ORIGINAL ARTICLE

Evaluation of Histologic Changes of the Skin

in Postmortem Period
Rajesh V. Bardale, MD, Nilesh K. Tumram, MD, Pradeep G. Dixit, MD(Path), MD(FM),
and Ashutosh Y. Deshmukh, MD

intracranial hemorrhage, n = 2; pulmonary consolidation, n = 2;

Abstract: Determination of the time of death is an important consid- ischemic heart disease, n = 1; electrocution, n = 1; pulmonary
eration in forensic practice. Many methods have been attempted to ac- tuberculosis, n = 1; and rheumatic heart disease, n = 1). The
curately and systematically determine the postmortem interval (PMI). bodies were kept at room temperature in the waiting room of the
Histologic examination of the skin or appendages is one of the methods mortuary. The average ambient temperature during the period
tried by few researchers. However, no attempt had been made to analyze varied from 23-C to 37-C, and the average humidity ranged from
the histologic changes in the skin and appendages simultaneously and to 17% to 87%. At autopsy, skin samples from the anterior chest
compare them with PMI. We sequentially studied the histologic changes were collected, parallel to the longitudinal midline incision. The
of the skin and appendages in the early PMI. The results of the present cases were divided into 3-, 6-, 9-, 12-, 15-, 18-, 21-, and 24-hour
study show that the skin undergoes progressive morphological changes intervals after death. Only 1 skin sample from each cadaver
in the postmortem period. The epidermis and the dermis appeared nor- was taken. The skin samples were carefully taken to avoid trau-
mal for 6 hours after death, and after this period, degenerative changes matic artifact. No person had any skin disease. The samples were
began. By 6 to 9 hours after death, degeneration began in the dermis, and collected from those bodies where actual time of death was
by the end of 18 hours, the dermis began to disintegrate. The sweat glands known. All samples were subjected to light microscopy. Tissues
appeared normal for approximately 3 to 4 hours. For 18 hours after death, were fixed in 10% formalin, embedded in paraffin wax, and
the sebaceous glands and hair follicles appeared normal, and after this stained with hematoxylin-eosin (H and E) and alcianYperiodic
period, degeneration began. acid-Schiff (alcian-PAS). The skin samples were observed for
Key Words: death, postmortem interval, skin, sweat glands, the following: epidermis, dermis, sweat glands, sebaceous glands,
hair follicle, sebaceous glands, forensic, autopsy and hair follicles.
(Am J Forensic Med Pathol 2012;33: 357Y361)
RESULTS
The results of the present study show that the skin under-
D etermination of the time of death is an important consider-
ation in forensic practice. Many methods have been attempted
to accurately and systematically determine the postmortem in-
goes progressive morphological changes in the postmortem pe-
riod. These changes can be observed on gross examination and
terval (PMI) with varied success.1,2 Histologic examinations of at cellular level by light microscopy.
tissues or body fluid cells for time-related morphological changes
is one of the methods tried by few researchers.3Y6 Except for very Skin
few studies,7Y9 examinations of the skin and its appendages have The skin is composed of the epidermis and the dermis. The
not been systematically analyzed. Moreover, environmental fac- epidermis is made up of stratified squamous epithelium and
tors such as temperature and humidity, as well as insect activity, consists of 5 layers: stratum basale, stratum spinosum, stratum
seem to be the most important factor affecting the skin in post- granulosum, stratum lucidum, and stratum corneum. The der-
mortem period.9 The present study was undertaken to evaluate moepidermal junction is not straight but wavy because of the
the histologic changes of the skin in hot weather and to identify presence of cone-shaped projections of the dermis upward into
morphological parameters that will help in determining the PMI the epidermis. These projections are called dermal papillae. The
in early postmortem period. dermis is made up of connective tissue and consists of 2 parts:
the superficial pars papillaris and the deeper pars reticularis. The
MATERIALS AND METHODS pars papillaris includes the connective tissue of the dermal pa-
The study was conducted at the Department of Forensic pillae. The pars reticularis consists mainly of bundles of collagen
Medicine, Government Medical College Hospital, Nagpur, from fibers and elastic fibers.
March to April 2010. The study consists of 30 human cadavers For an approximately 6-hour PMI, no morphological
comprising 24 male and 6 female, and their age ranged from 15 changes were observed in the epidermis and the dermis. After
to 64 years (mean age, 39.43 years). The death was due to var- this period, the vacuoles begin to appear in the spinous and basal
ious causes (head injury, n = 16; hanging, n = 3; poisoning, n = 3; layer, and cells appeared distended and balloon shaped (Fig. 1A).
With increasing PMI, the basal layer cells begin to distort with
incontinence of melanin pigment. These changes can be observed
Manuscript received March 15, 2011; accepted June 23, 2011. at approximately 9 to 12 hours after death. By this time, the der-
From the Department of Forensic Medicine, Government Medical College,
Nagpur, Maharashtra, India.
moepidermal junction became indistinct and focal separation
The authors report no conflicts of interest. begins to appear (Fig. 1B). At an approximately 12- to 18-hour
Reprints: Rajesh V. Bardale, MD, Department of Forensic Medicine, PMI, there was separation of the epidermis from the dermis.
Government Medical College, Hanuman Nagar, Nagpur 440003, By 6- to 9-hour PMI, focal degeneration appears in the
Maharashtra, India. E-mail: [email protected].
Copyright * 2012 by Lippincott Williams & Wilkins
dermis (Fig. 2A), and with increasing PMI, the degeneration
ISSN: 0195-7910/12/3304Y0357 becomes generalized. By the end of 18 hours, the dermis begins
DOI: 10.1097/PAF.0b013e31822c8f21 to disintegrate (Fig. 2B; Table 1).

