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The Detection of Microscopic Markers of Hemorrhaging and Wound Age on Dry Bone: A Pilot Study

Cattaneo, Cristina PhD, MD*; Andreola, Salvatore MD; Marinelli, Eloisa MD*; Poppa, Pasquale BSc*; Porta, Davide BSc*; Grandi, Marco MD*

The American Journal of Forensic Medicine and Pathology: March 2010 - Volume 31 - Issue 1 - p 22-26
doi: 10.1097/PAF.0b013e3181c15d74
Original Article

An example of the barriers and conceptual differences between forensic anthropology and pathology can be seen in determining the vitality of a wound. Pathology can make use of skin color and microscopic techniques; anthropology (as concerns the study of dry bone) needs different criteria. The diagnosis of the vitality of a wound (whether it is produced antemortem or postmortem) as well as determination of the time elapsed between the production of the wound and death is a crucial issue in forensic pathology. In fresh skin, the red-purplish coloration of a cut or bruise will reveal its vitality, whereas the change in coloration, from a macroscopic perspective, will reveal the time of survival. In more difficult cases, microscopic analyses can be performed. Bone follows similar “laws” as concerns the evolution of the histologic picture, but even if the beginning of healing processes (periosteal bone production and callus formation) can be detected macroscopically and radiologically, these processes require a long time.

The scope of this pilot study was therefore to collect bone fractures from cadavers with a known time of survival, have them undergo a simulated putrefaction procedure until they became “dry or macerated bone” and perform macroscopic and microscopic analysis to verify the potential of histology in identifying “vital” processes in putrefied soft-tissue-free bone.

A total of 6 samples of fractured bone (cranium, rib, and tibia) were taken from cadavers with known time of survival between trauma and death. Time intervals ranged from a few seconds after the bone fracture had been inflicted, to several hours, days, and weeks. A negative control was included (postmortem fracture). The bone was decalcified and stained with hematoxylin and eosin, Perls' (for the demonstration of hemosiderin deposits), Periodic Acid Schiff, phosphotungstic acid-hematoxylin, and Weigert (for the demonstration of fibrin). Immunohistochemistry was performed using a monoclonal antibody antihuman Glycophorin A.

Results show the presence of clots and red blood cell residues on the fractured margins, strongly indicative of vital reaction.

This study, though certainly not conclusive, shows that it may be worth pursuing the study of bone fractures from a histopathological point of view even on “dry bone” to verify whether the fracture is vital or not, and, if so, if its time of production can be verified.

From the *Laboratorio di Antropologia ed Odontologia Forense, Istituto di Medicina Legale, Università degli Studi, Milano, Italy; and †Dipartimento d Anatomia Pathologica, Istituto per la Ricerca e la Cura dei Tumori, Milano, Italy.

Manuscript received March 2, 2007; accepted May 13, 2007.

Figures can be viewed in color at

Reprints: Cristina Cattaneo, PhD, MD, Istituto di Medicina Legale, Università degli Studi, via Mangiagalli 37, 20133 Milano, Italy. E-mail:

© 2010 Lippincott Williams & Wilkins, Inc.