Anthrop. Anz. 69/4, pp.

439–460 Article
J. Biol. Clinic. Anthrop.
published online September 2012

The temperature of cremation and its effect on the micro-

structure of the human rib compact bone

Karolı́na Absolonová1, Miluše Dobisı́ková2, Michal Beran3, Jarmila

Zocová4, and Petr Velemı́nský2
1
Department of Anthropology and Human Genetics, Faculty of Science, Charles University,
Praha, Czech Republic
karolina.absolonova@seznam.cz
2
Department of Anthropology, National Museum, Praha, Czech Republic
3
Institute of Legal Medicine, 2nd Faculty of Medicine and Faculty Hospital Na Bulovce in
Prague, Charles University, Praha, Czech Republic
4
Retired person, formerly employed in Faculty of Science, Charles University, Czech Repub-
lic

With 5 figures and 8 tables

Summary: The presented study deals with the effect of the cremation temperature on the
microstructure and morphology of the human compact bone. The biological material con-
sisted of samples from ribs of recent Central European origin belonging to individuals of
known age, sex and cause of death. Each bone sample was divided into several sections. One
section remained unburned and the rest were burned at 700, 800 and 1000 °C. A few samples
were burned also at the temperature of 600 °C. The undecalcified unstained ground cross-
sections were made from burned and unburned bones; photographed and analysed using the
SigmaScan Pro 5 programme. During burning, both the macroscopic and microscopic
dimensions of the bone shrink, including the measures of the individual microstructures. The
percentual representation of the area of individual microstructures on the area of the cross-
section decreases. The number of individual microstructures per mm2 of the compact bone
cross-section increases. Most microstructural variables demonstrated statistically significant
differences at the individual temperatures of cremation. The burned bones showed a large
scale of the colours, especially at 700 °C.

Key words: histomorphometry, human rib, compact bone, burned bone, cremation tempera-
ture.

Introduction
In the physical or forensic anthropology, osteology and archaeology, we frequently
come across burned bones. Their analysis is very difficult as they undergo heat-
induced changes on both the macroscopic and microscopic level. Many chemical and
physical changes occur in the bones during burning, and it leads to the shrinkage of
the bones, changes in histological structure, colour, firmness and other characteris-
tics (Table 1). The bones become fragile, deformed and cracked. Shrinkage of burned
bones may lead to incorrect conclusions if methods designed for unburned bones are

쏘 2012 E. Schweizerbart’sche Verlagsbuchhandlung, Stuttgart, Germany www.schweizerbart.de

DOI: 10.1127/0003-5548/2012/0213 0003-5548/12/0213 $ 5.50
eschweizerbart_xxx
 440

Table 1. Changes in bone tissue depending on the temperature of cremation (Herrmann 1977, Schutkowski 1991, Harsányi 1993, Holden et al. 1995b,
Dokládal 1999, Thompson 2004, 2005, Hanson & Cain 2007, Walker et al. 2008, Ubelaker 2009, Harbeck et al. 2011).
Temperature Bone colour Mechanical Chemical composition Bone appearance Bone shrinkage Degree of
strength of bone burning
100 °C Yellow, Unchanged Shortening of collagen Appearance nearly like un- Approx. by 1 % I
200 °C brownish-grey, fibrils, loss of the ma- burned bone, first shrinkage Imperfect
Karolı́na Absolonová et al.

dark brown jority of collagen, out- and fractures, accumulation

flow of crystallic water of carbon in the osteocyte la-
from the bone mineral cunae, most microstructures
component, organic unchanged

eschweizerbart_xxx
300 °C Brown to black Decreases matter persists Carbonisation of organic II
400 °C Dark brown, Carbon oxidation, loss substances, the bone is im- Transitory Partly imper-
black of water content perfectly burned, changes in expansion fect
bone porosity, brownish-
black discolouration of the
lamellae, fissures extend
from the Haversian canals
500°C Greyish, milky Crystal size begins to The internal surface of the 0% III
light grey, grey- increase, most of the compact bone is still black in Perfect
blue carbon is oxidised some cases, on cross-section
it is light with many fissures
 Table 1. Continued.
Temperature Bone colour Mechanical Chemical composition Bone appearance Bone shrinkage Degree of
strength of bone burning
600 °C Milky white, Carbon oxidation and Chalky surface, the bone is 0% IV
chalky dull, grey CO2 emission continue, poorly mechanically resis- Perfect to
loss of the rest of colla- tant, many fissures are pre- chalky
gen, mineral recrystalli- sent on cross-section, micro-
sation begins structures still visible
700 °C Ranging from Chalky Apatite crystals lose wa- Smooth surface, the bone is From 750 °C,
dark grey to structure ter, crystals enlarge hard and fragile, develop- shrinkage continues
white ment of parabolic cracks
800 °C White, pink Increases Cremation is complete, Maximal shrinkage, the mi- 10–30% V

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CO2 emission ends, or- crostructures of the compact Chalky
ganic tissue has been bone are gradually lost as a
burned, formation of result of changes incurred by
new crystals crystals
900 °C Intense white, Chemical reactions of
1000 °C white even on solid particles, beta-tri-
breakage; calcium phosphate
purple tint forms, fusion of crys-
when burned in tals, recrystallisation of
air minerals, sintering
1600 °C Melting of bone miner-
al, recrystallisation after
Effect of cremation temperature on human compact bone

cooling
441
442 Karolı́na Absolonová et al.

