1. |
- Prittinen, Juha, 1989-, et al.
(author)
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Effect of centrifugal force on the development of articular neocartilage with bovine primary chondrocytes
- 2019
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In: Cell and Tissue Research. - New York : Springer. - 0302-766X .- 1432-0878. ; 375:3, s. 629-639
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Journal article (peer-reviewed)abstract
- A lot has been invested into understanding how to assemble cartilage tissue in vitro and various designs have been developed to manufacture cartilage tissue with native-like biological properties. So far, no satisfactory design has been presented. Bovine primary chondrocytes are used to self-assemble scaffold-free constructs to investigate whether mechanical loading by centrifugal force would be useful in manufacturing cartilage tissue in vitro. Six million chondrocytes were laid on top of defatted bone disks placed inside an agarose well in 50-ml culture tubes. The constructs were centrifuged once or three times per day for 15 min at a centrifugal force of 771×g for up to 4 weeks. Control samples were cultured under the same conditions without exposure to centrifugation. The samples were analysed by (immuno)histochemistry, Fourier transform infrared imaging, micro-computed tomography, biochemical and gene expression analyses. Biomechanical testing was also performed. The centrifuged tissues had a more even surface covering a larger area of the bone disk. Fourier transform infrared imaging analysis indicated a higher concentration of collagen in the top and bottom edges in some of the centrifuged samples. Glycosaminoglycan contents increased along the culture, while collagen content remained at a rather constant level. Aggrecan and procollagen α1(II) gene expression levels had no significant differences, while procollagen α2(I) levels were increased significantly. Biomechanical analyses did not reveal remarkable changes. The centrifugation regimes lead to more uniform tissue constructs, whereas improved biological properties of the native tissue could not be obtained by centrifugation.
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2. |
- Berg, B., et al.
(author)
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Development of osteoarthritis in patients with degenerative meniscal tears treated with exercise therapy or surgery : a randomized controlled trial
- 2020
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In: Osteoarthritis and Cartilage. - : Elsevier BV. - 1063-4584. ; 28:7, s. 897-906
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Journal article (peer-reviewed)abstract
- Objective: To evaluate progression of individual radiographic features 5 years following exercise therapy or arthroscopic partial meniscectomy as treatment for degenerative meniscal tear. Design: Randomized controlled trial including 140 adults, aged 35–60 years, with a magnetic resonance image verified degenerative meniscal tear, and 96% without definite radiographic knee osteoarthritis. Participants were randomized to either 12-weeks of supervised exercise therapy or arthroscopic partial meniscectomy. The primary outcome was between-group difference in progression of tibiofemoral joint space narrowing and marginal osteophytes at 5 years, assessed semi-quantitatively by the OARSI atlas. Secondary outcomes included incidence of radiographic knee osteoarthritis and symptomatic knee osteoarthritis, medial tibiofemoral fixed joint space width (quantitatively assessed), and patient-reported outcome measures. Statistical analyses were performed using a full analysis set. Per protocol and as treated analysis were also performed. Results: The risk ratios (95% CI) for progression of semi-quantitatively assessed joint space narrowing and medial and lateral osteophytes for the surgery group were 0.89 (0.55–1.44), 1.15 (0.79–1.68) and 0.77 (0.42–1.42), respectively, compared to the exercise therapy group. In secondary outcomes (full-set analysis) no statistically significant between-group differences were found. Conclusion: The study was inconclusive with respect to potential differences in progression of individual radiographic features after surgical and non-surgical treatment for degenerative meniscal tear. Further, we found no strong evidence in support of differences in development of incident radiographic knee osteoarthritis or patient-reported outcomes between exercise therapy and arthroscopic partial meniscectomy.
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3. |
- Hellberg, I., et al.
(author)
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3D analysis and grading of calcifications from ex vivo human meniscus
- 2023
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In: Osteoarthritis and Cartilage. - : Elsevier BV. - 1063-4584. ; 31:4, s. 482-492
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Journal article (peer-reviewed)abstract
- Objective: Meniscal calcifications are associated with the pathogenesis of knee osteoarthritis (OA). We propose a micro-computed tomography (μCT) based 3D analysis of meniscal calcifications ex vivo, including a new grading system. Method: Human medial and lateral menisci were obtained from 10 patients having total knee replacement for medial compartment OA and 10 deceased donors without knee OA (healthy references). The samples were fixed; one subsection was imaged with μCT, and the adjacent tissue was processed for histological evaluation. Calcifications were examined from the reconstructed 3D μCT images, and a new grading system was developed. To validate the grading system, meniscal calcification volumes (CVM) were quantitatively analyzed and compared between the calcification grades. Furthermore, we estimated the relationship between histopathological degeneration and the calcification severity. Results: 3D μCT images depict calcifications in every sample, including diminutive calcifications that are not visible in histology. In the new grading system, starting from grade 2, each grade results in a CVM that is 20.3 times higher (95% CI 13.3–30.5) than in the previous grade. However, there was no apparent difference in CVM between grades 1 and 2. The calcification grades appear to increase with the increasing histopathological degeneration, although histopathological degeneration is also observed with small calcification grades. Conclusions: 3D μCT grading of meniscal calcifications is feasible. Interestingly, it seems that there are two patterns of degeneration in the menisci of our sample set: 1) with diminutive calcifications (calcification grades 1–2), and 2) with large to widespread calcifications (calcification grades 3–5).
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4. |
- Karjalainen, V. P., et al.
(author)
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Quantitative three-dimensional collagen orientation analysis of human meniscus posterior horn in health and osteoarthritis using micro-computed tomography
- 2021
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In: Osteoarthritis and Cartilage. - : Elsevier BV. - 1063-4584. ; 29:5, s. 762-772
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Journal article (peer-reviewed)abstract
- Objective: Knee osteoarthritis (OA) is associated with meniscal degeneration that may involve disorganization of the meniscal collagen fiber network. Our aims were to quantitatively analyze the microstructural organization of human meniscus samples in 3D using micro-computed tomography (μCT), and to compare the local microstructural organization between OA and donor samples. Method: We collected posterior horns of both medial and lateral human menisci from 10 end-stage medial compartment knee OA patients undergoing total knee replacement (medial & lateral OA) and 10 deceased donors without knee OA (medial & lateral donor). Posterior horns were dissected and fixed in formalin, dehydrated in ascending ethanol concentrations, treated with hexamethyldisilazane (HMDS), and imaged with μCT. We performed local orientation analysis of collagenous microstructure in 3D by calculating structure tensors from greyscale gradients within selected integration window to determine the polar angle for each voxel. Results: In donor samples, meniscus bundles were aligned circumferentially around the inner border of meniscus. In medial OA menisci, the organized structure of collagen network was lost, and main orientation was shifted away from the circumferential alignment. Quantitatively, medial OA menisci had the lowest mean orientation angle compared to all groups, −24° (95%CI -31 to −18) vs medial donor and −25° (95%CI -34 to −15) vs lateral OA. Conclusions: HMDS-based μCT imaging enabled quantitative analysis of meniscal collagen fiber bundles and their orientations in 3D. In human medial OA menisci, the collagen disorganization was profound with overall lower orientation angles, suggesting collagenous microstructure disorganization as an important part of meniscus degradation.
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5. |
- Kestilä, I., et al.
