Reconstructing smoking history through dental cementum analysis - a preliminary investigation on modern and archaeological teeth

Valentina Perrone; Anna M Davies-Barrett; Mario Migliario; Patrick Randolph-Quinney; Sarah A Inskip; Edward C Schwalbe

doi:10.1371/journal.pone.0323812

. 2025 May 27;20(5):e0323812. doi: 10.1371/journal.pone.0323812

Reconstructing smoking history through dental cementum analysis - a preliminary investigation on modern and archaeological teeth

Valentina Perrone ^1,^2,^*, Anna M Davies-Barrett ², Mario Migliario ³, Patrick Randolph-Quinney ^4,⁵, Sarah A Inskip ², Edward C Schwalbe ^1,^*

Editor: Francisco Wanderley Garcia de Paula-Silva⁶

PMCID: PMC12111438 PMID: 40424319

Abstract

Acellular extrinsic fibre cementum (AEFC) has been widely utilised in cementochronology to estimate age at death, seasonality, and for life-history reconstruction. Smoking has been commonplace in the UK since the 17th century and is known to compromise oral health and to modulate physiological processes. This study aimed to investigate whether AEFC analysis could identify smoking activity in both modern and archaeological populations. A modern sample (70 teeth from 46 donors) with known age, sex, and smoking status was compared with an archaeological sample (18 teeth from 18 individuals), dating from the 18th/19th centuries in Coventry, UK, whose biographical information was recorded from coffin plates where available. Smoking status for the archaeological individuals was inferred from pipe notches and dental staining. AEFC analysis that was blinded to smoking status measured increment count, overall width and the presence of irregularities within the cementum microstructure in both samples. Results demonstrated that the AEFC width was significantly lower (p = 0.008) in current smokers compared to ex-smokers. Additionally, individuals with a history of smoking were significantly more likely to display disrupted incremental patterns within their AEFC (p < 0.001). This research suggests an association between smoking and periodontal ligament health, which influences AEFC formation and shows that the AEFC provides a record of smoking-related oral health damage. This research expands the potential applications of cementochronology to forensic and archaeological investigations for life history reconstruction.

Introduction

Teeth have proved to be useful for biological anthropology. Their macro- and microscopic analysis has revealed critical information on diet and migratory movements [1–3], age-at-death [4–6] and can contribute towards life-history reconstruction [7–11].

Teeth consist of three main hard tissues: enamel, dentine and cementum. Unlike the first two, dental cementum – and more specifically, the acellular extrinsic fibre cementum (AEFC) – continually grows on the coronal third of the dental root in alternating dark and light increments that are deposited until tooth loss or death of the individual. The continuity of cemental deposition is connected to its main role of anchoring the teeth in their sockets, withstanding masticatory and para-masticatory stresses. Like bones and other dental tissues, dental cementum also shows birefringent properties when observed in polarised light.

The pattern of cemental deposition is referred to under numerous names (e.g., tooth cementum annulation or TCA [12]) and is defined as the seasonal formation of alternating light increments and dark increments, the latter clinically known as lines of Salter [13]. In the Northern hemisphere, light increments form between April and September, and the lines of Salter form between September and April (i.e., respectively summer and winter season) [14]. Due to this seasonal phenomenon, each annulus (i.e., a pair of light and dark increments) corresponds to one chronological year. By adding the total number of annuli to the tooth-specific age at eruption, the age of the individual can be calculated [15]. Stott et al. [16] were the first to use human dental cementum to estimate age in adults, effectively starting what is now known as cementochronology [4,17]. Methods commonly adopted in biological anthropology for assessment of age in adults rely on the degree of age-related degeneration of specific skeletal areas, such as the pelvis or sternal rib ends [18–20]. In comparison to these techniques, one advantage of cementochronology is that it is assessed on a tissue that grows throughout life and does not remodel, and on increments that seem to be internally regulated by a “circannual clock”. In fact, the biological mechanisms of deposition of the AEFC are not well understood. Previously, the optical difference of the increments was thought to be due to a slower and faster rate of deposition of the tissue respectively occurring in the winter and summer season, which consequently resulted in a more organised and mineralised layer in the winter (dark increment) and a less organised and mineralised layer in the summer (light-coloured increment) [21]. Contrary to this, more recent studies have now shown through environmental scanning electron microscopy and Raman spectroscopy imaging that light increments have, instead, a higher mineral content, effectively re-opening the debate on the biological mechanisms behind the incremental pattern of AEFC [22–25].

As well as being useful for estimating age and season at death, cementochronology has been shown to be useful for assessing and dating the occurrence of significant life events, such as pregnancies, weaning, skeletal traumas or diseases (especially if related to calcium metabolism) [7–10]. Specifically, disruptions in deposition have been correlated to the observation of considerably wider white increments in pregnancy [10,23], skeletal traumas and renal disorders [10]. In Kagerer and Grupe [10], variations in increment thickness were also observed for the lines of Salter and, in one specific case, were associated with the prolapse of two intervertebral discs followed by injuries to the vertebrae. In another report [8], a number of physiological stressors (not only pregnancy, but also menopause, systemic illness and relocation) were identified on the AEFC by the occurrence of changes in the birefringent properties of cementum. The onset of puberty was also identified using cementochronology in both females and males [8,23]. In these studies, it was reported that variations in sex hormones were important regulators of cementum deposition.

Smoking is known to have a systemic impact on the body’s physiology, leading to a cascade of effects that includes reduction in calcium intestinal absorption [26], alterations in vitamin D metabolism [26] and alterations to the thyroid, parathyroid, adrenal, pituitary glands and sex hormones [27]. Numerous studies [28–31] have also highlighted the correlation between smoking, periodontitis and tooth loss, suggesting that smoking activity could potentially be detected within the AEFC. However, to the authors’ knowledge, no investigation has previously examined whether tobacco use can be predicted from analysis of AEFC. Since smoking can directly affect bone metabolism and oral health, the aim of this study was to assess whether tobacco consumption alters the dental cementum and its incremental growth. The hypothesis was that intense smoking activity might affect cementum increment formation and that periods of tobacco usage could be dated with cementochronology.

To test this, we analysed teeth from modern individuals with known smoking histories, and archaeological samples where it was possible to assess smoking status. As tobacco has one of the longest global histories of consumption as an intoxicant, the inclusion of archaeological smokers allowed us to explore how different ways and types of tobacco consumption might affect the AEFC in comparison to modern cigarettes. The pattern of cemental deposition of smokers and ex-smokers was also visually and statistically compared to that of non-smokers in order to identify irregularities that were potentially connected to smoking activity. Similar to the abovementioned studies addressing pathologies and life-history reconstruction, the cemental annulation of smokers and ex-smokers was expected to present some form of variation or disruption in the thickness and/or regularity of the increments.

The interest in understanding the impact of smoking on the AEFC is manifold, as it has clear application to the fields of dentistry, forensics and archaeology respectively by a) addressing potential factors altering the biology of the tissue and contributing to the currently ongoing research on periodontal regeneration; b) highlighting potential limitations in the application of cementochronology for age estimation and by providing a further identification tool when using dental cementum for life history reconstruction; and c) by providing archaeological and historical research with bio-anthropological evidence on the spread of tobacco in a given population.

Materials & methods

A sample consisting of 88 human teeth (70 modern and 18 archaeological), representing a total of 64 individuals (46 modern and 18 archaeological), was selected for the analysis. Additional samples were collected for this study, but later excluded. Reasons for exclusion included dental roots being too pathologically compromised or damaged, and breakages occurring during sample preparation.

The modern subgroup was collected from living donors undergoing necessary dental treatment involving tooth extraction. Donors were all adult individuals (age range: 19–86 years), who were informed of the aims and objectives of the study and, if willing to participate, gave full informed consent on the use of their samples. Donors were asked to complete a questionnaire regarding their personal data (such as age, date of tooth extraction, medical and smoking record). The teeth donated for this study were collected with full informed consent from adult donors both at the University of Eastern Piedmont (UPO, Italy) and at Northumbria University (UK), where the study was conducted. Ethical approval to conduct this study was granted by the Northumbria University Research Ethics Committee on April 1^st, 2021 (Ethical Ref. 29546). Sample collection was from April 2^nd, 2021–31^st March, 2022.

The total cohort (comprising both modern and archaeological samples) consisted of non-smokers (n = 33), ex-smokers (n = 17), smokers (n = 25), and for archaeological samples, included the additional categories potential smokers (n = 5), and indeterminate smoking status (n = 8) (Table S1). Through the questionnaire collected for the modern cohort, donors could identify themselves as non-smokers (i.e., never smoked); ex-smokers (i.e., used to smoke but had quit at the time of assessment); and smokers (i.e., regularly smoking at the time of the assessment). The categories “potential smokers” and “indeterminate smoking status” were introduced to account for the uncertainty bound to archaeological specimens, whose smoking status relied on the interpretation of archaeological clues, such as dark dental staining and presence of pipe wear notches (alterations which are known to be associated with tobacco consumption (Fig 1), [33]). In this study, archaeological samples with clear evidence of pipe notches and stains were classified as “smokers”; samples with evidence potentially connected to smoking (e.g., weak dental stains; small enamel alterations that could have been the onset of a pipe notch) were classified as “maybe” (i.e., potential smokers); samples with unclear smoking evidence (e.g., buried with a smoking pipe but no sign of smoking was identified on their dentition) were classified as “indet” (i.e., indeterminate smoking status); samples with no evidence were classified as “non-smokers”.

Fig 1 — Left: Example of pipe notch (arrowed) [SK134130, St James’ Gardens Burial Ground, Euston, London (image reprinted from [33] under a CC BY license, with permission from Manchester University Press, original copyright 2024]. Right: Example of staining due to smoking [SK417, Holy Trinity Church, Coventry (image reprinted from [33], under a CC BY license, with permission from Manchester University Press, original copyright 2024)].

The archaeological samples were donated from the School of Archaeology and Ancient History at the University of Leicester (UK). These were sampled from skeletal remains excavated from Holy Trinity Church graveyard (on the site of St. Mary’s Cathedral Church (site code: SMC99), Coventry, UK [32]), now preserved at the School of Archaeology and Ancient History, University of Leicester.

