Bayesian Analysis of the Dental Maturational Sequences of Lagar

Dental maturational sequence and dental tissue proportions in the early Upper Paleolithic child from Abrigo do Lagar Velho, Portugal Priscilla Bayle, Roberto Macchiarelli, Erik Trinkaus, Cidália Duarte, Arnaud Mazurier and João Zilhão Supporting Information

Fig. S1. Lagar Velho 1. Virtual 3D reconstruction of four deciduous and one permanent teeth assessed for linear, surface, and volumetric tissue proportions. A. deciduous upper right central incisor (Ui1, labial view); B. deciduous lower right lateral incisor (Li2, labial view); C. deciduous lower right canine (Lc, labial view); D. deciduous lower right second molar (Lm2, oblique mesio-buccal view); E. permanent lower right first molar (LM1, oblique mesio-buccal view). Enamel thickness topographic variation is rendered by means of a chromatic scale increasing from dark blue (thinner) up to red (thicker); radicular dentine is rendered in semi-transparency and the virtual filling of the pulp cavities of the deciduous teeth is shown in orange. Scale bar is 1 mm.

1 Fig. S2. Comparative estimates of the Average Enamel Thickness (AET; see Methods) of four deciduous and one permanent teeth in Lagar Velho 1 (LaVe1), La Madeleine 4 (Mad4; no data are available for Ui1 and LM1), Neandertals (Neand), and extant humans (EH). AET is the average straight-line distance between the enamel-dentin junction (EDJ) and the outer enamel surface, calculated as the quotient of the enamel volume and the EDJ surface area.

2 Fig. S3. Comparative estimates of the Relative Enamel Thickness (RET; scale-free relative estimate; see Methods) of four deciduous and one permanent teeth in Lagar Velho 1 (LaVe1), La Madeleine 4 (Mad4; no data are available for Ui1 and LM1), Neandertals (Neand), and extant humans (EH). RET is the AET scaled by the cube root of the crown dentine+pulp volume and multiplied by 100.

3 Fig. S4. Comparative enamel thickness cartographies of the deciduous lower second molar (upper) and the permanent lower first molar (lower) in: (A) a Neandertal (Roc de Marsal 1, upper; Abri Suard S14-7, lower); (B) Lagar Velho 1; (C) an extant human (EH-U57, upper; EH-U21, lower). Crowns shown in oblique mesio-buccal view. Enamel thickness topographic variation rendered by means of a chromatic scale increasing from dark blue (thinner) up to red (thicker). Scale bar is 1 mm.

4 Table S1. Lagar Velho 1. Degree of maturation of the elements of the lower deciduous and permanent dentition based on the descriptions and the estimates by Hillson (1) and Hillson and Santos Coelho (2) assessed following the system developed by Moorrees et al. (3), integrated by independent estimates assessed on the original material (by PB, RM, and CD) according to the scoring system of Demirjian et al. (4) for the permanent teeth, revised by Liversidge and Molleson (5) for the deciduous teeth, modified and adapted by Bayle et al. (6).

dental element maturational stage 1 descriptive criteria of crown and root calcification Li1 h2 Apical dentine edge is sharp and apex is closed. Li2 h2 Apical dentine edge is sharp and apex is closed. Lc h2 Apical dentine edge is sharp and apex is closed. Lm1 h2 Apical dentine edge is sharp and apex is closed. Because of post-mortem damage, the assessment of the maturational Lm2 h1/h2/r degree in both teeth is difficult. Root length is at least complete and a small amount of resorption possibly affects the distal root of the left tooth. Crown formation is substantially complete but not enough of either tooth LI1 C/D can be seen to examine the forming surface on the labial and lingual sides. LI2 C Crown formation is almost complete. The enamel formation is complete at the occlusal surface and its extension LC C and convergence towards the cervical region is seen. The enamel formation is complete at the occlusal surface and its extension LP3 C and convergence towards the cervical region is seen. While the right element is missing, only the lingual half of the developing LP4 B/C left crown is preserved. The occlusal surface seems complete but the crown is less developed than in the first premolar. In the practically complete left tooth, a plate of dentine forming the root LM1 E bifurcation is seen. LM2 B The coalescence of cusp tips is seen.

