Proteomic analysis of the extracellular matrix in idiopathic pes equinovarus

Abstract

Idiopathic pes equinovarus is a congenital deformity of the foot and lower leg defined as a fixation of the foot in adduction, supination, and varus. Although the pathogenesis of clubfoot remains unclear, it has been suggested that fibroblasts and growth factors are involved. To directly analyze the protein composition of the extracellular matrix in contracted tissue of patients with clubfoot. A total of 13 infants with idiopathic clubfoot treated with the Ponseti method were included in the present study. Tissue samples were obtained from patients undergoing surgery for relapsed clubfeet. Contracted tissues were obtained from the medial aspect of the talonavicular joint. Protein was extracted after digestion and delipidation using zip-tip C18. Individual collagenous fractions were detected using a chemiluminescent assay. Amino acid analysis of tissue samples revealed a predominance of collagens, namely collagen types I, III, and VI. The high content of glycine and h-proline suggests a predominance of collagens I and III. A total of 19 extracellular matrix proteins were identified. The major result of the present study was the observation that the extracellular matrix in clubfoot is composed of an additional 16 proteins, including collagens V, VI, and XII, as well as the previously described collagen types I and III and transforming growth factor β. The characterization of the general protein composition of the extracellular matrix in various regions of clubfoot may help in understanding the pathogenesis of this anomaly and, thus, contribute to the development of more efficacious therapeutic approaches.

Integrated bioinformatics analysis of potential pathway biomarkers using abnormal proteins in clubfoot

“…Abnormal biological development, such as muscle, neurological, skeletal and vascular abnormalities, has been previously identified and confirmed (Basit & Khoshhal, 2018;Eckhardt et al, 2019;Herceg et al, 2006;Hester et al, 2009;Lovell & Morcuende, 2007;Ošt'ádal et al, 2015). The HOX and TBX families governed limb identity, and fibroblast growth factor participated in the formation of limb muscles (Ohuchi & Noji, 1999;Wang et al, 2008).…”

“…Select soft tissues in clubfoot are contracted, resulting in stiffness. Extracellular matrix proteins, such as asporin, collagen types III, V, and VI, versican, tenascin-C, and TGF-beta induced protein, are highly expressed in contracted tissues clubfoot (Eckhardt et al, 2019;Ošt'ádal et al, 2015). Additionally, the expression levels of growth factors TGF-beta and platelet-derived growth factor are high and a blockade of growth factors led to decreased collagen expression, proliferation, and chemotaxis (Li et al, 2001).…”

“…PPI analysis was performed using the MEx Interactome database. The parameters were referred Eckhardt et al (2019) and Ošt'ádal et al (2015) to the literature-curated comprehensive data from the Innate DB (Breuer et al, 2013). Gene-miRNA interactome analysis was carried out with comprehensive experimentally validated miRNA-gene interaction data collected from TarBase and miRTarBase (Chou et al, 2018;Vlachos et al, 2015).…”

“…Clubfoot is associated with neuromuscular lesions, heredity abnormalities, skeletal dysplasia, soft tissue contracture, vascular abnormalities, ECM abnormalities, and intrauterine growth retardation (Chesney, Barker & Maffulli, 2007;Eckhardt et al, 2019;Miedzybrodzka, 2003;Ošt'ádal et al, 2015;Poon, Li & Alman, 2009;Sodre et al, 1990;Wang et al, 2013a;Wang et al, 2013b). In addition, smoking and viral infections in pregnant women are also closely related to congenital clubfoot (Palma et al, 2013;Robertson Jr & Corbett, 1997).…”

Proteomic analysis of dentin–enamel junction and adjacent protein‐containing enamel matrix layer of healthy human molar teeth

“…For MS/MS analyses, auto MS/MS with active exclusion (after 1 spectrum and release after 0.3 min) was set on. All LC‐MS/MS analyses were performed in duplicate as in the previous study . Data were processed using proteinscape software (ver.…”