used in their analysis. This is especially important when estimating age at death of the
individual, determining sex, species affiliation etc. This is why in our work we set out
to study the macroscopic and especially the microscopic changes that are induced in
bones by heat and that may affect their successful analysis. Today, the analysis of
burned bones is important especially for the identification of victims of natural disas-
ters, traffic and aircraft accidents, terrorist attacks etc. Burned human bones also
occur frequently in archaeological finds, as in certain cultures the cremation was for
long periods of time the sole or predominant form of burial.
We studied the changes induced by burning on the compact bone of the normal
human rib using light microscopy. In this approach we followed some previous
works, in which the effect of experimentally controlled heating on a sample of mod-
ern human or animal bone was studied, and the burned bones were compared with
unburned specimens (Herrmann 1976, Herrmann 1977, Swegle 1979, Bradtmiller &
Buikstra 1984, Shipman et al. 1984, Hummel & Schutkowski 1986, 1993, Nelson
1992, Harsányi 1993, Cattaneo et al. 1999, Hanson & Cain 2007).
In these studies the bones were burned at various temperatures because the effects
of different temperatures on the bone tissue are diverse. For example, at temperatures
below 700 °C the bones are incompletely cremated and their histological structure is
nearly identical to that of native bone (Herrmann 1977, Holden et al. 1995b). On the
other hand, at temperatures of around 1000 °C the microstructures of compact bone
are often very difficult to recognize or disappear entirely (van Vark 1970, Heussner
1987, Heussner 1990a, Heussner 1990b, Harsányi 1993, Hummel & Schutkowski
1993, Stloukal et al. 1999, Hanson & Cain 2007, Harbeck et al. 2011, Squires et al.
2011). With regard to the state of the burned bones and the degree of burning, Herr-
mann determined the critical level of 700–800 °C, at which the greatest changes
occur in the bone and above it the cremation is complete (Herrmann 1977).
Some of the studies also analyse the shrinkage of bones caused by burning, which
considerably complicates the analysis of burned bones (van Vark 1970, Herrmann 1976,
Herrmann 1977, Swegle 1979, Bradtmiller & Buikstra 1984, Shipman et al. 1984, Hum-
mel & Schutkowski 1986, Schutkowski 1991, Nelson 1992, Cattaneo et al. 1999,
Thompson 2005, Ubelaker 2009). The shrinkage is affected by the temperature and
duration of heat exposure and also by the morphology of the bone (the proportion of
compacta and spongiosa in the bone). The main cause of shrinking is the transformation
of the crystal structure of the bone during burning (Herrmann 1977, Thompson 2005).
Under the critical level of 700–800 °C the shrinkage of the bone is only about 1 to 2 %,
but above this level it can reach up to 30 % (Herrmann 1976, Herrmann 1977, Hummel
& Schutkowski 1986). The amount of longitudinal shrinkage depends on the length of
the bone sample, i.e. the longer pieces of bones shrink to a lesser extent than the shorter
ones. The degree of transverse shrinkage of the compact bone is determined by its thick-
ness. The shrinkage of the thicker compacta is less than the shrinkage of the thinner one
(Hummel & Schutkowski 1986, Schutkowski 1991).
The colour of burned bones is also important, because it is frequently used to estimate
the degree of burning (van Vark 1970, Herrmann 1977, Shipman et al. 1984, Schutkow-
ski 1991, Holden et al. 1995a, Holden et al. 1995b, Stiner et al. 1995, Quatrehomme et
al. 1998, Hanson & Cain 2007, Walker et al. 2008, Ubelaker 2009, Harbeck et al. 2011).
The colour is influenced by the temperature and duration of fire, by the oxygen availabil-
ity during burning and by some other conditions in the cremation environment, for
example the presence of organic materials in the surrounding soil (Stiner et al. 1995,

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 Effect of cremation temperature on human compact bone 443

Walker et al. 2008). During burning at temperatures below 700 °C the bone changes
its colour from brown through black to grey depending on the combustion of organic
carbon, which is completely removed from the bone at 800 °C. At the same time the
hydroxyapatite crystal structure is converted into beta-tricalcium phosphate, and the
colour of the bone turns white (Herrmann 1977, Walker et al. 2008, Harbeck et al.
2011). The colour of burned bones can also be depicted by UV-fluorescence and it
can be used to estimate the cremation temperature (Harbeck et al. 2011).
In some studies the burned bones were analysed using scanning electron micros-
copy (Herrmann 1976, Herrmann 1977, Shipman et al. 1984, Holden et al. 1995a,
1995b, Quatrehomme et al. 1998, Ubelaker 2009). At this level of observation we can
estimate the duration of fire and the temperature of cremation with an accuracy of
within 200 °C based on the formation, morphology and the size of the crystals of
bone mineral, and the state and orientation of the collagen fibres. At this ultrastruc-
tural level some heat-induced changes also correspond to the age of the individual
(Holden et al. 1995a).
Some works describe the structural changes in the mineral portion of the bone
related to the degree of burning using infrared spectroscopy and X-ray diffraction
techniques (Shipman et al. 1984, Holden et al. 1995b, Stiner et al. 1995, Rogers &
Daniels 2002, Hiller et al. 2003, Hanson & Cain 2007, Piga et al. 2008, Thompson et
al. 2009, Harbeck et al. 2011). These methods can be used to estimate the tempera-
ture of the fire and duration of burning (Shipman et al. 1984) as well as to differenti-
ate burned bone from non-burned bone (Hanson & Cain 2007) based on the bone
crystal structural characteristics and composition, and the C/P and C/C ratios
(Thompson et al. 2009).
The heat-induced changes in bone tissue can also be visualised by Magnetic Reso-
nance Imaging (Thompson & Chudek 2007). This technique provides 3D pictures of
the entire bone, thus it is non-destructive but still allows the internal structure of the
bone to be studied. Moreover, with this technique we can study the same section of
bone subjected to continued burning rather than comparing images of different bone
sections burned at different temperatures.
Currently we can also come across trace analyses of burned bones such as DNA or
stable isotope analyses (Grupe & Hummel 1991, Brown et al. 1995, Cattaneo et al.
1999, Pusch et al. 2000, Walker et al. 2008, Ubelaker 2009, Harbeck et al. 2011).
These methods can yield additional information about past populations or help to
solve a lot of criminal and civil cases. Nevertheless, these methods also have certain
limitations, because a stable isotope analysis of the light elements (C, N and O) is
only possible up to a cremation temperature of 200 °C, and the DNA can only be ana-
lysed in bones burned at temperatures of up to 600–700 °C depending on the dura-
tion of heat exposure (Harbeck et al. 2011).

Material and methods

In this study we used bone samples from the sternal third of the 3rd to 10th rib from individuals
of known age, sex and cause of death (Table 2). The samples originated from a recent Central
European population. All the individuals died unexpectedly, having been completely healthy
before their death, and they did not suffer from any chronic disease affecting the bone micro-
structure. The skeletal material was kindly provided by the Institute of Legal Medicine, 2nd
Faculty of Medicine and Faculty Hospital Na Bulovce in Prague, Charles University, Prague,

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444 Karolı́na Absolonová et al.

Table 2. Study samples, variables: age at death (N – number of individuals, SD – standard

deviation, SE – standard error). Testing of normal distribution using the Shapiro-Wilk’s W
test, skewness and kurtosis.
N Mean Age Confi- Confi- Median Vari- SD SE
Age Range dence dence ance
–95,000 % +95,000 %
Unburned 49 59.86 19–95 53.89 65.82 63.00 431.500 20.773 2.97
700 °C 36 58.36 19–95 51.15 65.58 62.50 454.637 21.322 3.55
800 °C 60 62.72 20–95 58.12 67.31 63.50 316.715 17.796 2.30

Coefficient of Skewness Kurtosis Shapiro- Shapiro-Wilk’s

Variation (%) Wilk’s W test p
(α = 0.05)
Unburned 34.7 –0.42 –0.68 0.96 0.0620
700 °C 36.5 –0.28 –0.90 0.96 0.2766
800 °C 28.4 –0.42 –0.50 0.97 0.1629

