Anat Embryol (2003) 207:221–232
DOI 10.1007/s00429-003-0343-4
ORIGINAL ARTICLE
Ralf J. Radlanski · Herbert Renz ·
Marie C. Klarkowski
Prenatal development of the human mandible
3D reconstructions, morphometry and bone remodelling pattern, sizes 12–117 mm CRL
Accepted: 23 July 2003 / Published online: 10 September 2003

Springer-Verlag 2003
Abstract Human embryos and fetuses (n=25) ranging
from 12 to 117 mm CRL (crown-rump-length) were
serially sectioned and the mandibles were reconstructed in
3D. In addition, characteristic areas of apposition,
resorption and resting zones were projected onto the
surface of the mandibular reconstructions after histolog-
ical evaluation of the remodeling processes. Furthermore,
morphometric data were taken to describe growth
processes in horizontal views. In this way the changing
outlines as seen in 3D could be correlated with the
remodeling patterns and with the changes in growth. In
these stages the mandible showed a general appositional
growth, but resorption areas were found at the posterior
margins of the mental foramen and at the lateral and
medial posterior bony planes at concave surfaces. The
bulging of bone underneath and over Meckel’s cartilage
could be recognized as active appositional growth areas.
Meckel’s cartilage itself lay in a trough which could be
characterized by less apposition and even resorption.
Questions were raised in how much the gap between our
present knowledge of genetic expression of signaling
molecules and the precise morphologic description of the
mandibles can be bridged.
Keywords Human mandible · Mandibular prenatal
growth · 3D reconstructions · Bone remodeling ·
Morphometry
Introduction
In many regions of the developing human face the quality
of the morphological description of the developmental
R. J. Radlanski ()) · H. Renz · M. C. Klarkowski
Universittsklinikum Benjamin Franklin der FU Berlin Klinik und
Poliklinik fr Zahn-Mund- und Kieferheilkunde,
Department of Experimental Dentistry and Oral Structural Biology,
Aßmannshauser Strasse 4–6, 14197 Berlin-Wilmersdorf, Germany
e-mail: ralfj.radlanski@medizin.fu-berlin.de
Tel.: +49-30-8445-6271
Fax: +49-30-8445-6392
processes lags behind the general progress that has been
made in the field of molecular biological research.
Biochemically, we know what forms bone (Olsen et al.
2000), and we know that e.g. Runx2 is essential for bone
formation (Shui et al. 2003). Factors involved in the early
patterning of the face are as well known (Helms et al.
1997; Thorogood et al. 1998; Schneider et al. 1999, 2001)
as the fact that there is a certain interbranchial arch and an
intrabranchial arch identity established by the distribution
of different Dlx genes (Depew et al. 2002; Cobourne and
Sharpe 2003). Perturbation and transplantation experi-
ments have lead to formation of extra bony entities in the
face (Lee SH et al. 2001; Helms and Schneider 2003), but
an exact morphological description has not yet been
given. For the commonly used animal model (avian and
murine), there are some 3D reconstructions available
(Kaufman 1998), but these refer to earlier developmental
stages. For the relatively late stages of craniofacial
development only 2D outlines (Rossant and Tam 2002)
have been given so far. For the human prenatal develop-
ment, only very sparse knowledge in 3 dimensions has
been presented, but no comprehensive description of
normal prenatal development in 3D is available. This
paper is focused on giving a comprehensive description of
the prenatal development of the human mandible in 3D,
together with maps of the remodeling areas and with
morphometric data.
Several studies have been dedicated to the prenatal
development of the human mandible and Meckel’s
cartilage (Meckel 1820), in most cases they rely on
histological (serial) sections (Burdi 1968; Durst-Zivkovic
and Davila 1974; Doskocil 1988; Orliaguet et al. 1994;
Rodriguez-Vasquez et al. 1997; Lee SK et al. 2001) and
in some cases 3D reconstructions have been made
(Fawcett 1906, 1910; Low 1909; Macklin 1914, 1921;
Blechschmidt 1960, 1973; Reulen 1997; Radlanski et al.
