Evaluation of Femur Length in Rats Using Three Different Methods and the Reliability of the Anthropometric Measurement Method

Year 2025, Volume: 14 Issue: 2, 514 - 522, 30.08.2025

Ahmet Emrah Açan , Burcu Aslantekin , Oğuzhan Selçuk Aldemir , Emrah Özcan , Sinan Saraçlı , Özgür Bulmuş , Aslı Karakılıç , Gülay Turan

Abstract

Objective: The skeletal system evaluated in rodent models plays a crucial role in various experimental studies, including bone development, fracture healing, biomechanical testing, osteoporosis, drug efficacy, and safety. This study aims to assess the applicability of the anthropometric method, which is widely used in humans, for measuring femur length in rats. Additionally, the reliability of the anthropometric method was compared with dissection and radiological methods. Materials and Methods: A total of 66 male Wistar Albino rats (14–16 weeks old) were used. The right femur length was measured using three different methods: anthropometric measurement, digital caliper measurement after dissection, and radiological measurement via micro-CT. The obtained data were analyzed, and the agreement between methods was evaluated using the Bland-Altman test. Results: The mean femur length was measured as 37.57 mm anthropometrically, 33.55 mm using a digital caliper post-dissection, and 33.84 mm using micro-CT. The intraclass correlation coefficient (ICC) values at a %90 confidence interval were as follows: 0.987 (0.979–0.992) between the dissection and radiological groups, 0.709 (0.564–0.812) between the anthropometric and dissection groups, and 0.713 (0.569–0.814) between the anthropometric and radiological groups. The anthropometric method showed a significant agreement with both dissection and radiological methods, with a difference of approximately 4 mm. Conclusion: The anthropometric method has been demonstrated to be a reliable, easy-to-use, non-invasive, cost-effective, and time-saving alternative for measuring femur length in rats. Since this method allows for repeated measurements on the same animal, it can reduce the number of animals used in studies, thereby promoting more ethical and efficient research in bone studies.

Keywords

Keywords: Femur length , rat , anthropometry , dissection , micro-CT.

Project Number

2024/178

Sıçanlarda Femur Uzunluğunun 3 Farklı Yöntem ile Değerlendirilmesi ve Antropometrik Ölçüm Yönteminin Güvenilirliği

Abstract

Amaç: Kemirgen modellerinde değerlendirilen iskelet sistemi; kemik gelişimi, kırık iyileşmesi, biyomekanik testler, osteoporoz, ilaç etkinliği ve güvenliği gibi birçok deneyde önemli yer tutmaktadır. Bu çalışmada, sıçanlarda en çok değerlendirilen kemiklerden olan femur’un uzunluğunu ölçmek için insanlarda yaygın olarak kullanılan antropometrik yöntemin sıçanlardaki uygulanabilirliği, bunun yanında antropometrik yöntemin güvenilirliğinin diseksiyon ve radyolojik yöntemler ile karşılaştırılması amaçlanmıştır. Gereç ve Yöntem: 66 adet erkek Wistar Albino sıçan kullanılmıştır ve sağ femur uzunluğu antropometrik olarak, diseksiyon sonrası dijital kumpas ile ve mikroCT ile radyolojik olarak ölçülmüştür. Elde edilen veriler analiz edilmiş ve Bland-Altman testi ile yöntemler arasındaki uyum değerlendirilmiştir. Bulgular: Femur uzunluğu; antropometrik olarak 37.57 mm, diseksiyon sonrası dijital kumpas ile 33.55 mm, mikroCT ile 33.84 mm bulunmuştur. %90 güven aralığında ICC katsayıları; diseksiyon ve radyolojik grup arasında 0.987 (0.979-0.992), antropometrik ve diseksiyon grup arasında 0.709 (0.564-0.812), antropometrik ve radyolojik grup arasında 0.713 (0.569-0.814) şeklindedir. Antropometrik yöntem, diseksiyon sonrası ve radyolojik yöntemlerle arasında 4 mm’lik farkla önemli derecede uyumlu olarak bulunmuştur. Sonuç: Sonuç olarak sıçanlarda femur uzunluğunu ölçmek için antropometrik yöntem, güvenilir, kolay uygulanabilir, invaziv olmayan, maliyet ve zaman tasarrufu sağlayan alternatif bir yöntem olduğu ortaya koyulmuştur. Bu yöntem, kullanılan hayvan sayısını azaltıp tek bir hayvanın tekrar tekrar değerlendirilmesine olanak sağladığından kemik çalışmalarında daha etik ve daha verimli çalışmaların yapılmasına imkân sağlayabilir.

