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Despite their clinical relevance, variations in HSB patterns across different tooth surfaces remain poorly understood. The aim of this study wass to correlate the width and density of HSB with the surface (labial, lingual, mesial distal and occlusal) of the tooth and the type of tooth. Materials and Methods Eighty intact permanent human teeth (20 each: incisors, canines, premolars, molars) from the maxillary and mandibular arches were sectioned in mesiodistal or labiolingual planes. Reflected-light photomicrographs (4&times; objective; Olympus CX20i) were analysed with ImageJ. HSB density was defined as the number of HSB pairs per millimetre (one dark + one light = pair). Group differences were assessed via repeated-measures ANOVA and independent t tests; p&lt; 0.05. Statistical analyses Repeated-measures ANOVA and independent t tests were used to assess differences of HSB densities among surfaces and tooth types. A p value of of p&lt;0.05 was considered . Results Significant variations in HSB density were observed across tooth surfaces within each tooth type and between the maxillary and mandibular arches. Compared with molars, mandibular buccal surfaces presented greater mean HSB densities in premolars (p=0.001) (p=0.201),whereas lingual surfaces presented consistent patterns across all tooth types (p&lt;0.05). Notably, incisal/occlusal surfaces demonstrated the greatest variability, with mandibular incisors exhibiting greater densities (p&lt;0.001) than maxillary incisors (p&lt;0.001). Conclusion This micrometric analysis provides detailed insights into the microstructural variations in the HSB across tooth surfaces, highlighting functional and anatomical influences. These findings underscore the clinical importance of HSB in adhesive restorations and enamel-related conditions, suggesting tailored approaches on the basis of tooth type and surface location. Further research is needed to explore the biomechanical implications and refine the clinical applications in dental practice. \" } { \"@context\": \"http://schema.org\", \"@type\": \"BreadcrumbList\", \"itemListElement\": [ { \"@type\": \"ListItem\", \"position\": \"1\", \"item\": { \"@id\": \"https://f1000research.com/\", \"name\": \"Home\" } }, { \"@type\": \"ListItem\", \"position\": \"2\", \"item\": { \"@id\": \"https://f1000research.com/browse/articles\", \"name\": \"Browse\" } }, { \"@type\": \"ListItem\", \"position\": \"3\", \"item\": { \"@id\": \"https://f1000research.com/articles/15-354/v1\", \"name\": \"Correlating the Densities of Hunter-schreger Bands With Function and...\" } } ] } Home Browse Correlating the Densities of Hunter-schreger Bands With Function and... ALL Metrics - Views Downloads Get PDF Get XML Cite How to cite this article Agarwal R, Sehgal L, Prince J et al. Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] . F1000Research 2026, 15 :354 ( https://doi.org/10.12688/f1000research.175647.1 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. Close Copy Citation Details Export Export Citation Sciwheel EndNote Ref. Manager Bibtex ProCite Sente EXPORT Select a format first Track Share ▬ ✚ Research Article Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] Radhika Agarwal 1 , Lavangi Sehgal https://orcid.org/0009-0003-9067-5617 2 , Jefferson Prince 3 , Srikant Natarajan https://orcid.org/0000-0002-2686-0397 4 , Shreya Bhat 5 , Siambiakthang Valte 6 Radhika Agarwal 1 , Lavangi Sehgal https://orcid.org/0009-0003-9067-5617 2 , [...] Jefferson Prince 3 , Srikant Natarajan https://orcid.org/0000-0002-2686-0397 4 , Shreya Bhat 5 , Siambiakthang Valte 6 PUBLISHED 04 Mar 2026 Author details Author details 1 Post graduate student,Department of Orthodontics and Dentofacial Orthopedics, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 2 Post graduate student ,Department of Prosthodontics and Crown and Bridge, Manav Rachna Dental College, Faridabad, Haryana, 121010, India 3 Associate Professor,Department of Oral and Maxillofacial Surgery, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, 576 104, India 4 Professor, Department of Oral Pathology and Microbiology, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 5 Post graduate student ,Department of Paediatric and Preventive Dentistry, Manipal College of Dental Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, 576 104, India 6 Post graduate student, Department of Conservative Dentistry and Endodontics, KLE Society's Institute of Dental Sciences, Belagavi, Karnataka, 590010, India Radhika Agarwal Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Resources, Software, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Lavangi Sehgal Roles: Data Curation, Investigation, Methodology, Resources, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Jefferson Prince Roles: Conceptualization, Data Curation, Resources, Software, Supervision, Writing – Original Draft Preparation, Writing – Review & Editing Srikant Natarajan Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Project Administration, Resources, Software, Visualization, Writing – Original Draft Preparation Shreya Bhat Roles: Conceptualization, Data Curation, Formal Analysis, Methodology, Writing – Original Draft Preparation Siambiakthang Valte Roles: Conceptualization, Investigation, Methodology, Resources OPEN PEER REVIEW DETAILS REVIEWER STATUS This article is included in the Manipal Academy of Higher Education gateway. Abstract Background Hunter-Schreger bands (HSBs) are distinctive optical features observed in enamel and are characterized by alternating dark and light bands that reflect the orientation of the enamel prism. Despite their clinical relevance, variations in HSB patterns across different tooth surfaces remain poorly understood. The aim of this study wass to correlate the width and density of HSB with the surface (labial, lingual, mesial distal and occlusal) of the tooth and the type of tooth. Materials and Methods Eighty intact permanent human teeth (20 each: incisors, canines, premolars, molars) from the maxillary and mandibular arches were sectioned in mesiodistal or labiolingual planes. Reflected-light photomicrographs (4× objective; Olympus CX20i) were analysed with ImageJ. HSB density was defined as the number of HSB pairs per millimetre (one dark + one light = pair). Group differences were assessed via repeated-measures ANOVA and independent t tests; p < 0.05. Statistical analyses Repeated-measures ANOVA and independent t tests were used to assess differences of HSB densities among surfaces and tooth types. A p value of of p <0.05 was considered . Results Significant variations in HSB density were observed across tooth surfaces within each tooth type and between the maxillary and mandibular arches. Compared with molars, mandibular buccal surfaces presented greater mean HSB densities in premolars ( p =0.001) ( p =0.201),whereas lingual surfaces presented consistent patterns across all tooth types ( p <0.05). Notably, incisal/occlusal surfaces demonstrated the greatest variability, with mandibular incisors exhibiting greater densities ( p <0.001) than maxillary incisors ( p <0.001). Conclusion This micrometric analysis provides detailed insights into the microstructural variations in the HSB across tooth surfaces, highlighting functional and anatomical influences. These findings underscore the clinical importance of HSB in adhesive restorations and enamel-related conditions, suggesting tailored approaches on the basis of tooth type and surface location. Further research is needed to explore the biomechanical implications and refine the clinical applications in dental practice. READ ALL READ LESS Keywords Hunter–Schreger bands, enamel microstructure, tooth surface, micrometric analysis Corresponding Author(s) Srikant Natarajan ( [email protected] ) Close Corresponding author: Srikant Natarajan Competing interests: No competing interests were disclosed. Grant information: The author(s) declared that no grants were involved in supporting this work. Copyright: © 2026 Agarwal R et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. How to cite: Agarwal R, Sehgal L, Prince J et al. Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] . F1000Research 2026, 15 :354 ( https://doi.org/10.12688/f1000research.175647.1 ) First published: 04 Mar 2026, 15 :354 ( https://doi.org/10.12688/f1000research.175647.1 ) Latest published: 18 Apr 2026, 15 :354 ( https://doi.org/10.12688/f1000research.175647.2 )  There is a newer version of this article available. Suppress this message for one day. 1. Introduction Hunter-Schreger bands (HSBs) are optical phenomena in which alternating dark and light bands appear when light is reflected off an enamel surface . . 1 Microscopic studies reveal a complex arrangement of enamel rods, which group into bundles or bands with different orientations relative to the tooth’s longitudinal section. Some of these rod groups appear longitudinally sectioned, whereas others seem to be cut transversely or obliquely. 2 These longitudinally sectioned groups are referred to as parazones, and those sectioned transversely are known as diazones. The interplay between parazones and diazones has been the foundation for Hunter-Schreger bands in enamel. Some researchers attribute these bands to differences in the density of calcification between alternating zones, whereas others suggest variations in the organic content between the enamel zones. 3 , 4 Recent synchrotron imaging of developing molars has demonstrated that enamel prisms generally follow a straight path but exhibit irregular, wave-like deviations in three-dimensional space. 5 HSB in different teeth of human dentition differ in terms of pattern and packing density. These properties of the HSB have been poorly investigated thus far, and the literature offers little explanation for their distribution. 6 The clinical significance of this study is to the differences and how they hold significant relevance for contemporary dental practice, particularly in procedures such as adhesive restoration bonding and in understanding enamel-related conditions such as cracked tooth syndrome and abfraction. 1 The aim of this study was to correlate the width and density of HSB with the surface (labial, lingual, mesial distal and occlusal) of the tooth and the type of tooth. The objectives are to measure the thickness of HSB in cross sections of teeth via reflected light microscopy; to correlate the density of HSB with occlusal, labial, lingual, mesial, and distal aspects of teeth; and to list the patterns of arrangement of Hunter-Schreger bands on human teeth on the basis of masticatory and occlusal forces (type traits of the tooth). 2. Methodology 2.1 Sample calculation This study was approved by the Institutional Ethics Committee (IEC ref. no. 23094). The extracted human teeth were collected from the Department of Oral and Maxillofacial Surgery; all the samples were anonymized and used following institutional guidelines. Tooth samples that had intact crowns; no evidence of attrition, abrasion, or erosion; and were visually free from hypoplasia and fractures were included in the study. On the basis of the study by Lynch et al. (2010), 1 Table 1 shows the HSB densities on various tooth types. The highest standard deviation was noted in premolars, at 2.2 units. Sample size was determined from published variability in HSB density 1 where the largest observed standard deviation was approximately σ = 2.2. For a two-sample comparison, using a two-sided α = 0.05 and power = 0.80, the required sample size per group to detect a between-group difference of d = 4.0 units is approximately n ≈ 5 per group (calculation based on the standard two-sample formula). Given the study design (incisor/canine/premolar/molar × maxilla/mandible with two section orientations), we allocated 80 teeth in total (10 teeth per tooth class per arch, further subdivided into two sectioning orientations of n = 5 each). Because some subgroup cells are small, subgroup analyses and interaction effects should be interpreted cautiously; we therefore provide effect sizes and 95% confidence intervals alongside p-values. Table 1. Independent t tests to compare the densities of HSB between the maxilla and mandible. Tooth Aspect Maxilla (n=5) Mean ± SD Mandible (n=5) Mean ± SD t P value Incisor Buccal 0.91±0.32 1.04±0.15 -0.814 0.439 Lingual 1.05±0.29 0.96±0.16 0.589 0.572 Mesial 0.75±0.17 0.99±0.23 -1.786 0.112 Distal 0.83±0.22 0.98±0.12 -1.37 0.208 Incisal/occlusal 0.51±0.12 1.17±0.14 -7.944 <0.001 Canine Buccal 1.06±0.17 1.04±0.18 0.151 0.884 Lingual 1.01±0.12 0.92±0.18 0.956 0.367 Mesial 1.03±0.13 0.96±0.18 0.779 0.458 Distal 1.04±0.07 0.96±0.12 1.31 0.227 Incisal/occlusal 0.91±0.11 0.7±0.16 2.416 0.042 Premolar Buccal 1.06±0.1 1.5±0.16 5.276 0.001 Lingual 1.02±0.15 1.42±0.22 3.342 0.01 Mesial 0.98±0.12 1.24±0.24 2.134 0.065 Distal 0.94±0.11 1.11±0.13 2.307 0.05 Incisal/occlusal 0.86±0.11 1.03±0.15 2.079 0.071 Molar Buccal 1.29±0.11 1.42±0.19 1.395 0.201 Lingual 1.44±0.14 1.57±0.13 1.605 0.147 Mesial 1.42±0.11 1.29±0.12 1.864 0.099 Distal 1.5±0.24 1.32±0.08 1.561 0.157 Incisal/occlusal 1.34±0.11 1.37±0.18 0.32 0.757 (The formula used is N = 2 ( Z 1 − α 2 ∗ k + Z 1 − β ) 2 σ 2 d 2 where σ is the standard deviation. Eighty teeth were selected and segregated into incisors, canines, premolars, and molars. A total of 10 samples were included in each category for both jaws, maxillary and mandible. Furthermore, each category was subdivided into 2 groups of five each. One group was used for mesiodistal sectioning, and the other was used for labiolingual sectioning. The occlusal aspect was also studied in each of the above groups. The distribution of teeth is shown in Figure 1 . Figure 1. Tooth-class allocation and sectioning methods used in the study: Maxillary and mandibular arches. 2.2 Preparation of samples A total of eighty wax molds, each measuring 2 × 2 inches, were fabricated. A thin mix of dental stone was poured into each mold, and once the initial set had occurred, the tooth was positioned so that half of the root and crown were embedded while the remaining portion projected outward. Two sectioning orientations were followed: one aligning the mesiodistal plane with the study plane and the other aligning the labiolingual plane. After complete setting, the specimens were mounted on a lathe, and half of the exposed tooth surface was carefully reduced to achieve a sectioned half-profile, similar to the methodology described by Lynch et al., 1 who emphasized controlled reduction to preserve structural detail. To improve visualization, the samples were subsequently polished using ground sectioning followed by pumice paste. Finally, the cleaned and prepared specimens were subjected to imaging. 2.3 Imaging Photomicrographs were captured using the microscope camera (Olympus CX20i) at a 4× objective and exported as uncompressed TIFF files. Images were calibrated with a stage micrometer and analysed in ImageJ version 1.59 (published by the National Institutes of Health, NIH, USA) as depicted in Figure 2 . The number of HSB pairs was counted within defined ROIs (1 dark + 1 adjacent light band = 1 pair). Figure 2. Representative reflected-light photomicrographs showing HSBs on different tooth surfaces. Panels: (A) maxillary incisor labial; (B) maxillary incisor lingual; (C) maxillary canine labial; (D) maxillary canine lingual; (E) mandibular premolar buccal; (F) mandibular molar buccal; (G) mandibular molar lingual; (H) mandibular molar occlusal; (I) mandibular molar lingual; (J) the mandibular molar occlusal. Images captured with 4× objective (Olympus CX20i); scale bar = 100 μm. Original TIFFs archived at Zenodo DOI: 10.5281/zenodo.17919950. Original unprocessed TIFF photomicrographs are available in the Zenodo deposit (see Data Availability) — file names correspond to the samples reported in Table 1 . The density of the HSB was calculated as follows: [ Density = Number of bands total length of enamel in which the bands are present ] The density of the HSB was then tabulated in Microsoft Excel and coded for the surface or the type of tooth. 