Histological investigation for the effect of botulinum toxin B inoculation in the gluteal group muscles of the thigh on the femoral bone of albino rabbits

S. M. Abdulateef; R. H. Thamer; A. K. Obayes

doi:10.15421/0225083

S. M. Abdulateef Al-Nahrain University
R. H. Thamer Al-Nahrain University
A. K. Obayes Al-Nahrain University

Keywords: botulinum toxin B, femur bone, gluteal group muscles, inoculation, periosteum.

Abstract

Botulinum toxin B is an attenuated toxin produced by the bacterium Clostridium botulinum used in non-surgical cosmetic operation s and for correcting skin wrinkles. This article was designed to describe t he effect of denervated hind limb skeletal mu s cle impact on the femur bone of albino rabbits . Twenty-five male animals were enrolled in the five study groups . Five rabbits served as the control group, were given only distilled water and sacrificed at the end of the study for histological study. The re maining 20 rabbits were inoculat ed with a single dosage of b otulinum toxin B (8 IU) between the gluteal muscle group and the b iceps femoris muscle of the thigh, and each group of five w as sacrificed sequentially on day 10, day 15, day 30, and day 45 after the i noculation. Samples of femur bone were removed from the dissected animals. The microscopic examination of femur bone sections of the experimental 1st group showed an irregular endosteum border and narrowed bone collar. Animals from the experimental 2nd group showed remnant lacunae of hyaline cartilage and atrophied osteocytes. In the experimental 3rd group, results showed increased cavities within the bone matrix. In the experimental 4th group, the microscopic examination revealed the diminish ed periosteal membrane to the endosteum, atrophied lacunae, surrounded with irregular periosteum. The femoral bone displayed numerous histological changes due to the flaccid paralysis of the gluteal group muscles (disused muscles of the hi n d l imb) of the thigh caused by a single botulinum toxin B dose.

References

Ajitha, R., Preethi, M., Sree, K. L., & Anbukkarasi, K. (2024). A histomorphological study of urinary bladder lesions in a tertiary care hospital. Texila International Journal of Public Health, 12(3), 48.

Al-Bari, A. A., & Al Mamun, A. (2020). Current advances in regulation of bone homeostasis. FASEB BioAdvances, 2(11), 668–679.

Alencar, C. H. M. F., Silveira, C. R. S., Cavalcante, M. M., Vieira, C. G. M., Teixeira, M. J. D., Neto, F. A., de Abreu, A., & Chhabra, A. (2020). Periosteum: An imaging review. European Journal of Radiology Open, 7, 100249.

Ali, D., Abdelzaher, W., & Abdel-Hafez, S. (2018). Evaluation of the rivastigmine role against botulinum toxin-A-induced osteoporosis in albino rats: A biochemical, histological, and immunohistochemical study. Human and Experimental Toxicology, 37(12), 1323–1335.

Ali, N. K. (2018). Estimation of some mineral (calcium, phosphorous, vitamin 25(OH) D, and alkaline phosphatase) in osteoporosis patients in Kirkuk city. Journal of Osteoporosis and Physical Activity, 6(2), 2–4.

Ali, S., Sulaiman, E., & Dhiaa, S. (2024). Histological effects of co enzyme Q10 on doxorubicin-induced deficits of cardiopulmonary axis in white albino rats. Georgian Medical News, (349), 54–59.

Alizadeh, A. M., Hashempour-Baltork, F., Alizadeh-Sani, M., Maleki, M., Azizi-Lalabad, M., & Khosravi-Darani, K. (2020). Inhibition of Clostridium (C.) botulinum and its toxins by probiotic bacteria and their metabolites: An update review. Quality Assurance and Safety of Crops and Foods, 12(SP1), 59–68.

Alkhafaji, N., Ahmed, M., & Karem, B. (2024). The effect of vitamin D on the histological structure of liver and lung in mice treated with amphotericin B. Georgian Medical News, (355), 134–141.