Keywords

Anahtar kelimeler: Femur uzunluğu , sıçan , antropometri , diseksiyon , mikroCT.

Project Number

2024/178

References

• Bagi, C. M., Berryman, E., & Moalli, M. R. (2011). Comparative bone anatomy of commonly used laboratory animals: implications for drug discovery. Comparative medicine, 61(1), 76–85.
• Cao, A. B., McGrady, L. M., & Wang, M. (2023). Effect of age on femur whole-bone bending strength of mature rat. Clinical biomechanics (Bristol, Avon), 101, 105828. https://doi.org/10.1016/j.clinbiomech.2022.105828
• Castillo, E. J., Croft, S. M., Jiron, J. M., & Aguirre, J. I. (2022). Bone structural, biomechanical, and histomorphometric characteristics of the hindlimb skeleton in the marsh rice rat (Oryzomys palustris). Anatomical record (Hoboken, N.J. : 2007), 305(11), 3133–3149. https://doi.org/10.1002/ar.24876
• Cvetkovic, V., Najman, S., Rajkovic, J., Zabar, A., Vasiljevic, P., Djordjevic, L., & Trajanovic, M. (2013). A comparison of the microarchitecture of lower limb long bones between some animal models and humans: a review. Veterinární Medicína, 58(7), 339–351. https://doi.org/10.17221/6914-vetmed
• Dancause, K. N., Cao, X. J., Veru, F., Xu, S., Long, H., Yu, C., Laplante, D. P., Walker, C. D., & King, S. (2012). Brief communication: prenatal and early postnatal stress exposure influences long bone length in adult rat offspring. American journal of physical anthropology, 149(2), 307–311. https://doi.org/10.1002/ajpa.22117
• Ekong, M. B., Ekanem, T. B., Sunday, A. O., Aquaisua, A. N., & Akpanabiatu, M. I. (2012). Evaluation of calabash chalk effect on femur bone morphometry and mineralization in young wistar rats: A pilot study. International journal of applied & basic medical research, 2(2), 107–110. https://doi.org/10.4103/2229-516X.106352
• Freitas, L., Bezerra, A., Resende-Coelho, A., Maciel, L., Gomez-Lazaro, M., Amorim, T., Fernandes, R., & Fonseca, H. (2025). Effect of Eight Months of Swimming on Bone Quality of Different Anatomical Regions: A Study on Wistar Rat Models. Calcified tissue international, 116(1), 29. https://doi.org/10.1007/s00223-024-01333-x
• Foster A. D. (2019). The impact of bipedal mechanical loading history on longitudinal long bone growth. PloS one, 14(2), e0211692. https://doi.org/10.1371/journal.pone.0211692
• Gürgül, S., Erdal, N., Yilmaz, S. N., Yildiz, A., & Ankarali, H. (2008). Deterioration of bone quality by long-term magnetic field with extremely low frequency in rats. Bone, 42(1), 74–80. https://doi.org/10.1016/j.bone.2007.08.040
• Hernández-Becerra, E., Jímenez-Mendoza, D., Mutis-Gonzalez, N., Pineda-Gomez, P., Rojas-Molina, I., & Rodríguez-García, M. E. (2020). Calcium Deficiency in Diet Decreases the Magnesium Content in Bone and Affects Femur Physicochemical Properties in Growing Rats. Biological trace element research, 197(1), 224–232. https://doi.org/10.1007/s12011-019-01989-9
• Jäger, M., Sager, M., Lensing-Höhn, S., & Krauspe, R. (2005). The critical size bony defect in a small animal for bone healing studies (I): Comparative anatomical study on rats' femur. Biomedizinische Technik. Biomedical engineering, 50(4), 107–110. https://doi.org/10.1515/BMT.2005.015
• Järvinen, T. L., Sievänen, H., Kannus, P., & Järvinen, M. (1998). Dual-energy X-ray absorptiometry in predicting mechanical characteristics of rat femur. Bone, 22(5), 551–558. https://doi.org/10.1016/s8756-3282(98)00025-8
• Kim, M. Y., Kim, J. Y., Lim, D., Lee, D., Kim, Y., Chang, G. T., Choi, H. Y., & Kim, H. (2012). Skeletal growth and IGF levels in rats after HT042 treatment. Phytotherapy research : PTR, 26(12), 1771–1778. https://doi.org/10.1002/ptr.4642