2.4 Reliability A subset of images (n = 10) was re-analysed by the primary observer and independently analysed by a second observer. Intraclass correlation coefficients (ICC) were calculated using a two-way mixed-effects model for absolute agreement (single measures). 2.5 Statistical analysis Statistical analyses were conducted using SPSS version 20.0 (IBM Corp., Armonk, NY). Normality of continuous variables was assessed with the Shapiro–Wilk test. Between-arch comparisons used independent-samples t-tests (two-sided) with Cohen’s d reported as the effect size (Hedges’ g reported when n is small). Within-tooth surface differences were assessed with repeated-measures ANOVA (surface as the within-subject factor; arch and tooth class as between-subject factors). Mauchly’s test was used to assess sphericity; where violated, Greenhouse–Geisser or Huynh–Feldt corrections are reported. Partial eta-squared (ηp 2 ) is reported as the effect size for ANOVA. Post hoc pairwise comparisons used Bonferroni correction. All tests used α = 0.05. Exact p-values, degrees of freedom, F statistics, effect sizes and 95% CIs are reported in text and tables. 3. Results 3.1 Independent t tests to compare the densities of HSB The most pronounced differences in HSB density were observed on the incisal/occlusal surfaces, with mandibular incisors showing greater densities than their maxillary counterparts did (1.17 ± 0.14 vs. 0.51 ± 0.12, p < 0.001) (see Table 1 ). This finding suggests a stronger adaptive response to masticatory forces in mandibular incisors. Similarly, a substantial difference was noted in the incisal/occlusal surface of the canines, where maxillary canines presented higher HSB density than mandibular canines did (0.91 ± 0.11 vs. 0.70 ± 0.16, p = 0.042), possibly reflecting differences in functional demands. Premolars consistently demonstrated greater HSB densities in mandibular teeth, especially on the buccal and lingual surfaces, with a notable difference on the buccal side (1.50 ± 0.16 vs. 1.06 ± 0.10, p = 0.001). ( Table 1 ) In contrast, molars presented minimal variation, with no statistically significant differences across most surfaces, indicating a more uniform HSB distribution in this tooth class. 3.2 Measurement reliability Intra-rater reliability was good (ICC = 0.87, 95% CI 0.74–0.94), indicating consistent measurement of HSB density across repeated trials. Inter-rater reliability was good (ICC = 0.84, 95% CI 0.69–0.93), demonstrating strong agreement between the two observers. 3.3 Repeated-measures ANOVA comparing HSB density Repeated-measures ANOVA was used to assess the impact of the dental surface (Buccal, Lingual, Mesial, Distal, and Incisal/Occlusal) on Hunter-Schreger Band (HSB) density across different tooth classes (Incisor, Canine, Premolar, Molar) in both the maxilla and mandible. The analysis revealed several key findings: ( Table 2 ) Table 2. Repeated-measures ANOVA comparing HSB density across tooth surfaces, arches, and tooth types. Source Type III sum of squares df Mean square F P value ηp 2 Surface (Side) Huynh-Feldt 0.947 4.000 0.237 9.617 <0.001 0.231032 Surface x Arch Huynh-Feldt 0.172 4.000 0.043 1.744 0.144 0.051745 Surface x Tooth Type Huynh-Feldt 0.525 12.000 0.044 1.776 0.059 0.142779 Surface x Arch x Type Huynh-Feldt 1.009 12.000 0.084 3.413 <0.001 0.24249 Error (side) Huynh-Feldt 3.152 128.000 0.025 The main effect of arch (Surface × Arch) was not statistically significant (F(4,128) = 1.744, p = 0.144, partial ηp 2 = 0.052), indicating no consistent global arch difference when averaged across tooth types and surfaces. However, specific tooth×surface comparisons ( Table 1 ) revealed localized differences (for example, mandibular incisors and mandibular premolar buccal/lingual surfaces), indicating localized rather than global arch effects. 3.4 Effect of surface morphology/texture The analysis revealed noteworthy differences in the HSB density across the five surfaces (F = 9.617, p < 0.001; Table 2 ). This suggests that the HSB density is not uniformly distributed across all the tooth surfaces. Specifically, the incisal/occlusal surfaces generally presented lower densities than did the other surfaces. For example, the incisal/occlusal surface of mandibular incisors was denser than that of maxillary incisors (mean ± SD: 1.17 ± 0.14 vs. 0.51 ± 0.12, p < 0.001; Table 2 ). Similarly, mandibular premolars presented greater HSB density on the buccal and lingual surfaces than did maxillary premolars (buccal: 1.50 ± 0.16 vs. 1.06 ± 0.10, p = 0.001; lingual: 1.42 ± 0.22 vs. 1.02 ± 0.15, p = 0.010; Table 2 ). 3.5 Effect of arch form No noteworthy differences were observed between the maxilla and mandible regarding the surfaces of the teeth (F = 1.744, p = 0.144; Table 2 ). This indicates that, overall, the distribution of the HSB density is relatively consistent between the two arches when it is averaged across all tooth classes and surfaces. For example, while mandibular premolars and molars tended to show greater densities on the buccal and lingual surfaces, these differences were not statistically significant across all tooth classes (e.g., buccal:maxilla 1.06 ± 0.10 vs. mandible 1.50 ± 0.16, p = 0.001). 3.6 Interaction between tooth class and surface The interaction effect between tooth class and surface type was nearly significant (F = 1.776, p = 0.059). This suggests that variations in HSB density across different surfaces may differ depending on the tooth class. For example, premolars showed more pronounced differences in HSB density between the buccal and lingual surfaces than incisors did (buccal: 1.06 ± 0.10 for premolars vs. 0.91 ± 0.32 for incisors; lingual: 1.42 ± 0.22 for premolars vs. 1.05 ± 0.29 for incisors; Table 2 ), indicating that premolars exhibit more variation in response to functional forces. 3.7 Three-way interaction among the tooth class, arch, and surface A significant three-way interaction was found among tooth class, arch, and surface (F = 3.413, p < 0.001; Table 2 ). This major interaction highlights that the effect of the tooth class and surface on the HSB density is influenced by the arch in which the teeth are located. For example: Mandibular molars had similar HSB densities on the buccal and lingual surfaces to those of maxillary molars (buccal: 1.29 ± 0.11 vs. 1.42 ± 0.19; lingual: 1.44 ± 0.14 vs. 1.57 ± 0.13), but differences were observed in other surfaces, such as the incisor/Occlusal (mandibular molars: 1.37 ± 0.18 vs. maxillary molars: 1.34 ± 0.11, p = 0.757; Table 2 ). 4. Discussion Hunter–Schreger bands (HSBs) are alternating light and dark bands of enamel created by prism decussation. In addition to optical phenomena, they play a biomechanical role by enhancing the resistance of enamel to crack propagation and stress concentration during mastication. 1 The present study demonstrated variations in HSB density across tooth types, surfaces, and arches, reflecting the adaptive response of enamel to functional demands. The observed variations may also be explained by enamel development. Ameloblast displacement during enamel formation alters prism orientation, resulting in denser decussation in regions designed to resist greater mechanical forces. 