5 Table S2. Lagar Velho 1. Series of possible maturational sequences (3 deciduous and 4 permanent) assessed for the mandibular dentition (represented by 5 deciduous and 7 permanent elements) following the stages modified and adapted from Liversidge and Molleson (5) for the deciduous and Demirjian et al. (4) for the permanent teeth (rev. in 6).

deciduous teeth permanent teeth seq. number seq. number (Li1-Li2-Lc-Lm1-Lm2) (LI1-LI2-LC-LP3-LP4-LM1-LM2) 1 h2-h2-h2-h2-h1 C-C-C-C-B-E-B I 2 h2-h2-h2-h2-h2 C-C-C-C-C-E-B II 3 h2-h2-h2-h2-r D-C-C-C-B-E-B III D-C-C-C-C-E-B IV

6 Table S3. Linear, surface and volumetric estimates and dental tissue proportions of four deciduous and one permanent teeth comparatively assessed in Lagar Velho 1 (LaVe1), La Madeleine 4 (Mad4; no data are available for Ui1 and LM1; from 7–9), Neandertals (Neand; from 6–13, and original data), and extant humans (EH; 7–10, 12–14, and original data). Average Enamel Thickness (AET) = enamel volume / EDJ* surface area. Relative Enamel Thickness (RET) = [AET / (crown dentine+pulp volume)1/3]. RET is a scale-free estimate (see Methods).

% of the crown enamel crown dentine crown pulp crown dentine+pulp EDJ* surface volume that is Average Enamel Relative Enamel volume (mm3) volume (mm3) volume (mm3) volume (mm3) area (mm²) dentine+pulp Thickness (mm) Thickness Ui1 Neand N 4 4 4 4 4 4 4 4 mean 27.16 81.38 11.59 92.97 94.83 77.39 0.29 6.32 range 24.24-30.24 76.05-83.32 7.79-13.92 87.27-97.21 92.67-98.21 76.19-78.90 0.25-0.33 5.67-7.11 LaVe1 28.78 79.46 8.28 87.74 80.83 75.30 0.36 8.01 EH N 2 2 2 2 2 2 2 2 mean 26.15 46.02 5.67 51.69 68.05 66.41 0.38 10.32 range 24.87-27.43 40.91-51.13 3.32-8.01 48.92-54.46 67.60-68.50 66.30-66.50 0.36-0.41 9.93-10.71 Li2 Neand N 4 4 4 4 4 4 4 4 mean 18.79 42.36 4.44 46.80 64.11 71.36 0.29 8.13 range 16.48-20.91 40.27-48.08 3.92-5.65 44.47-53.73 59.23-70.72 68.05-73.60 0.27-0.33 7.22-9.23 LaVe1 21.92 33.50 3.77 37.27 49.41 62.96 0.44 13.28 Mad4 N 2 2 2 2 2 2 2 2 mean 15.34 22.73 1.67 24.39 44.75 61.39 0.34 11.79 range 13.72-16.96 21.33-24.12 1.60-1.74 22.93-25.86 41.93-47.58 60.39-66.56 0.33-0.36 11.52-12.05 EH N 4 4 4 4 4 4 4 4 mean 17.87 18.12 2.21 20.33 42.37 53.22 0.42 15.45 range 15.72-24.01 16.53-19.79 1.38-3.17 18.12-22.96 38.19-50.57 47.95-59.00 0.32-0.58 11.10-20.58

7 Table S3 (cont.)

% of the crown enamel crown dentine crown pulp crown dentine+pulp EDJ* surface volume that is Average Enamel Relative Enamel volume (mm3) volume (mm3) volume (mm3) volume (mm3) area (mm²) dentine+pulp Thickness (mm) Thickness Lc Neand N 5 5 5 5 5 5 5 5 mean 32.78 75.00 12.66 87.67 91.99 72.79 0.36 8.02 range 25.52-37.86 49.79-101.14 8.16-27.08 76.87-110.91 85.51-104.34 70.59-75.84 0.30-0.39 7.02-8.74 LaVe1 36.73 62.72 5.94 68.66 73.49 65.15 0.50 12.21 Mad4 32.87 56.16 3.60 59.76 70.76 64.51 0.46 11.88 EH N 4 4 4 4 4 4 4 4 mean 32.49 44.25 4.26 48.51 65.23 59.89 0.50 13.66 range 24.97-40.22 32.93-48.98 2.09-7.29 35.03-55.80 49.78-75.42 56.98-64.38 0.41-0.59 10.71-15.78 Lm2 Neand N 6 6 6 6 6 6 6 6 mean 116.16 195.61 31.57 227.17 189.90 66.17 0.61 10.03 range 101.27-131.16 176.54-209.02 26.05-37.11 202.58-244.45 174.50-206.93 63.96-67.76 0.58-0.67 9.49-10.92 LaVe1 143.74 180.57 19.15 199.73 172.11 58.15 0.84 14.29 Mad4 N 2 2 2 2 2 2 2 2 mean 129.94 161.07 14.54 175.61 154.28 57.47 0.84 15.00 range 125.00-134.89 158.18-163.96 14.06-15.02 173.20-178.02 152.85-155.70 56.89-58.08 0.82-0.87 14.67-15.40 EH N 7 7 7 7 7 7 7 7 mean 104.79 140.46 15.36 155.82 138.07 59.79 0.76 14.10 range 81.76-127.98 123.74-172.86 6.40-38.29 130.14-183-56 117.84-151.26 52.04-64.89 0.63-0.97 11.98-18.93

