where
x.gtoreq.1 when y=1 or where y.gtoreq.1 when x=1. Preferably, y=1
and x=1-3. More preferably, y=1 and x=1. Preferably the two
components of the formula are present in equal proportions.
Accordingly, in one embodiment, the basic phase of ACFP has the
formula Ca.sub.3(PO.sub.4).sub.2Ca.sub.2(PO.sub.4)F.
[0017] In one embodiment, the ACP complex consists essentially of
phosphopeptides, calcium, phosphate and hydroxide ions and
water.
[0018] In one embodiment, the ACFP complex consists essentially of
phosphopeptides, calcium, phosphate, fluoride and hydroxide ions
and water.
DETAILED DESCRIPTION OF THE INVENTION
[0019] Any suitable protein disrupting agent can be used in the
method of the present invention. The agent is required to reduce
the proteinaceous barrier formed over the surface to be treated,
such as the pellicle over teeth. Examples of suitable agents
include bleach, detergent, chaotropic agents such as urea, high
phosphate concentrations, cocktails of proteases (e.g.
endopeptidases, proteinases and exopeptidases) and any other
protein solubilizing, disrupting or hydrolysing agent.
[0020] Examples of suitable bleaches include sodium hypochlorite
(NaOCl), and cabamide peroxide bleaches. In a preferred embodiment,
the bleach is an alkaline bleach. In a further preferred embodiment
the alkaline bleach is NaOCl. The protein disrupting agent acts to
solubilize and partially or wholly remove proteins from the dental
surface, particularly proteins of the pellicle.
[0021] In a further aspect of the present invention there is
provided a method of mineralizing a dental surface comprising
providing a protein disrupting agent and a source of ACP or ACFP.
In a preferred embodiment the dental surface is enamel.
[0022] In a further aspect of the present invention there is
provided a method for treating fluorosis comprising contacting a
fluorotic lesion in tooth enamel with a protein disrupting agent
and stabilized ACP and/or ACFP.
[0023] In a further aspect of the present invention there is
provided a method for treating dental caries comprising contacting
a caries lesion in tooth enamel with a protein disrupting agent and
stabilized ACP and/or ACFP.
[0024] In a further aspect of the present invention there is
provided a method for treating dental erosion comprising contacting
a lesion in tooth enamel caused by erosion with a protein
disrupting agent and stabilized ACP and/or ACFP.
[0025] In a further aspect of the present invention there is
provided a method for reducing white spot lesions on the tooth
enamel comprising contacting a white spot lesion with a protein
disrupting agent and stabilized ACP and/or ACFP.
[0026] In a further aspect of the present invention there is
provided a method for remineralizing a lesion in tooth enamel
comprising contacting the lesion with a protein disrupting agent
and stabilized ACP and/or ACFP.
[0027] Preferably the ACP and/or ACFP is stabilized by a
phosphopeptide. In a preferred embodiment the phosphopeptide is a
casein phosphopeptides. Preferably, the ACP or ACFP is in the form
of a casein phosphopeptide stabilized ACP or ACFP complex.
[0028] In one embodiment, the protein disrupting agent is NaOCl. A
concentration of about 1 to 20% NaOCl may be used. Alternatively,
the concentration of NaOCl is 1 to 10%. In a preferred embodiment,
about 5% NaOCl is used.
[0029] The protein disrupting agent may be contacted with the
dental surface for a period of about 1 to 60 minutes, or for about
1 to 30 minutes. In one embodiment, the protein disrupting agent is
contacted with the dental surface for about 20 minutes.
[0030] Preferably the stabilized ACP and/or ACFP is contacted with
the dental surface for a period of about 1 minute to 2 hours, or 5
minutes to 60 minutes or about 10 minutes. The stabilized ACP
and/or ACFP may be repeatedly applied to the dental surface over a
period of 1 day to several months.
[0031] In one embodiment, the stabilized ACP and/or ACFP is
contacted with the dental surface after the dental surface has been
contacted with the protein disrupting agent.
[0032] In a preferred embodiment, the protein disrupting agent is
contacted with the dental surface 1 to 60 minutes, or 1 to 30
minutes, or 1 to 5 minutes prior to contacting the dental surface
with the stabilized ACP and/or ACFP.
[0033] In a further aspect of the present invention there is
provided a method for mineralizing a tooth surface comprising
applying an ACP and/or ACFP complex to a tooth surface that has
been pre-treated with a protein disrupting agent. Preferably the
tooth surface is tooth enamel. In a preferred embodiment, the tooth
surface is tooth enamel containing a lesion selected from the group
consisting of one or more of a white spot lesion; a fluorotic
lesion; a caries lesion; or a lesion caused by tooth erosion. In a
further preferred embodiment the protein disrupting agent is a
bleach.
[0034] In one embodiment, the dental surface is in need of such
treatment. The invention also includes a method of treating a
subject suffering fluorosis, dental caries, dentinal
hypersensitivity or dental calculus.
[0035] Without being bound by any theory or mode of action it is
understood that pre-conditioning tooth enamel with a protein
disrupting agent results in partial or complete enamel
de-proteination, enhancing the diffusion of calcium and phosphate
into subsurface enamel.