Czech Republic. The biological material was acquired in accordance with the valid laws of
the Czech Republic.
We selected the rib in view of the other analysis that may be conducted on it. Ribs are not
affected by the biomechanical loads during locomotion or by the childbirth (Stout 1992,
Dudar et al. 1993, Kim et al. 2007, Pavón et al. 2010, Cannet et al. 2011); they are not usually
used in osteology so samples for cross-sections may be taken without fear of the damage of
the bone (Stout 1992, Stout & Paine 1992, Kim et al. 2007); the ribs are easily accessed
thanks to their position beneath the surface of the body (Stloukal et al. 1999, Streeter 2010);
and they are also well identifiable when mixed with other burned bones (Stloukal et al. 1999).
The sequence of the individual ribs was not taken into consideration during the research,
because the compact bone remodelling dynamics is comparable amongst individual ribs and
thus there is no need for their exact identification (Pirok et al. 1966, Frost 1969, Crowder &
Rosella 2007, Pavón et al. 2010). This may be advantageous in the analysis of fragmented
archaeological findings or in some forensic cases.
Every bone sample was divided into several sections. One section remained unburned and
the rest were burned at ambient temperatures of 700, 800 and 1000 °C in the Linn Elektro
Therm TC 805 electric furnace (Linn Elektro Therm GmbH, Bad Frankenhausen, Germany).
Some bone samples (n = 6) were burned also at the temperature of 600 °C. The bones select-
ed for burning were neither dehydrated nor degreased, and there was no adherent soft tissue
on them. The bone samples were placed in a cold furnace, and the increase in temperature in
the furnace to the target value occurred over 30 minutes. Burning at the target temperature
lasted a further 30 minutes and the cooling of the furnace including the bones lasted approxi-
mately 24 hours.
We selected the temperatures of cremation of 700 and 800 °C as the lower and upper limit
of the critical level set by Herrmann, when the greatest changes should be incurred by bones
(Herrmann 1977). We also selected the range of temperature with regard to the analysis of the
skeletal remains of victims of domestic house fires and crimes, of remains from crematoria
and last but not least also of archaeological findings. Temperatures of 700, 800 and 1000 °C
occur during domestic house fires (Holden et al. 1995a, 1995b, Quatrehomme et al. 1998)
and are used in modern crematoria (Heussner 1990a, Dunlop 2004, Ubelaker 2009, Harbeck
et al. 2011). Prehistoric cremations could have involved temperatures from 500 to 900 °C and
exceptionally up to 1000 °C (Heussner 1987, Heussner 1990a, Heussner 1990b, Schutkowski
1991, Harsányi 1993, Holden et al. 1995a, 1995b, Dokládal 1999, Hanson & Cain 2007, Har-
beck et al. 2011). An outdoor fire lighted on a wooden pyre, which may be associated with
criminal activity leading to the disposal of the victim’s body reaches the temperatures

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 Effect of cremation temperature on human compact bone 445

between 400 and 1000 °C (Heussner 1990a, Stiner et al. 1995, Hanson & Cain 2007, Ubela-
ker 2009). Experimental burning of bones on an outdoor fire and in a laboratory furnace
demonstrated that both types of burning lead to similar types of changes of bone microstruc-
ture and that we may apply the findings acquired during experimental burning of bones on
cases from everyday practice (Holden et al. 1995a, Hanson & Cain 2007).
Undecalcified and unstained ground cross-sections were made from the burned as well as
unburned bones. The unburned bones were first degreased and dehydrated using ascending
concentrations of ethanol (70 %, 90 %, 96 %, 100 %). The samples were embedded in the
Dentacryl two-component methylmetacrylate casting resin (SpofaDental Inc., Prague, Czech
Republic) and the cross-sections were removed using the diamond IsoMet™ 1000 Precision
Saw (Buehler Instruments, Ltd., Lake Bluff, IL, USA). The sections were glued to the slide
using the SuperBond Liquid-Loctite superglue (Henkel AG & Co. KGaA, Düsseldorf, Ger-
many) and once the glue had dried manually ground using water-resistant abrasive grinding
papers to the required thickness of approximately 100 μm. The ground sections were further
dehydrated in ascending concentrations of ethanol (70 %, 96 %, 100 %), cleared in xylene
and mounted in Canada balsam.
The ground sections were photographed at 100× magnification (10× lens and 10× eye-
piece) in the Olympus BX-50 light microscope with the digital Olympus C-7070 Wide-
Zoom camera (Olympus Europa Holding GmbH, Hamburg, Germany). The photographed
microscopic field was square-shaped, measuring 1859 × 1394 μm and included the whole
thickness of the compact bone including the part of the medullar cavity. The photographs
were adjusted and processed using the QuickPHOTO CAMERA 2.2 programme (Pro-
micra, Ltd., Prague, Czech Republic). The microphotographs of the ground sections were
analysed using the SigmaScan Pro 5 image analyser (Systat, Inc., Richmond, CA, USA).
One to four microscopic fields were analysed in each bone sample depending on the state
of bone preservation. On each long side of the cross-section there were analysed maxi-
mally two fields of view. The macroscopic and microscopic appearance, colour, firmness
and state of preservation, all of which are known to change under the influence of heat
(Table 1), were studied in all the bones. The following microstructural variables were eval-
uated in the compact bone:
1. Compact bone thickness (TL_KOM; μm): Measured in the observed field five times at
regular intervals and averaged.
2. Number of intact osteons per mm² (POC_OST; #/mm²): Number of intact osteons per
field / area of compact bone within the field * 1,000,000. An intact osteon is surrounded
by the concentric lamellae with the reversal line on its periphery, and has 100 % of the cir-
cumference of its Haversian canal intact. This variable involves all intact osteons includ-
ing those located on the periphery of the field if their Haversian canals are completely vis-
ible in the observed field.
3. Percentage of the total osteon area (PR_OST; %): The total area of the osteons per field
/ area of the compact bone within the field * 100. This measurement includes the area of
all types of intact osteons within the field, including those visible only partially on the
periphery of the field.
4. Osteon area (PL_OST; μm²): This is the average area of the intact osteon, including the
area of the Haversian canal. This and the following variables (no. 4–11) relate only to
osteons whose whole surface is visible in the observed field, that have been cut transver-
sally and not obliquely, that are not too elongated or drifting and that have only one Haver-
sian canal. The other types of osteons including those that are only partially visible at the
periphery of the field are included only in the number of intact osteons per mm² (variable
no. 2) and the percentage of the total osteon area (variable no. 3).
5. Osteon perimeter (OBV_O; μm).
6. Maximal osteon axis (MAX_O; μm).
7. Minimal osteon axis (MIN_O; μm).
8. Osteon diameter (PRUM_O; μm): This variable is calculated from the previous two vari-
ables according to the formula (MAX_O + MIN_O)/2

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446 Karolı́na Absolonová et al.

