Proteomic Analysis of Differential Protein Expression of Achilles Tendon in a Rabbit Model by Two-Dimensional Polyacrylamide Gel Electrophoresis at 21 Days Postoperation

Abstract

Postoperative early kinesitherapy has been advocated as an optimal method for treating Achilles tendon rupture. However, an insight into the rationale of how early kinesitherapy contributes to healing of Achilles tendon remains to be achieved, and research in the area of proteomic analysis of Achilles tendon has so far been lacking. Forty-two rabbits were randomized into control group, immobilization group, and early motion group, and received postoperative cast immobilization and early motion treatments. Achilles tendon samples were prepared 21 days following microsurgery, and the proteins were separated with two-dimensional polyacrylamide gel electrophoresis. Differentially expressed proteins were first recognized by PDQuest software, and then identified using peptide mass fingerprinting, tandem mass spectrometry, and database searching. A total of 463 ± 12, 511 ± 39, and 513 ± 80 protein spots were successfully detected in the two-dimensional polyacrylamide gels for the Achilles tendon samples of rabbits in the control group, immobilization group, and early motion group, respectively. There were 15, 8, and 9 unique proteins in these three groups, respectively, and some differentially expressed proteins were also identified in each group. It was indicated that some of the differentially expressed proteins were involved in various metabolism pathways and may play an important role in healing of Achilles tendon rupture. Postoperative early kinesitherapy resulted in differentially expressed proteins in ruptured Achilles tendon compared with those treated with postoperative cast immobilization. These differentially expressed proteins may contribute to healing of Achilles tendon rupture through a mechanobiological mechanism due to the application of postoperative early kinesitherapy.

Proteomic Analysis Reveals Age-related Changes in Tendon Matrix Composition, with Age- and Injury-specific Matrix Fragmentation

“…Although proteomic analysis has been used to identify many novel proteins in other connective tissues such as cartilage (19, 20), a review of the current literature shows few studies that have undertaken a proteomic analysis of tendon. Considering the studies that have addressed tendon proteomics, some have assessed the proteins produced by tendon fibroblasts in vitro (21, 22), whereas others have investigated alterations in protein profile as a result of artificially induced injury (23, 24). Smith et al (25) investigated changes in pericellular proteins during development, and Dakin et al (18) studied normal and diseased tendons from horses with a wide age range but do not report any data regarding age-related alterations in protein content.…”

Proteomic Differences between Male and Female Anterior Cruciate Ligament and Patellar Tendon

“…Furthermore, in other tissues correlation of mRNA and protein levels may be poor and depends partly on the abundance of the proteins involved and on complex post-transcriptional regulation and post-transcriptional modification pathways [16], [17]. Previously, proteomic techniques including two-dimensional gel electrophoresis (2D SDS-PAGE) have been used to evaluate the response of tendon and ligament to various stimuli [18]–[22]. However, potential disadvantages of 2D SDS-PAGE for evaluation of these tissues include the presence of highly abundant insoluble proteins such as mature type I collagen and the presence of anionic proteoglycans [23].…”

Minimally Invasive Reconstruction of Chronic Achilles Tendon Ruptures Using the Ipsilateral Free Semitendinosus Tendon Graft and Interference Screw Fixation

“…The safety of such an approach may be questioned, but both basic science and clinical studies show that early motion avoiding stretching of the reconstructed areas is not deleterious, and most likely beneficial, to the reconstruction. 12,27,31 The present study employed a well-validated scoring system to provide a quantitative assessment of clinical benefits related to the procedure. The ATRS is a patient-reported instrument with high reliability, validity, and sensitivity for measuring outcomes after the management of patients with a complete Achilles tendon rupture.…”