[0036] It is further understood that treatment of tooth enamel with
stabilised ACFP produces fluorapatite, which is more resistant to
acid challenge than normal tooth enamel. This may result in tooth
enamel with superior caries resistant properties. Accordingly, in a
preferred embodiment the method of the present invention includes
stabilised ACFP.
[0037] "Phosphopeptide" in the context of the description of this
invention means an amino acid sequence in which at least one amino
acid is phosphorylated. Preferably, the phosphopeptide includes one
or more of the amino acid sequence -A-B-C-, where A is a
phosphoamino residue, B is any amino acyl residue including a
phosphoamino residue and C is selected from a glutamyl, aspartyl or
phosphoamino residue. Any of the phosphoamino residues may
independently be a phosphoseryl residue. B is desirably a residue
the side-chain of which is neither relatively large nor
hydrophobic. It may be Gly, Ala, Val, Met, Leu, Ile, Ser, Thr, Cys,
Asp, Glu, Asn, Gln or Lys.
[0038] In another embodiment, at least two of the phosphoamino
acids in the sequence are preferably contiguous. Preferably the
phosphopeptide includes the sequence A-B-C-D-E, where A, B, C, D
and E are independently phosphoserine, phosphothreonine,
phosphotyrosine, phosphohistidine, glutamic acid or aspartic acid,
and at least two, preferably three, of the A, B, C, D and E are a
phosphoamino acid. In a preferred embodiment, the phosphoamino acid
residues are phosphoserine, most preferably three contiguous
phosphoserine residues. It is also preferred that D and E are
independently glutamic or aspartic acid.
[0039] It will also be understood that the term "comprises" (or its
grammatical variants) as used in this specification is equivalent
to the term "includes" and may be used interchangeably and should
not be taken as excluding the presence of other elements or
features.
[0040] In one embodiment, the ACP or ACFP is stabilized by a casein
phosphopeptide (CPP), which is in the form of intact casein or
fragment of the casein, and the complex formed preferably has the
formula [CPP(ACP).sub.8].sub.n or [(CPP)(ACFP).sub.8].sub.n where n
is equal to or greater than 1, for example 6. The complex formed
may be a colloidal complex, where the core particles aggregate to
form large (eg 100 nm) colloidal particles suspended in water.
Thus, the PP can be a casein protein or a polyphosphopeptide.
[0041] The PP may be from any source; it may be present in the
context of a larger polypeptide, including a full length casein
polypeptide, or it may be isolated by tryptic or other enzymatic or
chemical digestion of casein, or other phosphoamino acid rich
proteins such as phosphitin, or by chemical or recombinant
synthesis, provided that it comprises the sequence -A-B-C- or
A-B-C-D-E as described above. The sequence flanking this core
sequence may be any sequence. However, those flanking sequences in
.alpha..sub.s1(59-79) [1], .beta.(1-25) [2], .alpha..sub.s2(46-70)
[3] and .alpha..sub.s2(1-21) [4] are preferred. The flanking
sequences may optionally be modified by deletion, addition or
conservative substitution of one or more residues. The amino acid
composition and sequence of the flanking region are not
critical.
[0042] Examples of conservative substitutions are shown in Table 1
below.
TABLE-US-00002 TABLE 1 Exemplary Conservative Preferred
Conservative Original Residue Substitution Substitution Ala Val,
Leu, Ile Val Asn Gln Lys His Phe Gln Gln Asn Asn Gly Pro Pro Ile
Leu, Val, Met, Ala, Phe Leu Leu Ile, Val, Met, Ala, Phe Ile Lys
Arg, Gln, Asn Arg Phe Leu, Val, Ile, Ala Leu Pro Gly Gly Ser Thr
Thr Val Ile, Leu, Met, Phe, Ala Leu Asp Glu Glu Thr Ser Ser Trp Tyr
Tyr Tyr Trp Phe Thr Ser Phe
[0043] The flanking sequences may also include non-naturally
occurring amino acid residues. Commonly encountered amino acids
which are not encoded by the genetic code, include: [0044] 2-amino
adipic acid (Aad) for Glu and Asp; [0045] 2-aminopimelic acid (Apm)
for Glu and Asp; [0046] 2-aminobutyric (Abu) acid for Met, Leu, and
other aliphatic amino acids; [0047] 2-aminoheptanoic acid (Ahe) for
Met, Leu and other aliphatic amino acids; [0048] 2-aminoisobutyric
acid (Aib) for Gly; [0049] cyclohexylalanine (Cha) for Val, and Leu
and Ile; [0050] homoarginine (Har) for Arg and Lys; [0051]
2,3-diaminopropionic acid (Dpr) for Lys, Arg and His; [0052]
N-ethylglycine (EtGly) for Gly, Pro, and Ala; [0053]
N-ethylasparigine (EtAsn) for Asn, and Gln; [0054] Hydroxyllysine
(Hyl) for Lys; [0055] allohydroxyllysine (AHyl) for Lys; [0056]
3-(and 4) hydroxyproline (3Hyp, 4Hyp) for Pro, Ser, and Thr; [0057]
alloisoleucine (AIle) for Ile, Leu, and Val; [0058]
.rho.-amidinophenylalanine for Ala; [0059] N-methylglycine (MeGly,
sarcosine) for Gly, Pro, Ala. [0060] N-methylisoleucine (MeIle) for
Ile; [0061] Norvaline (Nva) for Met and other aliphatic amino
acids; [0062] Norleucine (Nle) for Met and other aliphatic amino
acids; [0063] Ornithine (Orn) for Lys, Arg and His; [0064]
Citrulline (Cit) and methionine sulfoxide (MSO) for Thr, Asn and
Gln; [0065] N-methylphenylalanine (MePhe), trimethylphenylalanine,
halo (F, Cl, Br and I) phenylalanine, triflourylphenylalanine, for
Phe.