Al-Suhaimi, E. A. (2022). Bone remodeling physiology: Regulation of parathyroid glands, C cells, vitamin D, and bone as an endocrine organ. In: Al-Suhaimi, E. A. (Eds.). Emerging concepts in endocrine structure and functions. Springer Nature Singapore, Singapore. Pp. 161–199.

Ashley, Z., Sutherland, H., Lanmüller, H., Russold, M. F., Unger, E., Bijak, M., Mayr, W., Boncompagni, S., Protasi, F., Salmons, S., & Jarvis, J. C. (2007). Atrophy, but not necrosis, in rabbit skeletal muscle denervated for periods up to one year. American Journal of Physiology – Cell Physiology, 292(1), 440–451.

Atiyeh, B. S., Rubeiz, M. T., & Hayek, S. N. (2008). Aesthetic/cosmetic surgery and ethical challenges. Aesthetic Plastic Surgery, 44(4), 1364–1374.

Bartl, R. (2023). Secondary osteoporosis in medical disciplines. In: Bartl, R. (Ed.). Osteoporosis in clinical practice. Springer International Publishing, Cham. Pp. 165–187.

Bianco, P. (2020). Structure and mineralization of bone. In: Bonucci, E. (Ed.). Calcification in biological systems. CRC Press, Boca Raton. Pp. 243–268.

Bomeli, S. R., Desai, S. C., Johnson, J. T., & Walvekar, R. R. (2008). Management of salivary flow in head and neck cancer patients – A systematic review. Oral Oncology, 44(11), 1000–1008.

Brent, M. B., Brüel, A., & Thomsen, J. S. (2021). A systematic review of animal models of disuse – induced bone loss. Calcified Tissue International, 108(5), 561–575.

Brotto, M., & Bonewald, L. (2015). Bone and muscle: Interactions beyond mechanical. Bone, 80, 109–114.

Brown, A., & Teller, C. (2022). Identifying and managing complications caused by cosmetic neurotoxin treatment. Dermatological Reviews, 3(4), 247–256.

Carina, V., Della Bella, E., Costa, V., Bellavia, D., Veronesi, F., Cepollaro, S., Fini, M., & Giavaresi, G. (2020). Bone’s response to mechanical loading in aging and osteoporosis: Molecular mechanisms. Calcified Tissue International, 107(4), 301–318.

Chappard, D., Chennebault, A., Moreau, M., Legrand, E., Audran, M., & Basle, M. F. (2001). Texture analysis of X-ray radiographs is a more reliable descriptor of bone loss than mineral content in a rat model of localized disuse induced by the Clostridium botulinum toxin. Bone, 28(1), 72–79.

Chen, H., Tian, X., Liu, X., Setterberg, R. B., Li, M., & Jee, W. S. S. (2008). Alfacalcidol-stimulated focal bone formation on the cancellous surface and increased bone formation on the periosteal surface of the lumbar vertebrae of adult female rats. Calcified Tissue International, 82(2), 127–136.

Delgado-Calle, J., & Bellido, T. (2022). The osteocyte as a signaling cell. Physiological Reviews, 102(1), 379–410.

Deng, J., Cohen, D. J., Redden, J., McClure, M. J., Boyan, B. D., & Schwartz, Z. (2021). Differential effects of neurectomy and botox-induced muscle paralysis on bone phenotype and titanium implant osseointegration. Bone, 153, 116145.

Devadze, R., Gvenetadze, A., Burkadze, G., & Kepuladze, S. (2022). Features of distribution of intratumoral lymphocytes in ovarian epithelial tumours of different histological types and degree of malignancy. Georgian Scientists, 4(5), 383–393.

Dutra, E. H., O’ Brien, M. H., Lima, A., Kalajzic, Z., Tadinada, A., Nanda, R., & Yadav, S. (2016). Cellular and matrix response of the mandibular condylar cartilage to botulinum toxin. PLoS One, 11(10), e0164599.