3 In contrast, areas such as cusp tips and cervical enamel may present a reduced HSB density, possibly due to overcrowded ameloblasts or a reduced nutrient supply, resulting in aprismatic enamel or prisms lacking decussation. 8 Clinically, these variations in HSB density are highly relevant. Regions of high HSB density (mandibular incisors, mandibular premolars, and maxillary canines) exhibit superior resistance to crack propagation and may provide more reliable bonding substrates for adhesive restorations. Conversely, regions of low HSB density (maxillary incisal edges, cervical enamel, some cusp tips) are prone to enamel wear, abfraction, and crack initiation, as reported in studies on cracked tooth syndrome and enamel fracture behaviour. 9 , 10 , 11 , 12 These findings align with reports linking cracked tooth syndrome further corroborate that enamel regions with reduced microstructural reinforcement, particularly those with sparse HSB decussation, are more vulnerable to crack propagation under functional loading. 9 Compared with maxillary incisors, mandibular incisors presented significantly greater HSB densities on the incisal surface ( p < 0.001). This correlates well with its function as the lower incisors have the role of cutting and are directly subjected to shear and compressive forces during incising. A deeper understanding of these teeth functions as a crack-stopping mechanism, reducing fracture risk in thin enamel zones. 1 , 3 A recent meta-analysis confirmed that the HSB density is greater in functionally stressed enamel regions, particularly the middle enamel third, than in the cervical third. 7 In contrast, maxillary incisors act largely as guiding teeth, facing lower direct occlusal loads, hence their reduced density. Compared with mandibular canines, maxillary canines presented greater HSB densities at the cusp tip/incisal edge ( p = 0.042). Maxillary canines are central to canine guidance , dissipating large lateral and protrusive forces during mandibular excursions. Denser HSB decussation in this region enhances crack resistance under multidirectional stresses. 5 , 7 Mandibular canines, although important for aesthetics and occlusal stability, bear comparatively less excursive load, explaining the lower density observed. Mandibular premolars consistently presented higher HSB densities than did maxillary premolars, particularly on the buccal and lingual surfaces ( p = 0.001 and p = 0.010). Functionally, mandibular premolars play a dual role in shearing and grinding, especially in unilateral mastication cycles, which subject them to oblique functional stresses. 11 Comparative micromechanical assessments indicate that premolars tend to display higher enamel hardness and elastic modulus values compared to canines and incisors. This observation highlights the structural adaptation of premolar enamel to withstand greater functional demands within this tooth class. In contrast, molars presented relatively uniform HSB densities across arches and surfaces, with no substantial differences. This is consistent with their role as primary grinding units , where forces are widely distributed across cusps and thick enamel layers. 1 Evolutionary studies indicate that molars adopt consistent HSB configurations to maximize wear resistance, with enamel prisms frequently oriented perpendicularly to occlusal surfaces for optimal reinforcement. 12 , 13 Thus, unlike incisors and premolars, molars benefit more from uniform reinforcement across surfaces. Repeated-measures ANOVA confirmed noteworthy differences across surfaces ( p < 0.001). The functional surfaces (incisal, buccal, and lingual) presented greater HSB density than did the mesial and distal surfaces. This observation reflects enamel’s adaptive reinforcement of load-bearing surfaces . 1 , 3 The greatest variability was observed at incisal/occlusal surfaces, likely due to differences in stress distribution: mandibular incisors and premolars withstand higher shear loads, whereas maxillary incisors act more as guiding teeth. HSB packing and decussation patterns likely reflect long-term functional demands on enamel shaped by diet and masticatory behaviour across populations. While the present results are consistent with an adaptive response of enamel microstructure to mechanical loading, testing evolutionary hypotheses requires population-level comparative data and higher-resolution 3-D imaging—topics for future work. 7 , 14 , 15 This study was limited by its modest sample size and reliance on two-dimensional sectioning, which may not fully represent the three-dimensional complexity of the HSB architecture. Future research using high-resolution 3D imaging and Nano indentation testing may provide a deeper understanding of how HSB orientation correlates with enamel strength. Furthermore, exploring HSB adaptations in attached or older teeth across diverse populations could reveal age-related changes in enamel biomechanics. 5. Conclusion This study revealed significant differences in Hunter-Schreger Band density across various tooth types and surfaces, reflecting adaptations to their functional roles. Compared with maxillary incisors, mandibular incisors exhibit notably greater HSB densities on the incisal/occlusal surfaces, suggesting an adaptation to greater masticatory forces. Maxillary canines present higher HSB densities than mandibular canines do, potentially indicating different functional demands. Additionally, mandibular premolars consistently display higher HSB densities on the buccal and lingual surfaces than their maxillary counterparts do, whereas molars show relatively uniform HSB densities across surfaces. These variations in HSB density highlight the specialized adaptation of dental tissues to their biomechanical environments. Ethics approval statement This study was done in collaboration between Manipal College of Dental Science, Mangalore, Manipal Academy of Higher Education, Manipal, India. This study was approved by the Institutional Ethics Committee of Manipal College of Dental Science, Mangalore (IEC Reference number: 23094). The requirement for written informed consent was waived. All research was performed in accordance with the Declaration of Helsinki. Patient consent Written informed consent was obtained from all participants for the use of extracted teeth. The requirement for written informed consent was waived. All research was performed in accordance with the Declaration of Helsinki. Preregistered data analysis The study was not preregistered. No formal data analysis plan was registered in an independent registry prior to data collection or analysis. Declaration of generative AI and AI-assisted technologies in the manuscript preparation process During the preparation of this work the author(s) used ChatGPT (OpenAI) in order to assist with language refinement, clarity improvement, and editing suggestions. After using this tool, the author(s) carefully reviewed and edited the content as needed and take full responsibility for the content of the publication. Data availability Zenodo. Correlating the densities of Hunter–Schreger bands with function and surfaces of teeth: underlying data. https://doi.org/10.5281/zenodo.17919950 . 16 Notes on image files. The zip file in the Zenodo deposit contains the original, unprocessed reflected-light photomicrographs used to generate the measurements in this study. The images are provided to ensure full transparency and reproducibility of image cropping, ROI placement and measurement steps. This project contains the following underlying data: • values.xlsx (Raw HSB counts, ROI lengths, and calculated HSB density values for all samples). • Uncompressed TIFF photomicrographs used for quantitative analysis and representative figures). • TABLE 1 and 2 excel files for the statistical results. Data are available under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) license. Acknowledgements Not applicable. References 1. Lynch CD, O’Sullivan VR, Dockery P, et al. : Hunter-Schreger Band patterns in human tooth enamel. J. Anat. 2010 Aug; 217 (2): 106–115. PubMed Abstract | Publisher Full Text | Free Full Text 2. Rodriguez-Palomo A, Jacobsen MS, Christensen TE, et al. : Enamel-like stiffness achieved by poorly oriented nanocrystals in the capping tissue of Mexican beaded lizard osteoderms. Acta Biomater. 