Increasing the level of cytoskeletal protein Flightless I reduces adhesion formation in a murine digital flexor tendon model.

Abstract

BACKGROUND:

Surgical repair of tendons is common, but function is often limited due to the formation of flexor tendon adhesions which reduce the mobility and use of the affected digit and hand. The severity of adhesion formation is dependent on numerous cellular processes many of which involve the actin cytoskeleton. Flightless I (Flii) is a highly conserved cytoskeletal protein, which has previously been identified as a potential target for improved healing of tendon injuries. Using human in vitro cell studies in conjunction with a murine model of partial laceration of the digital flexor tendon, we investigated the effect of modulating Flii levels on tenocyte function and formation of adhesions.

METHODS:

Human tenocyte proliferation and migration was determined using WST-1 and scratch wound assays following Flii knockdown by siRNA in vitro. Additionally, mice with normal and increased levels of Flii were subjected to a partial laceration of the digital flexor tendon in conjunction with a full tenotomy to immobilise the paw. Resulting adhesions were assessed using histology and immunohistochemistry for collagen I, III, TGF-β1and -β3 RESULTS: Flii knockdown significantly reduced human tenocyte proliferation and migration in vitro. Increasing the expression of Flii significantly reduced digital tendon adhesion formation in vivo which was confirmed through significantly smaller adhesion scores based on collagen fibre orientation, thickness, proximity to other fibres and crimping. Reduced adhesion formation was accompanied with significantly decreased deposition of type I collagen and increased expression of TGF-β1 in vivo.

CONCLUSIONS:

These findings suggest that increasing the level of Flii in an injured tendon may be beneficial for decreasing tendon adhesion formation.

• Authors: Jessica E Jackson, Zlatko Kopecki, Peter J Anderson, Allison J Cowin
• Journal: Journal of orthopaedic surgery and research (J Orthop Surg Res) Vol. 15 Issue 1 Pg. 362 (Aug 27 2020) ISSN: 1749-799X [Electronic] England
• PMID: 32854733 (Publication Type: Journal Article)

Chemical References

• FLII protein, human
• Fibril-Associated Collagens
• FlII protein, mouse
• Microfilament Proteins
• Trans-Activators
• Transforming Growth Factor beta

Topics

• Animals
• Cell Movement (genetics)
• Cell Proliferation (genetics)
• Disease Models, Animal
• Fibril-Associated Collagens (metabolism)
• Gene Expression
• Humans
• Immunohistochemistry
• Mice, Inbred BALB C
• Mice, Transgenic
• Microfilament Proteins (genetics, metabolism)
• Tendon Injuries (genetics, metabolism, pathology)
• Tenocytes (physiology)
• Tissue Adhesions (genetics, metabolism)
• Trans-Activators (genetics, metabolism)
• Transforming Growth Factor beta (metabolism)