Duchenne muscular dystrophy is caused by the absence of dystrophin. This study aimed to investigate femoral morphological characteristics of lack of dystrophin in MDX mice, considering that this model, different from DMD patient, is not influenced by corticosteroids administration and limited ambulation.
Materials and methods
Proximal femur of male 16-week-old Control and MDX mice were submitted to histological, morphometric (volume density of articular cartilage, compact bone, trabecular bone and bone marrow; articular cartilage layers area; articular cartilage cell area), and immunohistochemistry analysis for RUNX-2, RANK-L, MMP-2, MMP-9, Caspase-3 and KI-67.
MDX showed loss of linearity of articular cartilage with subchondral bone transition and elevation of this subchondral bone to the articular surface when compared with control. In addition, MDX presented morphological difference in the pantographic network of collagen fibers. Volume density of trabecular bone tissue was higher in the MDX than Control, but volume density of articular cartilage was lower in MDX (p < 0.05). The articular cartilage layers and chondrocytes area were significantly smaller in MDX than Control. These results associated to MMPs and osteogenic markers of proximal femur revealed an adaptation process as a consequence of lack of dystrophin.
The morphological changes observed in the bone tissue of the MDX may be not only secondary to muscle weakness or chronic use of corticosteroids but also our results indicate connections between decrease of cartilage thickness, collagen network alteration and consequent subchondral changes that may lead to articular cartilage degeneration and bone adaptation mechanism in MDX mice.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price includes VAT (USA)
Tax calculation will be finalised during checkout.
Lovering RM, Porter NC, Bloch RJ (2005) The muscular dystrophies: from genes to therapies. Phys Ther 85:1372–1388
Mendell JR, Rodino-Klapac LR, Sahenk Z, Roush K, Bird L, Lowes LP et al (2013) Eteplirsen for the treatment of Duchenne muscular dystrophy. Ann Neurol 74:637–647
Nakagaki WR, Bertran CA, Matsumura CY, Santo-Neto H, Camilli JA (2011) Mechanical, biochemical and morphometric alterations in the femur of mdx mice. Bone 48:372–379
Slemenda CW, Reister TK, Hui SL, Miller JZ, Christian JC, Johnston CC Jr (1994) Influences on skeletal mineralization in children and adolescents: evidence for varying effects of sexual maturation and physical activity. J Pediatr 125:201–207
Crabtree NJ, Roper H, McMurchie H, Shaw NJ (2010) Regional changes in bone area and bone mineral content in boys with duchenne muscular dystrophy receiving corticosteroid therapy. J Pediatr 156:450–455
McDonald DG, Kinali M, Gallagher AC, Mercuri E, Muntoni F, Roper H, Jardine P, Jones DH, Pike MG (2002) Fracture prevalence in Duchenne muscular dystrophy. Dev Med Child Neurol 44:695–698
Söderpalm AC, Magnusson P, Ahlander AC, Karlsson J, Kroksmark AK, Tulinius M, Swolin-Eide D (2007) Low bone mineral density and decreased bone turnover in Duchenne muscular dystrophy. Neuromuscul Disord 17:919–928
Janssen PM, Murray JD, Schill KE, Rastogi N, Schultz EJ, Tran T, Raman SV, Rafael-Fortney JA (2014) Prednisolone attenuates improvement of cardiac and skeletal contractile function and histopathology by lisinopril and spironolactone in the mdx mouse model of Duchenne muscular dystrophy. PLoS One 9:e88360
King WM, Kissel JT, Visy D, Goel PK, Matkovic V (2014) Skeletal health in Duchenne dystrophy: bone-size and subcranial dual-energy X-ray absorptiometry analyses. Muscle Nerve 49:512–519