8 Table S3 (cont.)

9 Table S4. Origin, reference group, age (oxygen isotope stage), and tooth-specific sample size of the microtomographic-based dental sample. Data sources: Abri Suard (France): 7, 11, 12, and original data; Abri Bourgeois-Delaunay (France): 7, 10; Roc de Marsal (France): 6–8; El Sidrón (Spain): 12; Engis (Belgium): 12, and original data; Spy (Belgium): 13; Lagar Velho (Portugal): present data; La Madeleine (France): 7–9; extant humans (worldwide): 6–10, 12, 14, and original data.

group site/sample age Ui1 Li2 Lc Lm2 LM1 total

Neandertal Abri Suard OIS 6 1 2 4 3 10

Abri Bourgeois-Delaunay OIS 5e 1 1

Roc de Marsal OIS 5a 2 2 2 2 8

El Sidrón OIS 3 2 2

Engis OIS 3 1 1 2

Spy OIS 3 1 1 1 3

Upper Paleolithic Lagar Velho OIS 2 1 1 1 1 1 5

La Madeleine OIS 2 2 1 2 5

extant EH present 2 4 4 7 18 35

10 Table S5. Maturational stage [modified and adapted from Liversidge and Molleson (5) for the deciduous and from Demirjian et al. (4) for the permanent teeth (in 6)], degree of occlusal wear (15), and general preservation quality of the enamel [ranging from "very bad" (complete enamel loss), to "very good" (no enamel loss)] directly assessed on a number of original fossil and extant (EH) teeth considered in the present analysis. Information concerning additional tooth material from other studies used in the comparative analysis of dental tissue proportions (12, 14) is not available here. Within the factors listed in Table S5, only the degree of dental wear and the enamel preservation state may affect the results presented in Table S3. Because all specimens reached at least the degree of "crown completion" (stage>d or D, for the deciduous and permanent elements, respectively), the mineralization stage has no effect on the virtual assessment of the crown variables. Conversely, as previously noted for the dentition of the late Upper Paleolithic child from La Madeleine (9), both Lagar Velho 1 upper central and lower lateral deciduous incisors exhibit a relatively advanced degree of wear in comparison with the extant specimens used in the comparative analysis. In Lagar Velho 1, besides the minor influence on the volumetric virtual estimates of Ui1, Li2, and Lc of artifacts related to some punctual enamel detachments at the cervix, the mesio-lower labial aspect of the crown of the deciduous canine also displays localized enamel hypoplasia (Fig. S1C), responsible for a spot-like thinning of the dental tissue.

tooth group site/sample spec. maturational stage wear degree enamel preservation quality Ui1 Neandertal Roc de Marsal RdM1 left h1 3/4 good Neandertal Roc de Marsal RdM1 right h1 3/4 good Neandertal Engis Engis 2 h1 4/5 pretty good Neandertal Spy Spy 589a h1 2 pretty good Upper Paleolithic Lagar Velho LaVe1 h2 4 pretty good extant EH EH-U21 r 2/3 good extant EH EH-U63 h1 1 good

11 Table S5 (cont.) tooth group site/sample spec. maturational stage wear degree enamel preservation quality Li2 Neandertal Abri Suard S14-3 r 4/5 very good Neandertal Roc de Marsal RdM1 left h1 3 pretty good Neandertal Roc de Marsal RdM1 right h2 2 good Neandertal Spy Spy 594a g 1 very good Upper Paleolithic Lagar Velho LaVe1 h2 3/4 pretty good Upper Paleolithic La Madeleine Mad4 left h2 2/3 good Upper Paleolithic La Madeleine Mad4 right r 2 very good extant EH EH-Sbg1 h1 1 very good extant EH EH-Sbg3 h1 1 good extant EH EH-U21 r 1 very good extant EH EH-UdP h1 1 very good Lc Neandertal Abri Suard S37 f 1 good Neandertal Abri Suard S45 g 1 good Neandertal Roc de Marsal RdM1 left g 1 good Neandertal Roc de Marsal RdM1 right g 1 good Neandertal Spy Spy 645a e 1 very good Upper Paleolithic Lagar Velho LaVe1 h2 2 pretty good Upper Paleolithic La Madeleine Mad4 g 1 very good extant EH EH-Sbg1 f 1 good extant EH EH-Sbg2 g 2/3 very good extant EH EH-U21 h1 1 pretty good extant EH EH-UdP g 1 good