No permits were required for the analyses of these remains for the described study, which complied with all relevant regulations. These samples belonged to individuals dating to 1776–1890 and consisted of adults (n = 16) and sub-adults (n = 2) (age range: 2–65 years), for whom thirteen had recorded biological sex, age and date of death on their respective coffin plates. Five, however, did not have this information recorded so age was estimated to be between 35 and 50 years of age through observation of the pelvic region [18] by an experienced osteoarchaeologist.

In accordance with guidance for destructive analysis, each tooth was imaged digitally prior to sampling [34]. For the archaeological sample, teeth were re-imaged after the removal of dental calculus. Dental roots were embedded in a mixture of epoxy resin and hardener (ratio 5:2.5, EpoThin, Buheler©) and cast in cylindrical moulds of solid polytetrafluoroethylene (PTFE), which made the roots more stable and resistant for the sectioning procedure. The PTFE casts were internally covered in a thin layer of soft paraffin to ease the cast out once dried. After removal of the crown, the dental roots were positioned in the casts standing on their flat coronal surface and completely covered with the resin. The casts were then dried in a vacuum desiccator for 15–20 minutes and left to cure at room temperature for 24 hours. As the casts dried, the sample could be processed for sectioning. According to the main protocols for cementochronology [17,35–37], five transverse non-demineralised ground sections of 80 µm were produced from the coronal half of each tooth (where the AEFC is normally found) by using a diamond wire saw (DWS.175, Diamond WireTech©) equipped with a micrometer. The thickness of the sections was set to a minimum of 80 µm to avoid unnecessary breakages during the sectioning procedure. This thickness is consistent with what is recommended for ideal visualisation of cementum annulations [17,35–37].

The same mixture of epoxy resin and hardener was then used to mount the sections on microscope slides which were covered with coverslips. This choice also followed the directions of previously successful studies [12] and the warnings that other mounting media (such as MMA) could hinder the correct visualisation of the increments [8].

Sections were analysed with a transmitted light microscope (Leica DM750©) fitted with an ICC50W camera module that permitted digital imaging of the sections as seen from the microscope. A 530nm retardation plate was also used to investigate the birefringent properties of the tissue. One region of interest (ROI) was selected for clarity and integrity of the AEFC per each section (n = 5) and recorded in a digital image. A total of five digital pictures were therefore recorded per each tooth. Digital images were saved in LAS X software at 20x and 40x magnification, for better visualisation of the cementum’s features. Counts and measurements were carried out in ImageJ software (version Java 1.8.0_172). Increment counting was performed using the multipoint tool, which records the number of the increments while counting (therefore limiting potential mistakes). Measurements of the AEFC’s width were taken at three equidistant points of each digital image. Counts of the increments and width measurements were then averaged by tooth in Excel. Assessment of age by cementochronology was performed according to previously published protocols [17,35–37] by adding the total pairs of the increments to the tooth-specific age at alveolar eruption (based on [38]). The term “eruption” refers more generally to the process of emergence of teeth in a functional (i.e., occlusal) position within the oral cavity [39]. This process often is divided into three steps: alveolar emergence (tooth is over the alveolar bone); clinical emergence (tooth breaks through the gingiva); and functional occlusion (tooth is occluding with its opposing tooth) [40]. Since clinical emergence and functional occlusion could be more subjective to each individual [40], estimation of age through cementochronology was here based on the age at alveolar emergence.

Presence/absence of smoking damage was firstly assessed by visual observations of the sections and then further investigated by statistical analysis. The visual assessment consisted in recording whether structural variations and irregularities in the microstructure of the cementum (e.g., alteration of the pattern; wider increments) were visible and could potentially be related to smoking activity. To further investigate the nature of this alteration, sections presenting damaged areas were also observed with the retardation plate. This allowed the comparison of the regular birefringent properties of the cemental increments with the areas of disruption. Once identified, the areas of smoking damage could also be aged through cementochronology.

Importantly, analyses of the sections were performed blind, without knowledge of smoking status; only during the final phase of the investigation was the smoking status of the donors disclosed.

Statistical analyses

Statistical analysis was performed in R software (version 4.2.3) and visualised using ggplot2 (version 4.2.3). Association between the smoking habits and the smoking damage with the overall AEFC’s width was assessed with Kruskal-Wallis (>2 groups) and Wilcoxon test (2 groups). The level of significance (p value) was set at 0.05. The association between true smoking status and predicted smoking status was tested using Fisher’s Exact test. The accuracy of the predictions was further addressed by calculating the overall accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Since this was a pilot study, no correction for multiple testing was undertaken.

Results

A visual assessment of the AEFC was carried out and compared between smokers, non-smokers and ex-smokers. The AEFC of smokers presented the usual cemental pattern (N = 16; Fig 2A), although it was of lower quality than that of non-smokers (Fig 2B). This manifested as less definition of the increments or, as in this case, in the presence of “dark areas” that spanned regions of the annulations (red arrow in Fig 2A). In contrast, the AEFC of ex-smokers presented strongly disrupted areas of tissue, in which the alternating cemental pattern was interrupted (black arrows in Fig 2C). These areas could be described as granular, structureless and “TV static”-like in appearance.

Fig 2 — Representative examples of acellular cementum (AC) in a 46-year-old smoker (A); in a 35-year-old non-smoker (B); and 58-year-old ex-smoker showing the smoking damage (black arrows and dashed line) (C). Teeth were affected by periodontal diseases (A, C) and one was impacted (B). [D = Dentin; Images taken with transmitted light microscopy at 20x, scale bar: 100µm].

When observed with a retardation plate, areas of smoking damage appeared purple in colour despite orientating the sample in parallel or perpendicular to the axis of the plate (Fig 3). This showed that these areas did not have birefringent properties (unlike the rest of the tissue).

Fig 3 — Examples of the effects of the retardation plate on acellular cementum (AC) when perpendicular to the sample (column A) and parallel to the sample (column B). No birefringence property is shown on the smoking damage (dashed lines). Sample in top row is sample shown in Fig. 2C [D = Dentine; White arrow = orientation of the retardation plate; images taken with transmitted light microscopy at 40x, scale bar: 50µm].

One of the teeth presenting smoking damage was from a donor who reported the exact age at which they started and ceased smoking. Cementochronology was performed to age the extent of the smoking damage observed on the section of this individual (Fig 2C). The total of annuli up to the start of the smoking-dependent damage was counted and added to the tooth-specific age at eruption (based on AlQathani [38]). To estimate the end of smoking-dependent damage, the total number of annuli counted from the external (most recent) side of the AEFC was subtracted from the known age of the individual. By doing so, the smoking damage observed on this individual was estimated to have occurred between age 22 and 41, which is congruent with the information collected by the donor, who reported to have started smoking at 28 years old and stopped at 38 years old (Table 1).

Table 1. Analyses on the smoking damage for prediction of individual’s smoking timeline. Data on sample VP_H_026 (shown in Fig 2C), indicating: type of tooth (FDI); Sex; Smoking status; Real age; Tooth-specific occlusion age; total count of the increments (IL Count); count of the increments up to the damage (smoking damage start) and from external border of the tissue (smoking damage end); prediction of age range at which the smoking damage occurred; and prediction of age of the individual.

Sample	FDI	Sex	Smoking status	Real Age (years)	Occlusion Age (years)	IL Count	Smoking Damage start (IL)	Smoking Damage end (IL)	Age of smoking damage (years)	Predicted Age (years)
VP_H_026	1.3	M	Ex Smoker (from age 28–38)	58	12.5	41.70	10	17	22.5–41	54.20

Open in a new tab

Data on sample VP_H_026 (shown in Fig 2C), indicating: type of tooth (FDI); Sex; Smoking status; Real age; Tooth-specific occlusion age; total count of the increments (IL Count); count of the increments up to the damage (smoking damage start) and from external border of the tissue (smoking damage end); prediction of age range at which the smoking damage occurred; and prediction of age of the individual.

Because the smoking damage was, in this case, close to the chronological time at which the donor reported to be a smoker, it was hypothesised to be caused by smoking activity and therefore, hereafter referred to as “smoking damage”. As a result, each tooth of the cohort was examined for evidence of smoking damage and placed into one of three categories: Yes/ No/ indeterminate (indet). For donors, self-reported smoking status was classified as smokers, non-smokers and ex-smokers. For the archaeological samples, smoking status was estimated as described in the methods. The occurrence of smoking damage and smoking habits was then assessed in the context of chronological age and cementum width (Fig 4).

There was a significant association between presence of smoking damage with smoking habits of the donors (p < 0.001, Fisher’s Exact test; Table 2). Evidence of smoking damage was found in 8/24 (33%) of current smokers and in 7/10 (70%) of ex-smokers; this was rarely (1/32 (3%)) observed in non-smokers..

Table 2. Association between occurrence of smoking damage by smoking habits. Smoking damage is associated with smoking status (p < 0.0001, Fisher’s Exact test).

	Smoking Damage
	No	Yes
Non-Smoker	31	1
Ex Smoker	3	7
Smoker	16	8

Open in a new tab

Smoking damage is associated with smoking status (p < 0.0001, Fisher’s Exact test)

The AEFC was also analysed in terms of overall width in relation to smoking habits of the donors and to the presence/absence of smoking damage. From this comparison, it was observed that in the modern subsample (Fig 5A), ex-smokers presented a thicker total AEFC than non-smokers and smokers (p = 0.05, Kruskal-Wallis). Interestingly, when comparing the overall width with the presence/absence of smoking damage (Fig 5B), it was found that sections presenting smoking damage also showed a thicker AEFC width (p = 0.01, Kruskal-Wallis). There was a significant difference between smokers and ex-smokers (p = 0.008, Wilcoxon test) (Fig 5A), and between sections showing smoking damage and those not showing smoking damage (p = 0.004, Wilcoxon test) (Fig 5B). These differences were not as easily identifiable in the archaeological subsample (Fig 5C-D), and were not significant, possibly because of the smaller sample size (n = 18), and/or the number of teeth (n = 13) with indeterminate (i.e., indet) or potential smoking evidence (i.e., maybe). However, also in the archaeological subsample, sections presenting smoking damage were on average thicker than sections with no smoking damage (p = 0.02, Kruskal-Wallis; p = 0.006, Wilcoxon test; Fig 5D).

