The contribution of collagen fibers to the mechanical compressive properties of the temporomandibular joint disc  S. Fazaeli, S. Ghazanfari, V. Everts,

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The contribution of collagen fibers to the mechanical compressive properties of the temporomandibular joint disc  S. Fazaeli, S. Ghazanfari, V. Everts, T.H. Smit, J.H. Koolstra  Osteoarthritis and Cartilage  Volume 24, Issue 7, Pages 1292-1301 (July 2016) DOI: 10.1016/j.joca.2016.01.138 Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Schematic view of the TMJ. The TMJ disc (blue), and the articular cartilage of the condyle and mandibular fossa (green) are shown from sagittal view. Osteoarthritis and Cartilage 2016 24, 1292-1301DOI: (10.1016/j.joca.2016.01.138) Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Indentation test on TMJ disc of porcine sample by the custom-made indentation device. Osteoarthritis and Cartilage 2016 24, 1292-1301DOI: (10.1016/j.joca.2016.01.138) Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Effect of collagenase treatment on gross morphology of the TMJ disc. TMJ discs are shown from a superoinferior view after PBS/collagenase incubation (A–B) and after harvesting 4 mm specimens from their five different regions (C–D). The loose gel-like stratum on the surface of treated (collagenase) sample has covered the punched areas on the five regions of the disc (D). Osteoarthritis and Cartilage 2016 24, 1292-1301DOI: (10.1016/j.joca.2016.01.138) Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Effect of collagenase treatment on the stress response during cyclic loading. (A) stress vs time and (B) hysteresis in stress-strain response. The control (PBS) data (black) and treated (collagenase) data (red) indicate the significant reduction of stress and energy dissipation capacity of the TMJ disc after the collagenase treatment. Osteoarthritis and Cartilage 2016 24, 1292-1301DOI: (10.1016/j.joca.2016.01.138) Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Effect of collagenase treatment on mechanical compressive properties of the TMJ discs. A: Collagenase treatment had a substantial effect on the instantaneous modulus of TMJ discs in all regions. The regional variation was observed for control (PBS) samples. B: Both collagenase treatment and disc region factors had a significant effect on energy dissipation capabilities of the TMJ discs. Collagenase treatment substantially reduced the maximum energy dissipation capability within all regions, which shows similar tendency observed for the instantaneous modulus data. All data are presented as mean ± 95% confidence interval. *indicates significant difference (n = 5, P < 0.05). Osteoarthritis and Cartilage 2016 24, 1292-1301DOI: (10.1016/j.joca.2016.01.138) Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Effect of collagenase treatment on collagen and GAG content of TMJ discs. (A) Approximately 80% of the dry weight of TMJ disc is composed of collagen fibers. Despite overall decrease of collagen content across the TMJ disc, the collagenase treatment did not induce significant collagen loss within single regions of the treaded (collagenase) sample. (B) GAG only consists less than 1% of TMJ disc's dry weight. Collagenase treatment induced GAG reduction in all regions. Also as a heterogeneous tissue, control (PBS) samples showed GAG regional variation. All data are presented as mean ± 95% confidence interval. *indicates significant difference (n = 5, P < 0.05). Osteoarthritis and Cartilage 2016 24, 1292-1301DOI: (10.1016/j.joca.2016.01.138) Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

Fig. 7 Effect of collagenase treatment on the integrity of the collagenous network. PLM images of representative regions of TMJ disc seen from above. The region-dependent collagen fiber alignment with its typical ring-like structure is clearly shown across all regions. Comparison of control (PBS) samples with treated (collagenase) samples in each region shows the lack of structural integrity in the collagenous network of the latter group. Scale bar: 2 mm. Osteoarthritis and Cartilage 2016 24, 1292-1301DOI: (10.1016/j.joca.2016.01.138) Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

Fig. 8 Effect of collagenase treatment on the integrity of the collagenous network. PLM images of representative regions of TMJ disc seen in the sagittal plane. The collagen fibers are aligned anteroposteriorly perpendicular to the mediolaterally oriented fibers in the PB and AB (not shown in this view). Comparison of control (PBS) samples with treated (collagenase) samples shows the lack of structural integrity in the collagenous network of the latter group. Scale bar: 2 mm. Osteoarthritis and Cartilage 2016 24, 1292-1301DOI: (10.1016/j.joca.2016.01.138) Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions