JOURNAL Of NUCLEAR And Related TECHNOLOGIES, Volume 6, No.

1, Special Edition, 2009

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AN EXPLORATORY STUDY OF HUMAN TEETH ENAMEL BY USING FT-RAMAN SPECTROSCOPY

Afishah Alias1 , Siti Rahayu Mohd Hashim1, Judith Mihaly2 , Julynnie Wajir1 & Fauziah Abdul Aziz1
1

School of Science and Technology, Universiti Malaysia Sabah, Kota Kinabalu, 88502, Sabah, MALAYSIA 2 Chemical Research Center, Hungarian Academy of Sciences, H-1525, Budapest, P.O. Box 17, Hungary

ABSTRACT
Unaffected, affected and heavily affected teeth enamel were studied by using FT-Raman spectroscopy. The 14 permanent teeths enamel surface were measured randomly, resulting in total n=43 FT-Raman spectra. The results obtained from FT-Raman spectra of heavily affected, affected and unaffected tooths enamel surfaces did not show any significant difference. In this study, Kruskal-Wallis and Wilcoxon rank sum tests were used to compare the intensity between the categories of enamel as well as the surfaces of teeth samples.

ABSTRAK
Tidak terjejas, terjejas dan terjejas dengan teruk enamel gigi telah dipelajari dengan menggunakan spektroskopi FT-Raman. 14 enamel gigi kekal permukaan telah diukur secara rawak, mengakibatkan dalam jumlah n=43 spektrum FT-Raman. Keputusan-keputusan itu diperolehi dari spektrum FT-Raman terjejas dengan teruk, gigi terjejas dan tidak terjejas permukaan-permukaan enamel melakukan tak tunjuk mana-mana perbezaan penting. Dalam kajian ini, Kruskal-Wallis dan Wilcoxon ujian-ujian jumlah lebat adalah sudah biasa bandingkan keamatan antara kategori-kategori enamel serta permukaan-permukaan contohcontoh gigi. Keywords: human teeth enamel, FT- Raman spectroscopy, Kruskal-Wallis, Wilcoxon

INTRODUCTION Dental caries is the common oral disease in the developed country. Carious lesions are regularly found at area between adjacent teeth. The detection of the caries lesions in these areas is difficult. These characteristics of the caries problem magnify the limitations of caries detection through conventional diagnostic methods that involve subjective clinical criteria and the use of diagnostic tools such as the dental explorer and dental radiographs (Alex et al., 2006). These conventional methods are adequate for detection of larger, possibly cavitated lesions, but due to poor specify and sensitivity, they are not suitable for detection of early stage, non-cavitated lesions (Alex et al., 2006). Therefore, better diagnostic tools are needed to detect early noncavitated lesions and to monitor their activity (Alex et al., 2006). Polarized Raman spectroscopy and optical coherence tomography have been used to detect these changes and potentially offer a means to detect and monitor early caries development (Michael et al., 2006). 141

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In this study, unaffected, affected and heavily affected teeth enamel were studied by using FTRaman spectroscopy and the first attempt was to analyze the data by using General Linear Modeling (GLM) method, two-way ANOVA. Unfortunately, the tests failed to identify significant differences of intensity amongst the three conditions. The results could be due to the critical violation of the normality assumption. This study has carried out non-parametric test as an alternative, as the data failed to follow the normality distribution after several transformations.

EXPERIMENTAL METHOD The samples of human adult teeth were obtained from Dr. Rashid Dental Clinic. From these teeth samples obtained, its have been categorized to 3 types of teeth: - heavily affected, affected and unaffected enamel. The enamel that has more than 3 affected surfaces was classified as heavily affected enamel, 3 or less affected surfaces as affected enamel and has non affected surface as unaffected enamel. They were then sterilized with sodium-hypochlorite of 5% water solution for 4 hours. These were then treated in a 6.85 M KOH water solution for 50 hour while changing the solution for every 2-3 hour (Fattibene et al., 2005). The enamel samples were then investigated by FT-Raman spectroscopic method at Chemical Research Center, Hungarian Academy of Sciences, Budapest, Hungary. The measurements were performed by BioRad (Digilab) dedicated FT-Raman spectrometer equipped with liquid nitrogen cooled germanium detector and Nd:YAG excitation laser with radiation line at 1064 nm. No sample preparation was required and the measurements were completely nondestructive. All spectra were collected using 500mW laser power, 4 cm-1 resolution and coaddition of 512 individual spectra. The spectra were than baseline corrected and normalized to the strongest 958 cm-1 peak, using Grams32 spectral evaluation program. The Kruskal-Wallis test is applied to compare two or more populations with the data are either ordinal or interval but non-normal (Keller and Warrack, 2003). In this study, the populations referred to are the classification of enamels and the position of enamel during the spectroscopy test. The test statistic of Kruskal-Wallis test is;
k T2 12 j H= 3(n + 1) n(n + 1) j =1 n j

(1)

Where,

n = the number of observations k = the number of populations compared j = 3 ( classification of enamels) @ 2 (enamel positions) T = the total of ranks

However, the Kruskal-Wallis can determine only whether a difference exists. To determine whether one population is larger than another, this study applied the Wilcoxon rank sum test. The standardized test statistic is; Z= Where,

T E (T )

(2)

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JOURNAL Of NUCLEAR And Related TECHNOLOGIES, Volume 6, No. 1, Special Edition, 2009

E (T) = and,

n1 (n1 + n2 + 1) 2
n1 n2 (n1 + n2 + 1) 12

(3)

T =

(4)

Both n1 and n2 is the number of observations belonged to two groups, lower ranks group and higher ranks group respectively.