Fig 5 — Boxplots show AEFC width grouped by smoking habits for modern samples (A) and for archaeological samples (C). The relationship between presence/absence of smoking damage and AEFC width is shown for modern samples (B) and archaeological samples (D); further details in Table S2.

Overall, in the modern cohort, smoking damage was observed mostly on sections belonging to ex-smokers, rather than current smokers (Fig 6).

Finally, the power of prediction of smoking status through the observed presence of smoking damage was also calculated (Table 3). This showed that if smoking damage is observed, there is an 85–92% chance that the individual was either a smoker or ex-smoker (PPV); and that if smoking damage is not observed, there is a 96% chance that the individual was a non-smoker (specificity).

Table 3. Prediction of smoking habits. Calculations of the power of prediction of smoking status based on the observation of smoking damage, in terms of overall accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV).

Predicting for:	Overall accuracy	Sensitivity	Specificity	PPV	NPV
All (Ex-Smokers & Smokers)	0.42	0.42	0.96	0.92	0.61
Smokers	0.35	0.28	0.96	0.85	0.65
Ex-Smokers	0.36	0.7	0.97	0.87	0.90

Open in a new tab

Calculations of the power of prediction of smoking status based on the observation of smoking damage, in terms of overall accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV).

A closer look into the archaeological subsample showed that, along with the observed pipe notches and dental staining associated with smoking, observation of smoking damage on the cementum could provide further evidence on the smoking habit of the individual (Table 4).

Table 4. Evidence of smoking in archaeological samples. Comparison between the assessment of smoking status through archaeological examination and by presence of smoking damage on the AEFC. Indeterminate = “indet”.

Sample ID	Repository Number	Sex	Smoking
Sample ID	Repository Number	Sex	Archaeological Record	Cementum
VP_ARC_001	SMC 99_SK978	M	indet	no
VP_ARC_002	SMC 99_SK978	M	indet	no
VP_ARC_003	SMC 99_SK417	F	possible staining	no
VP_ARC_004	SMC 99_SK672	F	indet	indet
VP_ARC_005	SMC 99_SK50	F	indet	no
VP_ARC_006	SMC 99_SK516	F	staining	no
VP_ARC_007	SMC 99_SK1248	M	indet	no
VP_ARC_008	SMC 99_SK1041	M	indet	no
VP_ARC_009	SMC 99_SK808	M	indet	yes
VP_ARC_010	SMC 99_SK1195	F	indet	no
VP_ARC_011	SMC 99_SK1522	M	staining	no
VP_ARC_013	SMC 99_SK144	M	indet	no
VP_ARC_014	SMC 99_SK32	NA	no	no
VP_ARC_015	SMC 99_SK836	M	staining	yes
VP_ARC_016	SMC 99_SK1089	M	staining	yes
VP_ARC_017	SMC 99_SK400	M	pipe notch	no
VP_ARC_018	SMC 99_SK1225	F	wear & staining	yes
VP_ARC_020	SMC 99_SK85	M	pipe notch & staining	yes

Open in a new tab

Comparison between the assessment of smoking status through archaeological examination and by presence of smoking damage on the AEFC. Indeterminate = “indet”.

Fig 7 shows three archaeological samples in which smoking damage was most evident. Cementochronology was applied to attempt an estimation of the age at which smoking damage first occurred, resulting in age: 25.5 years old in sample ARC_009 (Fig 7A); 18 years old in sample ARC_015 (Fig 7B) and 24.5 years old in sample ARC_018 (Fig 7C).

Discussion

The incremental non-remodelling nature of the AEFC allows for detection of illnesses and stressful events that, through cementochronology, can also be aged. Previously, events involving alterations of sex hormones and calcium metabolism have been positively identified on the AEFC [7,8,10,23]. In this study, the AEFC was investigated to determine whether smoking status could also be detected using the tissue, since it is known not only to carry a deeply disruptive action on the physiology of the body by altering its metabolism and hormone regulation, but also to have a direct effect on oral health.

The results of this preliminary investigation showed a significant association between smoking activity and the regular deposition of the AEFC, which seemed to be affected by smoking with different degrees of severity. These effects could be summarised overall with a) variations in the width of the tissue and b) presence of disrupted areas in which the typical annulation pattern is not present (herein referred to as “smoking damage”). Interestingly, smoking damage was more often observed in ex-smokers, who also showed a thicker AEFC than non-smokers and smokers. Smoking damage appears as isolated areas of ground substance with an absence of increments. As the AEFC is known to be avascular and acellular, its non-remodelling nature could also explain a) why smoking damage is more often found in ex-smokers (i.e., it becomes redefined by new incremental apposition once individuals stop smoking); and b) why the AEFC in ex-smokers is thicker than in other groups (smoking damage is not reabsorbed by the tissue, which instead resumes its regular deposition on top of it, resulting in a thicker cementum). These, however, are currently hypotheses that will need to be further investigated in future studies.

While the variations of the AEFC could be positively associated with the smoking status of modern donors, that association was not significant in the archaeological cohort (Fig 5C). This could be because the smoking status of these individuals was not certain but inferred, which led to most of the archaeological subsample having an indeterminate status (i.e., “indet”; “maybe”, n = 9). However, smoking damage observed in the AEFC of the archaeological samples was consistent with modern samples – smoking damage was significantly associated with a thicker cementum in both subgroups (Fig 5B–5D). When analysed with the retardation plate, smoking damage did not show any birefringent property, unlike the rest of the incremental pattern of the tissue, confirming that the increments do not form in these areas, and that they resume their regular pattern (along with their birefringent properties) once an individual quits smoking.

Unfortunately, the lack of consensus on the biology of the increments does not currently allow for a deeper understanding of what is being altered by smoking. According to Lieberman [41], for example, the two primary causes of the alternating pattern in cementum are variations in degree of mineralisation and in collagen orientation. While birefringence in cementum is normally associated with collagen orientation [41], Cool and colleagues [42] claimed, instead, that its birefringence and annulation are more closely related to the size and orientation of its crystals. More recent studies [22–25] found that the alternating pattern is indeed, as said by Lieberman [41], caused by both factors. In fact, they found that there is a difference in the degree of mineralisation, for which the bright increments show higher amounts of minerals (such as calcium, strontium, phosphorus, and zinc), but that the alternating increments also reflect peaks and drops in carbonate (hydroxy)apatite (cAp) as well as differences in collagen orientation.

The structureless and non-birefringent areas here associated with smoking suggest that smoking affects the AEFC both at its mineral and collagen level. Similar damage was also reported in two studies, where it was described as an area of high X-ray intensity, with no radial structure in the AEFC [43] and as a hypo-mineralised area of “amorphous appearance” in the cellular cementum [44]. Both studies looked at the effects of chronic inflammation of the PDL in acellular and cellular cementum and, in both cases, the damage observed was associated with the occurrence of severe periodontal diseases. Since a strong association between smoking and periodontal diseases [28–30,45,46] has been found, this would suggest that the occurrence of this specific area of damage could be directly caused by the development of periodontal diseases following intense smoking activity.

Amongst the localised side effects of smoking, there is, in fact, severe periodontitis, which is a major cause of tooth loss. Since periodontitis is a chronic inflammation of the PDL, it would frame smoking damage as more closely associated with the inflammatory-dependent damage already described by Yamamoto et al. and Simon et al. [43,44]. The AEFC is in fact, as the name suggests, acellular, and its growth is dependent on a layer of cells (cementoblasts) that lie on its external surface and are supported by the vascularised PDL [47]. Factors impacting the PDL would therefore impact the regular deposition of the cementum. Another study [48] also identified that estimates of age from teeth with untreated periodontitis and several degrees of alveolysis (resorption of the alveolar bone) were considerably underestimated with cementochronology, which is consistent with the idea that periodontitis, whether smoking-dependent or not, strongly affects cementum deposition.

The fact that the smoking damage also appeared on archaeological specimens with pipe notches and dental staining is interesting because, while the presence of smoking damage suggests that these individuals (similarly to modern samples showing smoking damage) were ex-smokers rather than smokers, pipe notches and staining would tend to indicate a regular and persistent smoking activity (requiring about 4 years of regular pipe-smoking to leave a well-defined pipe notch, according to historical sources [49]). However, as shown in Fig 6, smoking damage is not always present in all ex-smokers and often its presence is not clear. This could indicate that the occurrence of smoking damage might be influenced by the quantity and ways (i.e., smoking; snuff; chewing; etc.) in which tobacco was consumed rather than its frequency. Additionally, the presence of pipe notches on some of the archaeological specimens does not necessarily indicate a daily consumption and/or high quantities of tobacco but could have been caused by the use of a kind of clay pipe stem that was particularly abrasive [33]. To further complicate the matter, it is also unclear how pollution might have affected the evidence collected from the archaeological sample. In fact, recent studies regarding heavily polluted areas of South Korea and China [50,51] highlighted the relationship between air pollutants and the development of periodontitis. In particular, it was observed that the inhalation or ingestion of particulate matter (PM) (i.e., ash and smoke) triggered an inflammatory response at both systemic (i.e., metabolic, respiratory, cardiovascular diseases) and local levels (i.e., oral tissues) [50]. Presence of heavy metals (such as iron, manganese and copper) in drinking water was also recently associated with the occurrence of periodontitis [52]. Since our archaeological samples belonged to individuals that lived during the Industrial Revolution (circa 1760–1850 [53]), it cannot be excluded that the behaviour of AEFC in relation to the presence of the smoking damage may have been further modulated by environmental pollution. The industrial revolution in England was a period characterised by high levels of pollution both in the air and drinking water, especially connected to the waste products of the growing industries related to coal and textiles [53].

