Name: a friction testing-bioreactor device for study of synovial joint biomechanics, mechanobiology, and physical regulation
Uploaded: 2022-06-02T13:50:00
Description: Watch this Scientific Journal Video about A Friction Testing-Bioreactor Device for Study of Synovial Joint Biomechanics, Mechanobiology, and Physical Regulation at JoVE.com

This work was supported by the Orthopaedic Scientific Research Foundation, NIH 5R01 AR068133, NIH TERC 5P41EB027062, and NIGMS R01 692 GM083925 (Funder ID: 10.13039/100000057).

Juvenile bovine knee joints, obtained from a local abattoir, were used for the present study. Studies with such bovine specimen samples are exempted from Columbia Institutional Animal Care and Use Committee (IACUC).
1. Designing the friction testing device
NOTE: A schematic representation of the friction testing device is shown in Figure 1. The device is built on a rigid base plate (not shown), which serves as a platform ...

A dynamic mechanical environment exists within the joint as cartilage is subjected to compressive, tensile, and shear forces, and hydrostatic and osmotic pressures44,45. Although cartilage is the main load-bearing tissue of the joint, the synovium also undergoes frictional interactions with the cartilage surface and with itself in regions where the tissue folds. The physical interactions between cartilage and synovium are likely responsible for transferring cells...

Osteoarthritis (OA) is a debilitating, degenerative joint disease that affects more than 32 million American adults, with a healthcare and socio-economic cost of over $16.5 billion1. The disease has classically been characterized by the degradation of articular cartilage and subchondral bone; however, changes to the synovium have recently garnered appreciation as synovitis has been linked to OA symptoms and progression2,3,4. In primary (idiopathic) OA, normal &#39;wear and tear&#39; associated with aging inhibits cartilage&#39;s ability to sustain its load-bearing and lubrication functions. The stresses generated by prolonged sliding contact of articular cartilage layers or sliding contact of cartilage against implant materials have been shown to facilitate delamination wear through subsurface fatigue failure5,6. As a dynamic mechanical environment exists within the joint7,8, the frictional interactions of articular cartilage and synovium may influence joint homeostasis through tissue level wear and cellular mechanotransduction. To study these mechanical and mechanobiological processes, a device has been designed to replicate the motion of the joint with tight control over compressive and frictional loading5,6,9,10,11,12,13.
The present protocol describes a friction testing device that delivers reciprocal, translating motion and compressive load to contacting surfaces of living tissue explants. The computer-controlled device permits user control of the duration of each test, applied load, range of motion of the translation stage, and translation speed. The device is modular, allowing for testing of various counterfaces, such as tissue-on-tissue (cartilage-on-cartilage and synovium-on-cartilage) and tissue-on-glass. In addition to the functional measurements obtained by the tester, tissue and lubricating bath components can be assessed before and after testing to evaluate the biological changes imparted by a given experimental regimen.
Studies of cartilage tribology have been performed for decades, and several techniques have been developed to measure friction coefficients between cartilage and glass and cartilage on cartilage14,15. The different approaches are motivated by the joint and/or the lubrication mechanism of interest. There is often a tradeoff between the control of experimental variables and the recapitulation of physiologic parameters. Pendulum-style devices utilize intact joints as the fulcrum of a simple pendulum where one joint surface translates freely over the second surface14,16,17,18. Instead of using intact joints, friction measurements may be obtained by sliding cartilage explants over desired surfaces14,19,20,21,22,23,24,25. Reported friction coefficients of articular cartilage have varied over a wide range (from 0.002 to 0.5) depending on the operating conditions14,26. Devices have been created to replicate rotary motion23,27,28. Gleghorn et al.26 developed a multi-well custom tribometer to observe cartilage lubrication profiles using Stribeck curve analysis, and a linear oscillatory sliding motion was applied between cartilage against a flat glass counterface.
This device aims to isolate frictional responses and explore the mechanobiology of living tissues under various loading conditions. The device employs a simplified test set-up simulating joint articulation through compressive sliding, which can approximate both rolling and sliding motion with the understanding that the resistance in pure rolling motion is negligible relative to the measured friction coefficient of articular cartilage29. Originally built to study the effects of interstitial fluid pressurization on the frictional response of articular cartilage9, the tester has since been used to explore topics such as frictional effects of removing the superficial zone of cartilage10, lubricating effects of synovial fluid11, cartilage wear hypotheses5,6,30, and synovium-on-tissue friction measurements13. The friction-testing bioreactor can conduct friction experiments under sterile conditions, providing a novel mechanism to explore how frictional forces affect the mechanobiological responses of living cartilage and synovium. This design can be used as a biomimetic bioreactor to study the physical regulation of living joint tissues in response to applied physiologic loading associated with diarthrodial joint articulation.
This study presents a configuration for synovium-on-cartilage friction testing over a range of contact stresses and in different lubricating baths. The articulating surface area of most joints is, to a great extent, synovial tissue31. While synovium-on-cartilage sliding does not occur at primary load-bearing surfaces, the frictional interactions between the two tissues may still have important implications for tissue level repair and cell mechanotransduction. It has previously been shown that fibroblast-like synoviocytes (FLS) residing on the intimal layer of the synovium are mechanosensitive, responding to fluid-induced shear stress32. It has also been demonstrated that stretch33,34 and fluid-induced shear stress35 modulate FLS lubricant production. As such, direct sliding contact between synovium and cartilage may provide another mechanical stimulus to resident cells in the synovium.
Only a few reports on synovium friction coefficients have been published31,36. Estell et al.13 sought to expand on the previous characterization by utilizing biologically relevant counterfaces. With the friction testing device&#39;s ability to test living tissues, it is possible to mimic physiologic tissue interactions during joint articulation to elucidate the role of contact shear stress on synoviocyte function and its contribution to the crosstalk between synovium and cartilage. The latter has been implicated in mediating synovial joint inflammation in arthritis and post-injury. Due to the physical proximity of cartilage to synovium and synovial fluid, which contain synoviocytes that exhibit multipotent capacity, including chondrogenesis, it is postulated that synoviocytes play a role in cartilage homeostasis and repair by engrafting to the articular surface. In this context, physical contact and reciprocal shearing of cartilage-synovium and synovium-synovium may increase the accessibility of synoviocytes to regions of cartilage damage37,38,39,40. Studies utilizing synovium-on-cartilage configurations will not only provide insights into joint gross tissue mechanics and tribology, but they may also lead to new strategies for maintaining joint health.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
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			<td>DMEM (Cat No. 11-965-118; Gibco) supplemented with 50 &#956;g/mL L-proline (Cat. No. P5607; Sigma), 100 &#956;g/mL sodium pyruvate (Cat. No. S8636; Sigma), 1% ITS (Cat. No. 354350; Corning), and 1% antibiotic&#8211;antimycotic (Cat. No. 15-240-062, Gibco)</td>
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			<td>Excelta</td>
			<td>3C-S-PI</td>
			<td>Tissue mounting</td>
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			<td>MSA 670.63</td>
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			<td>JR3 Inc.</td>
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			<td>Custom made</td>
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			<td>Tissue mounting</td>
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			<td>Parker</td>
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			<td>Falcon</td>
			<td>351007</td>
			<td>Tissue mounting</td>
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			<td>PL325</td>
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			<td>Removable base</td>
			<td>ThorLabs, Inc.</td>
			<td>SB1B</td>
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			<td>FSC22</td>
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			<td>No. 4</td>
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			<td>Virtual Industries, Inc.</td>
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			<td></td>
			<td>Tissue mounting</td>
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			<td>Animal Technologies, Inc.</td>
			<td></td>
			<td>Friction testing bath</td>
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			<td>Custom made</td>
			<td></td>
			<td>Phosphate-Buffered Saline (PBS) with protease inhibitors: 0.04% isothiazolone-base biocide (Proclin 950 Cat. No. 46878-U; Sigma) and 0.1% protease inhibitor - 0.05 M ethylenediaminetetraacetic acid, EDTA (Cat. No. 0369; Sigma)</td>
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			<td>Fisher Scientific</td>
			<td>Isotemp</td>
			<td>Sterile culture</td>
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			<td>Vertical linear encoder (for loading stage)</td>
			<td>Renishaw</td>
			<td>T1031-30A</td>
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			<td>Voice coil actuator</td>
			<td>H2W Technologies</td>
			<td>NCC20-15-027-1RC</td>
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		</tr>
	</tbody>
</table>