[0066] In one embodiment, the PP is one or more phosphopeptides
selected from the group consisting of .alpha..sub.s1(59-79) [1],
.beta.(1-25) [2], .alpha..sub.s2(46-70) [3] and
.alpha..sub.s2(1-21) [4].
[0067] In another embodiment of the invention, the stabilised ACFP
or ACP complex is incorporated into oral compositions such as
toothpaste, mouth washes or formulations for the mouth to aid in
the prevention and/or treatment of dental caries, tooth decay,
dental erosion or fluorosis. The ACFP or ACP complex may comprise
0.01-50% by weight of the composition, preferably 1.0-50%. For oral
compositions, it is preferred that the amount of the CPP-ACP and/or
CPP-ACFP administered is 0.01-50% by weight, preferably 1.0%-50% by
weight of the composition. In a particularly preferred embodiment,
the oral composition of the present invention contains about 2%
CPP-ACP, CPP-ACFP or a mixture of both. The oral composition of
this invention which contains the above-mentioned agents may be
prepared and used in various forms applicable to the mouth such as
dentifrice including toothpastes, toothpowders and liquid
dentifrices, mouthwashes, troches, chewing gums, dental pastes,
gingival massage creams, gargle tablets, dairy products and other
foodstuffs. The oral composition according to this invention may
further include additional well known ingredients depending on the
type and form of a particular oral composition.
[0068] In certain preferred forms of the invention the oral
composition may be substantially liquid in character, such as a
mouthwash or rinse. In such a preparation the vehicle is typically
a water-alcohol mixture desirably including a humectant as
described below. Generally, the weight ratio of water to alcohol is
in the range of from about 1:1 to about 20:1. The total amount of
water-alcohol mixture in this type of preparation is typically in
the range of from about 70 to about 99.9% by weight of the
preparation. The alcohol is typically ethanol or isopropanol.
Ethanol is preferred.
[0069] The pH of such liquid and other preparations of the
invention is generally in the range of from about 5 to about 9 and
typically from about 5.0 to 7.0. The pH can be controlled with acid
(e.g. phosphoric acid, citric acid or benzoic acid) or base (e.g.
sodium hydroxide) or buffered (as with sodium citrate, benzoate,
carbonate, or bicarbonate, disodium hydrogen phosphate, sodium
dihydrogen phosphate, etc).
[0070] In other desirable forms of this invention, the stabilised
ACP or ACFP composition may be substantially solid or pasty in
character, such as toothpowder, a dental tablet or a toothpaste
(dental cream) or gel dentifrice. The vehicle of such solid or
pasty oral preparations generally contains dentally acceptable
polishing material. Examples of polishing materials are
water-insoluble sodium metaphosphate, potassium metaphosphate,
tricalcium phosphate, dihydrated calcium phosphate, anhydrous
dicalcium phosphate, calcium pyrophosphate, magnesium
orthophosphate, trimagnesium phosphate, calcium carbonate, hydrated
alumina, calcined alumina, aluminium silicate, zirconium silicate,
silica, bentonite, and mixtures thereof. Other suitable polishing
material include the particulate thermosetting resins such as
melamine-, phenolic, and urea-formaldehydes, and cross-linked
polyepoxides and polyesters. Preferred polishing materials include
crystalline silica having particle sizes of up to about 5 microns,
a mean particle size of up to about 1.1 microns, and a surface area
of up to about 50,000 cm.sup.2/g., silica gel or colloidal silica,
and complex amorphous alkali metal aluminosilicate.
[0071] When visually clear gels are employed, a polishing agent of
colloidal silica, such as those sold under the trademark SYLOID as
Syloid 72 and Syloid 74 or under the trademark SANTOCEL as Santocel
100, alkali metal aluminosilicate complexes are particularly useful
since they have refractive indices close to the refractive indices
of gelling agent-liquid (including water and/or humectant) systems
commonly used in dentifrices.
[0072] Many of the so-called "water insoluble" polishing materials
are anionic in character and also include small amounts of soluble
material. Thus, insoluble sodium metaphosphate may be formed in any
suitable manner, for example as illustrated by Thorpe's Dictionary
of Applied Chemistry, Volume 9, 4th Edition, pp. 510-511. The forms
of insoluble sodium metaphosphate known as Madrell's salt and
Kurrol's salt are further examples of suitable materials. These
metaphosphate salts exhibit only a minute solubility in water, and
therefore are commonly referred to as insoluble metaphosphates
(IMP). There is present therein a minor amount of soluble phosphate
material as impurities, usually a few percent such as up to 4% by
weight. The amount of soluble phosphate material, which is believed
to include a soluble sodium trimetaphosphate in the case of
insoluble metaphosphate, may be reduced or eliminated by washing
with water if desired. The insoluble alkali metal metaphosphate is
typically employed in powder form of a particle size such that no
more than 1% of the material is larger than 37 microns.