Tian C, Wong BL, Hornung L, Khoury JC, Miller L, Bange J, Rybalsky I, Rutter MM (2016) Bone health measures in glucocorticoid-treated ambulatory boys with Duchenne muscular dystrophy. Neuromuscul Disord 26:760–767
Buckner JL, Bowden SA, Mahan JD (2015) Optimizing bone health in Duchenne muscular dystrophy. Int J Endocrinol 2015:928385
Martel-Pelletier J, Boileau C, Pelletier JP, Roughley PJ (2008) Cartilage in normal and osteoarthritis conditions. Best Pract Res Clin Rheumatol 22:351–384
Akkiraju H, Nohe A (2015) Role of chondrocytes in cartilage formation, progression of osteoarthritis and cartilage regeneration. J Dev Biol 3:177–192
Isaac C, Wright A, Usas A, Li H, Tang Y, Mu X, Greco N, Dong Q, Vo N, Kang J, Wang B, Huard J (2013) Dystrophin and utrophin “double knockout” dystrophic mice exhibit a spectrum of degenerative musculoskeletal abnormalities. J Orthop Res 31:343–349
Larcher T, Lafoux A, Tesson L, Remy S, Thepenier V, François V, Le Guiner C, Goubin H, Dutilleul M, Guigand L, Toumaniantz G, De Cian A, Boix C, Renaud JB, Cherel Y, Giovannangeli C, Concordet JP, Anegon I, Huchet C (2014) Characterization of dystrophin deficient rats: a new model for Duchenne muscular dystrophy. PLoS One 9:e110371
Vainzof M, Ayub-Guerrieri D, Onofre PC, Martins PC, Lopes VF, Zilberztajn D, Maia LS, Sell K, Yamamoto LU (2008) Animal models for genetic neuromuscular diseases. J Mol Neurosci 34:241–248
Manning J, O’Malley D (2015) What has the mdx mouse model of Duchenne muscular dystrophy contributed to our understanding of this disease? J Muscle Res Cell Motil 36:155–167
Pelosi L, Berardinelli MG, Forcina L, Spelta E, Rizzuto E, Nicoletti C, Camilli C, Testa E, Catizone A, De Benedetti F, Musarò A (2015) Increased levels of interleukin-6 exacerbate the dystrophic phenotype in mdx mice. Hum Mol Genet 1:6041–6053
Abou-Khalil R, Yang F, Mortreux M, Lieu S, Yu YY, Wurmser M, Pereira C, Relaix F, Miclau T, Marcucio RS, Colnot C (2014) Delayed bone regeneration is linked to chronic inflammation in murine muscular dystrophy. J Bone Miner Res 29:304–315
Junqueira LC, Bignolas G, Bretani RR (1979) Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochemistry 11:447–455
Mandarim-de-Lacerda CA (2003) Stereological tools in biomedical research. An Acad Bras Ciênc 75:469–486
De Oliveira BC, de Oliveira F, Martini DT, Prisco CR, da Silva Riguetti MM, Liberti EA, de Campos BS (2010) The relative effects of severe burn injury and pre- and post-natal protein deprivation on mandibular condyle morphology. Histol Histopathol 25:45–54
Novotny SA, Warren GL, Lin AS, Guldberg RE, Baltgalvis KA, Lowe DA (2011) Bone is functionally impaired in dystrophic mice but less so than skeletal muscle. Neuromuscul Disord 21:183–193
Rufo A, Del Fattore A, Capulli M, Carvello F, De Pasquale L, Ferrari S, Pierroz D, Morandi L, De Simone M, Rucci N, Bertini E, Bianchi ML, De Benedetti F, Teti A (2011) Mechanisms inducing low bone density in Duchenne muscular dystrophy in mice and humans. J Bone Miner Res 26:1891–1903
Yoon SH, Grynpas MD, Mitchell J (2019) Growth hormone increases bone toughness and decreases muscle inflammation in glucocorticoid-treated Mdx mice, model of Duchenne muscular dystrophy. J Bone Miner Res 34:1473–1486
Maldonado M, Nam J (2013) The role of changes in extracellular matrix of cartilage in the presence of inflammation on the pathology of osteoarthritis. Biomed Res Int 2013:284873
Becerra J, Andrades JA, Guerado E, Zamora-Navas P, López-Puertas JM, Reddi AH (2010) Articular cartilage: structure and regeneration. Tissue Eng Part B Rev 16:617–627