9. Osteon feret diameter (F_PR_O; μm): This is the diameter of the osteon if for the same
area it was shaped like a circle. This variable is calculated directly by the SigmaScan Pro
5 programme.
10. Osteon shape factor (SFAC_OST): This variable describes the shape of the object. A cir-
cle has a shape factor of 1; a straight line has a shape factor of 0. This variable is also cal-
culated directly by the SigmaScan Pro 5 programme.
11. Osteon compactness (C_OST): This value also describes the shape of the object. A cir-
cle has a compactness of 12.57, while the compactness of a straight line approaches
infinity. This variable is calculated directly by the SigmaScan Pro 5 programme as well.
12. Percentage of the total Haversian canal area (PR_KAN; %): Total Haversian canal area
in the field / compact bone area * 100.
13. Haversian canal area (PL_KAN; μm²): This and the following variables (no. 13–20)
relate only to those Haversian canals that are not twice or more longer than wider and
that are not deformed by the outlets of several Volkmann canals.
14. Haversian canal perimeter (OBV_K; μm).
15. Maximal Haversian canal axis (MAX_K; μm).
16. Minimal Haversian canal axis (MIN_K; μm).
17. Haversian canal diameter (PRUM_K; μm).
18. Haversian canal feret diameter (F_PR_K; μm).
19. Haversian canal shape factor (SFAC_KAN).
20. Haversian canal compactness (C_KAN).
21. Haversian canal perimeter / osteon perimeter (OK_OO).
22. Haversian canal area / osteon area (PK_PO).
23. Haversian canal diameter / osteon diameter (PRK_PRO).
Three groups of individuals were analysed in order to describe the microstructural
changes induced by burning: a group of unburned bones and two groups of bones burned at
700 and 800 °C, respectively (Table 2). In view of the considerable damage incurred by the
compact bone, it was not possible to conduct an adequate histological analysis of bones
burned at 1000°C, and these samples were not included into the statistical analysis. Statistical
analysis was performed using the STATISTICA analysis software system, version 6.0 (Stat-
Soft, Inc., Tulsa, OK, USA). For each sample, data relating to the histomorphological vari-
ables were averaged from one to four microscopic fields according to the state of bone preser-
vation. T-tests (T-test for independent samples by groups, T-test for dependent samples)
were used for the comparison of samples (Hill & Lewicki 2007). Normality of sample distri-
bution was tested using a histogram, the Shapiro-Wilk’s W test, skewness and kurtosis (Hill
& Lewicki 2007). Unless stated otherwise, all statistical operations were conducted at the 5 %
level of significance.

Results
We evaluated the burned and unburned bones at the macroscopic and microscopic
level with respect to their overall state, colour, deformation, shrinkage and changes of
the microscopic structure. All changes induced in the bones by burning play a key
role when applying methods leading to the identification of individuals (Table 1).
The burned bones were quite well preserved, but their degree of damage increased
proportionally with the temperature of cremation (Figs 1–5). The bones were shrunk,
contorted, deformed and cracked. In some cases, the compact bone was peeling and
its edges were sometimes upturned. The bones were fragile but sufficiently firm for
the production of ground cross-sections. The burned bones also showed an extensive
scale of colours. Bones burned at 600 °C were intermediately dark grey with no dis-
tinctive bluish hue. Bones burned at 700 °C showed the greatest range of colours;
they were bright white, greyish-white, bluish-grey and dark grey, sometimes with a

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 Effect of cremation temperature on human compact bone 447

Fig. 1. Unstained undecalcified ground cross-section of human rib at 100× magnification;

20-year-old man; unburned bone.

Fig. 2. Unstained undecalcified ground cross-section of human rib at 100× magnification;

20-year-old man; bone burned at 600°C.

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448 Karolı́na Absolonová et al.

Fig. 3. Unstained undecalcified ground cross-section of human rib at 100× magnification;

20-year-old man; bone burned at 700°C.

Fig. 4. Unstained undecalcified ground cross-section of human rib at 100× magnification;

20-year-old man; bone burned at 800°C.

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 Effect of cremation temperature on human compact bone 449

Fig. 5. Unstained undecalcified ground cross-section of human rib at 100× magnification;

20-year-old man; bone burned at 1000 °C.

blue hue. Some of them had grey streaks, which at times were very dark or even
black. The spongiosa was usually without the orange hue but often it contained the
remnants of the soft tissues. The position of the samples in the furnace did not affect
the bone colour, as variously coloured bones were situated side by side in the furnace.
Bones burned at 800 and 1000 °C were white, sometimes with a yellow tint, and their
spongiosa in some cases had a shade of orange. Bones burned at 800 °C sometimes
had the small black and grey dots on their surface, which have never occurred in the
bones burned at 1000 °C.
At the microscopic level, we analysed individual microstructures and their
changes induced by burning (Tables 3–5). Microstructures in the bones were appar-
ent at all employed temperatures of cremation, including the temperature of 1000 °C.
Nevertheless, the microstructures in burned bones were difficult to identify and dif-
ferentiate from each other. The only microstructures that could be reliably identified
were the intact osteons and the Haversian canals. From this reason we focused our
analysis only on these two microstructures. Certain other researchers working with
burned bones came up with similar findings (Hummel & Schutkowski 1993). Most
structures, such as osteon fragments, merged and it was not possible to evaluate them.
The circumferential lamellae were often completely destroyed or could not be identi-
fied. It was difficult to differentiate some microstructures, mainly non-Haversian
canals and resorption cavities, from heat-induced damage. The concentric lamellae
of the osteons merged and fused and could not be clearly distinguished from each
other. The microscopic appearance of the samples burned at 600 °C was more or less
similar to that of unburned bones, and the heat-induced damage was not too great.

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450 Karolı́na Absolonová et al.

Table 3. Descriptive statistics of histological variables in unburned bones (N = 49). For