Nonetheless, the simultaneous presence of dental staining, pipe notches and smoking damages on some of these samples suggests that individuals in 18^th and 19^th century Coventry (UK) were regularly smoking, but with varying quantities of tobacco. In some cases, more than one patch of smoking damage was observed at different points in the cementum (as in Fig 7B). Similarly, this could indicate periods of more and less intense smoking activity, potentially related to tobacco availability or their personal habits and methods of tobacco consumption. The principles of cementochronology were applied to age the smoking damage (similar to other studies for reconstruction of life-history events). Forensically, this provides a further aid to the identification of human remains, while archaeologically it gives a clue into the trends and habits of tobacco consumption in a historical context. In this study, three archaeological individuals were estimated to have started consuming tobacco between the age of 18 and 25 years old. This would be consistent with the findings observed in a study of 234 individuals from three different locations of 1700-1800s England (one of which, Coventry, UK was the same population of our archaeological sample) that, from the analysis of dental staining and pipe notches, suggested that individuals were already smoking during (and even before) their adolescence [33].

The analysis of pathological samples extracted from individuals that were not only smokers but also had other underlying tooth pathology might have limited our understanding of the impact of smoking on the AEFC. Similarly, the unknowns concerning the urban environment in which the archaeological populations lived as well as the ways and frequency of their tobacco consumption, also represent another limitation. Despite these limitations, the preliminary findings of this study offer a foundation for future studies into the biology of the AEFC and the application of cementochronology for age estimation and life history reconstruction. Future studies should aim to a) understand the biological alteration of the smoking damage to further explore the biology of the increments; b) clarify how and when the smoking damage occurred, by accurately recording the number of cigarettes smoked and investigating other ways of smoking (e.g., vaping); c) study the smoking trends of past populations by selecting a larger cohort presenting archaeological evidence of smoking and the comparison with evidence from historical records regarding type of tobacco consumed, trade routes and rates of consumption.

Conclusions

Overall, this investigation into the effects of smoking on the dental cementum shows how the time-keeping and non-remodelling nature of the AEFC can be useful when reconstructing life-history events of past and modern individuals, adding to personal lifestyles and to population trends. Smoking habits were here investigated and analysed with cementochronology to investigate whether teeth from smokers and ex-smokers could be distinguished from never smokers. Further questions were raised about the occurrence of the smoking damage: how the smoking damage changes according to quantity of tobacco consumed, and type of smoking (e.g., vaping; snuff), and whether similar damage occurs in non-smokers that are affected by severe periodontitis. Future research could address these questions in modern populations, while, for archaeological specimens, a wider comparison between individuals living in rural and urban areas should be carried out, to better account for the potential effects of pollution on the AEFC.

This study also highlighted the level of physiological disruption that smoking causes to the body, and more specifically to the dental cementum. The smoking damage is not commonly addressed in the current literature and was only described in two studies published in the 1960s and 1980s that did not associate it with smoking but with high levels of inflammation of the PDL [43,44]. The occurrence of a similar kind of damage in ex-smokers would point at the strict interconnection between cemental increments and PDL, adding to the ongoing research directed at understanding the biology of AEFC increments.

Supporting information

Table S1. Study dataset.

Complete data and metadata of modern and archaeological samples. The table summarizes samples metadata regarding the time period (archaeological or modern); tooth type (following FDI identification system); dental pathologies; smoking habits; and real age of the samples (Real_Age and Real_Age_1 – the latter has been added for the archaeological samples, whose age was estimated and whose age range was averaged for the purpose of this study). The table also shows the outcome of the analyses carried out in this study (occurrence of the smoking damage; the total count of the increments counted (IL_Count); the measurement of cementum width (Width); and prediction of age (Predicted_Age)). “Occlusion_Age” refers to the standardised age at which teeth come into occlusion, according to AlQahtani et al., 2010 [38].

(XLSX)

pone.0323812.s001.xlsx^{(17.6KB, xlsx)}

Table S2. Kruskal-Wallis rank sum test.

Summary of the chi-squared and p-values between cementum width and smoking habits and damage, in overall cohort, modern and archaeological subsamples.

(DOCX)

pone.0323812.s002.docx^{(14.4KB, docx)}

Acknowledgments

The authors would like to thank Dr Sebastian Breitenbach for training on the use of the diamond wire saw, and Mr Philip Donnelly for manually producing the moulds and samples holder fitting the diamond wire saw at the department of Geography and Engineering, Northumbria University (UK). Finally, our thanks go to Dr Ralf Kist at Newcastle Dental School (UK) for their recommendations on this paper.

Data Availability

All relevant data are within the article and its supporting information files.

Funding Statement

SI reports funding from UK Research and Innovation – Future Leaders Fellowships grant MR/T022302/1. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