2001). In order to gain a better understanding in 3
dimensions, oil clearing techniques (Gantz 1922), frac-
tioned microradiography (Kjaer 1990; Kjaer and Graem
1990), or computed tomography of fetal material (Vi-
rapongse and Shapiro 1985; Neumann et al. 1997;
222

Radlanski et al. 1999b) have been performed. Although RDO (Eurobio, Paris, France) and EDTA for 2–30 days. Paraffin
our detailed knowledge about the formation of Meckel’s embedding was carried out according to standard histological
procedures, and the specimens were cut as 10 m-thick serial
cartilage and mandibular bone is far from being sparse, sections (Leica, Reichert-Jung RM 2065) in frontal, horizontal, and
we lack a continuous row of stages in order to describe sagittal planes. Routine staining was performed with hematoxylin-
the dynamic process of development with special refer- eosin, in addition, staining methods according to Masson-Goldner,
ence to possible alterations in shape and size. In order to Gieson modified according to Domagk, trichrome elastica staining
(Romeis 1989; Presnell and Schreibman 1997), and TRAP (tartate-
determine the localization of the cells that differentiate resistant acid phospatase) staining (Sigma, Deideshofen, Germany)
with processes on the cellular and subcellular levels, it is were applied according to Jsten et al. (1993).
essential to know the frame of the fetal “gross” anatomy.
This is necessary to contribute to bridging the gap
between molecular biology and morphologically oriented 3D reconstruction
embryological research. For this reason we have under- In each section, the contours to be reconstructed were determined
taken this study. The material may serve as reference for under a light microscope (Zeiss Universal, Oberkochen, Germany)
further studies. equipped with a camera lucida which provided magnification in the
range 1.25–10. Contours of the tissues from each section were
traced on transparent paper, superimposed on a light box and
correctly aligned using the contours of well characterized land-
Material and methods marks (e.g. ectoderm, eyes, skull base structures) as fiducial
markers (Gaunt and Gaunt 1978).
Material and staining Using the software Analysis (SIS, Mnster, Germany), com-
puter generated three-dimensional reconstructions were created.
A total of 25 human embryos and fetuses ranging from 12 to The description of the morphological findings is based on views
117 mm CRL (crown-rump-length) were used for this study from multiple perspectives and in stereoscopic mode.
(Table 1). The embryos of 12.0–16.8 mm CRL were provided from A fully automatic 3D reconstruction from scanned microscop-
the Steding collection (Dept. of Embryology, Zentrum Anatomy, ical images could not be applied, because they were too complex
Georg-August-University of Gttingen, Germany), the other em- and a differential diagnosis between different tissue types had to be
bryos and fetuses were taken from the Radlanski collection, Berlin, delivered by a trained morphologist in many cases.
Germany. All specimens were obtained from legal or spontaneous
abortions, and no part of the material gave indications of possible
malformation. Prior to histological processing, the heads of all Tracing of remodeling surfaces
specimens were documented by means of stereoscopic photography
which served as an aid for alignment of single sections to form the The histomorphological differentiation of the cells at the bone
reconstructions. surface was analyzed at magnifications between 100 and 400
The specimens were fixed in Bouin’s solution and, according to with reference to morphological and staining-specific criteria. A
standard histological procedures, the specimens were dehydrated in counting grid (1010 mm, line distance 0.5 mm; Zeiss, Oberko-
alcohol with increasing concentrations up to 100%. Depending on chen, Germany) in the ocular permitted quantification of cell
size and gross preparation, the specimens were decalcified using numbers.
In order to evaluate the validity of our analyses, we repeated the
measurements in 20 randomly selected sections after a 10-week
interval. The deviation ranged between 2.01 and 3.05%; in addition,
Table 1 Details of specimens examined in this study
comparison of right and left halves of the mandibles showed a high
Catalog CRL Section Estimated fertilization degree of correspondence. After a period of orientation and
number (mm) plane age (weeks) coordination of the diagnostic methods, all microscopic cell
characterizations were performed by one person.
STE 080787 12.0 HOR 6 The bony surfaces were characterized as follows:
STE 040647 14.0 FRO 6
STE 290687 15.6 HOR 6 1. Major apposition: more than 5 osteoblasts per grid square
STE 170452 16.8 SAG 6 (0.25 mm2) at a magnification of 160
GUS 110785 18.0 SAG 6 2. Minor apposition: up to 5 osteoblasts per grid square
CHR 220687 19.0 SAG 7 (0.25 mm2) at a magnification of 160
NEL 120989 21.0 HOR 7 3. Resorption: presence of osteoclasts
ALI 070785 21.0 HOR 7 4. Inactive regions, “lining cells” (Schenk et al. 1973; Jger 1996)
DID 110688 22.0 HOR 7 5. Acellular bony surface
EMM 150787 22.0 SAG 7 6. Dense mesenchymal cells, undifferentiated.