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Bardale et al Am J Forensic Med Pathol & Volume 33, Number 4, December 2012

FIGURE 1. Microphotograph showing the epidermis. A, Six hours after death, appearance of vacuoles in basal and spinous layer.
B, Nine hours after death, focal dermoepidermal separation (H and E, original magnification 40).

FIGURE 2. Microphotograph showing the dermis. A, Nine hours after death, rarefaction of the dermis. B, Eighteen hours after death,
dermal disintegration (H and E, original magnification 40).

Sweat Glands
The eccrine sweat glands are of tubular type. The gland TABLE 1. Time Course of Histologic Variations in the
tubule may be divided into 2 main parts: secretory tubule and Epidermis and the Dermis
duct. The secretory tubule is not branched and has a narrow
lumen. It is composed of 3 types of cells surrounded by a thin PMI, h Features
basal membrane. The outer myoepithelial cells form a basket 0Y6 Normal morphology of the epidermis and the dermis
surrounding the inner row of secretory cells. The secretory cells
6Y9 Appearance of vacuoles in cytoplasm of basal layer
are of 2 types: clear cells and dark cells. The duct is formed of and spinous layer
2 rows of cells. The internal cells are small, cube shaped with
Rarefaction of the dermis, elastic fibers get prominent
a high nucleus-cytoplasm ratio. The external cells are thin, in with focal fragmentation
contact with the basal membrane, and rich in mitochondria.
9Y12 Distorted basal layer cells with incontinence of
With H and E stain, the sweat glands appear normal up to 3 melanin pigment, focal epidermo-dermal separation
to 4 hours after death. After this period, degeneration begins in
918 Fragmentation of the dermis
form of appearance of empty vacuoles in the cytoplasm of the

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Am J Forensic Med Pathol & Volume 33, Number 4, December 2012 Histologic Skin Changes in Postmortem Period

FIGURE 3. Microphotograph showing the sweat glands. A, Six hours after death, appearance of vacuoles in cytoplasm (black arrows)
(H and E, original magnification 10). B, Fifteen hours after death, ruptured cells (H and E, original magnification 40).