References

1.Obertová Z, Skrzypek G, Danišík M, Rankenburg K, Cummaudo M, Olivieri L, et al. Stable Isotope Provenance of Unidentified Deceased Migrants-A Pilot Study. Biology (Basel). 2023;12(11):1371. doi: 10.3390/biology12111371 [DOI] [PMC free article] [PubMed] [Google Scholar]
2.Copeland SR, Grimes V, Neveling J. Isotopic evidence for the geographic origin, movement and diet of the Hofmeyr individual. Title of the Book or Collection. 2022. p. 47–68. [Google Scholar]
3.Paul KS, Feezell R, Hughes T, Brook AH. Integrating genealogy and dental variation: contributions to biological anthropology. American Journal of Biological Anthropology. 2022;181(S76):145–79. doi: 10.1002/ajpa.24662 [DOI] [Google Scholar]
4.Naji S, Rendu W, Gourichon L. Dental cementum in anthropology. Cambridge: Cambridge University Press, 2022. [Google Scholar]
5.Garizoain G, Parra RC, Aranda C, Zorba E, Moraitis K, Escalante-Flórez K, et al. Root dentin translucency and age at death estimation in adults using single rooted teeth: Update of the Forensic International Dental Database. Forensic Sci Int. 2023;343:111564. doi: 10.1016/j.forsciint.2023.111564 [DOI] [PubMed] [Google Scholar]
6.El-Bakary AA, Hammad SM, Mohammed F. Dental age estimation in Egyptian children, comparison between two methods. J Forensic Leg Med. 2010;17(7):363–7. doi: 10.1016/j.jflm.2010.05.008 [DOI] [PubMed] [Google Scholar]
7.Cerrito P, Cerrito L, Hu B, Bailey SE, Kalisher R, Bromage TG. Weaning, parturitions and illnesses are recorded in rhesus macaque (Macaca mulatta) dental cementum microstructure. Am J Primatol. 2021;83(3):e23235. doi: 10.1002/ajp.23235 [DOI] [PubMed] [Google Scholar]
8.Cerrito P, Bailey SE, Hu B, Bromage TG. Parturitions, menopause and other physiological stressors are recorded in dental cementum microstructure. Sci Rep. 2020;10(1):5381. doi: 10.1038/s41598-020-62177-7 [DOI] [PMC free article] [PubMed] [Google Scholar]
9.Cerrito P, Nava A, Radovčić D, Borić D, Cerrito L, Basdeo T, et al. Dental cementum virtual histology of Neanderthal teeth from Krapina (Croatia, 130-120 kyr): an informed estimate of age, sex and adult stressors. J R Soc Interface. 2022;19(187):20210820. doi: 10.1098/rsif.2021.0820 [DOI] [PMC free article] [PubMed] [Google Scholar]
10.Kagerer P, Grupe G. Age-at-death diagnosis and determination of life-history parameters by incremental lines in human dental cementum as an identification aid. Forensic Sci Int. 2001;118(1):75–82. doi: 10.1016/s0379-0738(00)00382-0 [DOI] [PubMed] [Google Scholar]
11.Panezic K, Porcic M, Urban P. Stressful times for women - increased physiological stress in Neolithic females detected in tooth cementum. J Archaeol Sci. 2020;122:105217. [Google Scholar]
12.Perrone V, Gocha TP, Randolph-Quinney P, Procopio N. Tooth Cementum Annulation: A Literature Review. Forensic Sciences. 2022;2(3):516–50. doi: 10.3390/forensicsci2030038 [DOI] [Google Scholar]
13.Berkovitz B, Holland G, Moxham B. Oral anatomy, histology & embryology. 5th ed. Elsevier. 2018. [Google Scholar]
14.Wedel VL. Determination of season at death using dental cementum increment analysis. J Forensic Sci. 2007;52(6):1334–7. doi: 10.1111/j.1556-4029.2007.00546.x [DOI] [PubMed] [Google Scholar]
15.Colard T, Bertrand B, Naji S, Delannoy Y, Bécart A. Toward the adoption of cementochronology in forensic context. Int J Legal Med. 2018;132(4):1117–24. doi: 10.1007/s00414-015-1172-8 [DOI] [PubMed] [Google Scholar]
16.Stott GG, Sis RF, Levy BM. Cemental annulation as an age criterion in forensic dentistry. J Dent Res. 1982;61(6):814–7. doi: 10.1177/00220345820610063401 [DOI] [PubMed] [Google Scholar]
17.Colard T, Bertrand B, Naji S, Delannoy Y, Bécart A. Toward the adoption of cementochronology in forensic context. Int J Legal Med. 2018;132(4):1117–24. doi: 10.1007/s00414-015-1172-8 [DOI] [PubMed] [Google Scholar]
18.Brooks S, Suchey J. Skeletal age determination based on the os pubis: a comparison of the Acsádi-nemeskéri and Suchey-brooks methods. Hum Evol. 1990;5:227–38. [Google Scholar]
19.Iscan M, Loth S, Wright R. Age estimation from the rib by phase analysis: white females. J Forensic Sci. 1985;29:853–63. [PubMed] [Google Scholar]
20.Işcan MY, Loth SR, Wright RK. Age estimation from the rib by phase analysis: white males. J Forensic Sci. 1984;29(4):1094–104. doi: 10.1520/jfs11776j [DOI] [PubMed] [Google Scholar]
21.Klezeval G. Recording structures of mammals: determination of age and reconstruction of life history. Rotterdam: A. A. Balkema. 1996. [Google Scholar]
22.Cabec Al, Garrevoet J, Spiers K. Incremental elemental distribution in chimpanzee cellular cementum: insights from synchrotron x-ray fluorescence and implications for life-history inferences. In: Naji S, Rendu W, Gourichon L, editors. Dental cementum in anthropology. Cambridge University Press. 2022. p. 138–54. [Google Scholar]
23.Colard T, Naji S, Debroucker A. Preliminary protocol to identify parturitions lines in acellular cementum. In: Naji S, Rendu W, Gourichon L, editors. Dental cementum in anthropology. Cambridge University Press. 2022. p. 234–48. [Google Scholar]
24.Naji S, Rendu W, Gourichon L. Recent advances on acellular cementum increments composition using synchrotron x-radiation. Dental cementum in anthropology. Cambridge University Press. 2022. p. 110–37. [Google Scholar]
25.Dean C, Le Cabec A, Spiers K, Zhang Y, Garrevoet J. Incremental distribution of strontium and zinc in great ape and fossil hominin cementum using synchrotron X-ray fluorescence mapping. J R Soc Interface. 2018;15(138):20170626. doi: 10.1098/rsif.2017.0626 [DOI] [PMC free article] [PubMed] [Google Scholar]
26.Cusano NE. Skeletal effects of smoking. Curr Osteoporos Rep. 2015;13:302–9. [DOI] [PubMed] [Google Scholar]
27.Kapoor D, Jones TH. Smoking and hormones in health and endocrine disorders. Eur J Endocrinol. 2005;152(4):491–9. doi: 10.1530/eje.1.01867 [DOI] [PubMed] [Google Scholar]
28.Obeid P, Bercy P. Effects of smoking on periodontal health: a review. Adv Ther. 2000;17(5):230–7. doi: 10.1007/BF02853162 [DOI] [PubMed] [Google Scholar]
29.Leite F, Nascimento G, Scheutz F. Effect of smoking on periodontitis: A systematic review and meta-regression. Am J Prev Med. 2018;54:831–41. [DOI] [PubMed] [Google Scholar]
30.Fiorini T, Musskopf ML, Oppermann RV, Susin C. Is there a positive effect of smoking cessation on periodontal health? A systematic review. J Periodontol. 2014;85(1):83–91. doi: 10.1902/jop.2013.130047 [DOI] [PubMed] [Google Scholar]
31.Bezerra Ferreira JD, Rodrigues JA, Piattelli A, Iezzi G, Gehrke SA, Shibli JA. The effect of cigarette smoking on early osseointegration of dental implants: a prospective controlled study. Clin Oral Implants Res. 2016;27(9):1123–8. doi: 10.1111/clr.12705 [DOI] [PubMed] [Google Scholar]
32.Soden I. Excavations at the cathedral church of St Mary, Coventry. 1999 -2000. Summary report. 2000. [Google Scholar]
33.Davies-Barrett A, Inskip S. Who smokes anymore? documentary, archaeological and osteological evidence for tobacco consumption and its relationship to social identity in industrial England, 1700–1850. In: Craig-Atkins E, Harvey K, editors. The material body. Manchester, England: Manchester University Press. 2024. p. 133–69. [Google Scholar]
34.Mays S, Elders J, Humphrey L, et al. Science and the Dead. Destructive sampling of archaeological human remains for scientific analysis. 2nd ed. Advisory Panel on the Archaeology of Burials in England, 2023. (accessed 12 June 2024). [Google Scholar]
35.Bertrand B, Cunha E, Bécart A, Gosset D, Hédouin V. Age at death estimation by cementochronology: Too precise to be true or too precise to be accurate?. Am J Phys Anthropol. 2019;169(3):464–81. doi: 10.1002/ajpa.23849 [DOI] [PubMed] [Google Scholar]
36.Wittwer-Backofen U, Gampe J, Vaupel JW. Tooth cementum annulation for age estimation: results from a large known-age validation study. Am J Phys Anthropol. 2004;123(2):119–29. doi: 10.1002/ajpa.10303 [DOI] [PubMed] [Google Scholar]
37.Wittwer-Backofen U. Age estimation using tooth cementum annulation. Methods Mol Biol. 2012;915:129–43. doi: 10.1007/978-1-61779-977-8_8 [DOI] [PubMed] [Google Scholar]
38.AlQahtani SJ, Hector MP, Liversidge HM. Brief communication: The London atlas of human tooth development and eruption. Am J Phys Anthropol. 2010;142(3):481–90. doi: 10.1002/ajpa.21258 [DOI] [PubMed] [Google Scholar]
39.Liversidge HM. Tooth Eruption and Timing. In: Irish J.D, Richard Scott G. (eds) A Companion to Dental Anthropology. Chichester, UK: Wiley Blackwell, John Wiley & Sons, Inc., pp. 159–71. [Google Scholar]
40.Harris E. Odontogenesis. A companion to dental anthropology. Hoboken NJ: Wiley. 2015. p. 142–58. [Google Scholar]
41.Lieberman DE. The Biological Basis for Seasonal Increments in Dental Cementum and Their Application to Archaeological Research. Journal of Archaeological Science. 1994;21(4):525–39. doi: 10.1006/jasc.1994.1052 [DOI] [Google Scholar]
42.Cool SM, Forwood MR, Campbell P, Bennett MB. Comparisons between bone and cementum compositions and the possible basis for their layered appearances. Bone. 2002;30(2):386–92. doi: 10.1016/s8756-3282(01)00686-x [DOI] [PubMed] [Google Scholar]
43.Yamamoto H, Sugahara N, Yamada N. Histopathological and microradiographic study of the exposed cementum from periodontally diseased human teeth. Bull Tokyo Med Dent Univ. 1966;13(3):407–21. [PubMed] [Google Scholar]
44.Simon JH, Yonemoto GS, Bakland LK. Comparison of cellular cementum in normal and diseased teeth - a scanning electron microscopic study. J Endod. 1981;7(8):370–5. doi: 10.1016/S0099-2399(81)80058-1 [DOI] [PubMed] [Google Scholar]
45.Duarte PM, Nogueira CFP, Silva SM. Impact of smoking cessation on periodontal tissues. Int Dent J. 2022;72:31–6. [DOI] [PMC free article] [PubMed] [Google Scholar]
46.Souto MLS, Rovai ES, Villar CC, Braga MM, Pannuti CM. Effect of smoking cessation on tooth loss: a systematic review with meta-analysis. BMC Oral Health. 2019;19(1):245. doi: 10.1186/s12903-019-0930-2 [DOI] [PMC free article] [PubMed] [Google Scholar]
47.Nanci A, Bosshardt DD. Structure of periodontal tissues in health and disease. Periodontol 2000. 2006;40:11–28. doi: 10.1111/j.1600-0757.2005.00141.x [DOI] [PubMed] [Google Scholar]
48.Broucker A de, Colard T, Penel G, Blondiaux J, Naji S. The impact of periodontal disease on cementochronology age estimation. Int J Paleopathol. 2016;15:128–33. doi: 10.1016/j.ijpp.2015.09.004 [DOI] [PubMed] [Google Scholar]
49.Effects of bagpipe which mentions tobacco pipe damage to teeth. Lancet. 1890;135:1438. [Google Scholar]
50.Marruganti C, Shin H-S, Sim S-J, Grandini S, Laforí A, Romandini M. Air Pollution as a Risk Indicator for Periodontitis. Biomedicines. 2023;11(2):443. doi: 10.3390/biomedicines11020443 [DOI] [PMC free article] [PubMed] [Google Scholar]
51.Huang K, Feng L-F, Liu Z-Y, Li Z-H, Mao Y-C, Wang X-Q, et al. The modification of meteorological factors on the relationship between air pollution and periodontal diseases: an exploration based on different interaction strategies. Environ Geochem Health. 2023;45(11):8187–202. doi: 10.1007/s10653-023-01705-6 [DOI] [PubMed] [Google Scholar]
52.Zhou S, Li W, Wan J, Fu Y, Lu H, Li N, et al. Heavy metals in drinking water and periodontitis: evidence from the national oral health survey from China. BMC Public Health. 2023;23(1):1706. doi: 10.1186/s12889-023-16391-3 [DOI] [PMC free article] [PubMed] [Google Scholar]
53.McGuire SA. Products of Industry: Pollution, Health, and England’s Industrial Revolution. 2020, pp. 203–231.

PLoS One. 2025 May 27;20(5):e0323812. doi: 10.1371/journal.pone.0323812.r001

Author response to Decision Letter 0

23 Jan 2025

PLoS One. doi: 10.1371/journal.pone.0323812.r002

Decision Letter 0

Francisco de Paula-Silva

25 Feb 2025

PONE-D-25-00796Reconstructing smoking history through dental cementum analysis - a preliminary investigation on modern and archaeological teeth.PLOS ONE

Dear Dr. Schwalbe,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by Apr 11 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Francisco Wanderley Garcia de Paula-Silva, DDS, MSc, PhD

Academic Editor

PLOS ONE

Journal requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf .

2. In your manuscript, please provide additional information regarding the specimens used in your study. Ensure that you have reported human remain specimen numbers and complete repository information, including museum name and geographic location.

If permits were required, please ensure that you have provided details for all permits that were obtained, including the full name of the issuing authority, and add the following statement:

'All necessary permits were obtained for the described study, which complied with all relevant regulations.'

If no permits were required, please include the following statement:

'No permits were required for the described study, which complied with all relevant regulations.'

For more information on PLOS ONE's requirements for paleontology and archeology research, see https://journals.plos.org/plosone/s/submission-guidelines#loc-paleontology-and-archaeology-research .

3. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match.

When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section.

4. Thank you for stating the following financial disclosure:

[Dr. Inskip reports funding from UK Research and Innovation – Future Leaders Fellowships grant MR/T022302/1].

Please state what role the funders took in the study. If the funders had no role, please state: ""The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.""

If this statement is not correct you must amend it as needed.

Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf.