RESULT AND DISCUSSION The results obtained from FT-Raman spectra of heavily affected, affected and unaffected tooth enamels surfaces did not show any significant difference. Figure 1 showed FT-Raman spectra of heavily affected (upper spectrum), affected (middle spectrum) and unaffected (lower spectrum) tooths enamel surfaces. For discriminating the 3 types of tooth enamel with FTRaman spectral features, Kruskal-Wallis was applied. The result in Table 1 showed that there is significant difference between the types of enamel and the positions of enamel with p-value < 0.001 for both tests. Figure 2 showed that the mean rank of intensity for side enamel surface, 5326.33, is significantly higher than the occlusal enamel which is 5066.67. The mean rank of intensity of heavily affected enamel, 5528.72, is found much higher than the affected and unaffected enamels with the mean rank are 5047.75 and 5013.03 respectively. But to identify which type of enamel is significantly difference with the other type of enamel, the Wilcoxon rank sum tests are carried out and the results can be found in Figure 3. From table 2, the p-values (both < 0.001) showed that the mean rank of intensity for heavily affected enamel is significantly difference with unaffected and affected enamel. Whereas, the mean rank of intensity of affected and unaffected enamel are not differ significantly with pvalue = 0.360 (> 0.05). It showed that the difference of the mean rank of intensity is depends on the categories of the teeth.

A CO3

3PO4

1PO4

1.2

958

.8 .6 1664 1448 .4 .2 0 1800 1600 1400

1099

4PO4
877 800 600
606

1200

1000

606 588 578

445 429 400

1068

1042

2PO4

200

Figure

1: Spektra FT-Raman of heavily affected (upper spectrum), affected (middle spectrum) and unaffected (lower spectrum) enamel teeth. 143

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Table 1: The Kruskal-Wallis results on determining a significant difference of intensity between the types of enamel and the surface.

Intensity by enamel types Chi-square statistic 63.951

Intensity by surface 19.463

Degree of freedom Significant value, p

2 < 0.001

1 < 0.001

Mean ranks between side enamel surface and occlusal enamel

5350 5300 5250 5200 5150 5100 5050 5000 4950 4900 Surface

Intensity

side enamel surface occlusal surface

Figure 2: The mean ranks between side enamel surface and occlusal enamel.

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Mean ranks of three types of enamel from the Kruskal Wallis test
5600 5500 5400 5300 5200 5100 5000 4900 4800 4700 category

In ten sity

heavily affected affected unaffected

Figure

3: The mean ranks of three types of enamel from the Kruskal_Wallis test. Table 2: The result of the Wilcoxon rank sum tests.

category intensity heavily affected unaffected Total intensity heavily affected affected Total intensity affected unaffected Total

N 3464 3464 6928 3464 3464 6928 3464 3464 6928

Mean Rank 3625.98 3303.02

Sum of Ranks 12560391.50 11441664.50

p-value

-6.720

< 0.001

3635.24 3293.76

12592487.00 11409569.00 -7.106 < 0.001

3486.49 3442.51

12077205.00 11924851.00 -0.915 0.360

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CONCLUSIONS The results obtained from FT-Raman spectra of heavily affected, affected and unaffected tooths enamel surfaces did not show any significant difference. However, by using KruskalWallis and Wilcoxon rank sum tests, the intensity between the categories of enamel as well as the surfaces of teeth samples can be discriminated. The results showed that Wilcoxon rank sum tests can be an indicator to characterize the conditions of the teeth.

ACKNOWLEGMENT The authors acknowledge the Hungarian Academy of Sciences and Universiti Malaysia Sabah for full support and cooperation.

REFERENCES Alex C.-T. Ko, Lin-P'ing Choo-Smith, Mark Hewko, Michael G. Sowa, Cecilia C. S. Dong, and Blaine Cleghorn., (2006), Detection of early dental caries using polarized Raman spectroscopy Optics Express, Vol. 14, 1: 203-215. Michael, G. S., Dan P. Popescu, Jeffrey Werner, Mark Hewko, Alex C.-T. Ko, Jeri Payette, Cecilia C. S. Dong, Blaine Cleghorn and Lin-Ping Choo-Smith., (2006), Precision of Raman depolarization and optical attenuation measurements of sound tooth enamel. Analytical and Bioanalytical Chemistry. ISSN 1618 - 2642. Vol. 387, 5: 1613-1619. Keller, G. and Warrack, B., (2003), Statistics for Management and Economics. ISBN 0-53439186-9, 6th edition, Thomson Learning.

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