BAL 110787 24.0 HOR 8
JOS 080289 25.0 HOR 8 These differently characterized bony surfaces were color-coded
OLL 080785 29.0 SAG 8 at the bony contour of the histological sections. Using the graphics
PIP 161189 31.0 SAG 8 function, these data were entered into a computer and reconstruc-
DOR 100785 37.0 SAG 9 tion showing the mandibles in lateral and medial projections were
KUR 030389 41.0 HOR 9 produced using the software Histol (LIST Electronics, Darmstadt-
HAN 040389 53.0 SAG 9 Eberstadt, Germany).
THE 230494 53.0 HOR 9
GIS 210789 66.0 HOR 10
XAN 060289 68.0 FRO 10 Morphometry
ART 270694 68.0 HOR 10
ARI 160589 76.0 SAG 11 In horizontal projections, a series of measurements describing the
FUL 140489 76.0 HOR 11 outline of mandibular bone formations (length, width, opening
FLO 010689 95.0 SAG 13 angle, and the position of the mental foramen) were taken (Fig. 1).
HUL 110589 117.0 FRO 14 Definition of points of measurement:
 223
– ML: the most prominent bulging of the bony mandible in
lingual direction, determined by raising a perpendicular to the
straight line between MA–MP
– ML’: projection of the point ML to the straight line MA–MP
– C: intersection between the perpendicular projection through
the point Lmand/2 with the medial mandibular contour
– F: center of the mental foramen
– F’: projection of the point F to the straight line MA–MP.

Length measurements at the mandible:

– MA–MP: length of one half of the mandible

Fig. 1 Schematic illustration of a mandible in the horizontal plane
with landmarks and measuring sections. For detailed information
see text
– MA: the most anterior point of bony formation of the mandible
– MP: the most posterior point of bony formation of the mandible.
Constructed points of measurement:
– MB: the most prominent bulging of the bony mandible in buccal
direction, determined by raising a perpendicular to the straight
line between MA–MP
– MB’: projection of the point MB to the straight line MA–MP
– Lmand: length of the mandible, projected to the midsagittal plane
– Lmand/2: half the length of Lmand
– MP–MP: distance between the posterior ends of the mandible
– MA–MA: distance between the anterior ends of the mandible
– MB–MB’: measurement of the most prominent bulging of the
bony mandible in buccal direction
– ML–ML’: measurement of the most prominent bulging of the
bony mandible in lingual direction
– MA–MB’: position of the buccal bulging of the mandibular
bone, projected to the straight line MA–MP, determined as the
distance from the anterior end of the bone (MA)
– MA–F’: position of the mental foramen, projected to the straight
line MA–MP, determined as the distance from the anterior end
of the bone (MA)
– F–F: bilateral distance between the mental foraminae
– C–C: bilateral width of mandible at half length
– <) MPMA–MAMP: opening angle of the mandible.
Wherever possible, the measurements (in mm) were taken from
the left and the right halves of the mandible. All measurements
were taken 3 times, mean values are given and the maximum
difference between left and right measurements was 4.78%.

Fig. 2a–k Mandibles of human

embryos and fetuses in lateral
and 45 anterior views arranged
according to maturation and at
the same scales. Scale bar for
a–f, and for f–k 2000 m, CRL
(crown-rump-length) sizes: a
22 mm, b 19 mm, c 22 mm, d
25 mm, e 24 mm, f 41 mm, g
53 mm, h 66 mm, i 76 mm, and
j 117 mm. k 117 mm, left half
of the mandible, medial view.
Bone brown, Meckels cartilage
blue, ossicula mentalia (carti-
lage stage in j, k) dark blue,
cartilaginous core of condylar
process (in i–k) light blue, car-
tilaginous core of coronoid
process (in i) turquoise, inferior
alveolar nerve yellow, lingual
nerve (where depicted) yellow,
inferior alveolar artery (in i–h)
red, dental primordia (in j) gray
224
Statistics
Because of the number of specimens, it seemed sensible to apply
mainly descriptive statistics. A possible correlation between the
size of the specimens (CRL) and the measured variables was
evaluated in scattergrams and the correlation with the size of the
specimens was tested after Pearson. The paired Wilcoxon-test was
used to differentiate the significance of values describing the
changing outline of the mandible (anterior width, half-length,
interforaminar width, and posterior width).