5. We are unable to open your Figure file [Fig_1.eps to Fig_7.eps]. Please kindly revise as necessary and re-upload.

6. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

**********

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

**********

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

**********

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

**********

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Reconstructing the life-history of people who lived in the past is one of the most exciting fields in bioarchaeology. Often, such reconstructions are made using only paleoanthropological data, such as estimating smoking prevalence based on tooth wear and dental plague studies, supplemented by historical records. This study is notable not only because it uses a new method for estimating smoking effects on an organism by analyzing increments of dental cementum. It also allows for a comparison of the use of this method in different populations: a modern one with a known human smoking history and an archaeological one, where only indirect evidence is used.

The article is written in fluent language. The problem and the relevance of the research are well stated. The methodology is clearly described, and appropriate statistical methods are applied. The results are presented clearly, and the tables and figures are informative. The discussion covers not only possible interpretations of the results but also the limitations of this study and future directions.

There are only a few clarifying questions that need to be addressed:

1. How was it established that the damages found in the study were related precisely to the effects of smoking? The authors cite the coincidence between the actual smoking period reported by one of the respondents and the time at which the cementum lesions developed. Is this coincidence the only basis for the argument?

2. The authors discuss the possibility that other causes, such as air pollution and metabolic or infectious diseases, could have contributed to the damages found in the archaeological material. Identifying the specific cause of damage in skeletal material in archaeological populations is usually impossible. However, have the links between smoking and comorbidities or injuries in the modern population not been investigated? An analysis of these associations would help better interpret the results obtained. Moreover, the authors provide a questionnaire in Supplements with questions on malnutrition or trauma.

3. The questionnaire form provided in the Supplements implies that extensive additional information was collected, but only data on smoking habits were used in the survey. Table S1 also includes information on dental diseases, but these are barely addressed in the text, except to mention the link between smoking and periodontitis. Are there plans to use the other information collected, and how?

However, the above comments do not detract from the value of the paper. The paper is certainly worth publishing.

Reviewer #2: The study provided a preliminary investigation using a sound methodology that has the potential to be applied to other future studies. The statistic analysis was rigorous and all data underlying the findings described in their manuscript were fully available. The language used was appropriate for the audience and the topic. A few structural modifications are suggested to improve the overall presentation and some of the interpretations given to the results.

Reviewer #2:

The paper represents a novel approach to the study of smoking status in past populations with the use of cementochronology. The authors provided an in-depth literature review on the topic including the answers that AEFC can provide to specific osteoarchaeological questions. The study provided a preliminary investigation using a sound methodology that has the potential to be applied to other future studies. The Discussion provided a good interpretation of the results. It also addressed the issues found with the archaeological sample and suggested areas of future potential development. Overall, the paper represents a great contribution to the field although some aspects of the Materials-Methods and Results could be explained and/or presented better.

-The Materials and Methods defined the archaeological samples in Lines 121-122 (Smokers, Potential smokers, Indet, Non-smoker) but the modern samples were defined in the Results (smokers, non-smokers and ex-smokers). I have to admit that I had to read these sections more than once to understand what sample was being referred to and how. This is the reason I believe the Materials and Methods – and Results would be better presented had these terms been explained in one place (the methods). If the donors' self-reported status is listed/defined in the methods, then the results section could present the data according to the cementochronology.

-Also, the term ‘smoking damage’ is defined in Line 238 but sometimes ‘damage’ was used interchangeably to mean ‘smoking damage’. I’d recommend sticking to this term instead of just calling it ‘damage’ because the word ‘damage’ was used in other sections of the paper to mean different things. So, to not create confusion I’d recommend you to state the full word ‘smoking damage’ to make a clear distinction from other meanings.

Discussion:

-The discussion mentions that recent studies in South Korea and China highlighted the relationship between air pollutants and the development of periodontitis. The assumption that your archaeological samples belonged to individuals who lived during the Industrial Revolution implied that they would have been exposed to environmental pollution. Now, specifically to your archaeological sample from industrial Coventry, what kind of potential pollution would have been the individuals exposed to that could justify the behaviour of AEFC in relation to the presence of smoking damage?

Other minor comments:

-Figure legend for figure 1 needs to say what context number/skeleton number it is

-Line 244: although technically you can start a sentence with a number ‘8/24’ it doesn’t look appropriate for a formal paper to do it.

**********

6. PLOS authors have the option to publish the peer review history of their article (what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy .

Reviewer #1: No

Reviewer #2: No

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step.

Attachment

Submitted filename: Feedback.docx

pone.0323812.s003.docx^{(15.7KB, docx)}

PLoS One. 2025 May 27;20(5):e0323812. doi: 10.1371/journal.pone.0323812.r003

Author response to Decision Letter 1

1 Apr 2025

Please see attached point-by-point response to reviewers and to the Academic editor. [Response to Reviewers.docx]

Reviewer 1

Reconstructing the life-history of people who lived in the past is one of the most exciting fields in bioarchaeology. Often, such reconstructions are made using only paleoanthropological data, such as estimating smoking prevalence based on tooth wear and dental plague studies, supplemented by historical records. This study is notable not only because it uses a new method for estimating smoking effects on an organism by analyzing increments of dental cementum. It also allows for a comparison of the use of this method in different populations: a modern one with a known human smoking history and an archaeological one, where only indirect evidence is used.

We thank the reviewer for their positive feedback and for their support on this study. We are very glad that they have appreciated the value of this work and its contribution to the field of bioarchaeology.

We thank the reviewer for raising this point. This coincidence is not the only basis for the argument, but rather is what drew our attention to the occurrence of smoking damage in smokers and ex-smokers. Through this more in-depth investigation, we demonstrate a significant association between consumption of tobacco and presence of damage in the wider cohort. This was also supported by the fact that the presence of the damage was also highly predictive of the smoking habit of the individuals.

Unfortunately, information on the precise time period between commencing smoking and its cessation was only available for one donor. In follow up studies, we are currently working on a new cohort of modern samples, where smoking timelines are more clearly defined.

We thank the reviewer for highlighting such an important point and agree that a better insight into modern comorbidities and injuries would help elucidate the complex interplay of factors that may engender damage to the AEFC.

This is why the questionnaire included questions related to presence of pathologies and conditions affecting the metabolism of vitamin C and D, for example. The section of the questionnaire regarding presence of pathologies, smoking, etc. was left at the donor’s discretion (see “Part 2- Optional”). As a result, not all donors completed the second part of the questionnaire, leaving us with data that was not adequately powered to make statistical inferences. As mentioned above, we are now working towards rectifying the gaps we identified during this pilot study in future works.

We thank the reviewer for this question. Please refer to the answer for A.2 which discusses the limited nature of additional information.

While the link between periodontal diseases and cementum alteration was more clearly noticeable (a point already addressed and highlighted in the literature on cementochronology - as in Broucker, Colard, Penel et al., 2016 - https://doi.org/10.1016/j.ijpp.2015.09.004), once again, the small number of samples with specific dental pathologies (i.e. caries and impaction) did not allow statistical analysis. The information collected on these other dental pathologies were therefore excluded from this pilot, but will be integrated in following studies.

However, the above comments do not detract from the value of the paper. The paper is certainly worth publishing.

We thank the reviewer once more for their support in our work, and for giving us the opportunity to further clarify the rationale behind our study.

Reviewer 2

The study provided a preliminary investigation using a sound methodology that has the potential to be applied to other future studies. The statistic analysis was rigorous and all data underlying the findings described in their manuscript were fully available. The language used was appropriate for the audience and the topic. A few structural modifications are suggested to improve the overall presentation and some of the interpretations given to the results.

We thank the reviewer for their positive feedback on our manuscript and welcome the suggested improvements.

1. The Materials and Methods defined the archaeological samples in Lines 121-122 (Smokers, Potential smokers, Indet, Non-smoker) but the modern samples were defined in the Results (smokers, non-smokers and ex-smokers). I have to admit that I had to read these sections more than once to understand what sample was being referred to and how. This is the reason I believe the Materials and Methods – and Results would be better presented had these terms been explained in one place (the methods). If the donors' self-reported status is listed/defined in the methods, then the results section could present the data according to the cementochronology.

We thank you the reviewer and agree that the definitions could benefit from a more unified definition with greater clarity. We have provided further definition on the terms in the Methods section - please find our changes in lines 131-146.

“The total cohort (comprising both modern and archaeological samples) consisted of non-smokers (n=33), ex-smokers (n=17), smokers (n=25), and for archaeological samples, included the additional categories potential smokers (n=5), and indeterminate smoking status (n=8) (Table S1). Through the questionnaire collected for the modern cohort, donors could identify themselves as non-smokers (i.e. never smoked); ex-smokers (i.e. used to smoke but had quit at the time of assessment); and smokers (i.e., regularly smoking at the time of the assessment). The categories “potential smokers” and “indeterminate smoking status” were introduced to account for the uncertainty bound to archaeological specimens, whose smoking status relied on the interpretation of archaeological clues, such as dark dental staining and presence of pipe wear notches (alterations which are known to be associated with tobacco consumption (Fig 1), [33]). In this study, archaeological samples with clear evidence of pipe notches and stains were classified as “smokers”; samples with evidence potentially connected to smoking (e.g., weak dental stains; small enamel alterations that could have been the onset of a pipe notch) were classified as “maybe” (i.e., potential smokers); samples with unclear smoking evidence (e.g., buried with a smoking pipe but no sign of smoking was identified on their dentition) were classified as “indet” (i.e., indeterminate smoking status); samples with no evidence were classified as “non-smokers”.”

We hope this is now more clearly presented and satisfies the reviewer’s recommendations.

2. Also, the term ‘smoking damage’ is defined in Line 238 but sometimes ‘damage’ was used interchangeably to mean ‘smoking damage’. I’d recommend sticking to this term instead of just calling it ‘damage’ because the word ‘damage’ was used in other sections of the paper to mean different things. So, to not create confusion I’d recommend you to state the full word ‘smoking damage’ to make a clear distinction from other meanings.

We thank you the reviewer for their suggestions. We have apported the recommended change throughout the manuscript (please see the revised manuscript with tracked changes).