gland cells (Fig. 3A). With increasing PMI, the number of empty
vacuoles in the cytoplasm increased, and at an approximately 15-
hour PMI, the cells appear ruptured (Fig. 3B; Table 2).
TABLE 2. Showing Time Course Histologic Variations in The basal membrane, the cytoplasm of secretory tubule
Sweat Glands (H and E Stain) cells, and the apex of duct cells are stained by alcian-PAS. The
PMI, h Features glycogen particles appear purple. With passage of PMI, the
glycogen particle begins to disappear, and the secretory cell
0Y3 Normal morphology begins to lose PAS positivity (Fig. 4A). This change can be
4Y6 Vacuoles begins to appear observed between 2 and 3 hours after death. After 3 hours, the
915 Ruptured cells cells appear PAS negative (Fig. 4B). However, the basal mem-
brane appears magenta for approximately 18 to 24 hours after
death (Fig. 4, C and D; Table 3).

FIGURE 4. Microphotograph showing the sweat glands. A, Three hours after death, PAS positivity of cells. B, Six hours after death,
PAS negativity of cells with appearance of cytoplasmic vacuoles (black arrows) (alcian-PAS, original magnification 10). C, Twelve hours
after death, cellular degeneration. D, Fifteen hours after death, PAS-negative ruptured cells with PAS-positive basal membrane
(alcian-PAS, original magnification 40).

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Bardale et al Am J Forensic Med Pathol & Volume 33, Number 4, December 2012

TABLE 3. Showing Time Course Histologic Variations in TABLE 4. Showing Time Course Histologic Variations in
Sweat Glands (Alcian-PAS Stain) Sebaceous Glands and Hair Follicles

PMI, h Features Sebaceous Gland Hair Follicle

2Y3 Secretory and duct cells begin to loose PAS positivity, PMI, h Features Features
basal layer appears magenta 0Y18 Normal and foamy Normal morphology
3Y6 PAS-negative secretory and duct cells, basal layer 918 Degeneration Layers begins to separate,
appears magenta begins in the cells disintegration of hair papilla,
918 Basal layer appears magenta PAS-positive basal lamina