• Kim, O. K., Yun, J. M., Lee, M., Park, S. J., Kim, D., Oh, D. H., Kim, H. S., & Kim, G. Y. (2020). A Mixture of Humulus japonicus Increases Longitudinal Bone Growth Rate in Sprague Dawley Rats. Nutrients, 12(9), 2625. https://doi.org/10.3390/nu12092625
• Lee, M. (1929). Determination of the surface area of the white rat with its application to the expression of metabolic results. American Journal of Physiology, 89(1), 24-33.
• Lezón, C. E., Olivera, M. I., Bozzini, C., Mandalunis, P., Alippi, R. M., & Boyer, P. M. (2009). Improved bone status by the beta-blocker propranolol in an animal model of nutritional growth retardation. The British journal of nutrition, 101(11), 1616–1620. https://doi.org/10.1017/s000711450811145x
• Maditz, K. H., Smith, B. J., Miller, M., Oldaker, C., & Tou, J. C. (2015). Feeding soy protein isolate and oils rich in omega-3 polyunsaturated fatty acids affected mineral balance, but not bone in a rat model of autosomal recessive polycystic kidney disease. BMC nephrology, 16, 13. https://doi.org/10.1186/s12882-015-0005-9
• Mahmoud, M., Kokozidou, M., Gögele, C., Werner, C., Auffarth, A., Kohl, B., Mrosewski, I., & Schulze-Tanzil, G. G. (2023). Does Vitamin K2 Influence the Interplay between Diabetes Mellitus and Intervertebral Disc Degeneration in a Rat Model?. Nutrients, 15(13), 2872. https://doi.org/10.3390/nu15132872
• Matuszewska, A., Nowak, B., Niżański, W., Eberhardt, M., Domrazek, K., Nikodem, A., Wiatrak, B., Zduniak, K., Olejnik, K., Merwid-Ląd, A., Tomkalski, T., Jędrzejuk, D., Szeląg, E., Podhorska-Okołów, M., Piotrowska, A., Jęśkowiak, I., Heinrich, A., Rutkowska, M., Dziewiszek, W., Sozański, T., … Szeląg, A. (2021). Long-Term Administration of Abacavir and Etravirine Impairs Semen Quality and Alters Redox System and Bone Metabolism in Growing Male Wistar Rats. Oxidative medicine and cellular longevity, 2021, 5596090. https://doi.org/10.1155/2021/5596090
• Novelli, E. L., Diniz, Y. S., Galhardi, C. M., Ebaid, G. M., Rodrigues, H. G., Mani, F., Fernandes, A. A., Cicogna, A. C., & Novelli Filho, J. L. (2007). Anthropometrical parameters and markers of obesity in rats. Laboratory animals, 41(1), 111–119. https://doi.org/10.1258/002367707779399518
• Nowińska, B., Folwarczna, J., Dusiło, A., Pytlik, M., Sliwiński, L., Cegieła, U., Kaczmarczyk-Sedlak, I., Pietryka, W., Hanke, T., & Trzeciak, H. I. (2012). Effects of vigabatrin on the skeletal system of young rats. Acta poloniae pharmaceutica, 69(2), 327–334.
• Padilla, C. J., Ferreyro, F. A., & Arnold, W. D. (2021). Anthropometry as a readily accessible health assessment of older adults. Experimental gerontology, 153, 111464. https://doi.org/10.1016/j.exger.2021.111464
• Paish, A. D. M., Nikolov, H. N., Welch, I. D., El-Warrak, A. O., Teeter, M. G., Naudie, D. D. R., & Holdsworth, D. W. (2020). Image-based design and 3D-metal printing of a rat hip implant for use in a clinically representative model of joint replacement. Journal of orthopaedic research : official publication of the Orthopaedic Research Society, 38(7), 1627–1636. https://doi.org/10.1002/jor.24706
• Pandit, A. P., Omase, S. B., & Mute, V. M. (2020). A chitosan film containing quercetin-loaded transfersomes for treatment of secondary osteoporosis. Drug delivery and translational research, 10(5), 1495–1506. https://doi.org/10.1007/s13346-020-00708-5
• Pazvant, G., & Kahvecioğlu, K. O. (2009). Tavşanlarda ön ve arka bacak uzun kemiklerinin homotipik varyasyonları üzerinde araştırmalar. İstanbul Üniversitesi Veteriner Fakültesi Dergisi, 35(2), 23–39. https://doi.org/10.16988/iuvfd.76069