3. The discussion mentions that recent studies in South Korea and China highlighted the relationship between air pollutants and the development of periodontitis. The assumption that your archaeological samples belonged to individuals who lived during the Industrial Revolution implied that they would have been exposed to environmental pollution. Now, specifically to your archaeological sample from industrial Coventry, what kind of potential pollution would have been the individuals exposed to that could justify the behaviour of AEFC in relation to the presence of smoking damage?

We thank the reviewer for raising a such an interesting point. We have hypothesised that the occurrence/visibility of the damage might be accentuated by the quantity of tobacco consumed by the individual. This hypothesis is being investigated in a new study we are currently finalising, which has demonstrated that heavy smokers are more likely to present damage compared to medium or occasional smokers.

Following this logic, we hypothesized that a greater exposure to environmental pollution might have worsened the dental cementum health in archaeological individuals (in the discussion, we have addressed this in lines 404-409). From historical sources, we know that, during the Industrial Revolution, individuals were exposed daily to pollutants in the water they were drinking and in the air they were breathing both outside (e.g., factories) and inside their houses (i.e., open fires for cooking/heating), smog produced by factories, of which there were many in Coventry. These would have included pollutants such as lead and carbon monoxide, that are also released and inhaled while smoking. This might have had an impact on the dental cementum, and in what we have observed in this study. Environmental pollution would have affected all individuals, regardless of their smoking habits. However, exposure to environmental pollution only would have probably not been enough to cause the smoking damage; whereas it could have added to the ongoing oral inflammation that smokers were subjected to due to their smoking habit.

However, since this was a pilot, we did not have enough data to investigate whether this is indeed the case, therefore, we have mentioned it as a hypothesis. In future work, we are seeking to address this point by comparing archaeological and modern smokers living in urban areas and rural areas.

4. Figure legend for figure 1 needs to say what context number/skeleton number it is

We thank the reviewer for bringing this to our attention. Please find the recommended change at line 206-209.

“Fig 1. Archaeological evidence of tobacco consumption. Left: Example of pipe notch (arrowed) [SK134130, St James’ Gardens Burial Ground, Euston, London (image reprinted from [33])]. Right: Example of staining due to smoking [SK417, Holy Trinity Church, Coventry (image reprinted from [33])].”

5. Line 244: although technically you can start a sentence with a number ‘8/24’ it doesn’t look appropriate for a formal paper to do it

We thank the reviewer for bringing this to our attention. Please find the recommended change at lines 257-259.

“Evidence of smoking damage was found in 8/24 (33%) of current smokers and in 7/10 (70%) of ex-smokers; this was rarely (1/32 (3%)) observed in non-smokers.”

Attachment

Submitted filename: Response to reviewers.docx

pone.0323812.s005.docx^{(28.3KB, docx)}

PLoS One. doi: 10.1371/journal.pone.0323812.r004

Decision Letter 1

Francisco de Paula-Silva

16 Apr 2025

Reconstructing smoking history through dental cementum analysis - a preliminary investigation on modern and archaeological teeth.

PONE-D-25-00796R1

Dear Dr. Schwalbe,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Francisco Wanderley Garcia de Paula-Silva, DDS, MSc, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

Reviewer #1: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

Reviewer #1: Yes

**********

6. Review Comments to the Author

Reviewer #1: I thank the authors for their detailed comments on the questions raised. The comments have been very valuable for a better understanding of the results presented in the paper and clearly outlined the way forward for future research.

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy .

Reviewer #1: No

**********

PLoS One. doi: 10.1371/journal.pone.0323812.r005

Acceptance letter

Francisco de Paula-Silva

PONE-D-25-00796R1

PLOS ONE

Dear Dr. Schwalbe,

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team.

At this stage, our production department will prepare your paper for publication. This includes ensuring the following:

* All references, tables, and figures are properly cited

* All relevant supporting information is included in the manuscript submission,

* There are no issues that prevent the paper from being properly typeset

You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps.

Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

If we can help with anything else, please email us at customercare@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Prof. Dr. Francisco Wanderley Garcia de Paula-Silva

Academic Editor

PLOS ONE

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Table S1. Study dataset.

(XLSX)

pone.0323812.s001.xlsx^{(17.6KB, xlsx)}

Table S2. Kruskal-Wallis rank sum test.

Summary of the chi-squared and p-values between cementum width and smoking habits and damage, in overall cohort, modern and archaeological subsamples.

(DOCX)

pone.0323812.s002.docx^{(14.4KB, docx)}

Attachment

Submitted filename: Feedback.docx

pone.0323812.s003.docx^{(15.7KB, docx)}

Attachment

Submitted filename: Response to reviewers.docx

pone.0323812.s005.docx^{(28.3KB, docx)}

Data Availability Statement

All relevant data are within the article and its supporting information files.

[pone.0323812.ref001] 1.Obertová Z, Skrzypek G, Danišík M, Rankenburg K, Cummaudo M, Olivieri L, et al. Stable Isotope Provenance of Unidentified Deceased Migrants-A Pilot Study. Biology (Basel). 2023;12(11):1371. doi: 10.3390/biology12111371 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0323812.ref002] 2.Copeland SR, Grimes V, Neveling J. Isotopic evidence for the geographic origin, movement and diet of the Hofmeyr individual. Title of the Book or Collection. 2022. p. 47–68. [Google Scholar]

[pone.0323812.ref003] 3.Paul KS, Feezell R, Hughes T, Brook AH. Integrating genealogy and dental variation: contributions to biological anthropology. American Journal of Biological Anthropology. 2022;181(S76):145–79. doi: 10.1002/ajpa.24662 [DOI] [Google Scholar]

[pone.0323812.ref004] 4.Naji S, Rendu W, Gourichon L. Dental cementum in anthropology. Cambridge: Cambridge University Press, 2022. [Google Scholar]

[pone.0323812.ref005] 5.Garizoain G, Parra RC, Aranda C, Zorba E, Moraitis K, Escalante-Flórez K, et al. Root dentin translucency and age at death estimation in adults using single rooted teeth: Update of the Forensic International Dental Database. Forensic Sci Int. 2023;343:111564. doi: 10.1016/j.forsciint.2023.111564 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref006] 6.El-Bakary AA, Hammad SM, Mohammed F. Dental age estimation in Egyptian children, comparison between two methods. J Forensic Leg Med. 2010;17(7):363–7. doi: 10.1016/j.jflm.2010.05.008 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref007] 7.Cerrito P, Cerrito L, Hu B, Bailey SE, Kalisher R, Bromage TG. Weaning, parturitions and illnesses are recorded in rhesus macaque (Macaca mulatta) dental cementum microstructure. Am J Primatol. 2021;83(3):e23235. doi: 10.1002/ajp.23235 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref008] 8.Cerrito P, Bailey SE, Hu B, Bromage TG. Parturitions, menopause and other physiological stressors are recorded in dental cementum microstructure. Sci Rep. 2020;10(1):5381. doi: 10.1038/s41598-020-62177-7 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0323812.ref009] 9.Cerrito P, Nava A, Radovčić D, Borić D, Cerrito L, Basdeo T, et al. Dental cementum virtual histology of Neanderthal teeth from Krapina (Croatia, 130-120 kyr): an informed estimate of age, sex and adult stressors. J R Soc Interface. 2022;19(187):20210820. doi: 10.1098/rsif.2021.0820 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0323812.ref010] 10.Kagerer P, Grupe G. Age-at-death diagnosis and determination of life-history parameters by incremental lines in human dental cementum as an identification aid. Forensic Sci Int. 2001;118(1):75–82. doi: 10.1016/s0379-0738(00)00382-0 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref011] 11.Panezic K, Porcic M, Urban P. Stressful times for women - increased physiological stress in Neolithic females detected in tooth cementum. J Archaeol Sci. 2020;122:105217. [Google Scholar]

[pone.0323812.ref012] 12.Perrone V, Gocha TP, Randolph-Quinney P, Procopio N. Tooth Cementum Annulation: A Literature Review. Forensic Sciences. 2022;2(3):516–50. doi: 10.3390/forensicsci2030038 [DOI] [Google Scholar]

[pone.0323812.ref013] 13.Berkovitz B, Holland G, Moxham B. Oral anatomy, histology & embryology. 5th ed. Elsevier. 2018. [Google Scholar]

[pone.0323812.ref014] 14.Wedel VL. Determination of season at death using dental cementum increment analysis. J Forensic Sci. 2007;52(6):1334–7. doi: 10.1111/j.1556-4029.2007.00546.x [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref015] 15.Colard T, Bertrand B, Naji S, Delannoy Y, Bécart A. Toward the adoption of cementochronology in forensic context. Int J Legal Med. 2018;132(4):1117–24. doi: 10.1007/s00414-015-1172-8 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref016] 16.Stott GG, Sis RF, Levy BM. Cemental annulation as an age criterion in forensic dentistry. J Dent Res. 1982;61(6):814–7. doi: 10.1177/00220345820610063401 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref017] 17.Colard T, Bertrand B, Naji S, Delannoy Y, Bécart A. Toward the adoption of cementochronology in forensic context. Int J Legal Med. 2018;132(4):1117–24. doi: 10.1007/s00414-015-1172-8 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref018] 18.Brooks S, Suchey J. Skeletal age determination based on the os pubis: a comparison of the Acsádi-nemeskéri and Suchey-brooks methods. Hum Evol. 1990;5:227–38. [Google Scholar]

[pone.0323812.ref019] 19.Iscan M, Loth S, Wright R. Age estimation from the rib by phase analysis: white females. J Forensic Sci. 1985;29:853–63. [PubMed] [Google Scholar]

[pone.0323812.ref020] 20.Işcan MY, Loth SR, Wright RK. Age estimation from the rib by phase analysis: white males. J Forensic Sci. 1984;29(4):1094–104. doi: 10.1520/jfs11776j [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref021] 21.Klezeval G. Recording structures of mammals: determination of age and reconstruction of life history. Rotterdam: A. A. Balkema. 1996. [Google Scholar]

