International Journal of Legal Medicine

, Volume 133, Issue 3, pp 711–718 | Cite as

Histomorphometric analysis of osteocyte lacunae in human and pig: exploring its potential for species discrimination

  • Marco CummaudoEmail author
  • Annalisa Cappella
  • Francesca Giacomini
  • Caterina Raffone
  • Nicholas Màrquez-Grant
  • Cristina Cattaneo
Original Article


In recent years, several studies have focused on species discrimination of bone fragments by histological analysis. According to literature, the most consistent distinguishing features are Haversian canal and Haversian system areas. Nonetheless, there is a consistent overlap between human and non-human secondary osteon dimensions. One of the features that have never been analyzed for the purpose of species discrimination is the osteocyte lacuna, a small oblong cavity in which the osteocyte is locked in. The aim of this study is to verify whether there are significant quantitative differences between human and pig lacunae within secondary osteons with similar areas. Study sample comprises the midshaft of long bones (humerus, radius, ulna, femur, tibia, and fibula) of a medieval human adult and a juvenile pig. Sixty-eight secondary osteons with similar areas have been selected for each species and a total of 1224 osteocyte lacunae have been measured. For each osteon, the total number of lacunae was counted, and the following measurements were taken: minimum and maximum diameter, area, perimeter, and circularity of nine lacunae divided between inner, intermediate, and outer lacunae. Statistical analysis showed minimal differences between human and pig in the number of lacunae per osteons and in the minimum diameter (P > 0.05). On the contrary, a significant difference (P < 0.001) has been observed in the maximum diameter, perimeter, area, and circularity. Although there is the need for further research on different species and larger sample, these results highlighted the potential for the use of osteocyte lacunae as an additional parameter for species discrimination. Concerning the difference between the dimensions of osteocyte lacunae based on their position within the osteon (inner, intermediate, and outer lacunae), results showed that their size decreases from the cement line towards the Haversian canal both in human and pig.


Forensic anthropology Bone histology Bone lacunae Bioarchaeology Human vs non-human Sus scrofa 