2025 Aug 14; 204 : 457–469. PubMed Abstract | Publisher Full Text 3. Francischone CE, Maenososno RM: ABSTRACTS OF THE WORKS PRESENTED AT THE 22nd Academic Conference of the Bauru School of Dentistry “Dr. Waldyr Antonio Janson”. J. Appl. Oral Sci. 2009; 17 (spe): 139–195. Publisher Full Text 4. Thompson VP: The tooth: An analogue for biomimetic materials design and processing. Dent. Mater. 2020 Jan 1; 36 (1): 25–42. PubMed Abstract | Publisher Full Text 5. Yang D, Bharatiya M, Grine FE: Hunter-Schreger Band configuration in human molars reveals more decussation in the lateral enamel of ‘functional’cusps than ‘guiding’cusps. Arch. Oral Biol. 2022 Oct 1; 142 : 105524. PubMed Abstract | Publisher Full Text 6. Tafforeau P, Zermeno JP, Smith TM: Tracking cellular-level enamel growth and structure in 4D with synchrotron imaging. J. Hum. Evol. 2012 Mar 1; 62 (3): 424–428. PubMed Abstract | Publisher Full Text 7. Koldehoff J, Swain MV, Schneider GA: The geometrical structure of interfaces in dental enamel: A FIB-STEM investigation. Acta Biomater. 2020 Mar 1; 104 : 17–27. PubMed Abstract | Publisher Full Text 8. Barnhart EC, Campbell PM, Noureldin A, et al. : The quality of etched enamel in different regions and tooth types and its significance in bonding and the development of white spot lesions. Angle Orthod. 2021 Sep 1; 91 (5): 576–582. PubMed Abstract | Publisher Full Text | Free Full Text 9. Lynch CD, McConnell RJ: The cracked tooth syndrome. Journal-Canadian Dental Association. 2002 Sep 1; 68 (8): 470–475. PubMed Abstract 10. Dioguardi M, Lo Muzio E, Illuzzi G, et al. : Analysis of the packing density of Hunter-Schreger bands (HSB): a systematic review and meta-analysis. Eur. J. Med. Res. 2025 Apr 15; 30 (1): 292. PubMed Abstract | Publisher Full Text | Free Full Text 11. Barri AB, Mahmoud BF, Elwan AS, et al. : Differences in the Buccolingual Inclinations of Premolars Among Class II Division I Subjects With Different Vertical Facial Patterns. Cureus. 2024 Oct 29; 16 (10). Publisher Full Text 12. Von Koenigswald W, Holbrook LT, Rose KD: Diversity and evolution of Hunter-Schreger Band configuration in tooth enamel of perissodactyl mammals. Acta Palaeontol. Pol. 2011 Mar; 56 (1): 11–32. Publisher Full Text 13. Koenigswald W, et al. : Diversity and evolution of Hunter–Schreger Band configuration in tooth enamel of perissodactyl mammals. Acta Palaeontol. Pol. 2011; 56 (1): 3–16. 14. Ungar PS: Evolution of the human diet: the known, the unknown, and the unknowable. Oxford University Press; 2007. 15. Kaifu Y, Kasai K, Townsend GC, et al. : Tooth wear and subsistence in prehistoric hunter–gatherers. Am. J. Phys. Anthropol. 2003; 120 (3): 233–251. 16. Agarwal R, Sehgal L, Bhat S, et al. : Correlating the densities of Hunter–Schreger bands with function and surfaces of teeth: underlying data. Zenodo. 2025. Publisher Full Text Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 04 Mar 2026 ADD YOUR COMMENT Comment Author details Author details 1 Post graduate student,Department of Orthodontics and Dentofacial Orthopedics, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 2 Post graduate student ,Department of Prosthodontics and Crown and Bridge, Manav Rachna Dental College, Faridabad, Haryana, 121010, India 3 Associate Professor,Department of Oral and Maxillofacial Surgery, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, 576 104, India 4 Professor, Department of Oral Pathology and Microbiology, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India 5 Post graduate student ,Department of Paediatric and Preventive Dentistry, Manipal College of Dental Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, 576 104, India 6 Post graduate student, Department of Conservative Dentistry and Endodontics, KLE Society's Institute of Dental Sciences, Belagavi, Karnataka, 590010, India Radhika Agarwal Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Resources, Software, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Lavangi Sehgal Roles: Data Curation, Investigation, Methodology, Resources, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Jefferson Prince Roles: Conceptualization, Data Curation, Resources, Software, Supervision, Writing – Original Draft Preparation, Writing – Review & Editing Srikant Natarajan Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Project Administration, Resources, Software, Visualization, Writing – Original Draft Preparation Shreya Bhat Roles: Conceptualization, Data Curation, Formal Analysis, Methodology, Writing – Original Draft Preparation Siambiakthang Valte Roles: Conceptualization, Investigation, Methodology, Resources Competing interests No competing interests were disclosed. Grant information The author(s) declared that no grants were involved in supporting this work. Article Versions (2) version 2 Revised Published: 18 Apr 2026, 15:354 https://doi.org/10.12688/f1000research.175647.2 version 1 Published: 04 Mar 2026, 15:354 https://doi.org/10.12688/f1000research.175647.1 Copyright © 2026 Agarwal R et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Download Export To Sciwheel Bibtex EndNote ProCite Ref. Manager (RIS) Sente metrics Views Downloads F1000Research - - PubMed Central info_outline Data from PMC are received and updated monthly. - - Citations open_in_new 0 open_in_new 0 open_in_new SEE MORE DETAILS CITE how to cite this article Agarwal R, Sehgal L, Prince J et al. Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] . F1000Research 2026, 15 :354 ( https://doi.org/10.12688/f1000research.175647.1 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS track receive updates on this article Track an article to receive email alerts on any updates to this article. TRACK THIS ARTICLE Share Open Peer Review Current Reviewer Status: ? Key to Reviewer Statuses VIEW HIDE Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Version 1 VERSION 1 PUBLISHED 04 Mar 2026 Views 0 Cite How to cite this report: Abdulla R. Reviewer Report For: Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] . F1000Research 2026, 15 :354 ( https://doi.org/10.5256/f1000research.193650.r471054 ) The direct URL for this report is: https://f1000research.com/articles/15-354/v1#referee-response-471054 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 11 Apr 2026 Riaz Abdulla , Yenepoya Dental College, Mangalore, India Approved VIEWS 0 https://doi.org/10.5256/f1000research.193650.r471054 The manuscript is well written and presents a scientifically sound study with an appropriate statistical analysis. The topic is relevant, and the findings contribute valuable insights into the distribution and functional significance of the Hunter–Schreger bands. The data availability ... Continue reading READ ALL The manuscript is well written and presents a scientifically sound study with an appropriate statistical analysis. The topic is relevant, and the findings contribute valuable insights into the distribution and functional significance of the Hunter–Schreger bands. The data availability and transparency were commendable. However, minor clarifications in the methodology, particularly regarding the occlusal surface evaluation and justification of the sectioning techniques, are necessary to ensure full reproducibility. Addressing these issues will further strengthen the manuscript. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: oral pathology,oral cancer I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Abdulla R. Reviewer Report For: Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] . F1000Research 2026, 15 :354 ( https://doi.org/10.5256/f1000research.193650.r471054 ) The direct URL for this report is: https://f1000research.com/articles/15-354/v1#referee-response-471054 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 29 Apr 2026 Srikant Natarajan , Professor, Department of Oral Pathology and Microbiology, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, 576104, India 29 Apr 2026 Author Response We sincerely thank the reviewer for the positive evaluation of our manuscript and for recognizing its scientific rigor, relevance, and transparency. We appreciate the suggestion for further methodological clarification. In ... Continue reading We sincerely thank the reviewer for the positive evaluation of our manuscript and for recognizing its scientific rigor, relevance, and transparency. We appreciate the suggestion for further methodological clarification. In response, the technique of evaluation of the occlusal aspect has now been more explicitly described in the methodology section. Additionally, the rationale for dividing samples into mesiodistal and buccolingual sectioning groups has been clarified with the additon with a figure (Figure 2). With this approach a comprehensive assessment of enamel banding patterns in two orthogonal planes was possible. i.e in the mesiodistal sections mesial, distal, and occlusal aspects and in the buccolingual sections buccal, lingual, and occlusal surfaces were evaluated. This dual-plane evaluation was essential to account for variations in enamel microstructure in all aspects of the tooth surface, associated with functional loading and stress distribution, particularly in primary load-bearing cusps and proximal regions. These revisions have been incorporated into the manuscript to enhance clarity and ensure reproducibility. We sincerely thank the reviewer for the positive evaluation of our manuscript and for recognizing its scientific rigor, relevance, and transparency. We appreciate the suggestion for further methodological clarification. In response, the technique of evaluation of the occlusal aspect has now been more explicitly described in the methodology section. Additionally, the rationale for dividing samples into mesiodistal and buccolingual sectioning groups has been clarified with the additon with a figure (Figure 2). With this approach a comprehensive assessment of enamel banding patterns in two orthogonal planes was possible. i.e in the mesiodistal sections mesial, distal, and occlusal aspects and in the buccolingual sections buccal, lingual, and occlusal surfaces were evaluated. This dual-plane evaluation was essential to account for variations in enamel microstructure in all aspects of the tooth surface, associated with functional loading and stress distribution, particularly in primary load-bearing cusps and proximal regions. These revisions have been incorporated into the manuscript to enhance clarity and ensure reproducibility. Competing Interests: no competing interests Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 29 Apr 2026 Srikant Natarajan , Professor, Department of Oral Pathology and Microbiology, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, 576104, India 29 Apr 2026 Author Response We sincerely thank the reviewer for the positive evaluation of our manuscript and for recognizing its scientific rigor, relevance, and transparency. We appreciate the suggestion for further methodological clarification. In ... Continue reading We sincerely thank the reviewer for the positive evaluation of our manuscript and for recognizing its scientific rigor, relevance, and transparency. We appreciate the suggestion for further methodological clarification. In response, the technique of evaluation of the occlusal aspect has now been more explicitly described in the methodology section. Additionally, the rationale for dividing samples into mesiodistal and buccolingual sectioning groups has been clarified with the additon with a figure (Figure 2). With this approach a comprehensive assessment of enamel banding patterns in two orthogonal planes was possible. i.e in the mesiodistal sections mesial, distal, and occlusal aspects and in the buccolingual sections buccal, lingual, and occlusal surfaces were evaluated. This dual-plane evaluation was essential to account for variations in enamel microstructure in all aspects of the tooth surface, associated with functional loading and stress distribution, particularly in primary load-bearing cusps and proximal regions. These revisions have been incorporated into the manuscript to enhance clarity and ensure reproducibility. We sincerely thank the reviewer for the positive evaluation of our manuscript and for recognizing its scientific rigor, relevance, and transparency. We appreciate the suggestion for further methodological clarification. In response, the technique of evaluation of the occlusal aspect has now been more explicitly described in the methodology section. Additionally, the rationale for dividing samples into mesiodistal and buccolingual sectioning groups has been clarified with the additon with a figure (Figure 2). With this approach a comprehensive assessment of enamel banding patterns in two orthogonal planes was possible. i.e in the mesiodistal sections mesial, distal, and occlusal aspects and in the buccolingual sections buccal, lingual, and occlusal surfaces were evaluated. This dual-plane evaluation was essential to account for variations in enamel microstructure in all aspects of the tooth surface, associated with functional loading and stress distribution, particularly in primary load-bearing cusps and proximal regions. These revisions have been incorporated into the manuscript to enhance clarity and ensure reproducibility. Competing Interests: no competing interests Close Report a concern COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Raj LSM. Reviewer Report For: Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] . F1000Research 2026, 15 :354 ( https://doi.org/10.5256/f1000research.193650.r469099 ) The direct URL for this report is: https://f1000research.com/articles/15-354/v1#referee-response-469099 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 25 Mar 2026 L S Makesh Raj , TN Dr. MGR Medical University, Tagore Dental College and Hospital, Chennai, Tamil Nadu, India Approved VIEWS 0 https://doi.org/10.5256/f1000research.193650.r469099 The article is well written and presents a strong statistical analysis. However, a few points in the Methodology section require clarification to ensure reproducibility: Criteria for evaluating the occlusal aspect The manuscript mentions that ... Continue reading READ ALL The article is well written and presents a strong statistical analysis. However, a few points in the Methodology section require clarification to ensure reproducibility: Criteria for evaluating the occlusal aspect The manuscript mentions that “the occlusal aspect was also studied in each of the above groups,” but the specific criteria or parameters used for this evaluation are not described. Please clarify what features were assessed or standards followed. Justification for sectioning approach The rationale behind dividing the samples into two groups—one for mesiodistal sectioning and the other for labiolingual sectioning—is not clearly explained. Kindly provide the reasoning for this methodological choice and how it contributes to the study objectives or improves the analysis. Providing these details will enhance the clarity and reproducibility of the study. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: oral Pathology and Forensic Odontology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Raj LSM. Reviewer Report For: Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] . F1000Research 2026, 15 :354 ( https://doi.org/10.5256/f1000research.193650.r469099 ) The direct URL for this report is: https://f1000research.com/articles/15-354/v1#referee-response-469099 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 18 Apr 2026 Srikant Natarajan , Professor, Department of Oral Pathology and Microbiology, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, 576104, India 18 Apr 2026 Author Response 1. We thank the reviewer for highlighting this point. The occlusal aspect was evaluated to assess the appearance and orientation of Hunter–Schreger bands at the cusp tips and occlusal enamel ... Continue reading 1. We thank the reviewer for highlighting this point. The occlusal aspect was evaluated to assess the appearance and orientation of Hunter–Schreger bands at the cusp tips and occlusal enamel surface. Specifically, the following features were considered: (1) visibility and clarity of banding patterns, (2) orientation of bands relative to the cusp slopes, and (3) continuity or disruption of bands from the subsurface enamel toward the occlusal surface. This assessment was performed qualitatively under magnification to correlate surface expression with underlying band patterns observed in sectional planes. The corresponding details have now been added to the revised manuscript. (Figure 2) 2. We appreciate the reviewer’s observation. The division of samples into mesiodistal and buccolingual sectioning groups was undertaken to enable comprehensive evaluation of enamel banding patterns in two orthogonal planes. Mesiodistal sectioning allowed visualization of mesial, distal, and occlusal surfaces; similarly, buccolingual sectioning enabled evaluation of buccal, lingual, and occlusal aspects, providing complementary information on band orientation across the crown as mentioned in Figure 2. This approach was essential because enamel microstructure, including Hunter–Schreger bands, varies with functional loading and stress distribution. Maxillary palatal cusps and mandibular buccal cusps serve as primary load-bearing areas, while proximal surfaces experience different stress patterns during mastication. Assessing specimens in both planes allowed for a more accurate interpretation of band orientation and distribution in relation to these biomechanical factors, thereby strengthening the study’s analytical framework. 1. We thank the reviewer for highlighting this point. The occlusal aspect was evaluated to assess the appearance and orientation of Hunter–Schreger bands at the cusp tips and occlusal enamel surface. Specifically, the following features were considered: (1) visibility and clarity of banding patterns, (2) orientation of bands relative to the cusp slopes, and (3) continuity or disruption of bands from the subsurface enamel toward the occlusal surface. This assessment was performed qualitatively under magnification to correlate surface expression with underlying band patterns observed in sectional planes. The corresponding details have now been added to the revised manuscript. (Figure 2) 2. We appreciate the reviewer’s observation. The division of samples into mesiodistal and buccolingual sectioning groups was undertaken to enable comprehensive evaluation of enamel banding patterns in two orthogonal planes. Mesiodistal sectioning allowed visualization of mesial, distal, and occlusal surfaces; similarly, buccolingual sectioning enabled evaluation of buccal, lingual, and occlusal aspects, providing complementary information on band orientation across the crown as mentioned in Figure 2. This approach was essential because enamel microstructure, including Hunter–Schreger bands, varies with functional loading and stress distribution. Maxillary palatal cusps and mandibular buccal cusps serve as primary load-bearing areas, while proximal surfaces experience different stress patterns during mastication. Assessing specimens in both planes allowed for a more accurate interpretation of band orientation and distribution in relation to these biomechanical factors, thereby strengthening the study’s analytical framework. Competing Interests: The authors declare no competing interests. Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 18 Apr 2026 Srikant Natarajan , Professor, Department of Oral Pathology and Microbiology, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, 576104, India 18 Apr 2026 Author Response 1. We thank the reviewer for highlighting this point. The occlusal aspect was evaluated to assess the appearance and orientation of Hunter–Schreger bands at the cusp tips and occlusal enamel ... Continue reading 1. We thank the reviewer for highlighting this point. The occlusal aspect was evaluated to assess the appearance and orientation of Hunter–Schreger bands at the cusp tips and occlusal enamel surface. Specifically, the following features were considered: (1) visibility and clarity of banding patterns, (2) orientation of bands relative to the cusp slopes, and (3) continuity or disruption of bands from the subsurface enamel toward the occlusal surface. This assessment was performed qualitatively under magnification to correlate surface expression with underlying band patterns observed in sectional planes. The corresponding details have now been added to the revised manuscript. (Figure 2) 2. We appreciate the reviewer’s observation. The division of samples into mesiodistal and buccolingual sectioning groups was undertaken to enable comprehensive evaluation of enamel banding patterns in two orthogonal planes. Mesiodistal sectioning allowed visualization of mesial, distal, and occlusal surfaces; similarly, buccolingual sectioning enabled evaluation of buccal, lingual, and occlusal aspects, providing complementary information on band orientation across the crown as mentioned in Figure 2. This approach was essential because enamel microstructure, including Hunter–Schreger bands, varies with functional loading and stress distribution. Maxillary palatal cusps and mandibular buccal cusps serve as primary load-bearing areas, while proximal surfaces experience different stress patterns during mastication. Assessing specimens in both planes allowed for a more accurate interpretation of band orientation and distribution in relation to these biomechanical factors, thereby strengthening the study’s analytical framework. 1. We thank the reviewer for highlighting this point. The occlusal aspect was evaluated to assess the appearance and orientation of Hunter–Schreger bands at the cusp tips and occlusal enamel surface. Specifically, the following features were considered: (1) visibility and clarity of banding patterns, (2) orientation of bands relative to the cusp slopes, and (3) continuity or disruption of bands from the subsurface enamel toward the occlusal surface. This assessment was performed qualitatively under magnification to correlate surface expression with underlying band patterns observed in sectional planes. The corresponding details have now been added to the revised manuscript. (Figure 2) 2. We appreciate the reviewer’s observation. The division of samples into mesiodistal and buccolingual sectioning groups was undertaken to enable comprehensive evaluation of enamel banding patterns in two orthogonal planes. Mesiodistal sectioning allowed visualization of mesial, distal, and occlusal surfaces; similarly, buccolingual sectioning enabled evaluation of buccal, lingual, and occlusal aspects, providing complementary information on band orientation across the crown as mentioned in Figure 2. This approach was essential