• Prodinger, P. M., Foehr, P., Bürklein, D., Bissinger, O., Pilge, H., Kreutzer, K., von Eisenhart-Rothe, R., & Tischer, T. (2018). Whole bone testing in small animals: systematic characterization of the mechanical properties of different rodent bones available for rat fracture models. European journal of medical research, 23(1), 8. https://doi.org/10.1186/s40001-018-0307-z
• Prodinger, P. M., Bürklein, D., Foehr, P., Kreutzer, K., Pilge, H., Schmitt, A., Eisenhart-Rothe, R. V., Burgkart, R., Bissinger, O., & Tischer, T. (2018). Improving results in rat fracture models: enhancing the efficacy of biomechanical testing by a modification of the experimental setup. BMC musculoskeletal disorders, 19(1), 243. https://doi.org/10.1186/s12891-018-2155-y
• Ponyrko, A. O., Bumeister, V. I., Dmytruk, S. M., Yarmolenko, O. S., Teslyk, T. P., Riabenko, T. V., & Shkolna, I. (2021). Structural Changes Of Long Tubular Bones Of Mature Rats Under The Hyperglycemıa. Wiadomosci lekarskie (Warsaw, Poland : 1960), 74(9 cz 1), 2052–2059.
• Rumbo-Rodríguez, L., Sánchez-SanSegundo, M., Ferrer-Cascales, R., García-D'Urso, N., Hurtado-Sánchez, J. A., & Zaragoza-Martí, A. (2021). Comparison of Body Scanner and Manual Anthropometric Measurements of Body Shape: A Systematic Review. International journal of environmental research and public health, 18(12), 6213. https://doi.org/10.3390/ijerph18126213
• Sato, D., Takahata, M., Ota, M., Fukuda, C., Tsuda, E., Shimizu, T., Okada, A., Hiruma, Y., Hamano, H., Hiratsuka, S., Fujita, R., Amizuka, N., Hasegawa, T., & Iwasaki, N. (2018). Siglec-15-targeting therapy increases bone mass in rats without impairing skeletal growth. Bone, 116, 172–180. https://doi.org/10.1016/j.bone.2018.07.026
• Sato, Y., Tagami, T., Akimoto, T., Takiguchi, T., Endo, Y., Tsukamoto, T., Hara, Y., & Yokobori, S. (2024). Development and validation of a novel overhead method for anteroposterior radiographs of fractured rat femurs. Scientific reports, 14(1), 5536. https://doi.org/10.1038/s41598-024-56238-4
• Shantanu, K., Singh, S., Kumar, D., Singh, A., Tewari, P. G., & Gupta, P. (2023). Anatomical Variation in Quadriceps Angle With Regard to Different Anthropometric Parameters in a Tertiary Care Center in Northern India: A Descriptive Study. Cureus, 15(1), e34224. https://doi.org/10.7759/cureus.34224
• Silveira, E. M. S., Santos, M. C. Q., da Silva, T. C. B., Silva, F. B. O., Machado, C. V., Elias, L., Kolberg, A., Kroth, A., & Partata, W. A. (2020). Aging and low-intensity exercise change oxidative biomarkers in brain regions and radiographic measures of femur of Wistar rats. Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas, 53(6), e9237. https://doi.org/10.1590/1414-431x20209237
• Soon, G., Quintin, A., Scalfo, F., Antille, N., Williamson, G., Offord, E., & Ginty, F. (2006). PIXImus bone densitometer and associated technical measurement issues of skeletal growth in the young rat. Calcified tissue international, 78(3), 186–192. https://doi.org/10.1007/s00223-005-0191-8
• Williams, J. A., Windmill, J. F. C., Tanner, K. E., Riddell, J. S., & Coupaud, S. (2019). Global and site-specific analysis of bone in a rat model of spinal cord injury-induced osteoporosis. Bone reports, 12, 100233. https://doi.org/10.1016/j.bonr.2019.100233
• Wong, K. L., Zhang, S., Tan, S. S. H., Cheow, Y. A., Lai, R. C., Lim, S. K., Hui, J. H. P., & Toh, W. S. (2022). Mesenchymal Stem Cell Exosomes Promote Growth Plate Repair and Reduce Limb-Length Discrepancy in Young Rats. The Journal of bone and joint surgery. American volume, 104(12), 1098–1106. https://doi.org/10.2106/JBJS.21.00789