abbreviations of variables see the Material and methods section.
Mean Median Minimum Maxi- Coefficient Standard Standard
mum of Variation Deviation Error
(%)
TL_KOM 517.60 481.26 241.80 1054.93 37.7 195.251 27.89
POC_OST 14.09 14.05 4.88 22.11 27.8 3.913 0.56
PL_OST 28 089.85 27 520.24 14 521.66 49 045.42 31.3 8802.393 1257.49
PR_OST 39.80 39.92 18.70 59.41 26.7 10.639 1.52
OBV_O 654.18 655.97 472.82 926.80 16.3 106.778 15.25
MAX_O 230.32 227.42 165.45 355.10 16.9 39.013 5.57
MIN_O 157.00 159.91 101.96 207.73 16.3 25.617 3.66
PRUM_O 193.66 193.94 139.43 267.93 16.1 31.166 4.45
F_PR_O 179.30 178.97 128.77 243.24 15.9 28.496 4.07
PL_KAN 2172.26 1922.08 1027.87 7645.06 49.1 1066.018 152.29
PR_KAN 3.24 2.95 1.19 6.58 36.9 1.194 0.17
OBV_K 166.12 163.01 120.13 249.67 16.5 27.398 3.91
MAX_K 59.25 57.86 44.04 88.05 17.1 10.119 1.45
MIN_K 39.97 39.13 26.45 63.76 17.2 6.860 0.98
PRUM_K 49.61 49.01 35.25 75.77 16.5 8.207 1.17
F_PR_K 46.92 46.31 32.95 71.78 16.4 7.702 1.10
OK_OO 0.26 0.25 0.20 0.41 15.4 0.040 0.01
PK_PO 0.08 0.07 0.05 0.28 48.8 0.039 0.01
PRK_PRO 0.26 0.26 0.20 0.41 15.4 0.040 0.01
C_OST 16.93 16.87 15.38 18.91 4.0 0.681 0.10
C_KAN 16.00 15.94 15.22 17.98 3.6 0.583 0.08
SFAC_OST 0.75 0.75 0.69 0.82 3.6 0.027 0.00
SFAC_KAN 0.79 0.80 0.71 0.83 3.0 0.024 0.00

As we can see in Tables 3 to 5, some variables had a higher standard deviation than
others, i.e. compact bone thickness (TL_KOM), the osteon area (PL_OST), the oste-
on perimeter (OBV_O) and the Haversian canal area (PL_KAN). Therefore we
excluded these four variables from further statistical analyses, because we cannot
reliably say, whether the differences between these variables in burned and unburned
bones are caused by temperature or by their high standard deviations. Nevertheless,
not taking into account the reasons for this behaviour, these four variables decreased
with an increasing cremation temperature.
The groups of unburned bones and bones burned at 700 and 800 °C were com-
pared using T-tests for independent samples by groups (Tables 6–8). We used this
type of T-test because the proportion of individuals in the analysed groups was not
completely identical because of the damage incurred by the bones during burning
and the subsequent processing (Table 2).
The groups of unburned bones and bones burned at 700 °C differed in 13 variables
(Table 6). The both groups did not differ in the percentage of the total Haversian
canal area, Haversian canal perimeter / osteon perimeter, Haversian canal area / oste-
on area, Haversian canal diameter / osteon diameter, Haversian canal compactness
and Haversian canal shape factor.

eschweizerbart_xxx
 Effect of cremation temperature on human compact bone 451

Table 4. Descriptive statistics of histological variables in bones burned at 700 °C (N = 36).

For abbreviations of variables see the Material and methods section.
Mean Median Minimum Maxi- Coefficient Standard Standard
mum of Variation Deviation Error
(%)
TL_KOM 372.85 356.39 147.26 826.81 39.9 148.933 24.82
POC_OST 16.85 16.53 3.80 33.95 33.7 5.684 0.95
PL_OST 21 504.71 20 328.15 11 154.08 46 046.16 34.4 7395.518 1232.59
PR_OST 35.18 34.57 14.51 52.90 29.8 10.468 1.75
OBV_O 591.78 582.30 427.87 900.72 17.7 104.927 17.49
MAX_O 209.93 201.39 157.19 316.06 18.1 37.970 6.33
MIN_O 133.42 130.74 91.41 195.44 17.7 23.659 3.94
PRUM_O 171.67 168.55 124.30 255.75 17.1 29.421 4.90
F_PR_O 157.41 155.20 112.88 231.79 16.7 26.285 4.38
PL_KAN 1659.02 1448.45 787.91 3906.77 42.0 697.335 116.22
PR_KAN 2.99 2.80 0.82 6.19 39.6 1.183 0.20
OBV_K 148.14 144.72 104.17 226.49 18.3 27.179 4.53
MAX_K 54.05 51.82 39.37 84.55 19.1 10.348 1.73
MIN_K 34.17 33.08 21.95 55.97 21.0 7.189 1.20
PRUM_K 43.98 42.83 30.66 67.16 18.5 8.126 1.35
F_PR_K 41.74 40.71 28.79 64.63 18.7 7.799 1.30
OK_OO 0.25 0.25 0.20 0.32 14.0 0.035 0.01
PK_PO 0.08 0.07 0.04 0.14 28.8 0.023 0.00
PRK_PRO 0.26 0.25 0.20 0.33 13.9 0.036 0.01
C_OST 17.89 17.81 16.25 19.88 5.3 0.945 0.16
C_KAN 16.02 15.93 14.78 19.41 5.8 0.936 0.16
SFAC_OST 0.72 0.72 0.66 0.78 4.7 0.034 0.01
SFAC_
KAN 0.80 0.80 0.68 0.85 4.6 0.037 0.01

The groups of bones burned at 700 and 800 °C also differed in 13 variables
(Table 7). These groups did not differ in the percentage of the total Haversian canal
area, Haversian canal perimeter / osteon perimeter, Haversian canal area / osteon
area, Haversian canal diameter / osteon diameter, osteon compactness and osteon
shape factor. Apart from compactness and shape factor of osteons and Haversian
canals, bones burned at 700 and 800 °C did not differ in the same variables that also
did not differ between unburned bones and bones burned at 700 °C.
The groups of unburned bones and bones burned at 800 °C differed in 16 variables
(Table 8), and did not differ only in Haversian canal perimeter / osteon perimeter,
Haversian canal area / osteon area and Haversian canal diameter / osteon diameter.
From these data we can deduce that the greatest changes in the bones occur around
the temperature of 800 °C, i.e. around the upper limit of Hermann’s critical level
(Herrmann 1977).
The groups of unburned bones and bones burned at 700 °C and 800 °C were also
compared using the T-test for dependent samples (data not shown in the tables), in
such a way that pairs of the same individuals represented in both compared groups
were selected for testing. This test described the effect of burning in the same individ-
uals, and the influence of the group’s varied composition was eliminated.

eschweizerbart_xxx
452 Karolı́na Absolonová et al.

Table 5. Descriptive statistics of histological variables in bones burned at 800 °C (N = 60).

