Simulated impact trauma and osteoarthritis: the role of cell volume and mechanotransduction

Nedelcheva, Y. 2013. Simulated impact trauma and osteoarthritis: the role of cell volume and mechanotransduction. Thesis University of Westminster School of Life Sciences

TitleSimulated impact trauma and osteoarthritis: the role of cell volume and mechanotransduction
AuthorsNedelcheva, Y.
Abstract

Health and emotional benefits are linked to participation in exercise, however

single-impact load (e.g. trauma) and altered joint loading can cause bone

fracture(s) resulting in permanent cartilage damage and increased risk of

osteoarthritis (OA).

This study investigated the role of single mechanical load and the effects of

chondroprotective agents over short periods of time (2-30 min) post impact.

Mechanical load (force 1.14 N) induced by a drop tower device caused cell

death from as early as 2 min. Pre-incubation in hypertonic media protected

chondrocytes from cell death, whereby at 30 min, death was decreased from

9.22 % to 3.42 % (p<0.01), thus implicating volume regulatory changes as a

potential key mechanism for chondroprotection, with in situ chondrocytes

altering their cell volume in response to hypertonicity by 20 %. Investigation

of the cell cytoskeleton, showed that hypertonicity increased cortical actin by

29 % within the superficial zone (SZ) only (p<0.01).

Volume and actin polymerisation regulation are governed by intracellular

Ca2+ and the regulatory volume decrease (RVD) inhibitor REV5901 has been

linked to both and was therefore tested for its potential chondroprotective

properties. Impact led to cell death, which was significantly reduced by

REV5901 from 10.92 % to 5.44 % (p<0.001) and initial cell volume was

reduced by 25%. Cortical actin staining was increased by 20 % within the SZ

of articular cartilage only (p<0.01). GdCl3 (blocker for stretch-activated

calcium channels), uridine-5’-triphosphate (uridine; calcium mobilizing

mediator) and the phosphoinositide 3-kinases (PI3K) inhibitor wortmannin

were used to determine its mechanism of action. Wortmannin alone

increased cell death and inhibited REV5901 chondroprotective effects with

no alteration in cell volume compared to REV5901 alone. Whilst uridine

reduced cell volume with 20 % (p<0.05) it did not reduce cell death

significantly from 10.92 % to 8.10 % (p>0.05) compared to control. GdCl3

inhibited REV5901 chondroprotective effects by increasing cell death by ~ 5

% compared to control (p<0.05), F-actin staining appeared reduced 72.84 AU

and not significantly different from control (p>0.05).

The role of the cell cytoskeleton is important for cell mechanotransduction

and for maintaining integrity as actin microfilaments are recognized to bear

tension therefore, alterations of the actin-binding proteins responsible for

actin treadmilling cofilin, profilin and gelsolin mRNA was compared to

untreated chondrocytes. REV5901 was observed to reduce cofilin (known

actin depolymerizing factor) with 30 % (p<0.05) and increase profilin

significantly by 75 % (p<0.001) respectively. Western blot analysis showed

that only cofilin and gelsolin were expressed in all samples with no detection

of profilin. REV5901 was observed to significantly reduce cofilin by 28 %

(p<0.05).

This data highlights that REV5901 exhibits chondroprotective properties in

part due to the polymerisation of the actin cytoskeleton via PI3Ks pathway.

This could offer a novel therapeutic opportunity for perention of irreversible

cartilagedamage following acute impact trauma.

Year2013
FileYanitsa_NEDELCHEVA_2013.pdf
Publication dates
Completed2013

Related outputs

REV5901: the protective effect on bovine articular chondrocytes following single impact trauma
Nedelcheva, Y., Getting, S.J. and Kerrigan, M.J.P. 2010. REV5901: the protective effect on bovine articular chondrocytes following single impact trauma. Physiological Society Meeting 19. University of Manchester

REV5901: an investigation on the chondroprotective effect in bovine chondrocytes following single impact trauma
Nedelcheva, Y., Getting, S.J. and Kerrigan, M.J.P. 2010. REV5901: an investigation on the chondroprotective effect in bovine chondrocytes following single impact trauma. Physiological Society Meeting 21. University of Durham

Making assessment count – integrating feedback and reflection through eReflect
Kerrigan, M.J.P., Clements, M.O., Bond, A., Oradini, F., Nedelcheva, Y. and Saunders, G. 2009. Making assessment count – integrating feedback and reflection through eReflect. in: Terry, E.B., Jefferies, A.L. and Bracq, A. (ed.) Proceedings of the Fourth International Blended Learning Conference "Engaging Students in the Curriculum" Hatfield University of Hertfordshire.

University of westminster, Making assessment count: a JISC funded project
Kerrigan, M.J.P., Clements, M.O., Bond, A., Nedelcheva, Y. and Saunders, G. 2009. University of westminster, Making assessment count: a JISC funded project. 6th International Conference of the Association for Learning Technology. Manchester, UK 08 - 10 Sep 2009 Association for Learning Technology.

University of Westminster, Making assessment count: a JISC funded project
Kerrigan, M.J.P., Clements, M.O., Bond, A., Nedelcheva, Y. and Saunders, G. 2009. University of Westminster, Making assessment count: a JISC funded project. SSHL: Teaching and Learning Day. University of Westminster, London 2009 Association for Learning Technology.

University of Westminster, Making assessment count: a JISC funded project
Kerrigan, M.J.P., Clements, M.O., Bond, A., Nedelcheva, Y. and Saunders, G. 2009. University of Westminster, Making assessment count: a JISC funded project. University of Westminster Teaching & Learning Symposium. University of Westminster, London 2009 Association for Learning Technology.

The role of the actin cytoskeleton in chondrocyte protection following single impact
Nedelcheva, Y., Getting, S.J. and Kerrigan, M.J.P. 2008. The role of the actin cytoskeleton in chondrocyte protection following single impact. pA2 Online. From the University of Brighton, Winter 2008 Meeting: Proceedings of the British Pharmacological Society. 6 (4).

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