Sebaceous Glands these studies were conducted in cool to temperate region where
Sebaceous glands are seen most typically in relation to hair the bodies are well preserved for quite a long time than the bodies
follicles but can also be seen in a few areas devoid of hairs as found in tropical countries like India. No such study was reported
modified sebaceous glands. Sebaceous glands are composed of from this part of the region. Therefore, there is a need for such
lobules of sebaceous cells containing small round nuclei and study. Similarly, previous studies had noted the findings of the
abundant fatty, network-like cytoplasm. skin alone or sweat glands or hair follicles.7Y10 In this study, an
The cells of sebaceous glands appear normal and foamy for attempt had been made to analyze the histologic changes in the
an approximately 18-hour PMI (Fig. 5A). Afterward, as the PMI skin and appendages simultaneously and compare them with PMI.
increases, the cells begins to degenerate (Fig. 5B; Table 4) in So far as English literature is concerned, only 3 studies were
form of nuclear disintegration and loss of cellular architecture. reported on this subject. Cingolani et al7 studied the morphology
of sweat glands for a period of 12 hours after death. On light
Hair Follicle microscopy, they had noted normal morphology of sweat glands
Each hair contains of a part seen on the surface of body for 6 hours after death, and after this period, vacuoles in the
(called shaft) and a part anchored in the thickness of the skin cytoplasm began to appear. On alcian-PAS stain, the secretory
(called root). The root has an extended lower end called the bulb. tubule cells progressively lost their PAS positivity within 6 hours
The bulb is invaginated from below by part of the dermis that after death. Brzezinski et al,8 while studying the epidermis in
constitutes the hair papilla. The root of each hair is surrounded refrigerated bodies, had noted the development of crescent-
by a tubular sheath called the hair follicle. shaped nuclei in spinous layer. These crescent-shaped nuclei
The hair follicle appears normal, and by 18-hour PMI, the were surrounded by a hallow area after 1 day after death, and the
degenerative change begins to appear. The hair follicle layers number of such cells increased significantly each day during the
begin to separate with formation of vacuoles in between them. first 8 days postmortem. Similarly, in the basal layer, balloon-
The papillary part of hair begins to disintegrate (Fig. 6, A and B). shaped cells with pyknotic nuclei appeared, and with increasing
By the end of 21 to 24 hours, the basal lamina separating the PMI, focal dermoepidermal separation was observed. Kovarik
outer root sheath was PAS positive (Table 4). et al9 evaluated gross and histologic changes in 3 cadavers in
early postmortem period (G1 week) in outdoor, shaded and cool
DISCUSSION to temperate climate (38-FY77-F). On gross examination, the
Gross changes occurring in the skin in postmortem period is skin appeared normal from day 1 to 7 except for increased
a known phenomenon used to estimate death interval. The gross wrinkling in the acral skin. On microscopic examination, focal
changes that are frequently used include skin slippage, blister separation of the epidermis from the dermis was evident from
formation, skin discoloration, dehydration and mummification, day 1 to 7. Degeneration of the sweat glands was observed by
and/or adipocere formation. However, the timing of development day 5, whereas dermal degeneration was evident by day 2.
is unpredictable. To overcome this problem, few researchers had Considering the present study, we have sequentially studied
used histologic changes of the skin or appendages.7Y9 Most of the histologic appearance of the skin, sweat glands, sebaceous

FIGURE 5. Microphotograph showing the sebaceous gland. A, Six hours after death, normal sebaceous gland cells. B, Twenty-one hours
after death, degeneration of sebaceous gland cells (H and E, original magnification 40).

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Am J Forensic Med Pathol & Volume 33, Number 4, December 2012 Histologic Skin Changes in Postmortem Period

FIGURE 6. Microphotograph showing the hair follicle. A, Twenty-one hours after death, hair follicle layers begin to separate and
appearance of vacuoles (H and E, original magnification 40). B, Twenty-one hours after death, disintegration of hair papilla
(H and E, original magnification 10).

glands, and hair follicles in same environmental condition with death interval, degeneration began. The degeneration was better
average ambient temperature varied from 23-C to 37-C and appreciated in sebaceous glands than the hair follicles. Because
average humidity ranged from 17% to 87%. On microscopic of the nonavailability of samples more than 24 hours, further
examination, the epidermis and the dermis appeared normal for a studies were not done. Currently, there is very limited scientific
6-hour PMI, and after this period, degenerative changes began. information available regarding the process of sebaceous gland
At an approximately 12- to 18-hour PMI, there was separation of and hair follicle degeneration.10 Further studies are required to
the epidermis from the dermis. By 6- to 9-hour PMI, degener- evaluate these changes and characterize the postmortem chan-
ation began in the dermis, and by the end of 18 hours, the dermis ges. The study is limited by the small sample size and evaluation
began to disintegrate. By H and E stain, the sweat glands ap- of skin changes in a common environmental setting.
peared normal for approximately 3 to 4 hours, and after this pe- In conclusion, the study may be used to assist in the esti-
riod, empty vacuoles began to appear in the cytoplasm of the mation of time of death in early postmortem period. Figure 7
gland cells. On alcian-PAS stain, the glycogen particles dis- summarizes the potential for determining PMI by various para-
appeared, and the secretory cells progressively lose their PAS meters. Meaningful conclusions in respect to PMI are perhaps
positivity by approximately 2 to 3 hours after death. deducible by considering all the variables discussed thus far, not
When the results of the present study were compared with in isolation but in combination.
published literature, the histologic pattern and findings are more
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