For abbreviations of variables see the Material and methods section.
Mean Median Minimum Maxi- Coefficient Standard Standard
mum of Variation Deviation Error
(%)
TL_KOM 286.18 269.44 127.38 570.93 36.4 104.254 13.46
POC_OST 20.99 20.35 7.69 42.48 32.5 6.811 0.88
PL_OST 14 926.05 14 051.97 8175.84 30 398.46 31.6 4710.255 608.09
PR_OST 30.82 31.22 11.84 58.52 33.6 10.340 1.34
OBV_O 491.44 478.27 349.82 718.98 15.3 75.304 9.72
MAX_O 178.55 176.85 126.13 254.32 15.3 27.397 3.54
MIN_O 109.34 106.23 76.96 170.45 16.9 18.507 2.39
PRUM_O 143.94 140.49 103.45 210.21 15.2 21.935 2.83
F_PR_O 131.73 128.79 97.53 190.17 15.1 19.921 2.57
PL_KAN 1211.19 1095.66 487.59 2895.07 42.1 510.410 65.89
PR_KAN 2.58 2.53 0.82 6.03 43.4 1.120 0.15
OBV_K 126.32 126.21 86.99 183.23 16.7 21.061 2.72
MAX_K 46.87 47.00 32.42 65.43 16.6 7.785 1.01
MIN_K 27.43 26.91 17.71 42.18 19.1 5.249 0.68
PRUM_K 37.15 36.80 25.32 53.81 16.9 6.288 0.81
F_PR_K 34.67 34.82 23.48 51.17 17.2 5.973 0.77
OK_OO 0.26 0.26 0.17 0.40 18.5 0.048 0.01
PK_PO 0.08 0.08 0.03 0.20 43.8 0.035 0.00
PRK_PRO 0.26 0.26 0.17 0.41 18.9 0.049 0.01
C_OST 17.76 17.69 15.92 20.71 5.7 1.006 0.13
C_KAN 16.99 16.84 15.50 18.92 4.8 0.823 0.11
SFAC_OST 0.72 0.72 0.62 0.79 5.0 0.036 0.01
SFAC_
KAN 0.75 0.76 0.69 0.81 3.9 0.029 0.00

The groups of unburned bones and bones burned at 700 °C (n = 36; average age
58.47; age range 19–95; standard deviation 22.361; standard error 3.73; coefficient
of variation 38.24) differed at the 0.01 % level of significance (p < 0.0001) in a total
of 11 variables. Maximal osteon axis (p = 0.0002) and the percentage of the total
Haversian canal area (p = 0.0003) differed at the 0.1 % level of significance. The
number of intact osteons per mm² (p = 0.0141) differed in both groups at the 5 % lev-
el of significance. The groups did not differ in Haversian canal perimeter / osteon
perimeter, Haversian canal area / osteon area, Haversian canal diameter / osteon
diameter, Haversian canal compactness and Haversian canal shape factor. Apart from
the percentage of the total Haversian canal area, the variables in which these groups
do not differ are the same as when using the T-test for independent samples by
groups.
The groups of bones burned at 700 and 800 °C (n = 28; average age 63.1; age
range 20–95; standard deviation 20.024; standard error 3.78; coefficient of variation
31.71) differed at the 0.01 % level of significance (p < 0.0001) in the number of intact
osteons per mm², minimal osteon axis, Haversian canal compactness and Haversian
canal shape factor. At the 0.1 % level of significance, they differed in osteon diameter
(p = 0.0003), osteon feret diameter (p = 0.0002), minimal Haversian canal axis

eschweizerbart_xxx
 Effect of cremation temperature on human compact bone 453

Table 6. T-test for independent samples by groups for unburned bones versus bones burned
at 700°C; df = 83. (SD – standard deviation; * – significant at p < 0.05; ** – significant at p
< 0.01; *** – significant at p < 0.001; **** – significant at p < 0.0001). For abbreviations of
variables see the Material and methods section.
Mean Mean Difference p SD SD F-ratio p
Un- 700 °C in % com- Un- 700 °C Vari- Vari-
burned N = 36 pared to burned N = 36 ances ances
N = 49 unburned N = 49
AGE 59.86 58.36 – 0.7464 20.773 21.322 1.05 0.8557
POC_OST 14.09 16.85 +19.6 0.0095** 3.913 5.684 2.11 0.0166
PR_OST 39.80 35.18 –11.6 0.0498* 10.639 10.468 1.03 0.9313
MAX_O 230.32 209.93 –8.9 0.0183* 39.013 37.970 1.06 0.8770
MIN_O 157.00 133.42 –15.0 0.0000**** 25.617 23.659 1.17 0.6284
PRUM_O 193.66 171.67 –11.4 0.0015** 31.166 29.421 1.12 0.7289
F_PR_O 179.30 157.41 –12.2 0.0005*** 28.496 26.285 1.18 0.6229
PR_KAN 3.24 2.99 –7.7 0.3319 1.194 1.183 1.02 0.9668
OBV_K 166.12 148.14 –10.8 0.0036** 27.398 27.179 1.02 0.9726
MAX_K 59.25 54.05 –8.8 0.0229* 10.119 10.348 1.05 0.8745
MIN_K 39.97 34.17 –14.5 0.0003*** 6.860 7.189 1.10 0.7539
PRUM_K 49.61 43.98 –11.3 0.0023** 8.207 8.126 1.02 0.9630
F_PR_K 46.92 41.74 –11.0 0.0031** 7.702 7.799 1.03 0.9237
OK_OO 0.26 0.25 –3.8 0.4417 0.040 0.035 1.32 0.3921
PK_PO 0.08 0.08 0.0 0.6345 0.039 0.023 2.77 0.0022
PRK_PRO 0.26 0.26 0.0 0.7540 0.040 0.036 1.22 0.5380
C_OST 16.93 17.89 +5.7 0.0000**** 0.681 0.945 1.93 0.0347
C_KAN 16.00 16.02 +0.1 0.9011 0.583 0.936 2.58 0.0024
SFAC_OST 0.75 0.72 –4.0 0.0000**** 0.027 0.034 1.66 0.1009
SFAC_
KAN 0.79 0.80 +1.3 0.6401 0.024 0.037 2.37 0.0056

(p = 0.0002) and Haversian canal feret diameter (p = 0.0005). At the 1 % level of sig-
nificance, both groups differed in maximal osteon axis (p = 0.0011), Haversian canal
perimeter (p = 0.0022), maximal Haversian canal axis (p = 0.0092) and Haversian
canal diameter (p = 0.0014). The groups did not differ in the percentage of the total
osteon area, the percentage of the total Haversian canal area, Haversian canal perime-
ter / osteon perimeter, Haversian canal area / osteon area, Haversian canal diameter /
osteon diameter, osteon compactness and osteon shape factor. Except for the percent-
age of the total osteon area, this group of variables is identical to that in the case of
the T-test for independent samples by groups.
The groups of unburned bones and bones burned at 800 °C (n = 38; average age
64.0; age range 20–95; standard deviation 17.717; standard error 2.87; coefficient of
variation 27.71) differed at the 0.01 % level of significance (p < 0.0001) in a total of
14 variables. The percentage of the total osteon area (p = 0.0006) differed at the 0.1 %
level of significance, while the percentage of the total Haversian canal area (p =
0.0021) differed in both groups at the 1 % level of significance. The groups did not
differ in Haversian canal perimeter / osteon perimeter, Haversian canal area / osteon
area and Haversian canal diameter / osteon diameter, as when using the T-test for
independent samples by groups.

eschweizerbart_xxx
454 Karolı́na Absolonová et al.