12 Table S5 (cont.) tooth group site/sample spec. maturational stage wear degree enamel preservation quality Lm2 Neandertal Abri Suard S14-5 r 3 very good Neandertal Abri Suard S37 left f 2 very good Neandertal Abri Suard S37 right f 2 good Neandertal Abri Suard S42 g 2 very good Neandertal Roc de Marsal RdM1 left g 2 very good Neandertal Roc de Marsal RdM1 right g 2 very good Upper Paleolithic Lagar Velho LaVe1 h1/h2/r 2 very good Upper Paleolithic La Madeleine Mad4 left h1 2 good Upper Paleolithic La Madeleine Mad4 right h1 2 good extant EH EH-U21 h1 3 pretty good extant EH EH-U56 e 1/2 very good extant EH EH-U57 e 1/2 very good extant EH EH-U63 g 2/3 good extant EH EH-UdP f 2 very good extant EH EH-UTP r 3/4 very good extant EH EH-Sbg2 h1 2/3 very good LM1 Neandertal Abri Suard S5 D/E 1 very good Neandertal Abri Suard S14-7 D/E 1 very good Neandertal Abri Suard S49 D 1 good Neandertal Abri Bourgeois-Delaunay BD-J4-C9 G 3 good Upper Paleolithic Lagar Velho LaVe1 E 1 good extant EH EH-Sbg4 E 1 very good extant EH EH-U21 D/E 1 very good extant EH EH-UdP D 1 very good extant EH EH-UdP2 G 2/3 very good

13 Supporting Information References 1. Hillson SW (2002) in Portrait of the artist as a child - The Gravettian human skeleton from the Abrigo do Lagar Velho and its archeological context, eds Zilhão J, Trinkaus E (Instituto Português de Arqueologia, Lisbon), pp 242−245. 2. Hillson SW, Santos Coelho JM (2002) in Portrait of the artist as a child - The Gravettian human skeleton from the Abrigo do Lagar Velho and its archeological context, eds Zilhão J, Trinkaus E (Instituto Português de Arqueologia, Lisbon), pp 342−355. 3. Moorrees CFA, Fanning EA, Hunt EE (1963) Age variation of formation stages for ten permanent teeth. J Dent Res 42:1490−1502. 4. Demirjian A, Goldstein H, Tanner JM (1973) A new system of dental age assessment. Hum Biol 45:211−227. 5. Liversidge HM, Molleson T (2004) Variation in crown and root formation and eruption of human deciduous teeth. Am J Phys Anthropol 123:172−180. 6. Bayle P, Braga J, Mazurier A, Macchiarelli R (2009) Dental developmental pattern of the Neanderthal child from Roc de Marsal: a high-resolution 3D analysis. J Hum Evol 56:66−75. 7. Bayle P (2008) PhD thesis (Université Toulouse III − Paul Sabatier). 8. Bayle P (2008) Proportions des tissus des dents déciduales chez deux individus de Dordogne (France): l’enfant néanderthalien du Roc de Marsal et le spécimen du Paléolithique supérieur final de La Madeleine. Bull Mém Soc Anthropol Paris 20:151−163. 9. Bayle P, Braga J, Mazurier A, Macchiarelli R (2009) High-resolution comparative analysis of the dental development in the Late Paleolithic child from La Madeleine. Am J Phys Anthropol 138:493−498. 10. Macchiarelli R et al. (2006) How Neanderthal molar teeth grew. Nature 444:748−751. 11. Macchiarelli R, Mazurier A, Volpato V (2007) in Les Néandertaliens. Biologie et cultures, eds Vandermeersch B, Maureille B (Editions du CTHS, Paris), pp 169−179. 12. Olejniczak AJ et al. (2008) Dental tissue proportions and enamel thickness in Neandertal and modern human molars. J Hum Evol 55:12−23. 13. Bayle P, Macchiarelli R (2009) in Spy cave. State of 120 years of pluridisciplinary research on the Betche-aux-Rotches from Spy (Jemeppe-sur-Sambre, Province of Namur, Belgium), eds Semal P, Hauzeur A (Royal Belgian Institute of Natural Sciences, Brussels), in press. 14. Kono R (2004) Molar enamel thickness and distribution patterns in extant great apes and humans: new insights based on a 3-dimensional whole crown perspective. Anthropol Sci 112:121−146. 15. Smith BH (1984) Patterns of molar wear in hunter-gatherers and agriculturalists. Am J Phys Anthropol 63:39−56.