Results
Morphology
Meckel’s cartilage
Meckel’s cartilage was first found in the embryo of
14 mm CRL, and it ran almost straight in the embryo of
22 mm CRL (Figs. 2a, 3a, 5a) where its anterior ends
were 500 m apart. As soon as both sticks of Meckel’s
cartilage approximated each other in the midline, a
vertical extension, called the hamulus, could be seen
(Figs. 3b–j, 4a–c, 5b–g). A smaller extension in the caudal
direction could also be seen on either anterior end of
Meckel’s cartilage (Fig. 4a). At its posterior end, Meck-
Fig. 3 Mandibles of human embryos and fetuses in cranial views
arranged according to maturation and at the same scales. Scale bar
for a–j, and for k–m 2000 m. CRL sizes a 22 mm, b 19 mm, c
21 mm, d 22 mm, e 25 mm, f 24 mm, g 29 mm, h 41 mm, i 53 mm,
j: 66 mm, k: 76 mm, and l: 117. m same as l, but transparent bone
to view the outline of Meckels cartilage. Bone brown, Meckels
el’s cartilage developed a different sized extension, the
manubrium mallei, which could be seen albeit weakly,
even in a very early stage in the 22 mm CRL embryo
(Fig. 3a). During development Meckel’s cartilage did not
remain straight, but became U-shaped (Fig. 3d, g), V-
shaped (Fig. 3f, i, j), and even lyrate (Fig. 3e).
Three curvatures in space could be described: we
found curvatures in lateral and cranial directions that
occurred in almost every specimen at typical places: the
most accentuated curve was seen at the posterior border
of the mandibular bone, where Meckel’s cartilage would
swing out laterally and cranially (Fig. 3e). The second
curve was a bulging to the medial plane and into a caudal
direction, most prominent in the fetus 66 mm CRL
(Fig. 2h). Another more or less sharp bend to the medial
was found in the region of the mental foramen (Fig. 3j).
The extent of the curvatures, however, varied from
relatively straight to extremely curved, almost zig-zagged,
as seen in the specimen 117 mm CRL (Figs. 2k, 3l, m).
Resorption of Meckel’s cartilage started at its anterior
ends at 117 mm CRL, and the remnant of its most anterior
end described an extremely meandering outline (Fig. 3l,
m). In the very anterior part new cartilaginous formations
were found which are called ossicula mentalia in the
cartilage blue, ossicula mentalia (cartilage stage in l, m) dark blue,
cartilaginous core of condylar process (in i, k–m) light blue,
cartilaginous core of coronoid process (in k) turquoise, inferior
alveolar nerve yellow, lingual nerve (where depicted) yellow,
inferior alveolar artery (in k–m) red, inferior alveolar vein (in l, m)
dark blue

Fig. 4 Mandibles of human embryos and fetuses in anterior views
arranged according to maturation and at the same scales, except a,
which shows a close-up of the symphyseal region. Scale bar for a
500 m, for b–: 2000 m. CRL sizes a 21 mm, b 25 mm, c 24 mm,
d 66 mm, e 76 mm, f 117 mm. Bone brown, Meckels cartilage blue,
cartilaginous core of condylar process (in e, f) light blue,
cartilaginous core of coronoid process (in d) turquoise, inferior
alveolar nerve yellow, lingual nerve (where depicted) yellow,
inferior alveolar artery (in e, f) red
literature (Goret-Nicaise 1982), but they were by far not
ossified in our stages.
Another cartilaginous formation was found in the
center of the condylar process (Figs. 2i–j, 3i, k–m, 4e, f,
5d, f, g), and in the coronoid process also (Figs. 2i, 4e, 5f).
Mandibular bone
The first mandibular ossification was found in the embryo
of 15.6 mm CRL at the branching of the mandibular nerve
in the future region of the mental foramen. From there the
flat bony plate spread out into anterior and posterior
directions, at a distance of ca. 50 m lateral to Meckel’s
cartilage (Figs. 2a, 3a, 5a). Mandibular bone followed the
changing curved outline of Meckel’s cartilage (Fig. 3a–
m), and not before a size of 53 mm CRL was reached, a
condylar and a coronoid process was clearly discernible
(Figs. 2g, 5d). There were some single ossifications in the
corners underneath the peg-shaped posterior end of
Meckel’s cartilage in the embryo of 37 mm CRL (not
shown) and in some fetuses of later stages (Figs. 2h, 5e).