Table 7. T-test for independent samples by groups for bones burned at 700 °C versus bones
burned at 800 °C; df = 94. (SD – standard deviation; * – significant at p < 0.05; ** – signifi-
cant at p < 0.01; *** – significant at p < 0.001; **** – significant at p < 0.0001). For abbrevi-
ations of variables see the Material and methods section.
Mean Mean Difference p SD SD F-ratio p
700 °C 800 °C in % 700 °C 800 °C Vari- Vari-
N = 36 N = 60 compared N = 36 N = 60 ances ances
to 700°C
AGE 58.36 62.72 – 0.2843 21.322 17.796 1.44 0.2177
POC_OST 16.85 20.99 +24.6 0.0029** 5.684 6.811 1.44 0.2516
PR_OST 35.18 30.82 –12.4 0.0493* 10.468 10.340 1.02 0.9151
MAX_O 209.93 178.55 –14.9 0.0000**** 37.970 27.397 1.92 0.0265
MIN_O 133.42 109.34 –18.0 0.0000**** 23.659 18.507 1.63 0.0947
PRUM_O 171.67 143.94 –16.2 0.0000**** 29.421 21.935 1.80 0.0458
F_PR_O 157.41 131.73 –16.3 0.0000**** 26.285 19.921 1.74 0.0593
PR_KAN 2.99 2.58 –13.7 0.0943 1.183 1.120 1.12 0.6961
OBV_K 148.14 126.32 –14.7 0.0000**** 27.179 21.061 1.67 0.0827
MAX_K 54.05 46.87 –13.3 0.0002*** 10.348 7.785 1.77 0.0529
MIN_K 34.17 27.43 –19.7 0.0000**** 7.189 5.249 1.88 0.0325
PRUM_K 43.98 37.15 –15.5 0.0000**** 8.126 6.288 1.67 0.0812
F_PR_K 41.74 34.67 –16.9 0.0000**** 7.799 5.973 1.71 0.0695
OK_OO 0.25 0.26 +4.0 0.4851 0.035 0.048 1.87 0.0496
PK_PO 0.08 0.08 0.0 0.6098 0.023 0.035 2.22 0.0128
PRK_PRO 0.26 0.26 0.0 0.8137 0.036 0.049 1.82 0.0595
C_OST 17.89 17.76 –0.7 0.5399 0.945 1.006 1.13 0.7005
C_KAN 16.02 16.99 +6.1 0.0000**** 0.936 0.823 1.30 0.3741
SFAC_OST 0.72 0.72 0.0 0.4463 0.034 0.036 1.09 0.8058
SFAC_
KAN 0.80 0.75 –6.3 0.0000**** 0.037 0.029 1.65 0.0892

Unburned bones and bones burned at various temperatures differed in most of the
studied variables. When using the T-test for dependent samples or the T-test for inde-
pendent samples, the set of variables in which the groups did not differ was the same
except for a few exceptions. Both types of T-tests demonstrated particularly different
levels of significance in those variables that differed between the compared groups.
This may be explained by the various compositions of the compared groups when
using both types of T-tests.
The bone shrinks during burning and it induces in the bones many microstructural
changes, which influence almost all the studied variables. The dimensions of the
microstructures are intensively reduced, while the number of osteons per mm² of the
compact bone increases. At the same time, the percentual representation of the total
osteon area and the total Haversian canal area on the surface of the cross-section
decreases. The shape of the cross-section of the osteons and Haversian canals also
change. The differences between the groups become statistically more significant as
the temperature of cremation increases (Table 6–8).
All the groups together did not differ only in three variables – Haversian canal
perimeter / osteon perimeter, Haversian canal area / osteon area, and Haversian canal
diameter / osteon diameter. These three variables are thus independent on the degree
of burning. Haversian canal perimeter / osteon perimeter denotes how many percent

eschweizerbart_xxx
 Effect of cremation temperature on human compact bone 455

Table 8. T-test for independent samples by groups for unburned bones versus bones burned
at 800°C; df = 107. (SD – standard deviation; * – significant at p < 0.05; ** – significant at
p < 0.01; *** – significant at p < 0.001; **** – significant at p < 0.0001). For abbreviations
of variables see the Material and methods section.
Mean Mean Difference p SD SD F-ratio p
Un- 800 °C in % Un- 800 °C Vari- Vari-
burned N = 60 compared burned N = 60 ances ances
N = 49 to un- N = 49
burned
AGE 59.86 62.72 – 0.4407 20.773 17.796 1.36 0.2572
POC_OST 14.09 20.99 +49.0 0.0000**** 3.913 6.811 3.03 0.0001
PR_OST 39.80 30.82 –22.6 0.0000**** 10.639 10.340 1.06 0.8290
MAX_O 230.32 178.55 –22.5 0.0000**** 39.013 27.397 2.03 0.0101
MIN_O 157.00 109.34 –30.4 0.0000**** 25.617 18.507 1.92 0.0178
PRUM_O 193.66 143.94 –25.7 0.0000**** 31.166 21.935 2.02 0.0105
F_PR_O 179.30 131.73 –26.5 0.0000**** 28.496 19.921 2.05 0.0092
PR_KAN 3.24 2.58 –20.4 0.0035** 1.194 1.120 1.14 0.6341
OBV_K 166.12 126.32 –24.0 0.0000**** 27.398 21.061 1.69 0.0548
MAX_K 59.25 46.87 –20.9 0.0000**** 10.119 7.785 1.69 0.0556
MIN_K 39.97 27.43 –31.4 0.0000**** 6.860 5.249 1.71 0.0507
PRUM_K 49.61 37.15 –25.1 0.0000**** 8.207 6.288 1.70 0.0519
F_PR_K 46.92 34.67 –26.1 0.0000**** 7.702 5.973 1.66 0.0633
OK_OO 0.26 0.26 0.0 0.9962 0.040 0.048 1.41 0.2181
PK_PO 0.08 0.08 0.0 0.9853 0.039 0.035 1.25 0.4152
PRK_PRO 0.26 0.26 0.0 0.9602 0.040 0.049 1.49 0.1575
C_OST 16.93 17.76 +4.9 0.0000**** 0.681 1.006 2.19 0.0059
C_KAN 16.00 16.99 +6.2 0.0000**** 0.583 0.823 1.99 0.0151
SFAC_OST 0.75 0.72 –4.0 0.0000**** 0.027 0.036 1.81 0.0358
SFAC_
KAN 0.79 0.75 –5.1 0.0000**** 0.024 0.029 1.44 0.1931

of the osteon circumference corresponds to the circumference of the Haversian canal.