a friction testing-bioreactor device for study of synovial joint biomechanics, mechanobiology, and physical regulation

In primary osteoarthritis (OA), normal 'wear and tear' associated with aging inhibits the ability of cartilage to sustain its load-bearing and lubrication functions, fostering a deleterious physical environment. The frictional interactions of articular cartilage and synovium may influence joint homeostasis through tissue level wear and cellular mechanotransduction. To study these mechanical and mechanobiological processes, a device capable of replicating the motion of the joint is described. The friction testing device controls the delivery of reciprocal translating motion and normal load to two contacting biological counterfaces. This study adopts a synovium-on-cartilage configuration, and friction coefficient measurements are presented for tests performed in a phosphate-buffered saline (PBS) or synovial fluid (SF) bath. The testing was performed for a range of contact stresses, highlighting the lubricating properties of SF under high loads. This friction testing device can be used as a biomimetic bioreactor for studying the physical regulation of living joint tissues in response to applied physiologic loading associated with diarthrodial joint articulation.

A synovium-on-cartilage configuration was used to friction test juvenile bovine explants. The synovium was mounted on a 10 mm diameter acrylic loading platen such that the intimal layer would be in contact with the underlying cartilage. A tibial strip was used as the cartilage counterface (Figure 6A). Tibial strips were cut with a depth of approximately 1.4 mm and a size of 10 mm x 30 mm. The samples were tested for 1 h at 37 &#176;C in a phosphate-buffered saline (PBS) bath or a bovine syno...

Watch this Scientific Journal Video about A Friction Testing-Bioreactor Device for Study of Synovial Joint Biomechanics, Mechanobiology, and Physical Regulation at JoVE.com

A Friction Testing-Bioreactor Device for Study of Synovial Joint Biomechanics, Mechanobiology, and Physical Regulation

The present protocol describes a friction testing device that applies simultaneous reciprocal sliding and normal load to two contacting biological counterfaces.

In primary osteoarthritis (OA), normal 'wear and tear' associated with aging inhibits the ability of cartilage to sustain its load-bearing and lubrication ...

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