225
Fig. 5 Mandibles of human embryos and fetuses in dorsal views
arranged according to maturation and at the same scales. Scale bar
for a–g 2000 m. CRL sizes a 22 mm, b 19 mm, c 25 mm, d
53 mm, e 66 mm, f 76 mm, g 117 mm. Bone brown, Meckels
cartilage blue, ossicula mentalia (cartilage stage in g) dark blue,
cartilaginous core of condylar process (in d, f–g) light blue,
cartilaginous core of coronoid process (in f) turquoise, inferior
alveolar nerve yellow, lingual nerve (where depicted) yellow,
inferior alveolar artery (in f) red
The location of the mental foramen was recognizable
with the first ossification. An incomplete embracing of
the mental nerve by mandibular bone was seen in the
embryo of 22 mm CRL (Fig. 2a), and in all subsequent
stages it was completely encircled by bone, except in the
fetus of 117 mm CRL, where the dental primordium was
in such close vicinity that the right mental foramen was
cranially open again (Figs. 2j, 4f). During the stages of
development the mental foramen changed its outline, as
previously described in more detail elsewhere (Radlanski
et al. 2002).
In no case were the anterior ends of mandibular bone
approximated closer than 0.05 mm, which was seen in the
fetus of 117 mm CRL. The outline of the anterior ends of
the mandible was variable and not at all symmetrical
(Fig. 4a–f). In general, the bony formations followed the
cartilaginous formations given by Meckel’s cartilage.
In a region directly posterior to the mental foramen,
first signs of bone penetrating underneath and above
Meckel’s cartilage were found in an embryo of 29 mm
CRL. These bony protrusions extended further, however,
226
Fig. 6 a–d Embryo 25 mm CRL, horizontal section, mandibular
region, Meckels cartilage and adjacent bone in hematoxylin-eosin
staining. a Survey. Upper frame marks b, middle frame marks c, and
lower frame marks d. Scale bar 1 mm. b Magnified section from a.
Heavy osteoblastic activity around the most dorsal part of the
forming mandibular bone. c magnified section from a. Mental
foramen and inferior alveolar nerve passing through. Moderate
osteoblastic activity at adjacent bone (upper left and lower right). d
Magnified section from a. Heavy to moderate osteoblastic activity of
mandibular bone close to anterior end of Meckels cartilage. Scale
bars for b–d 100 m. e–h Fetus 53 mm CRL, horizontal section,
mandibular region, Meckels cartilage, adjacent bone and dental
primordia in Masson-Goldner (with light green) staining. e Survey.
Fig. 7 Bone remodeling patterns obtained from histological eval-
uation, characterized and color-coded in groups and projected to the
mandibular surface, and arranged at the same scales. a, c Right half
of mandible, buccal aspect, left half of mandible, lingual aspect. b,
d, e Right half of mandible, buccal aspect (left), right half of
mandible, lingual aspect (right). CRL sizes a 25 mm, b 41 mm, c
63 mm, d 76 mm, e 117 mm. Scale bar 2000 m
even in the fetus of 117 mm CRL, the medial aspects of
Meckel’s cartilage were still not covered by the bulging
bony protrusions (Fig. 5g). Only in the most anterior
region, anterior to the mental foramen, was the mandib-
ular body completely ossified after local resorption of
Meckel’s cartilage (Fig. 3i–m).
As early as in the 7th week (embryo of 19 mm CRL) a
bony gutter was formed to embed the inferior alveolar
nerve and its accompanying vessels. This gutter spread
out in a posterior direction, as the bony plates elevated to
either side (Fig. 2c–j), but it did not extend further than
the anterior half of the mandible of the 117 mm CRL fetus
(Fig. 3j–l).
Distances between bone and Meckel’s cartilage was
never less than 50 m (Fig. 6h), only in more posterior
parts was the distance between Meckel’s cartilage and the
bone swinging out into lateral direction increased (Fig. 3).
Frame marks position of f. Scale bar 1 mm. f Dental primordium of
lower left deciduous canine in cap stage, adjacent bone with
moderate osteoblastic activity (left) and osteoclastic activity (top).
Scale bar 100 m. g More caudal survey, frame marks position of h.