Haversian canal area / osteon area denotes how many percent of the osteon area is
occupied by the area of the Haversian canal, and Haversian canal diameter / osteon
diameter informs us about how many percent of the osteon diameter falls to the canal
diameter. The mutual spatial relationship between the osteon and Haversian canal is
thus not affected by burning.
Some microstructures behaved differently during burning between the sexes.
Osteon compactness increased in women under the influence of heat; the osteons
became more oval on the cross-section – “they elongated”. In men, osteon compact-
ness also increased at 700 °C but decreased at 800 °C. This means that the osteons
returned to their original shape. But they did not attain their original shape at the tem-
perature of 800 °C and remained “elongated” compared to the osteons in the
unburned bones. This process was confirmed by the corresponding change of the
osteon shape factor.

Discussion and conclusions

We used ribs in our research for some evident advantages, but during our research we
discovered that with respect to the burning, the ribs also have certain limitations.

eschweizerbart_xxx
456 Karolı́na Absolonová et al.

Thanks to their thin compact bone, the ribs are often seriously damaged after crema-
tion, and it has a negative influence to the state of their microstructures. The small
thickness of the compact bone also results in the bone shrinking transversely to a
greater degree than would have been achieved at the same temperature in a thick
compact bone such as in the femur. It was demonstrated experimentally that trans-
verse shrinkage of the compact bone is influenced by its thickness (Hummel &
Schutkowski 1986, Schutkowski 1991). Moreover, the combustion causes in the thin
compact bone of the rib the birth of many cracks, the loss of the superficial layers of
the compact bone, and at some sites the complete disintegration of the bone, which
can make its histological analysis impossible or very difficult.
A surprising discovery in our research was a wide spectrum of colours in bones
burned at 700 °C. At this temperature, the bones probably burn to various degrees so
that some of them are already completely burned (white colour) while others are not
(grey, blue etc.). The colour of cremated bone depends on the oxidation of organic
carbon, which is completely burned at temperatures around 800 °C (Harbeck et al.
2011). The range of colours present at 700 °C in our study shows that colour itself
cannot be used for the reliable estimation of cremation temperature. In our case we
could only determine whether the bone was burned completely or incompletely or
eventually state that the temperature probably reached 울 700 °C (grey, blue, etc.) or
욷 700 °C (white). This result of our study is in accordance with certain previous
works according to which neither colour nor hardness of the bone are useful criteria
for determination of the exact temperature of cremation (Herrmann 1976, Herrmann
1977, Thompson 2005, Walker et al. 2008, Harbeck et al. 2011, Squires et al. 2011).
The temperature of cremation induces shrinkage of burned bones in both the mac-
roscopic and microscopic level of the compact bone. Nevertheless, in some previous
studies we can read various data concerning the effect of temperature on the compact
bone. In the work of Bradtmiller & Buikstra (1984), it is stated that osteon dimen-
sions increase. In contrast, other works state that they decrease (Herrmann 1976,
1977, Nelson 1992, Hummel & Schutkowski 1993). Nelson (1992) and some other
authors (Squires et al. 2011) state that osteon dimensions decrease, but at the same
time the Haversian canal dimensions increase. In our study, we observed a decrease
in both osteon and Haversian canal dimensions, and this decrease intensified as the
temperature of cremation increased (Tables 3–8). The results of the work of Bradt-
miller & Buikstra (1984) were in all probability caused by the fact that in their study
the bones were burned at a temperature of 600 °C. At this temperature the bones may
transiently expand or the degree of shrinkage is so low that it is undetectable for the
measuring techniques used (Bradtmiller & Buikstra 1984, Schutkowski 1991, Nelson
1992, Thompson 2005). In some cases the changes in the size of the bones are influ-
enced also by the length of the cooling period (Thompson 2005). The difference
between our work and Nelson’s (1992) study is probably caused by the utilisation of
a different type of bone (femur) or the possibility of low temperature reached in the
bones in his study (537–815 °C).
In some previous works (Hummel & Schutkowski 1986, 1993, Schutkowski
1991), it is stated that as the shrinkage of the transverse section is linear, the relative
distribution of structural elements and their relative size ratios remain constant. In
our study, we observed certain changes in the relative representation of microstruc-
tures on the surface of the section. Specifically, there occurred a statistically signifi-
cant decrease in the percentual representation of osteon and Haversian canal areas on

eschweizerbart_xxx
 Effect of cremation temperature on human compact bone 457

the section area, with the decrease of the percentual osteon area being statistically
more significant (Tables 6–8). The shape of the cross-section of the osteons and
Haversian canals also changed, which was expressed as a change in their compact-
ness and shape factor. During burning only the mutual planar and dimensional rela-
tionship between the osteon and Haversian canal remained unchanged, which is
expressed with the aid of three indexes (OK_OO, PK_PO, PRK_PRO).
In conclusion, we must especially stress the fact that the shrinkage of burned
bones leads to a significant change in the size and number of microstructures per sur-
face area unit of the compact bone and that unburned bones and bones burned at vari-
ous temperatures differ statistically significantly in most studied variables. This is
why it is not possible in any case to use methods developed on unburned bones for the
analysis of burned bones, because this procedure leads to erroneous and misleading
results (Heussner 1987, Heussner 1990a, Bruchhaus 1990, Herrmann 1990, Ullrich
1990, Cattaneo et al. 1999, Thompson 2004, Thompson 2005). The differences
between the individual temperatures of cremation also indicate that the histomorpho-
logical methods developed for burned bones as a whole perhaps need not yield
completely reliable results. This question requires the further research, which would
comprise the larger range of the cremation temperatures. The next problem in the
analysis of burned bones is the fact that the temperature of cremation cannot be deter-
mined according to the colour of the burned bones, and its estimation is, despite the
continual progress in the natural sciences, still difficult (Squires et al. 2011). Never-
theless, the histological analysis of burned bones is still indispensable, because in
some cases it can give more accurate results than some modern methods, such as bio-
molecular analyses (Cattaneo et al. 1999).

Acknowledgements
The study was supported by The Charles University Grant Agency (grant 135 808) and by
Ministry of Culture of the Czech Republic for financial support under long-term institutional
funding of conceptual development of the Natural History Museum of the National Museum
in Prague. We thank the autopsy technicians of the Institute of Legal Medicine, 2nd Faculty of
Medicine and Faculty Hospital Na Bulovce in Prague, Charles University for the biological
material sampling. We also thank Prof. RNDr. Martin Mihaljevič, CSc. from the Institute of
Geochemistry, Mineralogy & Mineral Resources, Faculty of Science, Charles University in
Prague for providing the electric combustion furnace, and our colleagues from the Depart-
ment of Palaeontology, Natural History Museum of the National Museum in Prague for the
loan of the microscope with a digital camera.

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Submitted: 2011-December-15;
accepted: 2012-April-20.

Address for correspondence: Karolı́na Absolonová, Vyšehradská 7, Praha 2, 128 00, Czech
Republic.
karolina.absolonova@seznam.cz

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