  1. 1.
    Blau S, Briggs C (2011) The role of forensic anthropology in disaster victim identification (DVI). Forensic Sci Int 205(1–3):29–35. Google Scholar
  2. 2.
    Brits D, Steyn M, L’Abbe EN (2014) A histomorphological analysis of human and non-human femora. Int J Legal Med 128(2):369–377. Google Scholar
  3. 3.
    Cuijpers AGFM (2006) Histological identification of bone fragments in archaeology: telling humans apart from horses and cattle. Int J Osteoarchaeol 16:465–480. Google Scholar
  4. 4.
    Cuijpers AGFM (2009) Distinguishing between the bone fragments of medium-sized mammals and children. A histological identification method for archaeology. Anthropol Anz 67(2):181–203Google Scholar
  5. 5.
    Hillier ML, Bell LS (2007) Differentiating human bone from animal bone: a review of histological methods. J Forensic Sci 52(2):249–263. Google Scholar
  6. 6.
    Locke M (2004) Structure of long bones in mammals. J Morphol 262:546–565. Google Scholar
  7. 7.
    Mulhern DM, Ubelaker DH (2001) Differences in osteon banding between human and nonhuman bone. J Forensic Sci 46(2):220–222. Google Scholar
  8. 8.
    Sawada J, Nara T, Fukui J, Dodo Y, Hirata K (2014) Histomorphological species identification of tiny bone fragments from a paleolithic site in the northern Japanese archipelago. J Archaeol Sci 46:270–280. Google Scholar
  9. 9.
    Cattaneo C, Porta D, Gibelli D, Gamba C (2009) Histological determination of the human origin of bone fragments. J Forensic Sci 54:531–533. Google Scholar
  10. 10.
    Crescimanno A, Stout SD (2012) Differentiating fragmented human and nonhuman long bone using osteon circularity. J Forensic Sci 57(2):287–294. Google Scholar
  11. 11.
    Dominguez VM, Crowder CM (2012) The utility of osteon shape and circularity for differentiating human and non-human Haversian bone. Am J Phys Anthropol 149(1):84–91. Google Scholar
  12. 12.
    Martiniaková M, Grosskopf B, Omelka R, Vondráková M, Bauerová M (2006a) Differences among species in compact bone tissue microstructure of mammalian skeleton: use of a discriminant function analysis for species identification. J Forensic Sci 51(6):1235–1239. Google Scholar
  13. 13.
    Martiniaková M, Grosskopf B, Vondráková M, Omelka R, Fabĭs M (2006b) Differences in femoral compact bone tissue microscopic structure between adult cows (Bos taurus) and pigs (Sus scrofa domestics). Anat Histol Embryol 35:167–170. Google Scholar
  14. 14.
    Martiniaková M, Grosskopf B, Omelka R, Vondráková M, Bauerová M (2007a) Histological analysis of ovine compact bone tissue. J Vet Med Sci 69:409–411. Google Scholar
  15. 15.
    Martiniaková M, Grosskopf B, Omelka R, Dammers K, Vondráková M, Bauerová M (2007b) Histological study of compact bone tissue in some mammals: a method for species determination. Int J Osteoarchaeol 17:82–90. Google Scholar
  16. 16.
    Urbanová P, Novotný V (2005) Distinguishing between human and non-human bones: histometric method for forensic anthropology. Anthropologie 43:77–85Google Scholar
  17. 17.
    Mulhern DM, Ubelaker DH (2011) Differentiating human from nonhuman bone microstructure. In: Crowder C, Stout SD (eds) Bone histology: an anthropological perspective. CRC Press, Boca Raton, pp 109–134. Google Scholar
  18. 18.
    Cummaudo M, Cappella A, Biraghi M, Raffone C, Màrquez-Grant N, Cattaneo C (2018) Histomorphological analysis of the variability of the human skeleton: forensic implications. Int J Legal Med 132:1493–1503. Google Scholar
  19. 19.
    Albu I, Georgia R, Georoceneau M (1990) The canal system in the diaphyseal compacta of the femur in some mammals. Anat Anz 170(3–4):191–187Google Scholar
  20. 20.
    Dittman K (2003) Histomorphometrische untersuchung der knochenmikrostructur von primate and haustieren mit dem ziel der speziesdentifikaton unter berücksichtingung von domestikationseffekten. Anthropol Anz 61(2):175–188Google Scholar
  21. 21.
    Martin RB, Gibson VA, Stover SM, Gibeling JC, Griffin LV (1996) Osteonal structure in the equine third metacarpus. Bone 19(2):165–171Google Scholar
  22. 22.
    Zerwekh JE (1992) Bone metabolism. Semin Nephrol 12:79–90Google Scholar
  23. 23.
    Freemont AJ (1993) Basic bone cell biology: a review. Int J Exp Pathol 74:411–416Google Scholar
  24. 24.
    Stout SD, Crowder C (2011) Bone remodeling, histomorphology, and histomorphometry. In: Crowder C, Stout SD (eds) Bone histology: an anthropological perspective. CRC Press, Boca Raton, pp 1–21Google Scholar
  25. 25.
    Qiu S, Fyhrie DP, Palnitkar S, Rao DS (2003) Histomorphometric assessment of Haversian canal and osteocyte lacunae in different-sized osteons in human rib. Anat Rec 272a(2):520–525. Google Scholar
  26. 26.
    Nijweide PJ, Burger EH, Klein-Nulend J (2002) The osteocyte. In: Bilezikian JP, Raisz LG, Rodan GA (eds) Principles of bone biology, 2nd edn. Academic, San Diego, pp 93–107Google Scholar
  27. 27.
    Parfitt AM (2005) Targeted and nontargeted remodeling: relationship to basic multicellular unit organization and progression. Bone 30(1):5–7Google Scholar
  28. 28.
    Martin RB (2000) Does osteocyte formation cause the nonlinear refilling of osteons? Bone 26(1):71–78Google Scholar
  29. 29.
    Qiu S, Rao DS, Palnitkar S, Parfitt AM (2006) Differences in osteocyte and lacunar density between Black and White American women. Bone 38:130–135Google Scholar
  30. 30.
    Sissons HA, O’Connor P (1977) Quantitative histology of osteocyte lacunae in normal human cortical bone. Calcif Tissue Res 22(Suppl):530–533Google Scholar
  31. 31.
    Ascenzi MG, Gill J, Lomovtsev A (2008) Orientation of collagen at the osteocyte lacunae in human secondary osteons. J Biomech 41(16):3426–3435Google Scholar