Aho O-M, Finnilä M, Thevenot J, Saarakkala S, Lehenkari P (2017) Subchondral bone histology and grading in osteoarthritis. PLoS ONE 12:e0173726
Lu CH, Yeh TS, Yeh CL, Fang YH, Sung LY, Lin SY, Yen TC, Chang YH, Hu YC (2014) Regenerating cartilages by engineered ASCs: prolonged TGF-β3/BMP-6 expression improved articular cartilage formation and restored zonal structure. Mol Ther 22:186–195
Goldring MB, Marcu KB (2009) Cartilage homeostasis in health and rheumatic diseases. Arthritis Res Ther 11:224
Houard X, Goldring MB, Berenbaum F (2013) Homeostatic mechanisms in articular cartilage and role of inflammation in osteoarthritis. Curr Rheumatol Rep 15:375
Cruz-Guzmán Odel R, Rodríguez-Cruz M, Escobar Cedillo RE (2015) Systemic inflammation in Duchenne muscular dystrophy: association with muscle function and nutritional status. Biomed Res Int 2015:891972
Kondo M, Yamaoka K, Tanaka Y (2014) Acquiring chondrocyte phenotype from human mesenchymal stem cells under inflammatory conditions. Int J Mol Sci 17:21270–21285
Pearle AD, Warren RF, Rodeo SA (2005) Basic science of articular cartilage and osteoarthritis. Clin Sports Med 24:1–12
Wei B, Gu Q, Li D, Yan J, Guo Y, Mao F, Xu Y, Zang F, Wang L (2014) Mild degenerative changes of hip cartilage in elderly patients: an available sample representative of early osteoarthritis. Int J Clin Exp Pathol 15:6493–6503
Florencio-Silva R, Sasso GR, Sasso-Cerri E, Simões MJ, Cerri OS (2015) Biology of bone tissue: structure, function, and factors that influence bone cells. Biome Res Int 2015:421746
Galliera E, Randelli P, Dogliotti G, Dozio E, Colombini A, Lombardi G, Cabitza P, Corsi MM (2010) Matrix metalloproteases MMP-2 and MMP-9: are they early biomarkers of bone remodelling and healing after arthroscopic acromioplasty? Injury 41:1204–1207
Ben David D, Reznick AZ, Srouji S, Livne E (2008) Exposure to pro-inflammatory cytokines upregulates MMP-9 synthesis by mesenchymal stem cells-derived osteoprogenitors. Histochem Cell Biol 129:589–597
Ruiz PLM, Handan BA, de Moura CFG, Assis LR, Fernandes KR, Renno ACM, Ribeiro DA (2018) Protective effect of grape or apple juices in bone tissue of rats exposed to cadmium: role of RUNX-2 and RANK/L expression. Environ Sci Pollut Res Int 25:15785–15792
Ueta C, Iwamoto M, Kanatani N, Yoshida C, Liu Y, Enomoto-Iwamoto M, Ohmori T, Enomoto H, Nakata K, Takada K, Kurisu K, Komori T (2001) Skeletal malformations caused by overexpression of Cbfa1 or its dominant negative form in chondrocytes. J Cell Biol 2:87–100
Baum R, Gravallese EM (2014) Impact of inflammation on the osteoblast in rheumatic diseases. Curr Osteoporos Rep 12:9–16
Cardoso L, Herman BC, Verborgt O, Laudier D, Majeska RJ, Schaffler MB (2009) Osteocyte apoptosis controls activation of intracortical resorption in response to bone fatigue. J Bone Miner Res 24:597–605
Kennedy OD, Laudier DM, Majeska RJ, Sun HB, Schaffler MB (2014) Osteocyte apoptosis is required for production of osteoclastogenic signals following bone fatigue in vivo. Bone 64:132–137
Authors would like to thank Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) for the scholarship (grant number 14/22211-6).
Conflict of interest
The authors of the manuscript have no conflict of interest to declare.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
dos Santos, J.F., Lazzarin, M.C., Baptista, V.I.A. et al. Articular cartilage degeneration and bone adaptation due to lack of dystrophin in mice. J Bone Miner Metab (2021). https://doi.org/10.1007/s00774-021-01270-x
- MDX mice
- Duchenne muscular dystrophy
- Articular cartilage