Cryopreservation Of Porcine Articular Cartilage: Mri And Biochemical Results After Different Freezing Protocols
2-step procedure; articular-cartilage; biochemical; cryopreservation; culture cells frozen; diffusion; dimethyl sulfoxide; femoral condyles; fresh; Life Sciences & Biomedicine - Other Topics; magnetic-resonance microscopy; MRI; osteoarticular allografts; osteochondral allograft transplantation; Physiology; porcine; survival; tissue
The objective of this study was to investigate the effects of cryopreservation on the components of articular cartilage (AC) matrix by utilizing magnetic resonance imaging (MRI) and biochemical assessments. Porcine AC (10 mm osteochondral dowels) was collected into four groups - (1) phosphate buffered saline (PBS) control, (2) PBS snap frozen in liquid nitrogen, (3) slow-cooled in dimethyl sulfoxide (DMSO), and (4) slow cooled in PBS (in absence of DMSO). MRI results demonstrated three distinct zones in the cartilage. After exposure to ice formation during cryopreservation procedures, alterations in MRI determined matrix fixed charged density and magnetization transfer rate were noted. In addition, biochemical assays demonstrated significant alterations in chondroitin sulfate and hydroxyproline content over time without differences in hydration or DNA content. In conclusion, MRI was able to detect some changes in the intact cartilage matrix structure consistent with biochemical assessments after ice formation during cryopreservation of intact porcine AC. Furthermore, biochemical assessments supported some of these findings and changed significantly after incubating the cartilage matrix for 36-72 h in PBS in terms of chondroitin sulfate and hydroxyproline content. (c) 2006 Elsevier Inc. All rights reserved.
Laouar L; Fishbein K; McGann L E; Horton W E; Spencer R G; Jomha N M
Cryobiology
2007
2007-02
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1016/j.cryobiol.2006.10.193" target="_blank" rel="noreferrer noopener">10.1016/j.cryobiol.2006.10.193</a>
Fourier Transform Infrared Imaging Spectroscopic Analysis Of Tissue Engineered Cartilage: Histologic And Biochemical Correlations
Biochemistry & Molecular Biology; bioreactor; chondrocytes; collagen; Fourier transform infrared imaging spectroscopy; hollow fiber bioreactor; hollow-fiber; human articular-cartilage; magnetic-resonance microscopy; matrix; Medical Imaging; model; mri techniques; Nuclear Medicine &; Optics; osteoarthritis; proteoglycans; Radiology; tissue engineered cartilage; tissue engineering
The composition of cartilage is predictive of its in vivo performance. Therefore, the ability to assess its primary macromolecular components, proteoglycan (PG) and collagen, is of great importance. In the current study, we hypothesized that PG content and distribution in tissue engineered cartilage could be determined using Fourier-transform infrared imaging spectroscopy (FT-IRIS). The cartilage was grown from chondrocytes within a hollow fiber bioreactor (HFBR) system previously used extensively to study cartilage development. FT-IRIS analysis showed a gradient of PG content, with the highest content in the center near the nutritive fibers and the lowest near the interior surface of the HFBR. Further, we found significantly greater PG content in the region near culture medium inflow (45.0%) as compared to the outflow region (24.7%) (p < 0.001). This difference paralleled the biochemically determined glycosaminoglycan difference of 42.6% versus 27.8%. In addition, FT-IRIS-determined PG content at specific positions within the tissue sections correlated with histologically determined PG content (R=50.73, p=50.007). In summary, FT-IRIS determination of PG correlates with histological determination of PG and yields quantitatively similar results to biochemical determination of glycosaminoglycan in developing cartilage. (c) 2005 Society of Photo-Optical Instrumentation Engineers.