Scale bar 250 m. h Magnified section from g. Osteoclastic activity
at bone close to the anterior end of Meckels cartilage. Scale bar
50 m. i and j Fetus 117 mm CRL, frontal section, mandibular
region in Masson-Goldner (with anilin-blue) staining. i Meckels
cartilage (C), mandibular bone (B) and part of the dental primordium
of the lower right deciduous incisor (D). Frame to indicate position
of j. Scale bar 500 m. j Magnified section from i. Osteoclastic
activity towards dental primordium (right), moderate osteoblastic
activity towards mandibular surface (left). Scale bar 100 m

Fig. 8 Scattergram with width
measurements of the mandibles
Fig. 9 Scattergram with length
measurements of the mandibles
Histology and remodeling pattern
Major apposition, revealing more than 5 osteoblasts
counted per grid square (0.25 mm2) at a 160 magnifi-
cation was found in the major part of the mandibular
surface in all specimens (Figs. 6 and 7). No resorption at
all was seen in the embryo of 25 mm CRL (Fig. 6a–d),
with only some sites of lining cells at the medial aspect of
mandibular bone (Fig. 7a). Major apposition, and also
narrow fields of minor apposition (less than 5 osteoblasts
counted per grid square) were found at the buccal and at
the lingual aspects of the mandible in the fetus of 41 mm
CRL (Fig. 7b). In addition, very minor islands of
resorption were seen in the posterior half of the mandible
on either side (Fig. 7b). Although the major mass of the
mandibular body grew by major apposition in the fetus of
53 mm CRL, a more elaborate pattern of minor apposition
areas, resorption fields, and inactive regions was found on
227
the buccal and on the lingual aspects of the mandible.
Along the line where Meckel’s cartilage ran along at the
medial aspect of the mandible, a band of minor apposi-
tion, and even resorption in the most anterior (Fig. 6g, h)
and most posterior parts, could be seen (Figs. 2g, 7c).
Next to dental primordia, resorption and apposition could
be found (Fig. 6e, f). In the fetuses of 76 and 117 mm
CRL the buccal and lingual resorption fields could be
correlated with the buccal and the lingual concavities
leading into the area between the condylar and coronoid
processes (Figs. 1i–k, 7d, e). Bone remodeling was more
variable (Fig. 6i, j), and the areas of resorption became
more and more solitary, smaller, scattered and more
numerous.
From the embryo of 41 mm CRL, the mental foramen,
lying in appositional areas, was bordered by variable
small extensions of resorption fields and lining cells
(Fig. 7b–e).
228
Fig. 10 Scattergram for the
opening angle of the mandibles
Fig. 11 Scattergram for the po-
sition of the mental foramen
Morphometry
The overall shape of the mandible in the horizontal plane
showed a general trend to enlarge (Figs. 8 and 9). The
length of the mandible (measured as MA–MP, and as
Lmand) correlated with overall body growth, although
during the younger stages between 20 and 25 mm CRL
this connection was not quite obvious. The posterior
width (MP–MP) of the mandible increased, as well as the
distance between the mental foraminae on either side
(Fright–Fleft). The transversal diameter of the mandible in
its middle part (C–C) increased only when the older
stages were reached. The anterior distance between the
two halves of the mandibular bone (MA–MA) did not
increase. Up to this part, all the correlations were
significant at p=0.01. The opening angle of the mandible
could not be correlated with overall body growth
(Fig. 10). The extent of the opening angle of the V-
shaped mandibular arch was very variable in our recon-
structed specimens, and the outline changing between V-
shapes, U-shapes, and even a lyrate arrangement did not
reveal a straight developmental pattern. The mental
foramen remained in the anterior quarter of the mandib-
ular body (Fig. 11) and did not move.
Discussion
Crown-rump-length
When we arranged our specimens with increasing size, it
became obvious that in the younger stages maturity did
not coincide with an increase in body size. It is difficult to
extract the exact age from CRL (crown-rump-length)
data, but if one could do so, it would not help much either.
The individual variation of maturity is neither bound to
size nor to age (Kjaer 1989b). The use of the CRL is not
the most precise method, but the most commonly used
(Hinrichsen 1990).

Variety of forms
The relatively large number of fully 3-dimensionally
reconstructed mandibles enabled us to compare a variety
of forms that showed the different shapes during the
developmental stages. On the one hand this revealed the
changes from stage to stage, on the other hand the range
of individual expression of form could be shown as well.
At present, we are unable to distinguish between
“normal” development and individual variation. Our
reconstructions, however, show a variety of forms that
is far beyond the conventional brief and schematic
descriptions known thus far.
Meckel’s cartilage
We found initial formations of Meckel’s cartilage at a size
of 14 mm CRL, which relates to 6th–7th week (Hinrich-
sen 1990). This range is supported by Low (1909),
Glatthor (1963), Durst-Zivkovic and Davila (1974),
Kitamura (1989), Orliaguet et al. (1994), Ten Cate
(1998), and Berkovitz et al. (2002) and Lee SK et al.
(2001) reported early ossifications in the 5th week.
