Date Published: April 23, 2019
Publisher: Public Library of Science
Author(s): Moeed Akbar, Michael McLean, Emma Garcia-Melchor, Lindsay AN Crowe, Paul McMillan, Umberto G. Fazzi, David Martin, Angus Arthur, James H. Reilly, Iain B. McInnes, Neal L. Millar, Amin Mohamadi.
Frozen shoulder is a common, fibro-proliferative disease characterised by the insidious onset of pain and progressively restricted range of shoulder movement.
Shoulder capsule samples were collected from 10 patients with idiopathic frozen shoulder and 10 patients undergoing shoulder stabilisation surgery. Fibroblast activation marker expression (CD248, CD146, VCAM and PDPN, FAP) was quantified using immunohistochemistry. Control and diseased fibroblasts were cultured for in vitro studies from capsule biopsies from instability and frozen shoulder surgeries, respectively. The inflammatory profile and effects of IL-1β upon diseased and control fibroblasts was assessed using ELISA, immunohistochemistry and qPCR.
Immunohistochemistry demonstrated increased expression of fibroblast activation markers CD248, CD146, VCAM and PDPN in the frozen shoulder group compared with control (p < 0.05). Fibroblasts cultured from diseased capsule produced elevated levels of inflammatory protein (IL-6, IL-8 & CCL-20) in comparison to control fibroblasts. Exposing control fibroblasts to an inflammatory stimuli, (IL-1ß) significantly increased stromal activation marker transcript and protein expression (CD248, PDPN and VCAM). These results show that fibroblasts have an activated phenotype in frozen shoulder and this is associated with inflammatory cytokine dysregulation. Furthermore, it supports the hypothesis that activated fibroblasts may be involved in regulating the inflammatory and fibrotic processes involved in this disease.
The fibroproliferative disorder, frozen shoulder is a multifactorial disease in which patients present with limited active and passive shoulder movement. Despite a prevalence of 2–5% and known risk factors including diabetes, thyroid disease, stroke and autoimmune diseases there remains a significant lack of understanding of the molecular mechanisms underpinning this common fibrotic disorder which affects many recreational sporting individuals.
This study is the first to investigate the difference in expression of fibroblast activation in capsule tissue from control and adhesive capsulitis patients. We have demonstrated the presence of CD248, CD146, VCAM, PDPN, CD34 and FAP in shoulder capsule tissue of patients with frozen shoulder co existing with proinflammatory cytokine expression. Importantly inflammatory activation of healthy fibroblasts leads to overexpression of fibroblast activation markers in addition to secreting inflammatory proteins suggesting this may drive disease chronicity as illustrated in Fig 6.
This original study, is the first to have shown that there is elevation of fibroblast activation markers in the shoulder capsule of patients with frozen shoulder compared to control capsular tissue while furthermore confirming significantly increased proinflammatory signalling within frozen shoulder capsule. This data is consistent with the hypothesis that persistent fibroblast activation may play a role in the pathology of frozen shoulder and could explain the cellular mechanisms behind capsular fibrosis and persistent inflammation.