INDIVIDUAL VARIATION OF MUCIN CONCENTRATION IN HUMAN SALIVA

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INDIVIDUAL VARIATION OF MUCIN CONCENTRATION IN
HUMAN SALIVA

• P.A. Denny, W. Wu, and P.C. Denny
• School of Dentistry, University of Southern
  California
• Los Angeles, USA
• Data reported in part in A Blind Study of MG1
  and MG2 Mucin Concentrations in Repeated
  Collections of Whole Saliva from Young Subjects
  by W. Wu, a Master of Science degree thesis for
  the Craniofacial Biology Program, School of
  Dentistry, USC
• 1993

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I. Introduction

• The study presented in P.C. Denny, P.A. Denny,
  D.K. Klauser, S.H. Hong, M. Navazesh, and L.A.
  Tabak. Age-related Changes in Mucins from Human
  Whole Saliva, J Dent Res. 701320-1327 (1991)
  revealed that MUC7 mucin concentration in a
  single collection of unstimulated whole saliva
  from each individual ranged from lt6 units/ml to
  1500 units/ml in a group of young adults 18-33
  years old.
• We subsequently pursued the question whether this
  broad range of mucin concentrations was a
  reflection of the range of variation within each
  individual or a reflection of the range of
  individual differences.
• Our approach was to select six new individuals
  from the same age category whose MUC7 mucin
  concentrations in one collection of unstimulated
  saliva ranged from lt100 units/ml to 1500
  units/ml.
• Repeated collections of both unstimulated and
  stimulated whole saliva were obtained at a
  minimum of weekly intervals over a period of
  three months. Flow rates, and concentrations of
  MUC7 mucin and MUC5B mucin were obtained for each
  sample.

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II. Quantitation of MUC7 and MUC5B Mucins (from
Denny et al., J Dent Res. 701320-1327 (1991))
The two mucins in whole saliva were fractionated
by SDS-PAGE, stained with Stains-all,
photographed in black and white using a red
filter, and quantitated by densitometry.
Mouse Muc10
Whole Human Saliva
MUC5B
MUC7
MUC5B
MUC7
1.25 2.5 5 10 20 40 Mucin Units
Mucin Units
There was no detectable loss of either mucin
under our conditions for collection, storage,
assay and re-assay of the same sample. Neuraminid
ase removes all Stains-all reactivity with MUC7
mucin and approximately 50 of reactivity with
MUC5B mucin (L.W. Baughan, F.J. Robertello, D.C.
Sarrett, P.A. Denny, and P.C. Denny. Salivary
Mucin as Related to Oral Streptococcus mutans in
the elderly, Oral Micro. Immunol. 1510-14
(2000)).
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IV. Individual and Population Variation of MUC7
Mucin Concentrations in Repeated Collections of
Unstimulated Saliva
Subject Number of Collections Mean SEM (units/ml) Range Coefficient of Variation (CV)
1 6 400.021.3 141 13.0
2 7 1548.398.9 615 16.9
3 6 1072.769.2 300.5 15.8
4 6 720.578.0 501.0 26.5
5 3 114.013.3 42.5 20.3
6 5 378.538.5 239.5 22.7
All 33 794.989.4 1741.5 64.6
Two-level Nested Analysis of Variance (nANOVA)
Level DF FS P-value Percentage of Total Variation
Between Individuals 5 51.53 plt0.001 89.8
Within Individuals 27 50.93 plt0.001 9.8
Test Variation 33 0.4
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VI. Individual and Population Variation of MUC5B
Mucin Concentrations in Repeated Collections of
Unstimulated Saliva
Subject Number of Collections Mean SEM (units/ml) Range Coefficient of Variation (CV)
1 6 346.027.7 158.5 19.6
2 7 649.9112.0 805.0 45.6
3 6 621.958.4 364.0 23.0
4 7 535.673.3 497.0 36.2
5 3 122.831.9 109.5 45.0
6 5 283.553.5 301.0 42.2
All 34 467.441.8 1076.0 52.2
Two-level Nested Analysis of Variance (nANOVA)
Level DF FS P-value Percentage of Total Variation
Between Individuals 5 6.08 plt0.001 47.4
Within Individuals 28 145.28 plt0.001 51.9
Test Variation 34 0.7
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VII. Means Test for Repeated Collections of
Unstimulated Whole Saliva
MUC7 mucin
MUC5B mucin
0 250 500 750
1000 1250 1500 Individual Mean Mucin
Concentrations
Distance between arrows on same horizontal plane
is significant at plt0.03
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VIII. Individual and Population Variation of
MUC7and MUC5B Mucinsin Repeated Collections of
Stimulated Whole Saliva
Subject Number of Collections MUC7 mucin Mean SEM (units/ml) CV MUC5B mucin Mean SEM (units/ml) CV
1 7 75.59.3 32.3 436.065.2 39.5
2 7 120.818.3 40.0 267.034.6 34.3
3 6 206.794.6 112.1 487.831.0 15.5
4 6 215.654.7 62.2 450.748.3 26.3
5 3 16.55.6 58.5 198.323.6 20.6
6 5 14.58.5 131.1 339.979.0 51.9
All 34 118.522.4 110.0 377.825.9 39.9
Two-level Nested Analysis of Variance (nANOVA)
Level DF MUC7 FS MUC7 P-value MUC7 Variation MUC5B FS MUC5B P-value MUC5B Variation
Between Individuals 5 5.98 plt0.001 46.9 3.94 plt0.01 34.3
Within Individuals 28 159.1 plt0.001 52.4 110.5 plt0.001 64.5
Test 34 0.7 1.2
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X. Individual and Population Variation of
Salivary Flow Ratesin Repeated Collections of
Unstimulated and Stimulated Whole Saliva
Subject Number of Collections Unstimulated Mean SEM (gm/min) CV Stimulated Mean SEM (gm/min) CV
1 7 0.4670.034 19.4 1.0860.084 20.4
2 7 0.2330.028 32.3 1.7640.079 11.8
3 6 0.3860.030 18.9 0.8030.064 19.7
4 7 0.4450.044 25.9 0.9260.087 24.9
5 3 0.2790.048 29.8 0.7180.012 2.9
6 5 0.3400.042 27.9 1.3870.108 17.3
All 35 0.3680.020 32.7 1.1540.069 35.5
Two-level Nested Analysis of Variance (nANOVA)
Level DF Unstim. FS Unstim. P-value Unstim. Variation Stim. FS Stim. P-value Stim. Variation
Between Individuals 5 6.49 plt0.025 48.7 21.8 plt0.001 78.4
Within Individuals 29 51.3 21.6
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XI. Levels of Correlation Between Individual Mean
Mucin Concentrations and Mean Flow Rates
Comparison
r
Adjusted r2
p-value