because enamel microstructure, including Hunter–Schreger bands, varies with functional loading and stress distribution. Maxillary palatal cusps and mandibular buccal cusps serve as primary load-bearing areas, while proximal surfaces experience different stress patterns during mastication. Assessing specimens in both planes allowed for a more accurate interpretation of band orientation and distribution in relation to these biomechanical factors, thereby strengthening the study’s analytical framework. Competing Interests: The authors declare no competing interests. Close Report a concern COMMENT ON THIS REPORT Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 04 Mar 2026 ADD YOUR COMMENT Comment keyboard_arrow_left keyboard_arrow_right Open Peer Review Reviewer Status info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Reports Invited Reviewers 1 2 Version 2 (revision) 18 Apr 26 Version 1 04 Mar 26 read read L S Makesh Raj , Tagore Dental College and Hospital, Chennai, India Riaz Abdulla , Yenepoya Dental College, Mangalore, India Comments on this article All Comments (0) Add a comment Sign up for content alerts Sign Up You are now signed up to receive this alert Browse by related subjects keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2026 Abdulla R. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 11 Apr 2026 | for Version 1 Riaz Abdulla , Yenepoya Dental College, Mangalore, India 0 Views copyright © 2026 Abdulla R. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The manuscript is well written and presents a scientifically sound study with an appropriate statistical analysis. The topic is relevant, and the findings contribute valuable insights into the distribution and functional significance of the Hunter–Schreger bands. The data availability and transparency were commendable. However, minor clarifications in the methodology, particularly regarding the occlusal surface evaluation and justification of the sectioning techniques, are necessary to ensure full reproducibility. Addressing these issues will further strengthen the manuscript. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise oral pathology,oral cancer I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (1) Author Response 29 Apr 2026 Srikant Natarajan, Professor, Department of Oral Pathology and Microbiology, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, 576104, India We sincerely thank the reviewer for the positive evaluation of our manuscript and for recognizing its scientific rigor, relevance, and transparency. We appreciate the suggestion for further methodological clarification. In response, the technique of evaluation of the occlusal aspect has now been more explicitly described in the methodology section. Additionally, the rationale for dividing samples into mesiodistal and buccolingual sectioning groups has been clarified with the additon with a figure (Figure 2). With this approach a comprehensive assessment of enamel banding patterns in two orthogonal planes was possible. i.e in the mesiodistal sections mesial, distal, and occlusal aspects and in the buccolingual sections buccal, lingual, and occlusal surfaces were evaluated. This dual-plane evaluation was essential to account for variations in enamel microstructure in all aspects of the tooth surface, associated with functional loading and stress distribution, particularly in primary load-bearing cusps and proximal regions. These revisions have been incorporated into the manuscript to enhance clarity and ensure reproducibility. View more View less Competing Interests no competing interests reply Respond Report a concern Abdulla R. Peer Review Report For: Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] . F1000Research 2026, 15 :354 ( https://doi.org/10.5256/f1000research.193650.r471054) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/15-354/v1#referee-response-471054 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2026 Raj L. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 25 Mar 2026 | for Version 1 L S Makesh Raj , TN Dr. MGR Medical University, Tagore Dental College and Hospital, Chennai, Tamil Nadu, India 0 Views copyright © 2026 Raj L. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The article is well written and presents a strong statistical analysis. However, a few points in the Methodology section require clarification to ensure reproducibility: Criteria for evaluating the occlusal aspect The manuscript mentions that “the occlusal aspect was also studied in each of the above groups,” but the specific criteria or parameters used for this evaluation are not described. Please clarify what features were assessed or standards followed. Justification for sectioning approach The rationale behind dividing the samples into two groups—one for mesiodistal sectioning and the other for labiolingual sectioning—is not clearly explained. Kindly provide the reasoning for this methodological choice and how it contributes to the study objectives or improves the analysis. Providing these details will enhance the clarity and reproducibility of the study. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise oral Pathology and Forensic Odontology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (1) Author Response 18 Apr 2026 Srikant Natarajan, Professor, Department of Oral Pathology and Microbiology, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, 576104, India 1. We thank the reviewer for highlighting this point. The occlusal aspect was evaluated to assess the appearance and orientation of Hunter–Schreger bands at the cusp tips and occlusal enamel surface. Specifically, the following features were considered: (1) visibility and clarity of banding patterns, (2) orientation of bands relative to the cusp slopes, and (3) continuity or disruption of bands from the subsurface enamel toward the occlusal surface. This assessment was performed qualitatively under magnification to correlate surface expression with underlying band patterns observed in sectional planes. The corresponding details have now been added to the revised manuscript. (Figure 2) 2. We appreciate the reviewer’s observation. The division of samples into mesiodistal and buccolingual sectioning groups was undertaken to enable comprehensive evaluation of enamel banding patterns in two orthogonal planes. Mesiodistal sectioning allowed visualization of mesial, distal, and occlusal surfaces; similarly, buccolingual sectioning enabled evaluation of buccal, lingual, and occlusal aspects, providing complementary information on band orientation across the crown as mentioned in Figure 2. This approach was essential because enamel microstructure, including Hunter–Schreger bands, varies with functional loading and stress distribution. Maxillary palatal cusps and mandibular buccal cusps serve as primary load-bearing areas, while proximal surfaces experience different stress patterns during mastication. Assessing specimens in both planes allowed for a more accurate interpretation of band orientation and distribution in relation to these biomechanical factors, thereby strengthening the study’s analytical framework. View more View less Competing Interests The authors declare no competing interests. reply Respond Report a concern Raj LSM. Peer Review Report For: Correlating the Densities of Hunter-schreger Bands With Function and Surfaces of Teeth: A Micrometric Analysis [version 1; peer review: 2 approved] . F1000Research 2026, 15 :354 ( https://doi.org/10.5256/f1000research.193650.r469099) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/15-354/v1#referee-response-469099 Alongside their report, reviewers assign a status to the article: Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. 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