  32. 32.
    Carter Y, Thomas CDL, Clement JG, Peele AG, Hannah K, Cooper DML (2013) Variation in osteocyte lacunar morphology and density in the human femur - a synchrotron radiation micro-CT study. Bone 52(1):126–132. Google Scholar
  33. 33.
    Dong P, Haupert S, Hesse B, Langer M, Gouttenoire PJ, Bousson V, Peyrin F (2014) 3D osteocyte lacunar morphometric properties and distributions in human femoral cortical bone using synchrotron radiation micro-CT images. Bone 60:172–185. Google Scholar
  34. 34.
    Hannah KM, Thomas CDL, Clement JG, De Carlo F, Peele AG (2010) Bimodal distribution of osteocyte lacunar size in the human femoral cortex as revealed by micro-CT. Bone 47(5):866–871. Google Scholar
  35. 35.
    Teti A, Zallone A (2009) Do osteocytes contribute to bone mineral homeostasis? Osteocytic osteolysis revisited. Bone 44:11–16Google Scholar
  36. 36.
    Stern AR, Nicolella DP (2013) Measurement and estimation of osteocyte mechanical strain. Bone 54(2):191–195Google Scholar
  37. 37.
    Bach-Gansmo FL, Brüel A, Jensen MV, Ebbesen EN, Birkedal H, Thomsen JS (2016) Osteocyte lacunar properties and cortical microstructure in human iliac crest as a function of age and sex. Bone 91:11–19Google Scholar
  38. 38.
    Skedros JG, Grunander TR, Hamrick MW (2005) Spatial distribution of osteocyte lacunae in equine radii and third metacarpals: considerations for cellular communication, microdamage detection and metabolism. Cells Tissues Organs 180:215–236Google Scholar
  39. 39.
    Hobdell MH, Howe CE (1971) Variation in bone matrix volume associated with osteocyte lacunae in mammalian and reptilian bone. Isr J Med Sci 7:492–493Google Scholar
  40. 40.
    Mullender MG, van der Meer DD, Huiskes R, Lips P (1996) Osteocyte density changes in aging and osteoporosis. Bone 18:109–113Google Scholar
  41. 41.
    Mullender MG, Tan SD, Vico L, Alexandre C, Klein-Nulend J (2005) Differences in osteocyte density and bone histomorphometry between men and women and between healthy and osteoporotic subjects. Calcif Tissue Int 77:291–296Google Scholar
  42. 42.
    Qiu S, Rao DS, Palnitkar S, Parfitt AM (2003) Reduced iliac cancellous osteocyte density in patients with osteoporotic vertebral fracture. J Bone Miner Res 18:1657–1663Google Scholar
  43. 43.
    van Hove RP, Nolte PA, Vatsa A, Semeins CM, Salmon PL, Smit TH, Klein-Nulend J (2009) Osteocyte morphology in human tibiae of different bone pathologies with different bone mineral density—is there a role for mechanosensing? Bone 45:321–329Google Scholar
  44. 44.
    Marotti G, Favia A, Zallone A (1972) Quantitative analysis on the rate of secondary bone mineralization. Calcif Tissue Res 10(1):67–81Google Scholar
  45. 45.
    Marotti G (1979) Osteocyte orientation in human lamellar bone and its relevance to the morphometry of periosteocytic lacunae. Metab Bone Dis Relat 333:325–333Google Scholar
  46. 46.
    Pokines JT (2015) Identification of nonhuman remains received in a medical examiner setting. J Forensic Identif 65(3):223–246Google Scholar
  47. 47.
    Maat GJR, Van Den Bos RPM, Aarents MJ (2001) Manual preparation of ground sections for the microscopy of natural bone tissue: update and modification of Frost’s “rapid manual method”. Int J Osteoarchaeol 11(5):366–374. Google Scholar
  48. 48.
    Frasca P, Harper RA, Katz JL (1977) Collagen fibre orientations in human secondary osteons. Acta Anat (Basel) 98:1–13Google Scholar
  49. 49.
    Ardizzoni A (2001) Osteocyte lacunar size-lamellar thickness relationships in human secondary osteons. Bone 28(2):215–219. Google Scholar
  50. 50.
    Remaggi F, Canè V, Palumbo C, Ferretti M (1998) Histomorphometric study on the osteocyte lacuno-canalicular network in animals of different species. I. Woven-fibered and parallel-fibered bones. Ital J Anat Embryol 103(4):145–155Google Scholar
  51. 51.
    Ferretti M, Muglia MA, Remaggi F, Canè V, Palumbo C (1999) Histomorphometric study on the osteocyte lacuno-canalicular network in animals of different species. II. Parallel-fibered and lamellar bones. Ital J Anat Embryol 104(3):121–131Google Scholar
  52. 52.
    van Oers RFM, Wang H, Bacabac RG (2015) Osteocyte shape and mechanical loading. Curr Osteoporos Rep 13(2):61–66. Google Scholar
  53. 53.
    Vatsa A, Breuls RG, Semeins CM, Salmon PL, Smit TH, Klein-Nulend J (2008) Osteocyte morphology in fibula and calvaria—is there a role for mechanosensing? Bone 43:452–458Google Scholar
  54. 54.
    Britz HM, Thomas CDL, Clement JG, Cooper DML (2009) The relation of femoral osteon geometry to age, sex, height and weight. Bone 45:77–83Google Scholar
  55. 55.
    Currey JD (1964) Some effects of ageing in human Haversian systems. J Anat 98(1):69–75Google Scholar
  56. 56.
    Evans FG (1976) Mechanical properties and histology of cortical bone from younger and older men. Anat Rec 185(1):12Google Scholar
  57. 57.
    Mulhern DM, Van Gerven DP (1997) Patterns of femoral bone remodeling dynamics in a medieval Nubian population. Am J Phys Anthropol 104:133–146Google Scholar
  58. 58.
    Thompson DD (1980) Age changes in bone mineralization, cortical thickness and Haversian canal area. Calcif Tissue Int 31:5–11Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.LABANOF (Laboratorio di Antropologia e Odontologia Forense) Dipartimento di Scienze Biomediche per la SaluteUniversità degli Studi di MilanoMilanItaly
  2. 2.Cranfield Forensic Institute, Defence Academy of the United KingdomCranfield UniversityShrivenhamUK

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