[0073] The polishing material is generally present in the solid or
pasty compositions in weight concentrations of about 10% to about
99%. Preferably, it is present in amounts from about 10% to about
75% in toothpaste, and from about 70% to about 99% in toothpowder.
In toothpastes, when the polishing material is silicious in nature,
it is generally present in an amount of about 10-30% by weight.
Other polishing materials are typically present in amount of about
30-75% by weight.
[0074] In a toothpaste, the liquid vehicle may comprise water and
humectant typically in an amount ranging from about 10% to about
80% by weight of the preparation. Glycerine, propylene glycol,
sorbitol and polypropylene glycol exemplify suitable
humectants/carriers. Also advantageous are liquid mixtures of
water, glycerine and sorbitol. In clear gels where the refractive
index is an important consideration, about 2.5-30% w/w of water, 0
to about 70% w/w of glycerine and about 20-80% w/w of sorbitol are
preferably employed.
[0075] Toothpaste, creams and gels typically contain a natural or
synthetic thickener or gelling agent in proportions of about 0.1 to
about 10, preferably about 0.5 to about 5% w/w. A suitable
thickener is synthetic hectorite, a synthetic colloidal magnesium
alkali metal silicate complex clay available for example as
Laponite (e.g. CP, SP 2002, D) marketed by Laporte Industries
Limited. Laponite D is, approximately by weight 58.00% SiO.sub.2,
25.40% MgO, 3.05% Na.sub.2O, 0.98% Li.sub.2O, and some water and
trace metals. Its true specific gravity is 2.53 and it has an
apparent bulk density of 1.0 g/ml at 8% moisture.
[0076] Other suitable thickeners include Irish moss, iota
carrageenan, gum tragacanth, starch, polyvinylpyrrolidone,
hydroxyethylpropylcellulose, hydroxybutyl methyl cellulose,
hydroxypropyl methyl cellulose, hydroxyethyl cellulose (e.g.
available as Natrosol), sodium carboxymethyl cellulose, and
colloidal silica such as finely ground Syloid (e.g. 244).
Solubilizing agents may also be included such as humectant polyols
such propylene glycol, dipropylene glycol and hexylene glycol,
cellosolves such as methyl cellosolve and ethyl cellosolve,
vegetable oils and waxes containing at least about 12 carbons in a
straight chain such as olive oil, castor oil and petrolatum and
esters such as amyl acetate, ethyl acetate and benzyl benzoate.
[0077] It will be understood that, as is conventional, the oral
preparations will usually be sold or otherwise distributed in
suitable labelled packages. Thus, a jar of mouth rinse will have a
label describing it, in substance, as a mouth rinse or mouthwash
and having directions for its use; and a toothpaste, cream or gel
will usually be in a collapsible tube, typically aluminium, lined
lead or plastic, or other squeeze, pump or pressurized dispenser
for metering out the contents, having a label describing it, in
substance, as a toothpaste, gel or dental cream.
[0078] Organic surface-active agents may be used in the
compositions of the present invention to achieve increased
prophylactic action, assist in achieving thorough and complete
dispersion of the active agent throughout the oral cavity, and
render the instant compositions more cosmetically acceptable. The
organic surface-active material is preferably anionic, non-ionic or
ampholytic in nature and preferably does not interact with the
active agent. It is preferred to employ as the surface-active agent
a detersive material which imparts to the composition detersive and
foaming properties. Suitable examples of anionic surfactants are
water-soluble salts of higher fatty acid monoglyceride
monosulfates, such as the sodium salt of the monosulfated
monoglyceride of hydrogenated coconut oil fatty acids, higher alkyl
sulfates such as sodium lauryl sulfate, alkyl aryl sulfonates such
as sodium dodecyl benzene sulfonate, higher alkylsulfo-acetates,
higher fatty acid esters of 1,2-dihydroxy propane sulfonate, and
the substantially saturated higher aliphatic acyl amides of lower
aliphatic amino carboxylic acid compounds, such as those having 12
to 16 carbons in the fatty acid, alkyl or acyl radicals, and the
like. Examples of the last mentioned amides are N-lauroyl
sarcosine, and the sodium, potassium, and ethanolamine salts of
N-lauroyl, N-myristoyl, or N-palmitoyl sarcosine which should be
substantially free from soap or similar higher fatty acid material.
The use of these sarconite compounds in the oral compositions of
the present invention is particularly advantageous since these
materials exhibit a prolonged marked effect in the inhibition of
acid formation in the oral cavity due to carbohydrates breakdown in
addition to exerting some reduction in the solubility of tooth
enamel in acid solutions. Examples of water-soluble non-ionic
surfactants suitable for use are condensation products of ethylene
oxide with various reactive hydrogen-containing compounds reactive
therewith having long hydrophobic chains (e.g. aliphatic chains of
about 12 to 20 carbon atoms), which condensation products
("ethoxamers") contain hydrophilic polyoxyethylene moieties, such
as condensation products of poly (ethylene oxide) with fatty acids,
fatty alcohols, fatty amides, polyhydric alcohols (e.g. sorbitan
monostearate) and polypropyleneoxide (e.g. Pluronic materials).