The three typical curves we found could be seen in all
of the stages. The extent of the curvatures, however,
varied from rather straight to extremely zig-zagged in the
specimen 117 mm CRL. Here, we excluded a possible
alignment artifact by checking other structures of the head
in the same range of sections. From these curvatures the
developmental movements (Blechschmidt 1948) and the
pressure of the expanding Meckel’s cartilage (Blech-
schmidt 1973; Sperber 2001) into anterior direction were
concluded.
It is well known that the posterior pegs of Meckel’s
cartilage contribute to the formation of the auditory
ossicles of the middle ear (Moore 1977; Hinrichsen 1990;
Sperber 2001). Our reconstructions that include the
posterior end of Meckel’s cartilage reveal its variability
in size and outline of protrusions.
As long as the anterior ends of Meckel’s cartilage did
not meet, there was no formation of a hamular process.
These hamula were very variable in size, outline and
angle in the cranial direction. In no instance did we find a
fusion of Meckel’s cartilage or mandibular bone in the
midline of the face in our specimen sample, which was
supported and reviewed recently by Rodriguez-Vasquez
et al. (1997).
In the posterior processes of the mandible, carrot-
shaped cartilaginous centers were found. This holds true
for the condylar process in the 53 mm CRL fetus, and for
the coronoid process as well in the 66 mm CRL fetus, and
was confirmed in other studies devoted to the develop-
ment of the temporomandibular joint (Kitamura 1989;
Radlanski et al. 1999a).
There are different views concerning the origin of the
ossicula mentalia in the anterior region of the fetal
mandible. Rodriguez-Vasquez et al. (1997) suggested that
these cartilaginous formations are remnants of Meckels
229
cartilage, but according to Hinrichsen (1990) a special
chondroid tissue is found in this region of which the
ossification mode is not clear. But since it is cartilage
which ossifies, it should not be as desmal, as the
mandibular bone which is formed next to Meckels’s
cartilage. Our study, however, could not give any hint for
the origin of the ossicula mentalia, because they occurred
only in our latest stage in a region where Meckel’s
cartilage has been present in less mature mandibles.
In most regions mandibular bone ran rather close to
Meckel’s cartilage, which was corroborated by Lee SK et
al. (2001). At the posterior ends we observed an increase
in distance between the two tissues. With only few
exceptions was the mandible bent out laterally even
farther than Meckel’s cartilage deviated into a lateral
direction.
Up to now we do not know how the formation of that
specific appearance is controlled. So far we know which
genes are responsible to trigger formation of cartilage
within the first branchial arch (Plant et al. 2000), but
what, if not pure mechanical factors (Blechschmidt 1973),
controls the thickness of Meckel’s cartilage, what leads to
the curvature of Meckel’s cartilage and to the formation
of the posterior manubrium mallei and the anterior
hamulus?
Mandibular ossification and remodeling
The first mandibular ossification was observed by us in
the embryo of 15.6 mm CRL at the branching of the
mandibular nerve in the future region of the mental
foramen. In agreement with others, Berkovitz and Mox-
ham (1988), Kjaer (1990), Lee SK et al. (2001), Testut
(1893), Scott and Dixon (1978), and Ten Cate (1998)
maintain an initial ossification in the region of the later
gonial angle, and Mall (1906) found multiple ossification
centers. However, we found minor insular ossification
centers in embryos larger than 37 mm CRL, predomi-
nantly in the corners underneath the posterior pegs of
Meckel’s cartilage.
The formation of the mental foramen started as a bony
notch through which the inferior alveolar nerve crossed
into the lateral direction. It was found completely
encircled by bone in a specimen of 19 mm, but there
were other specimens in which its circumference re-
mained cranially open until 29 mm CRL. However, as we
could show in a more detailed study recently (Radlanski
et al. 2002), it was reopened in later stages because of
heavy resorption due to the expansion of the adjacent
dental primordium in a fetus of 76 mm CRL in that study,
and in the fetus of 117 mm CRL in this study, before it
was completely formed again in later stages. Maybe this
influence of the adjacent dental primordium is the reason
for the irregular data in the literature concerning the
completed formation that range from 18 mm CRL (Kjaer
1989a; Mller 1995) up to the 4th month (Kvinnsland
1969) and the 7th month (Kirsch 1955) in a radiographic
study.
230
The maximum vertical height of the mandibular bone
moved from the region of the mental foramen in younger
embryos to the posterior third of the mandibular bone, and
the formation of the condylar and coronoid processes at
the end of the mandible was clearly recognized in the
embryo DOR 37 mm CRL, although there were several
smaller specimens where one could interpret bony
lingulae such as primordia. Fawcett (1910) and Kitamura
(1989) described the first formation of the coronoid
process at day 47 and the condylar processes at day 60.
