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Research Papers

Mechanical Integrity of a Decellularized and Laser Drilled Medial Meniscus

[+] Author and Article Information
Emily H. Lakes, Peter S. McFetridge

J Crayton Pruitt Family Department
of Biomedical Engineering,
University of Florida,
Gainesville, FL 32610;
Institute for Cell and Tissue
Science and Engineering,
University of Florida,
Gainesville, FL 32610

Andrea M. Matuska

J Crayton Pruitt Family Department
of Biomedical Engineering,
University of Florida,
Gainesville, FL 32610

Kyle D. Allen

Assistant Professor
J Crayton Pruitt Family Department
of Biomedical Engineering,
University of Florida,
1275 Center Drive,
Biomedical Sciences Building,
Gainesville, FL 32610;
Institute for Cell and Tissue
Science and Engineering,
University of Florida,
Gainesville, FL 32610
e-mail: kyle.allen@bme.ufl.edu

1Corresponding author.

Manuscript received September 23, 2015; final manuscript received December 16, 2015; published online January 29, 2016. Assoc. Editor: Michael Detamore.

J Biomech Eng 138(3), 031006 (Jan 29, 2016) (12 pages) Paper No: BIO-15-1477; doi: 10.1115/1.4032381 History: Received September 23, 2015; Revised December 16, 2015

Since the meniscus has limited capacity to self-repair, creating a long-lasting meniscus replacement may help reduce the incidence of osteoarthritis (OA) after meniscus damage. As a first step toward this goal, this study evaluated the mechanical integrity of a decellularized, laser drilled (LD) meniscus as a potential scaffold for meniscal engineering. To evaluate the decellularization process, 24 porcine menisci were processed such that one half remained native tissue, while the other half was decellularized in sodium dodecyl sulphate (SDS). To evaluate the laser drilling process, 24 additional menisci were decellularized, with one half remaining intact while the other half was LD. Decellularization did not affect the tensile properties, but had significant effects on the cyclic compressive hysteresis and unconfined compressive stress relaxation. Laser drilling decreased the Young's modulus and instantaneous stress during unconfined stress relaxation and the circumferential ultimate strength during tensile testing. However, the losses in mechanical integrity in the LD menisci were generally smaller than the variance observed between samples, and thus, the material properties for the LD tissue remained within a physiological range. In the future, optimization of laser drilling patterns may improve these material properties. Moreover, reseeding the construct with cells may further improve the mechanical properties prior to implantation. As such, this work serves as a proof of concept for generating decellularized, LD menisci scaffolds for the purposes of meniscal engineering.

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Figures

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Fig. 2

Top: Hematoxylin and eosin staining for (a) native, (b) decellularized, and (c) LD menisci. Bottom: DAPI staining for (d) native, (e) decellularized, and (f) LD menisci. Slices for all staining were taken at 10 μm on a cryotome. (g) Macro view of a LD meniscus showing full thickness pores.

Grahic Jump Location
Fig. 1

Sample preparation where all menisci were dissected in half to allow for different treatments within the same meniscus (native/decellularized or decellularized/LD). (a) Tensile samples were taken as both radial and circumferential in relation to the circumferential collagen fibers of the meniscus. (b) For compression testing, two cylindrical samples were taken next to each other in the central portion of the meniscus. Samples were then cut with parallel blades.

Grahic Jump Location
Fig. 3

Native and decellularized tissue results from radial tensile testing showing the: (a) hysteresis area from cyclic loading, (b) loading energy from cyclic loading, (c) peak stress from cyclic loading, (d) representative curves from cyclic loading, (e) Young's modulus from the linear portion of the pull to failure curve, (f) strain at UTS, and (e) UTS. Lines connect samples from the same meniscus, and the legend indicates whether samples from the anterior or posterior region of the meniscus. * = significance from paired t-test (p < 0.05).

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Fig. 6

Decellularized and LD tissue results from radial tensile testing showing the: (a) hysteresis area from cyclic loading, (b) loading energy from cyclic loading, (c) peak stress from cyclic loading, (d) representative curves from cyclic loading, (e) Young's modulus from the linear portion of the pull to failure curve, (e) strain at UTS, and (g) UTS. Lines connect samples from the same meniscus, and the legend indicates whether samples from the anterior or posterior region of the meniscus.

Grahic Jump Location
Fig. 7

Decellularized and LD tissue results from circumferential tensile testing showing the: (a) hysteresis area from cyclic loading, (b) loading energy from cyclic loading, (c) peak stress from cyclic loading, (d) representative curves from cyclic loading, (e) Young's modulus from the linear portion of the pull to failure curve, (f) strain at UTS, and (g) UTS. Lines connect samples from the same meniscus, and the legend indicates whether samples from the anterior or posterior region of the meniscus. * = significance from paired t-test (p < 0.05).

Grahic Jump Location
Fig. 8

Decellularized and LD tissue results from compressive testing showing the: (a) hysteresis area from cyclic loading, (b) loading energy from cyclic loading, (c) peak stress from cyclic loading, (d) representative curves from cyclic loading, (e) Young's modulus from the linear portion of the loading phase, (f) instantaneous stress at the beginning of stress relaxation, and (g) steady-state stress after stress relaxation. Lines connect samples from the same meniscus, and the legend indicates whether samples from the anterior or posterior region of the meniscus. * = significance from paired t-test (p < 0.05).

Grahic Jump Location
Fig. 4

Native and decellularized tissue results from circumferential tensile testing showing the: (a) hysteresis area from cyclic loading, (b) loading energy from cyclic loading, (c) peak stress from cyclic loading, (d) representative curves from cyclic loading, (e) Young's modulus from the linear portion of the pull to failure curve, (f) strain at UTS, and (g) UTS. Lines connect samples from the same meniscus, and the legend indicates whether samples from the anterior or posterior region of the meniscus.

Grahic Jump Location
Fig. 5

Native and decellularized tissue results from compressive testing showing the: (a) hysteresis area from cyclic loading, (b) loading energy from cyclic loading, (c) peak stress from cyclic loading, (d) representative curves from cyclic loading, (e) Young's modulus from the linear portion of the loading phase, (f) instantaneous stress at the beginning of stress relaxation, and (g) steady-state stress after stress relaxation. Lines connect samples from the same meniscus, and the legend indicates whether samples from the anterior or posterior region of the meniscus. * = significance from paired t-test (p < 0.05).

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