(author)
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Three-dimensional microstructure of human meniscus posterior horn in health and osteoarthritis
- 2019
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In: Osteoarthritis and Cartilage. - : Elsevier BV. - 1063-4584. ; 27:22, s. 1790-1799
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Journal article (peer-reviewed)abstract
- Objective: To develop and perform ex vivo 3D imaging of meniscus posterior horn microstructure using micro-computed tomography (μCT), and to compare specimens from healthy references against end-stage osteoarthritis (OA) using conventional section-based histology and qualitative μCT. Design: We retrieved human medial and lateral menisci from 10 deceased donors without knee OA (healthy references) and medial and lateral menisci from 10 patients having total knee replacement for medial compartment OA. Meniscal posterior horns were dissected and fixed in formalin. One subsection underwent hexamethyldisilazane (HMDS) treatment and μCT imaging. Pauli's histopathological scoring was performed for 3 other subsections. The differences in histopathological scores were estimated using mixed linear regression, resulting in fixed effects estimates for within-knee comparisons and adjusted for age and body mass index for between-subjects comparisons. Results: 3D visualization with μCT qualitatively revealed similar microstructural changes in the posterior horns as conventional histology. The mean histopathological score was higher for medial menisci from OA knees vs both medial reference menisci (mean difference [95% CI], 3.9 [2.6,5.3]), and lateral menisci from OA knees (3.9 [2.9,5.0]). The scores were similar between lateral menisci from OA knees and lateral reference menisci (0.8 [−0.6,2.2]), and between medial and lateral reference menisci (0.8 [−0.3,1.9]). Conclusions: HMDS-based μCT protocol allows unique 3D visualization of meniscus microstructures. Posterior horns of medial menisci from medial compartment OA knees had higher histopathological scores than both the lateral posterior horns from the same OA knees and medial reference menisci, suggesting a strong association between meniscus degradation and unicompartmental knee OA.
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6. |
- Kobrina, Yevgeniya, et al.
(author)
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Clustering of infrared spectra reveals histological zones in intact articular cartilage
- 2012
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In: Osteoarthritis and Cartilage. - : Elsevier BV. - 1063-4584. ; 20:5, s. 460-468
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Journal article (peer-reviewed)abstract
- Objective: Articular cartilage (AC) exhibits specific zonal structure that follows the organization of collagen network and concentration of tissue constituents. The aim of this study was to investigate the potential of unsupervised clustering analysis applied to Fourier transform infrared (FIR) microspectroscopy to detect depth-dependent structural and compositional differences in intact AC. Method: Seven rabbit and eight bovine intact patellae AC samples were imaged using FTIR microspectroscopy and normalized raw spectra were clustered using the fuzzy C-means algorithm. Differences in mean spectra of clusters were investigated by quantitative estimation of collagen and proteoglycan (PG) contents, as well as by careful visual investigation of locations of spectral changes. Results: Clustering revealed the typical layered structure of AC in both species. However, more distinct clusters were found for rabbit samples, whereas bovine AC showed more complex layered structure. In both species, clustering structure corresponded with that in polarized light microscopic (PLM) images; however, some differences were also observed. Spectral differences between clusters were identified at the same spectral locations for both species. Estimated PG/collagen ratio decreased significantly from superficial to middle or deep zones, which might explain the difference in clustering results compared to PLM. Conclusion: FTIR microspectroscopy in combination with cluster analysis allows detailed examination of spatial changes in AC. As far as we know, no previous single technique could reveal a layered structure of AC without any a priori information. (C) 2012 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.
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7. |
- Nissilä, Tuukka, et al.
(author)
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Cellulose nanofiber aerogels impregnated with bio-based epoxy using vacuum infusion : Structure, orientation and mechanical properties
- 2018
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In: Composites Science And Technology. - : Elsevier. - 0266-3538 .- 1879-1050. ; 155, s. 64-71
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Journal article (peer-reviewed)abstract
- Cellulose nanofiber aerogels were used as preforms that were impregnated with a bio-epoxy resin via a widely used vacuum infusion process. The simple and straightforward nanocomposite processing approach resulted in an almost 70% improvement in the storage modulus of the polymer with only an 11.7 wt% cellulose nanofiber content. The nanofibers were well dispersed in the polymer matrix and the fiber structures were anisotropically aligned. The impregnation time of the aerogels was also significantly lower than that of the more commonly used nanopapers. It was thus shown that environmentally friendly and mechanically robust nanocomposites could be produced by impregnating cellulose nanofiber aerogels with a thermosetting resin using a processing approach that has potential to be scaled up for commercial use.
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