Lee SK et al. (2001), however, did not recognize the
coronoid process before the 14th week. The formation of
the condylar process with its cartilaginous core has been
described by us elsewhere (Radlanski et al. 1999a).
Our reconstructions revealed the first signs of man-
dibular bone penetrating underneath and above Meckel’s
cartilage in an embryo of 29 mm CRL, in a region directly
posterior to the mental foramen. The attachment of the
myelohyoid muscle was at Meckel’s cartilage in earlier
stages and it changed to a bony attachment when bone
emerged underneath Meckel’s cartilage. We could see
specimens that had an attachment of this muscle to
Meckel’s cartilage and to mandibular bone next to each
other (Radlanski et al. 2001). This embracing of Meckel’s
cartilage spreads out longitudinally into both directions,
but never covered the lingual aspect of Meckel’s carti-
lage, except in the anterior region where resorption of
Meckel’s cartilage was observed in the fetus 76 mm CRL
and later. Lee SK et al. (2001) reported that after a close
contact between bone and Meckel’s cartilage in earlier
stages, bone detached from Meckel’s cartilage at 12
weeks into a lateral direction.
The remodeling pattern was simple in earlier stages
where almost ubiquitously apposition was found (Mauser
et al. 1975; Radlanski and Klarkowski 2001). In later
stages, however, an elaborate pattern of apposition and
resorption was found, which could be matched with the
3D outline of bone in most cases. Resorption fields were
found at the posterior margins of the mental foramen and
at the lateral and medial posterior bony planes at concave
surfaces. The bulging of bone underneath and over
Meckel’s cartilage could be recognized as active appo-
sitional growth. Meckel’s cartilage itself lay in a trough
with could be characterized by less apposition and even
resorption from the 53-mm CRL stage onwards.
It might be true that Meckel’s cartilage acts as a frame
for mandibular bone (Sperber 2001) to spread out along
its lateral borders, but bony formations extend further
away from the framing cartilage, and the remodeling
pattern leads to the typical—and within certain limits—
individual appearance of the mandible. Today, we know
much about the chemical processes that lead to bone
formation at the cellular level (Helms et al. 1997;
Thorogood et al. 1998; Schneider et al. 1999, 2001;
Olsen et al. 2000; Depew et al. 2002; Cobourne and
Sharpe 2003; Helms and Schneider 2003; Shui et al.
2003), but what information do the bone forming cells
receive to create those different forms of the mandibles,
as seen in our 3D reconstructions with some being
asymmetrical with varying shapes deviating from the
known schematic pattern.
Morphometry
The overall shape of the mandible in the horizontal plane
showed a general trend to enlarge. Most of the length
measurements in different orientations could be correlated
with overall body growth, although during the younger
embryos between 20 and 25 mm CRL this connection was
not quite obvious. The opening angle of the mandible,
however, could not be correlated with overall body
growth and the extent of the opening angle of the V-
shaped mandibular arch was very variable in our recon-
structed specimens.
This observation no longer supports speculation raised
on a combined radiographic and morphometric study with
different samples that this might be due to a swinging out
and in of the mandible (Kjaer et al. 1999) in relation to the
palatal closure and the descending of the tongue and other
related tissues (Blechschmidt 1960; Sperber 2001). In
addition, the values obtained to describe the bulging in
and out did not show any clear correlation. So in this
region, a more individual developmental pattern seemed
to be obvious.
Future research
The shape of the mandible and its alterations during
development are typical and complicated, and it should be
very interesting to see the signaling patterns that foster
and inhibit growth of the adjacent tissues on the
molecular level. This could not be done with our human
material which is needed, however, as a reference for
comparative embryological studies. Further research in
the animal model should concentrate on bridging the gap
between morphologically oriented embryological re-
search and developmental molecular biology.
Acknowledgements We thank Prof. Dr. G. Steding (Gttingen) and
PD Dr. J. Mnner (Gttingen) for permission to use part of the
Gttingen collection and Mrs. B. Danielowski and Mrs. B. Schwarz
for their technical support in our histological and image analysis
laboratory. Concerning statistical questions we thank PD Dr. Dr.
W. Hopfenmller (Dept of Medical Informatics, University Clinic
Benjamin Franklin) for his assistance. We are indebted to Mrs. Hala
Zreiquat, PhD, for proofreading our manuscript as a native speaker.
Parts of this study have been supported by the Deutsche
Forschungsgemeinschaft (DFG) Ra 428/1–3.
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