Kim M; Bi X H; Horton W E; Spencer R G; Camacho N P
Journal of Biomedical Optics
2005
2005-05
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1117/1.1922329" target="_blank" rel="noreferrer noopener">10.1117/1.1922329</a>
Differences In Infrared Spectroscopic Data Of Connective Tissues In Transflectance And Transmittance Modes
BaF2 window; bone; cartilage; Chemistry; collagen orientation; consequences; Fourier transform infrared imaging spectroscopy; Low-e; proteoglycan; scattering; slide; Transflectance; Transmittance; variance
Fourier transform infrared imaging spectroscopy (FT-IRIS) has been used extensively to characterize the composition and orientation of macromolecules in thin tissue sections. Earlier and current studies of normal and polarized FT-IRIS data have primarily used tissues sectioned onto infrared transmissive substrates, such as salt windows. Recently, the use of low-emissivity ("low-e") substrates has become of great interest because of their low cost and favorable infrared optical properties. However, data are collected in transflectance mode when using low-e slides and in transmittance mode using salt windows. In the current study we investigated the comparability of these two modes for assessment of the composition of connective tissues. FT-IRIS data were obtained in transflectance and transmittance modes from serial sections of cartilage, bone and tendon, and from a standard polymer, polymethylmethacrylate. Both non-polarized and polarized FTIR data differed in absorbance, and in some cases peak position, between transflectance and transmittance modes. However, the FT-IRIS analysis of the collagen fibril orientation in cartilage resulted in the expected zonal arrangement of fibrils in both transmittance and transflectance. We conclude that numerical comparison of FT-IRIS-derived parameters of tissue composition should account for substrate type and data collection mode, while analysis of overall tissue architecture may be more invariant between modes. (c) 2013 Elsevier B.V. All rights reserved.
Hanifi A; McGoverin C; Ou Y T; Safadi F; Spencer R G; Pleshko N
Analytica Chimica Acta
2013
2013-05
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1016/j.aca.2013.03.053" target="_blank" rel="noreferrer noopener">10.1016/j.aca.2013.03.053</a>
Modification Of Osteoarthritis In The Guinea Pig With Pulsed Low-intensity Ultrasound Treatment
aggrecan; articular-cartilage; cartilage; Cartilage repair; degeneration; double-blind trial; electromagnetic-fields; gene-expression; in-vitro; knee osteoarthritis; matrix production; Orthopedics; osteoarthritis; placebo-controlled trials; Rheumatology; therapeutic ultrasound; tissue engineered cartilage; Ultrasound
Objective: The Hartley guinea pig develops articular cartilage degeneration similar to that seen in idiopathic human osteoarthritis (OA). We investigated whether the application of pulsed low-intensity ultrasound (PLIUS) to the Hartley guinea pig joint would prevent or attenuate the progression of this degenerative process. Methods: Treatment of male Hartley guinea pigs was initiated at the onset of degeneration (8 weeks of age) to assess the ability of PLIUS to prevent OA, or at a later age (12 months) to assess the degree to which PLIUS acted to attenuate the progression of established disease. PLIUS (30 mW/cm(2)) was applied to stifle joints for 20 min/day over periods ranging from 3 to 10 months, with contralateral limbs serving as controls. Joint cartilage histology was graded according to a modified Mankin scale to evaluate treatment effect. Immunohistochemical staining for interleukin-1 receptor antagonist (IL-1ra), matrix metalloproteinase (MMP)-3, MMP-13, and transforming growth factor (TGF)-beta 1 was performed on the cartilage to evaluate patterns of expression of these proteins. Results: PLIUS did not fully prevent cartilage degeneration in the prevention groups, but diminished the severity of the disease, with the treated joints showing markedly decreased surface irregularities and a much smaller degree of loss of matrix staining as compared to controls. PLIUS also attenuated disease progression in the groups with established disease, although to a somewhat lesser extent as compared to the prevention groups. Immunohistochemical staining demonstrated a markedly decreased degree of TGF-beta 1 production in the PLIUS-treated joints. This indicates less active endogenous repair, consistent with the marked reduction in cartilage degradation. Conclusions: PLIUS exhibits the ability to attenuate the progression of cartilage degeneration in an animal model of idiopathic human OA. The effect was greater in the treatment of early, rather than established, degeneration. Published by Elsevier Ltd on behalf of Osteoarthritis Research Society International.
Gurkan I; Ranganathan A; Yang X; Horton W E; Todman M; Huckle J; Pleshko N; Spencer R G
Osteoarthritis and Cartilage
2010
2010-05
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1016/j.joca.2010.01.006" target="_blank" rel="noreferrer noopener">10.1016/j.joca.2010.01.006</a>