[0079] The surface active agent is typically present in amount of
about 0.1-5% by weight. It is noteworthy, that the surface active
agent may assist in the dissolving of the active agent of the
invention and thereby diminish the amount of solubilizing humectant
needed.
[0080] Various other materials may be incorporated in the oral
preparations of this invention such as whitening agents,
preservatives, silicones, chlorophyll compounds and/or ammoniated
material such as urea, diammonium phosphate, and mixtures thereof.
These adjuvants, where present, are incorporated in the
preparations in amounts which do not substantially adversely affect
the properties and characteristics desired.
[0081] Any suitable flavouring or sweetening material may also be
employed. Examples of suitable flavouring constituents are
flavouring oils, e.g. oil of spearmint, peppermint, wintergreen,
sassafras, clove, sage, eucalyptus, marjoram, cinnamon, lemon, and
orange, and methyl salicylate. Suitable sweetening agents include
sucrose, lactose, maltose, sorbitol, xylitol, sodium cyclamate,
perillartine, AMP (aspartyl phenyl alanine, methyl ester),
saccharine, and the like. Suitably, flavour and sweetening agents
may each or together comprise from about 0.1% to 5% more of the
preparation.
[0082] The invention also provides an ACP or ACFP composition as
described above further including a protein disrupting agent. In
one embodiment, the protein disrupting agent is a bleach. In a
preferred embodiment the bleach is NaOCl.
[0083] The compositions of this invention can also be incorporated
in lozenges, or in chewing gum or other products, e.g. by stirring
into a warm gum base or coating the outer surface of a gum base,
illustrative of which are jelutong, rubber latex, vinylite resins,
etc., desirably with conventional plasticizers or softeners, sugar
or other sweeteners or such as glucose, sorbitol and the like.
[0084] In a further aspect, the invention provides compositions
including pharmaceutical compositions comprising any of the ACFP
and/or ACP complexes as described above together with a protein
disrupting agent and a pharmaceutically-acceptable carrier. Such
compositions may be selected from the group consisting of dental,
anticariogenic compositions and therapeutic compositions. Dental
compositions or therapeutic compositions may be in the form of a
gel, liquid, solid, powder, cream or lozenge. Therapeutic
compositions may also be in the form of tablets or capsules. In one
embodiment, the ACP and/or ACFP complexes are substantially the
only remineralizing active components of such a composition. For
example, a creme formulation may be employed containing: water;
glycerol; CPP-ACP; D-sorbitol; silicon dioxide; sodium
carboxymethylcellulose (CMC-Na); propylene glycol; titanium
dioxide; xylitol; phosphoric acid; guar gum; zinc oxide; sodium
saccharin; ethyl p-hydroxybenzoate; magnesium oxide; butyl
p-hydroxybenzoate and propyl p-hydroxybenzoate.
[0085] The invention further includes a formulation described above
provided together with instructions for its use to treat or prevent
any one or more of dental caries or tooth decay, dental erosion and
fluorosis.
[0086] In one embodiment, the active components of the composition
consist essentially of the protein disrupting agent and stabilised
ACP and/or ACFP. It is believed, without being bound by any theory
or mode of action, that the stabilised ACP and/or ACFP and the
protein disrupting agent are central to the therapeutic or
preventative effect of the above embodiments of the invention, and
thus embodiments consisting essentially of those components (with
carriers, excipients and the like as required) are included within
the scope of the invention.
[0087] The invention also relates to a kit for the treatment or
prevention of one or more of dental caries, fluorosis and dental
erosion including (a) a protein disrupting agent and (b) a CPP-ACP
or CPP-ACFP complex in a pharmaceutically acceptable carrier.
Desirably, the kit further includes instructions for their use for
the mineralization of a dental surface in a patent in need of such
treatment. In one embodiment, the agent and the complex are present
in suitable amounts for treatment of a patient.
[0088] In a further aspect, there is provided a method of treating
or preventing one or more of each of dental caries, tooth decay,
dental erosion and fluorosis, comprising the steps of administering
a protein disrupting agent to the teeth of a subject followed by
administering an ACP or ACFP complex or composition. Topical
administration of the complex is preferred. The method preferably
includes the administration of the complex in a formulation as
described above.
[0089] In a further aspect there is provided the use of a protein
disrupting agent in the manufacture of a first composition and use
of stabilized amorphous calcium phosphate (ACP) or amorphous
calcium fluoride phosphate (ACFP) in a manufacture of a second
composition, the first and second compositions being used for the
treatment and/or prevention of one or more of dental caries, tooth
decay, dental erosion and fluorosis, wherein the first composition
is applied to a dental surface prior to the second composition.
[0090] In a further aspect there is provided a first composition
including a protein disrupting agent and a second composition
including stabilized amorphous calcium phosphate (ACP) or amorphous
calcium fluoride phosphate (ACFP) for the treatment and/or
prevention of one or more of dental caries, tooth decay, dental
erosion and fluorosis, wherein the first composition is applied to
a dental surface prior to the second composition.