Griffiths, D., & Mullock, A. (2017). Cosmetic surgery: Regulatory challenges in a global beauty market. Health Care Analysis, 26(3), 220–234.

Gropp, K. E., & Varela, A. (2024). Bones, joints, and teeth. In: Haschek, W. M., Rousseaux, C. G., ... & Bolon, B. (Eds.). Haschek and Rousseaux’ s Handbook of toxicologic pathology. Academic Press. Pp. 249–360.

Hao, Y., Ma, Y., Wang, X., Jin, F., & Ge, S. (2011). Short-term muscle atrophy caused by botulinum toxin-A local injection impairs fracture healing in the rat femur. Journal of Orthopaedic Research, 30(4), 574–580.

Kelly, R. R., Sidles, S. J., & LaRue, A. C. (2020). Effects of neurological disorders on bone health. Frontiers in Psychology, 11, 612366.

Kim, H. J., Yum, K.-W., Lee, S.-S., Heo, M.-S., & Seo, K. (2003). Effects of botulinum toxin type A on bilateral masseteric hypertrophy evaluated with computed tomographic measurement. Dermatologic Surgery, 29(5), 484–489.

Kingery, W. (2003). A substance P receptor (NK1) antagonist enhances the widespread osteoporotic effects of sciatic nerve section. Bone, 33(6), 927–936.

Lanyon, L. E. (1996). Using functional loading to influence bone mass and architecture: objectives, mechanisms, and relationship with estrogen of the mechanically adaptive process in bone. Bone, 18(1), S37–S43.

Liu, S., Liu, S., Li, S., Liang, B., Han, X., Liang, Y., & Wei, X. (2023). Nerves within bone and their application in tissue engineering of bone regeneration. Frontiers in Neurology, 13, 1085560.

Nazzal, M. K., Morris, A. J., Parker, R. S., White, F. A., Natoli, R. M., Kacena, M. A., & Fehrenbacher, J. C. (2024). Do not lose your nerve, be callus: Insights into neural regulation of fracture healing. Current Osteoporosis Reports, 22(1), 182–192.

Nichat, P., Varadarajan, S., Balaji, T. M., Rao, N. N., Jayanandan, M., Govindarajan, S. (2024). Myofibroblasts and tumor micro-environment in oral squamous cell carcinomas – a histochemical and immunohistochemical analysis. Texila International Journal of Public Health, 12(3), 53.

Owen, M., Gray, B., Hack, N., Perez, L., Allard, R. J., & Hawkins, J. M. (2022). Impact of botulinum toxin injection into the masticatory muscles on mandibular bone: A systematic review. Journal of Oral Rehabilitation, 49(6), 644–653.

Park, M. Y., & Ahn, K. Y. (2021). Scientific review of the aesthetic uses of botulinum toxin type A. Archives of Craniofacial Surgery, 22(1), 1–10.

Plotkin, L. I., Mathov, I., Aguirre, J. I., Parfitt, A. M., Manolagas, S. C., & Bellido, T. (2005). Mechanical stimulation prevents osteocyte apoptosis: Requirement of integrins, Src kinases, and ERKs. American Journal of Physiology – Cell Physiology, 289(3), C633–C643.

Preethi, M., Roshan, A., Niranjan N., & Ajitha, R. (2024). A comparative study on rapid, simple and cost-effective method in determining the decalcification agent in bone tissue processing in hematoxylin and eosin staining procedure. Texila International Journal of Public Health, 12(3), 41.

Raman, S., Yamamoto, Y., Suzuki, Y., & Matsuka, Y. (2023). Mechanism and clinical use of botulinum neurotoxin in head and facial region. Journal of Prosthodontic Research, 67(4), 493–505.

Ravi, C., Manogaran, Y., & Ramasamy, P. (2025). Extraction, characterization and antioxidative potential of a bioactive polymeric material from the cuttlebone of Sepia brevimana. Texila International Journal of Public Health, 13(1), 69.