• Unstimulated MUC5B and MUC7 concentrations
  0.927 0.824 0.0078
• Unstimulated and stimulated MUC7 concentrations
  0.604 0.213 0.20
• Unstimulated and stimulated MUC5B concentrations
  0.501 0.064 0.31
• Stimulated MUC5B and MUC7 concentrations
  0.695 0.354 0.13
• Unstimulated MUC5B concentration and flow rate
  0.015 0.000 0.978
• Unstimulated MUC7 concentration and flow rate
  0.335 0.000 0.516
• Stimulated MUC5B concentration and flow rate
  0.269 0.000 0.606
• Stimulated MUC7 concentration and flow rate
  0.176 0.000 0.739

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XII. Conclusions and Implications from the Study
of Individual Variation of Mucin Concentrations
in Unstimulated Whole Saliva

• These studies suggest that each person within an
  age range of 23-31 years old has a distinctive
  signature mean MUC7 mucin concentration in their
  resting saliva. Limited individual variation
  indicates that these signature means range from
  very low to high mucin concentrations.
• The coefficients of variation are similar for all
  unstimulated MUC7 mucin concentration means in
  the test group. Thus as the mean mucin
  concentration decreases, the absolute range of
  variation narrows to the point that a single
  collection could identify with confidence those
  individuals who possess very low mean mucin
  concentrations in their resting saliva.
• There is the potential for detecting possible
  long-term impacts of mucin concentration on oral
  health if broader-based studies confirm that
  mucin levels in unstimulated saliva exhibit
  limited individual variation while still showing
  a wide range of population variation.
• When viewed as analytes for diagnostic or
  forecasting purposes, the two mucins illustrate
  that concentrations may be the same or very
  different in unstimulated and stimulated salivas
  and that each analyte may exhibit different but
  characteristic levels of individual variation.