Shear-Induced Formation of a Lamina Splendens-like Gel

from Synovial Fluid

Annie Fang, Sierra Cook, Delphine Gourdon
Department of Materials Science and Engineering, Cornell University
Project funded by: Engineering Learning Initiatives & Diversity Programs in Engineering

Abstract Objectives Results

Before Shear After Shear
Agglomeration of SF is
Recent research has reported of a gel-like layer found on Characterize how different shearing parameters affect the ability of SF to form a robust, long-lasting gel-layer responsible for the
the surface of articular cartilage called the Lamina Splendens Identify which SF components are involved in the gel-formation process. inhomogeneities in the
(LS) and hinted of the aggregation of Synovial Fluid (SF) as the fringes after shear.
Regardless of shearing
origin of LS. Thin films of different SF components were parameters, all films
sheared under loads (1.5 mN to 6 mN) to determine the ideal
load sustaining the most stable gel-like layer. To isolate the SF Materials & Methods increased in thickness
immediately as shearing
components responsible for aggregation, solutions of began.
hyaluronan (HA), lubricin (LUB), and serum albumin (SA) Clean surfaces must be prepared by cleaving and silvering mica. Individual mica flakes and other dust may contribute to particle Optimal Shearing Parameters
were sheared at the previously determined ideal load. The interaction, disrupt aggregation and film thickening. To prevent contamination:
longest-sustaining gels were sheared under low load and All surface preparation was performed under a vented hood. Low load (~1.68 mN) shearing at 3
Sheets were cleaved facing down to avoid any potential floating mica flakes from adhering m/s in SF leads to sustainable gel:
velocity. Using only the inner faces of two cleaved mica pieces as the clean top surface. large and stable film thickness over
The surface interactions (film thickening, agglomeration of
many shearing cycles
SF particles) of FN+HA and FN+SF films were noticeably Individual mica sheets were cleaved as thinly as possible, producing 3-4 fringes through the spectrometer. Fewer visible fringes
similar. Because HA composes the majority of LS, this means better resolution. The Surface Force Apparatus (SFA) characterizes the optimal shearing parameters and degree of
similarity thus bolsters the hypothesis that LS formation arises aggregation by measuring film thickness and normal & friction forces. In each experiment, normal force is first applied to bring
the two surfaces closer to contact before any shearing is performed. Two notable instances occur during a regular normal force Contribution of SF Components to Gel Formation
from the shear-induced aggregation of SF particles.
run: reaching the onset of interaction and hard-wall. The onset of interaction yields the film thickness unperturbed by any
surface force while the hard-wall value signifies the stable film thickness after a period of high applied load. FN+HA film
Film thickening much greater than
Basis of each experiment: FN+SF film

Background A thin layer of fibronectin anchored SF components in every experiment

Thin films of each solution of interest were deposited between the two surfaces
Similar agglomeration of particles to
FN+SF film

Cross-configured cylindrical
silica surfaces coated in
Two sets of experiments: FN+LUB film
back-silvered mica create an optical 1. Testing Optimal Conditions for 2. Determining which SF components are Much less film thickening than FN+SF
Lamina Splendens (LS) interferometer when light is shone
through. Passing Newtons Rings LS-like gel formation by varying involved in LS-like gel formation through film
A film-like substance located in the uppermost layer of through a spectrometer produces
articular cartilage Fringes of Equal Chromatic Order shearing velocity and load isolation Film thickness depletes quickly
(FECO). The shape of the fringes
Widely regarded to play an essential role in providing an correspond to the shape of the
Force = 1-2 mN
incredibly low friction surface for joint lubrication and wear surfaces and helps detect
agglomerates.
protection SF or 6-7 mN FN+SA film
Containing primarily hyaluronan (HA), phospholipids, and FN Film thickening much greater than
glycoprotein Shearing under previously-determined FN+SF film, decreases very gradually
Velocity = 30 m/s or 3m/s optimal gel-formation parameters over time

Discussion & Conclusion

Composition of LS is similar to Synovial Fluid (SF) found in the
cavities of moveable joints, giving rise to the hypothesis that
LS is sourced from SF. SF lubricates cartilage in articular joints
by reducing friction and providing wear protection.
Banquy (2014)s work focused on shear-induced synovial fluid
gels; the project featured the formation of a robust LS-like gel
from synovial fluid under shear.
References
A readily available reservoir of SF is necessary to sustain a lubricating gel-layer. The rate of new agglomerate formation
Banquy X, Lee DW, Das S, Hogan J, Israelachvili JN. 2014. Shear-induced aggregation of
must be greater than the rate of agglomerate expulsion in order for a gel-like layer to maintain its thickness. New mammalian synovial fluid components under boundary lubrication conditions. Adv Funct
agglomerates are thus able to be created by tapping into the large SF reservoir. In articular joints, this dependency on the ease Materials. 24(21):3152-3161.
of accessibility of SF means that cartilage-bound SF is an insufficient source to sustain a long-lasting gel-like layer. Segar, Todd. "Skeletal Joints - Lessons" Tes Teach with Blendspace. Tes Teach, n.d.
Ddinait, Andra. Biomimetic Lubrication. Soft Matter, The Royal Society of Chemistry, 21 Oct.
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Under low loads (~1.5 mN), the SF medium has greater fluidity while individual SF components experience less resistance
in reorienting during aggregation. Low loads also operate under a smaller contact area and allows greater exchange of material
Acknowledgements
between the contact junction and the SF reservoir. In combination with a low load, a low shearing velocity (~3 m/s) provided
the optimal conditions for sustaining a gel. Gels are not maintained when shearing at high load and velocity due to the
tendency for agglomerates to be expelled rapidly from the contact junction. The similarity between FN+HA and FN+SF films
shows that HA plays an essential role in SF aggregation. The prevalence of HA as the major component of LS is critical evidence Funding for this research was co-supported by Engineering Learning
in the link between the source of LS formation and the shear-induced aggregation of SF particles. Initiatives and Diversity Programs in Engineering