[pone.0323812.ref022] 22.Cabec Al, Garrevoet J, Spiers K. Incremental elemental distribution in chimpanzee cellular cementum: insights from synchrotron x-ray fluorescence and implications for life-history inferences. In: Naji S, Rendu W, Gourichon L, editors. Dental cementum in anthropology. Cambridge University Press. 2022. p. 138–54. [Google Scholar]

[pone.0323812.ref023] 23.Colard T, Naji S, Debroucker A. Preliminary protocol to identify parturitions lines in acellular cementum. In: Naji S, Rendu W, Gourichon L, editors. Dental cementum in anthropology. Cambridge University Press. 2022. p. 234–48. [Google Scholar]

[pone.0323812.ref024] 24.Naji S, Rendu W, Gourichon L. Recent advances on acellular cementum increments composition using synchrotron x-radiation. Dental cementum in anthropology. Cambridge University Press. 2022. p. 110–37. [Google Scholar]

[pone.0323812.ref025] 25.Dean C, Le Cabec A, Spiers K, Zhang Y, Garrevoet J. Incremental distribution of strontium and zinc in great ape and fossil hominin cementum using synchrotron X-ray fluorescence mapping. J R Soc Interface. 2018;15(138):20170626. doi: 10.1098/rsif.2017.0626 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0323812.ref026] 26.Cusano NE. Skeletal effects of smoking. Curr Osteoporos Rep. 2015;13:302–9. [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref027] 27.Kapoor D, Jones TH. Smoking and hormones in health and endocrine disorders. Eur J Endocrinol. 2005;152(4):491–9. doi: 10.1530/eje.1.01867 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref028] 28.Obeid P, Bercy P. Effects of smoking on periodontal health: a review. Adv Ther. 2000;17(5):230–7. doi: 10.1007/BF02853162 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref029] 29.Leite F, Nascimento G, Scheutz F. Effect of smoking on periodontitis: A systematic review and meta-regression. Am J Prev Med. 2018;54:831–41. [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref030] 30.Fiorini T, Musskopf ML, Oppermann RV, Susin C. Is there a positive effect of smoking cessation on periodontal health? A systematic review. J Periodontol. 2014;85(1):83–91. doi: 10.1902/jop.2013.130047 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref031] 31.Bezerra Ferreira JD, Rodrigues JA, Piattelli A, Iezzi G, Gehrke SA, Shibli JA. The effect of cigarette smoking on early osseointegration of dental implants: a prospective controlled study. Clin Oral Implants Res. 2016;27(9):1123–8. doi: 10.1111/clr.12705 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref032] 32.Soden I. Excavations at the cathedral church of St Mary, Coventry. 1999 -2000. Summary report. 2000. [Google Scholar]

[pone.0323812.ref033] 33.Davies-Barrett A, Inskip S. Who smokes anymore? documentary, archaeological and osteological evidence for tobacco consumption and its relationship to social identity in industrial England, 1700–1850. In: Craig-Atkins E, Harvey K, editors. The material body. Manchester, England: Manchester University Press. 2024. p. 133–69. [Google Scholar]

[pone.0323812.ref034] 34.Mays S, Elders J, Humphrey L, et al. Science and the Dead. Destructive sampling of archaeological human remains for scientific analysis. 2nd ed. Advisory Panel on the Archaeology of Burials in England, 2023. (accessed 12 June 2024). [Google Scholar]

[pone.0323812.ref035] 35.Bertrand B, Cunha E, Bécart A, Gosset D, Hédouin V. Age at death estimation by cementochronology: Too precise to be true or too precise to be accurate?. Am J Phys Anthropol. 2019;169(3):464–81. doi: 10.1002/ajpa.23849 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref036] 36.Wittwer-Backofen U, Gampe J, Vaupel JW. Tooth cementum annulation for age estimation: results from a large known-age validation study. Am J Phys Anthropol. 2004;123(2):119–29. doi: 10.1002/ajpa.10303 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref037] 37.Wittwer-Backofen U. Age estimation using tooth cementum annulation. Methods Mol Biol. 2012;915:129–43. doi: 10.1007/978-1-61779-977-8_8 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref038] 38.AlQahtani SJ, Hector MP, Liversidge HM. Brief communication: The London atlas of human tooth development and eruption. Am J Phys Anthropol. 2010;142(3):481–90. doi: 10.1002/ajpa.21258 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref039] 39.Liversidge HM. Tooth Eruption and Timing. In: Irish J.D, Richard Scott G. (eds) A Companion to Dental Anthropology. Chichester, UK: Wiley Blackwell, John Wiley & Sons, Inc., pp. 159–71. [Google Scholar]

[pone.0323812.ref040] 40.Harris E. Odontogenesis. A companion to dental anthropology. Hoboken NJ: Wiley. 2015. p. 142–58. [Google Scholar]

[pone.0323812.ref041] 41.Lieberman DE. The Biological Basis for Seasonal Increments in Dental Cementum and Their Application to Archaeological Research. Journal of Archaeological Science. 1994;21(4):525–39. doi: 10.1006/jasc.1994.1052 [DOI] [Google Scholar]

[pone.0323812.ref042] 42.Cool SM, Forwood MR, Campbell P, Bennett MB. Comparisons between bone and cementum compositions and the possible basis for their layered appearances. Bone. 2002;30(2):386–92. doi: 10.1016/s8756-3282(01)00686-x [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref043] 43.Yamamoto H, Sugahara N, Yamada N. Histopathological and microradiographic study of the exposed cementum from periodontally diseased human teeth. Bull Tokyo Med Dent Univ. 1966;13(3):407–21. [PubMed] [Google Scholar]

[pone.0323812.ref044] 44.Simon JH, Yonemoto GS, Bakland LK. Comparison of cellular cementum in normal and diseased teeth - a scanning electron microscopic study. J Endod. 1981;7(8):370–5. doi: 10.1016/S0099-2399(81)80058-1 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref045] 45.Duarte PM, Nogueira CFP, Silva SM. Impact of smoking cessation on periodontal tissues. Int Dent J. 2022;72:31–6. [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0323812.ref046] 46.Souto MLS, Rovai ES, Villar CC, Braga MM, Pannuti CM. Effect of smoking cessation on tooth loss: a systematic review with meta-analysis. BMC Oral Health. 2019;19(1):245. doi: 10.1186/s12903-019-0930-2 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0323812.ref047] 47.Nanci A, Bosshardt DD. Structure of periodontal tissues in health and disease. Periodontol 2000. 2006;40:11–28. doi: 10.1111/j.1600-0757.2005.00141.x [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref048] 48.Broucker A de, Colard T, Penel G, Blondiaux J, Naji S. The impact of periodontal disease on cementochronology age estimation. Int J Paleopathol. 2016;15:128–33. doi: 10.1016/j.ijpp.2015.09.004 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref049] 49.Effects of bagpipe which mentions tobacco pipe damage to teeth. Lancet. 1890;135:1438. [Google Scholar]

[pone.0323812.ref050] 50.Marruganti C, Shin H-S, Sim S-J, Grandini S, Laforí A, Romandini M. Air Pollution as a Risk Indicator for Periodontitis. Biomedicines. 2023;11(2):443. doi: 10.3390/biomedicines11020443 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0323812.ref051] 51.Huang K, Feng L-F, Liu Z-Y, Li Z-H, Mao Y-C, Wang X-Q, et al. The modification of meteorological factors on the relationship between air pollution and periodontal diseases: an exploration based on different interaction strategies. Environ Geochem Health. 2023;45(11):8187–202. doi: 10.1007/s10653-023-01705-6 [DOI] [PubMed] [Google Scholar]

[pone.0323812.ref052] 52.Zhou S, Li W, Wan J, Fu Y, Lu H, Li N, et al. Heavy metals in drinking water and periodontitis: evidence from the national oral health survey from China. BMC Public Health. 2023;23(1):1706. doi: 10.1186/s12889-023-16391-3 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0323812.ref053] 53.McGuire SA. Products of Industry: Pollution, Health, and England’s Industrial Revolution. 2020, pp. 203–231.

PERMALINK

Reconstructing smoking history through dental cementum analysis - a preliminary investigation on modern and archaeological teeth

Valentina Perrone

Anna M Davies-Barrett

Mario Migliario

Patrick Randolph-Quinney

Sarah A Inskip

Edward C Schwalbe

Roles

Abstract

Introduction

Materials & methods

Fig 1. Archaeological evidence of tobacco consumption.

Statistical analyses

Results

Fig 2. Visual comparison of the AEFC in smoker, non-smoker and ex-smoker individuals.

Fig 3. Smoking-damaged areas show no birefringence.

Fig 4. Distribution of samples according to presence/absence of the smoking damage (A) and to the smoking habits of the donors(B).

Table 2. Association between occurrence of smoking damage by smoking habits. Smoking damage is associated with smoking status (p < 0.0001, Fisher’s Exact test).

Smoking damage is associated with smoking status (p < 0.0001, Fisher’s Exact test)

Fig 5. Smoking status affects AEFC width.

Fig 6. Occurrence of smoking damage in non-smokers, ex-smokers and smokers.

Table 3. Prediction of smoking habits. Calculations of the power of prediction of smoking status based on the observation of smoking damage, in terms of overall accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV).

Table 4. Evidence of smoking in archaeological samples. Comparison between the assessment of smoking status through archaeological examination and by presence of smoking damage on the AEFC. Indeterminate = “indet”.

Fig 7. Examples of the occurrence of the smoking damage in the archaeological sample (black arrows).

Discussion

Conclusions

Supporting information

Acknowledgments

Data Availability

Funding Statement

References

Author response to Decision Letter 0

Decision Letter 0

Francisco de Paula-Silva

Roles

Author response to Decision Letter 1

Decision Letter 1

Francisco de Paula-Silva

Roles

Acceptance letter

Francisco de Paula-Silva

Roles

Associated Data

Supplementary Materials

Data Availability Statement

ACTIONS

PERMALINK

RESOURCES

Similar articles

Cited by other articles

Links to NCBI Databases