Immunohistochemistry
Indirect immunohistochemistry was performed for the specific
identification of ECM components and cell phenotype. In relation to the
ECM fibers, collagens types I, III and IV were determined using this
method. In addition, versican (VSC), a hyaluronan-binding proteoglycan
which interacts with other ECM components and contributes to control
tissue hydration was also studied (Godoy-Guzmán et al., 2018). To
identify specific cell phenotypes, immunohistochemical methods were used
for the proteins CD73 (marker of stemness and undifferentiation), S100
(marker of chondrogenic and neural crest lineage) and CD34 a well-known
marker of blood vessels (García-Martínez et al., 2017; Vela-Romera
et al., 2019). All immunohistochemical analyses were carried out using
routine methods, and were conducted using the same conditions for all
samples. Technical details are summarized in Supplementary Table S1.