[0091] It will be clearly understood that, although this
specification refers specifically to applications in humans, the
invention is also useful for veterinary purposes. Thus in all
aspects the invention is useful for domestic animals such as
cattle, sheep, horses and poultry; for companion animals such as
cats and dogs; and for zoo animals.
[0092] The invention will now be further described with reference
to the following non-limiting examples.
[0093] One example of a mineralizing composition is a composition
comprising the following (in decreasing order of proportion):
[0094] water [0095] glycerol [0096] CPP-ACP [0097] D-sorbitol
[0098] silicon dioxide [0099] sodium carboxymethylcellulose
(CMC-Na) [0100] propylene glycol [0101] titanium dioxide [0102]
xylitol [0103] phosphoric acid [0104] guar gum [0105] zinc oxide
[0106] sodium saccharin [0107] ethyl p-hydroxybenzoate [0108]
magnesium oxide [0109] butyl p-hydroxybenzoate [0110] propyl
p-hydroxybenzoate
[0111] Such a composition is available from GC corporation under
the name Tooth Mousse.TM.. This is suitable for use after a protein
disrupting agent, and is in the form of a paste or creme to
facilitate its retention on teeth for a suitable period.
Alternatively, this mineralizing composition may contain a protein
disrupting agent, such as sodium hypochlorite.
[0112] The effectiveness of the invention may be demonstrated as
follows.
[0113] Seven premolar teeth with FLE (Thylstrup Fejerskov Index,
TF=3) were selected from teeth extracted for orthodontic reasons
from healthy patients aged 10-28 years from the Royal Dental
Hospital of Melbourne, Australia. Informed patient consent was
obtained for the extracted teeth and the study protocol was
approved by the Human Research Ethics Committee of The University
of Melbourne. All specimens were debrided of adherent soft tissue
and stored in 18% w/v formalin acetate solution at room
temperature.
[0114] The teeth were cleaned with a rotating rubber cup and pumice
and rinsed in double de-ionized water (DDW) (Fejerskov et al.,
1988). The anatomical crowns were sectioned from the roots using a
water-cooled diamond blade. Each crown was sectioned to provide a
pair of enamel blocks each containing a FLE. A 4.times.4 mm.sup.2
window was created over each lesion by placing a rectangular piece
of Parafilm.RTM. (American National Can, Chicago, Ill., USA.) over
the lesion and covering the surrounding enamel with nail varnish
(Revlon.TM., N.Y., USA). The parafilm was then carefully removed to
reveal the enamel lesion window which was divided into halves as
control and test windows. The control window was covered with nail
varnish. The two lesions of each specimen were randomly assigned to
one of two remineralization groups; Group I--treatment with 5% w/v
CPP-ACFP and Group II--treatment with 5% w/v CPP-ACFP immediately
following pre-conditioning with 5.25% NaOCl.
[0115] CPP-ACFP was obtained from Recaldent Pty Ltd (Melbourne,
Australia) and contained 47.6% w/w CPP, 15.7% w/w Ca.sup.2+, 22.9%
w/w PO.sub.4.sup.3- and 1.2% w/w F.sup.-. The CPP-ACFP was
dissolved in distilled and deionized water at 5% w/v and adjusted
to pH 7.0 with HCl. For the first group, each specimen was placed
in 2 ml of 5% w/v, CPP-ACFP in a 5 ml plastic vial at 37.degree. C.
The CPP-ACFP solution was changed daily for 10 days. For the second
group, each specimen was placed in a 5.25% NaOCl solution for 20
mins, rinsed and then placed in 2 ml of 5% w/v CPP-ACFP in a 5 ml
plastic vial at 37.degree. C. The CPP-ACFP solution was changed
daily for 10 days.
[0116] A Chroma Meter (Minolta ChromaMeter CR241, Minolta, Japan)
was used to record surface reflectance. Surface reflectance
measurement was established in L*a*b* color space by the Commission
de L'Eclairage in 1978, and measurements relate to human colour
perception in three colour dimensions (Commision Internationale de
L'Eclaige, 1978). The L* values represent colour gradients from
white to black, a* values represent colour gradients from green to
red, and b* values represent colour gradients from blue to yellow
(Commision Internationale de L'Eclaige, 1978). Only L* value
measurements were used in this study with whiter colours having a
higher reading, and darker colours a lower reading. To ensure a
reproducible position of specimens in the Chroma Meter, a wax mold
for each sample was prepared and stored. All samples were air-dried
with a dental triplex syringe for 60 s before each measurement.
Individual specimens were repositioned ten times both before and
after treatment, and colour reflectance L* values were
recorded.
[0117] Each specimen was removed from the mineralizing solution and
rinsed in DDW for 60 s and blotted dry with blotting paper. The
nail varnish on the control window was removed gently with acetone.
The control and test windows were then separated by cutting through
the midline between the windows. The two half-slabs were then
placed with the lesion windows parallel and embedded in cold curing
methacrylate resin (Paladur, Heraus Kulzer, Germany). The two
paired enamel half-slabs were then sectioned, and subjected to
microradiography and microdensitometric image analysis to determine
mineral content exactly as described by Shen et al. (2001).
[0118] An area free of defects close to the midline of each
microradiographic image of each lesion (control and test) was
chosen and scanned six times (Shen et al., 2001). Each scan
comprised 200 readings, taken from the enamel surface to the
mid-enamel region to include the total fluorotic lesion. The test
(CPP-ACFP-treated) lesion was scanned to exactly the same depth as
the control (untreated) lesion. The gray values obtained from each
scan were converted to the equivalent thickness of aluminium (tA)
using the image of the aluminium stepwedge included with each
section (Shen et al., 2001). Using the formula of Angmar et al.
(1963), the percentage volume of mineral was obtained for each
reading as follows: V=(52.77(tA)-4.54)/tS. Where: V=volume of
mineral as a percentage; tA=the relative thickness of aluminium
obtained from the gray value scanned; and tS=section thickness (80
.mu.m).
[0119] From the densitometric profile of [(vol % min versus lesion
depth (mm)] for each lesion DZ values were calculated using
trapezoidal integration (Reynolds, 1997). The difference between
the area under the profile of the untreated fluorotic enamel in the
control window with adjacent normal enamel was designated DZf, and
the difference between the area under the CPP-ACFP-treated
fluorotic enamel in the test window and adjacent normal enamel was
designated DZr. Percentage mineralization (% M) of the fluorotic
lesion was therefore (1-DZr/(DZf).times.100 (Reynolds, 1997).
[0120] Following the microradiography the sections containing both
control and mineralized FLE were subjected to Energy Dispersive
X-ray Analysis (EDAX) as described previously (Reynolds, 1997).
[0121] Mean L* values were compared using a one way classification
analysis of variance (ANOVA) with a Scheffe multiple comparison.
The mean % M values were also compared using a one-way ANOVA.
Overall mean L* and % M values were analysed using a paired data
Student's t-test.
[0122] The L*values of the untreated fluorotic enamel lesions
ranged from 79.1 to 87.8 with a mean value of 83.6.+-.3.6 (Table
1). Treatment with 5% CPP-ACFP significantly reduced the L*value to
74.6.+-.4.1, which was not significantly different to normal enamel
(Table 1). Pre-conditioning with NaOCl followed by 5% CPP-ACFP
treatment significantly reduced the L*value to 72.6.+-.5.6, which
was also not significantly different to normal enamel (Table 1).
There was no significant difference in L*values for the two
post-treatment (CPP-ACFP and NaOCl/CPP-ACFP) groups. The appearance
of the surface enamel of both treatment groups had substantially
improved with both exhibiting the appearance of normal, translucent
enamel.
[0123] The difference between the mineral content of sound enamel
and that of the pre-treatment lesions (DZf) varied from 426 to
12,048 vol % min. mm (Table 2). No correlation was found between
surface reflectance (L*) and DZf of the untreated FLE. Treatment
with 5% CPP-ACFP alone substantially increased the mineral content
of the fluorotic lesions to restore 32.7% to 55.5% of the missing
mineral, with a mean value of 44.8.+-.10.6% (Table 2). Restoring
100% of the missing mineral would convert the entire lesion to
sound enamel with respect to mineral content. Pre-conditioning of
the enamel with NaOCl before CPP-ACFP treatment increased mineral
uptake from 73.6% to 92.8% of the missing mineral with a mean value
of 80.1.+-.7.8% (Table 2). Energy dispersive X-ray analysis of the
mineralized lesion of the transverse sections confirmed the mineral
formed by the CPP-ACFP treatment was a fluoride-containing
apatite.
TABLE-US-00003 TABLE 1 Effect of 5% CPP-ACFP with and without NaOCl
pre-conditioning on colour reflectance (L*) of fluorotic enamel
specimens Colour Reflectance (L*) Values Fluorotic enamel specimens
I II III IV V VI VII Overall Mean Pre-treatment 82.9 .+-. 0.9.sup.a
85.5 .+-. 1.8 84.3 .+-. 0.4 82.5 .+-. 1.3 87.8 .+-. 0.6 79.1 .+-.
0.9 83.0 .+-. 0.6 83.6 .+-. 3.6 Post-CPP-ACFP treatment 74.1 .+-.
0.7.sup.b 72.0 .+-. 0.5 78.2 .+-. 0.4 76.1 .+-. 0.7 79.5 .+-. 0.8
69.7 .+-. 1.5 72.3 .+-. 1.7 74.6 .+-. 4.1.sup.c Post-NaOCl/CPP-ACFP
treatment 69.2 .+-. 1.0.sup.b 72.3 .+-. 1.1 76.9 .+-. 1.4 72.2 .+-.
1.3 78.5 .+-. 1.4 61.6 .+-. 1.2 77.4 .+-. 1.0 72.6 .+-. 5.6.sup.c
.sup.an = 20 .sup.bn = 10 .sup.cPost-treatment mean value is
significantly different from pre-treatment mean value (paired
Student's t-test, p < 0.01) but not significantly different from
normal enamel 71.6 .+-. 3.1.
TABLE-US-00004 TABLE 2 Effect of 5% CPP-ACFP with and without NaOCl
pre-conditioning on mineral content of fluorotic enamel Specimens
Overall Treatment I II III IV V VI VII Mean Natural fluorotic
.DELTA.Zf 2331 .+-. 352.sup.a --.sup.c 3869 .+-. 70.sup.a 2468 .+-.
