4. Discussion
The present study aimed to compare the effectiveness of different formulas in estimating the age of a Brazilian subpopulation by calculating pulp/tooth ratio values in maxillary canines and mandibular second premolars using panoramic and periapical radiographs. The results revealed minimal agreement between the actual and estimated ages using the equations developed by Cameriere et al. [
5], Cameriere et al. [
2], Azevedo et al. [
11], Lee et al. [
12] and Anastácio et al. [
13].
Among the conventional radiographic methods for estimating age in adults, those proposed by Cameriere [
1,
2] and Kvaal [
3] are the most widely studied and utilized [
4]. The Cameriere method was chosen for this experiment due to its ease of reproducibility and the proportional nature of the obtained values [
2,
5], as opposed to the metric approach of Kvaal's method [
3,
6,
7]. Additionally, Cameriere’s method is less prone to geometric errors commonly found in radiographic images [8-9].
Previous studies employing the pulp/tooth ratio strategy typically selected single-rooted teeth, as they undergo fewer morphological changes over time. Among the dental groups investigated, maxillary canines and mandibular premolars were the most used [2,5,7,10-15]. It is noteworthy that Cameriere’s methodology [
5] recommends using a minimum of 20 points for outlining the tooth and 10 points for outlining the pulp. For the execution of this study, a pen and digitizing table were chosen to ensure ease, precision, and standardization of markings [
16]. The consistently high ICC values exceeding 0.99% across all measurements underscore the reproducibility and standardization achieved in this study.
In this study, the formulas utilizing the pulp/tooth ratio of canines showed the smallest difference between actual and estimated age when using the equation proposed by Cameriere et al [
5] with periapical radiographs (+7 years). Although originally developed for application in periapical radiographs, its application in panoramic radiographs resulted in a difference of +9.1 years. Interestingly, this value closely aligned with the difference obtained (+8.9 years) using a specific formula for the Brazilian population with periapical radiographs [
11].
For lower premolars, the smallest difference (+4.1 years) was observed on panoramic radiographs after applying the equation by Cameriere et al. [
2]. While this formula was initially developed for panoramic radiographs, its application to periapical radiographs yielded a difference of +6.1 years. Notably, this value closely resembled the actual age when compared to the values obtained using specific formulas for Korean [
12] and Portuguese [
13] populations produced in panoramic radiographs.
Although the original formulas developed for maxillary canines and mandibular premolars [
2,
5] yielded lower values when compared to other formulas [
11,
12,
13], all results demonstrated statistically significant differences (p < 0.001). In the case of maxillary canines, the estimated ages surpassed the actual ages. The standard deviation (SD) ranged from ±10 [
11] to ±12.6 years [
2], indicating considerable variability in the data. Additionally, the confidence interval (CI) exhibited wide values, signifying limited agreement.
Similarly, the estimated ages for mandibular second premolars also exceeded the actual ages, except when using the formula for the Portuguese population [
13]. The SD ranged from ±1.8 [
13] to ±10.5 years [
12]. Like the canines, the CI displayed large values, indicating limited agreement, despite the forensic literature accepting a margin of error between 6 to 8 years [
17], preferably less than 10 years [
4].
The measurements obtained in this study underwent linear regressions, resulting in the construction of four equations tailored for each type of radiograph (panoramic and periapical) (
Table 7). This approach is warranted due to specific geometric variations inherent in panoramic and periapical radiographs [
8,
9,
18]. Previous studies have often applied the same formula to both panoramic and periapical radiographs [
15,
19].
The number of exams/patients included in the studies exhibits considerable variation. The smallest sample size observed was 30 individuals [
20], while the largest included 606 individuals [
2]. Challenges with sample sizes are inherent in studies aimed at determining age in adults, particularly after applying stringent inclusion and exclusion criteria [
6,
17]. Similar challenges were encountered in the present study. Research in this field suggests incorporating formulas derived from diverse populations or augmenting the sample size [
7,
10,
15,
21,
22,
23,
24].
The absence of methodological standardization impedes or complicates the establishment of correlations and hinders the explanation of certain results [
4]. Some studies express data in terms of standard error (SE) [
5,
12,
25], while others use SD [
11,
26]. Additionally, some studies provide the R
2 values [
15,
27], while simultaneously reporting values for EP and mean error (EM) [
13]. In contrast, this research, alongside R
2, presented its results in terms of SD, mean absolute error (EMA), and EP. This approach facilitated comparisons with the original formulas [
4].
The linear regression analysis of maxillary canines on periapical radiographs revealed R
2= 0.35 and EP= 10.26. These values are notably discrepant and higher when compared to the original formula [
5], which reported R
2 = 0.86 and EP=5.45. Similarly, the linear regression values for mandibular premolars on panoramic radiographs showed R
2 = 0.32 and EP= 10.47, whereas the original formula indicated R
2 = 0.75; EP= 7.42 [
2].
The extreme quartile diagrams suggest a direct correlation between increasing age and secondary dentin deposition, as evidenced by the gradual decrease in pulp/tooth ratio values among individuals aged 20 to 70 years or older. However, these diagrams also exhibit wide ranges and lack concentration of data points, indicating a weak correlation between age group and pulp/tooth ratio. In essence, individuals across different age groups exhibit similar values, aligning with findings for mandibular second premolars in the Portuguese population [
13], but diverging from results reported in other studies [
1,
2,
5,
15], where this correlation appeared stronger.
The scatter plots did not reveal a linear correlation, consistent with the findings from the extreme quartile plots. This observation suggests the absence of sexual dimorphism, aligning with previous reports in the literature [
2,
5,
11,
12,
13].
The estimation of the age in living individuals requires higher accuracy compared to age estimation in cadavers or skeletons [
26]. However, the range of applicable methods is limited [
17], categorized into morphophysiological, radiographic, and biomechanical [
28], each with its indications and specificities [
29,
30]. Among the non-invasive methods, imaging tests are widely employed due to their ease, reproducibility, and applicability in both in vivo and ex vivo settings [
6,
28,
31,
32]. It’s evident that imaging tests utilized in children and adolescents, which classify or measure during the mineralization, formation, development, and eruption phases of the teeth present, yield more precise results compared to the methods used in adults, even when applying foreign formulas [
33,
34,
35,
36,
37,
38].
Periapical radiographs can exhibit distortions, particularly in the vertical angle, resulting in elongated or shortened images, despite the use of radiographic positioners [
8]. Panoramic radiographs, besides magnification, may also show vertical and horizontal distortion [
9], inherent to each device, individual anatomical variations, or arising from minor positioning errors [
8,
18,
39,
40,
41,
42,
43,
44]. Another limitation of conventional exams is the projection of two-dimensional images of three-dimensional structures [
45], restricting any analysis, quantitative or qualitative, to the vertical and mesiodistal dimensions of teeth or anatomical structures [
46]. The morphology of the root canal can change over time, with secondary dentin deposition occurring with aging [
47,
48]. Young patients exhibit greater variations compared to older patients [
49]. Inclusion specialists in Dental Radiology in the research team could enhance sample selection and measurement execution [
4].
Cone beam computed tomography represents a new technology with increased significance across various dental disciplines. Numerous studies investigating age estimation have emerged utilizing this tool [
48,
50,
51]. Despite promising prospects [
47], challenges akin to those encountered in conventional methods have been noted [
50,
51,
52,
53,
54,
55]. Human programming and artificial intelligence stand as potential techniques to mitigate sample selection and evaluation biases in the future [
56,
57,
58]. New investigations should be encouraged with larger sample sizes and the development of specific population formulas. It is pertinent for forensic dentistry that future studies compare the results obtained through different imaging modalities.