Ru, J., & Wang, Y. (2020). Osteocyte apoptosis: The roles and key molecular mechanisms in resorption-related bone diseases. Cell Death and Disease, 11(10), 846.

Srivastava, S., Kharbanda, S., Pal, U., & Shah, V. (2015). Applications of botulinum toxin in dentistry: A comprehensive review. National Journal of Maxillofacial Surgery, 6(2), 152–159.

Taha, M. M., Taha, E. M., & Mohammed, S. K. (2023). The role of testosterone level in women with osteopenia. Baghdad Science Journal, 21(2), 19.

Takata, S., & Yasui, N. (2001). Disuse osteoporosis. The Journal of Medical Investigation, 48(3–4), 147–156.

Tukeshov, S., Baysekeev, T., Choi, E., Kulushova, G., Nazir, M., Jaxymbayev, N., & Turkmenov, A. (2024). Osteosynthesis of complex comminuted hand bone fractures by applying the lacing method (a clinical case study). Georgian Medical News, 348, 40–43.

Vieira, A. E., Repeke, C. E., Ferreira Junior, S. de B., Colavite, P. M., Biguetti, C. C., Oliveira, R. C., Assis, G. F., Taga, R., Trombone, A. P. F., & Garlet, G. P. (2015). Intramembranous bone healing process subsequent to tooth extraction in mice: Micro-computed tomography, histomorphometric and molecular characterization. PLoS One, 10(5), e0128021.

Wang, P., Wang, C., Meng, H., Liu, G., Li, H., Gao, J., Tian, H., & Peng, J. (2022). The role of structural deterioration and biomechanical changes of the necrotic lesion in collapse mechanism of osteonecrosis of the femoral head. Orthopaedic Surgery, 14(5), 831–839.

Warner, S. E., Sanford, D. A., Becker, B. A., Bain, S. D., Srinivasan, S., & Gross, T. S. (2006). Botox induced muscle paralysis rapidly degrades bone. Bone, 38(2), 257–264.

Wawrzyniak, A., & Balawender, K. (2022). Structural and metabolic changes in bone. Animals, 12(15), 1946.

Wu, X., Xu, X., Liu, Q., Ding, J., Liu, J., Huang, Z., Huang, Z., Wu, X., Li, R., Yang, Z., Jiang, H., Liu, J., & Zhu, Q. (2021). Unilateral cervical spinal cord injury induces bone loss and metabolic changes in non-human primates (Macaca fascicularis). Journal of Orthopaedic Translation, 29, 113–122.

Xu, R., Li, S., Zheng, A., & He, L. (2024). Effect of Xiaoyao pills combined with alendronate on bone density in postmenopausal patients with osteoporosis. Georgian Medical News, 351, 100–101.

Yevchuk, Y., Rozhko, M., Pantus, A., Yarmoshuk, I., & Pantus, P. (2024). Analysis of the clinical effectiveness of using the created combined fibrin-bone scaffold for the reconstruction of bone tissue defects of the jaws. Georgian Medical News, 352–353, 6–13.

Zeng, Q. Q., Jee, W. S., Bigornia, A. E., King Jr., J. G., D'Souza, S. M., Li, X. J., Ma, Y. F., & Wechter, W. J. (1996). Time responses of cancellous and cortical bones to sciatic neurectomy in growing female rats. Bone, 19(1), 13–21.

Zhou, C., Zhang, W., Pang, W., Lin, H., Chen, M., & Wu, L. (2020). Dynamic changes of neuromuscular structure and function after local injection of botulinum toxin A. Revista Científica de la Facultad de Ciencias Veterinarias, 30(5), 2331–2341.

Zhu, S., Chen, M., Ying, Y., Wu, Q., Huang, Z., Ni, W., Wang, X., Xu, H., Bennett, S., Xiao, J., & Xu, J. (2022). Versatile subtypes of pericytes and their roles in spinal cord injury repair, bone development and repair. Bone Research, 10(1), 30.