323.sup.a 2706 .+-. 103.sup.a 3238 .+-. 194.sup.a --.sup.c lesion
(vol % min .mu.m) CPP-ACFP treated .DELTA.Zr 1203 .+-. 241.sup.a
--.sup.c 1723 .+-. 262.sup.a 1618 .+-. 427.sup.a 1270 .+-.
596.sup.a 2178 .+-. 216.sup.a --.sup.c (vol % min .mu.m) % M.sup.b
48.4 --.sup.c 55.5 34.5 53.1 32.7 --.sup.c 44.8 .+-. 10.6 Natural
fluorotic .DELTA.Zf 2199 .+-. 266.sup.a 6501 .+-. 441.sup.a
--.sup.c 1181 .+-. 261.sup.a 2461 .+-. 213.sup.a --.sup.c 12048
.+-. 512.sup.a lesion (vol % min .mu.m) NaOCl/CPP-ACFP .DELTA.Zr
581 .+-. 230.sup.a 471 .+-. 285.sup.a --.sup.c 211 .+-. 137.sup.a
552 .+-. 203.sup.a --.sup.c 3087 .+-. 723.sup.a treated (vol % min
.mu.m) % M.sup.b 73.6 92.8 --.sup.c 82.1 77.6 --.sup.c 74.4 80.1
.+-. 7.8 .sup.aMean .+-. SD (n = 6) .sup.b% M = percentage
mineralization (1 - .DELTA.Zr/.DELTA.Zf) .times. 100 .sup.cSample
lost during processing
[0124] In the clinic, as an example of a patient in need of
remineralizing treatment of the tooth enamel, the patient is
treated using the steps of: [0125] 1. Pretreating an enamel area in
need of treatment, isolated using a rubber dam, with a 5% solution
of NaOCl for 5 minutes. [0126] 2. Removing the NaOCl solution from
the area with a moist cotton bud. [0127] 3. Applying the
CPP-ACP-containing topical creme Tooth Mousse.TM. (GC Corporation)
to the enamel surface immediately for 5 minutes and then the
patient further applies the Tooth Mousse.TM. nightly without
rinsing for four weeks.
[0128] It will be understood that the invention disclosed and
defined in this specification extends to all alternative
combinations of two or more of the individual features mentioned or
evident from the text or drawings. All of these different
combinations constitute various alternative aspects of the
invention.
REFERENCES
[0129] Angmar B, Carlstrom D, Glas J E (1963). Studies on the
ultrastructure of dental enamel. IV. The mineralization of normal
human enamel. J Ultrastruct Res 8:12-23. [0130] Aoba T, Fejerskov O
(2002). Dental fluorosis: chemistry and biology. Crit Rev Oral Biol
Med 13:155-70. [0131] Black G, McKay F (1916). Mottled teeth--An
endemic developmental imperfection of the teeth heretofore unknown
in the literature of dentistry. Dent Cosmos 58:129-156. [0132]
Commision Internationale de L'Eclaige (1978). Recommendations on
uniform colour spaces, colour difference equations and psychometric
colour terms. Paris: Bureau Centrale de la DIE Suppl. 2:15. [0133]
Den Besten P K, Thariani H (1992). Biological mechanisms of
fluorosis and level and timing of systemic exposure to fluoride
with respect to fluorosis. J Dent Res 71:1238-43. [0134] Fejerskov
O, Baelum V, Manji F, Moller I (1988). Dental Fluorosis--A handbook
for health workers Copenhagen: Munksgard. [0135] Fejerskov O, Manji
F, Baelum V (1990). The nature and mechanisms of dental fluorosis
in man. J Dent Res 69 Spec No: 692-700; discussion 721. [0136]
Fejerskov O, Yanagisawa T, Tohda H, Larsen M J, Josephsen K, Mosha
H J (1991). Posteruptive changes in human dental fluorosis--a
histological and ultrastructural study. Proc Finn Dent Soc
87:607-19. [0137] Fejerskov O, Ekstrand J, Burt B (1996). Fluoride
in dentistry. 2nd ed. Copenhagen: Munksgard. [0138] Giambro N J,
Prostak K, Den Besten P K (1995). Characterization of fluorosed
human enamel by color reflectance, ultrastructure, and elemental
composition. Caries Res 29:251-7. [0139] Reynolds E C (1997).
Remineralization of enamel subsurface lesions by casein
phosphopeptide-stabilized calcium phosphate solutions. J Dent Res
76:1587-95. [0140] Reynolds E C (1998). Anticariogenic complexes of
amorphous calcium phosphate stabilized by casein phosphopeptides: a
review. Spec Care Dentist 18:8-16. [0141] Reynolds E C, Cai F, Shen
P, Walker G D (2003). Retention in plaque and remineralization of
enamel lesions by various forms of calcium in a mouthrinse or
sugar-free chewing gum. J Dent Res 82:206-11. [0142] Shen P, Cai F,
Nowicki A, Vincent J, Reynolds E C (2001). Remineralization of
enamel subsurface lesions by sugar-free chewing gum containing
casein phosphopeptide-amorphous calcium phosphate. J Dent Res
80:2066-70.
* * * * *