Fixation, decalcification, and tissue processing effects on articular cartilage proteoglycans

Kiviranta, J.; Tammi, Markku; Jurvelin, Jukka S.; Säämänen, Anna‐Marja; Helminen, H. J.

doi:10.1007/bf00553719

Cited by 13 publications

(15 citation statements)

References 16 publications

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“…Formalin fixation and EDTA acid decalcification used in the processing of the tissues in our study seem to preserve proteoglycan content 38. Other fixation and decalcification techniques may, however, effect proteoglycan content and staining influencing the reproducibility of the HHGS score.…”

Section: Discussionmentioning

confidence: 74%

Validity of histopathological grading of articular cartilage from osteoarthritic knee joints

Østergaard

Andersen

Petersen

et al. 1999

Annals of the Rheumatic Diseases

102

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Objectives-To determine the validity of the histological-histochemical grading system (HHGS) for osteoarthritic (OA) articular cartilage. Methods-Human articular cartilage was obtained from macroscopically normal (n = 13) and OA (n = 21) knee joints. Sections of central and peripheral regions of normal samples were produced. Sections of regions containing severe, moderate, and mild OA changes were produced from each OA sample. A total of 89 sections were graded by means of the HHGS (0-14) twice by three observers. Results-Average scores for regions designated severe (8.64) and moderate (5.83) OA were less than the expected (10-14 and 6-9, respectively) according to the HHGS, whereas average scores for the region designated mild (5.29) OA and central and peripheral regions (2.19) of normal cartilage were higher than expected (2-5 and 0-1, respectively). The HHGS was capable of diVerentiating between articular cartilage from macroscopically normal and OA joints and between the region designated severe OA and other regions. However, the HHGS did not adequately diVerentiate between regions designated mild and moderate OA. Values for sensitivity, specificity, and eYciency for all regions varied considerably. Conclusion-The HHGS is valid for normal and severe OA cartilage, but does not permit distinction between mild and moderate OA changes in articular cartilage. (Ann Rheum Dis 1999;58:208-213) The histopathology of osteoarthritis (OA) is generally graded using the histologicalhistochemical grading system (HHGS) proposed by Mankin et al 1 or a number of histopathological grading systems that are modifications of the original system. 2-7 The HHGS was initially developed for the grading of OA in human articular cartilage 1 and has been used extensively in human studies. More recently, the HHGS and modifications thereof have also become extensively used for the grading of articular cartilage from animal models of OA. We have previously investigated the intraobserver and interobserver reproducibilities of the HHGS based on human articular cartilage 30 and concluded that they were inadequate. Similar values regarding the intraobserver and interobserver reproducibilities of the HHGS have been reported in a study on materials from an experimental animal model of OA. 31 The results of our previous study also indicated that the validity of the HHGS was questionable. It is of paramount importance to determine if the HHGS is valid. A valid grading system with inadequate intraobserver and interobserver reproducibilities could and should be improved upon. However, a grading system that is not valid should be disregarded altogether and replaced by a completely new grading system.To evaluate the validity of a histopathological grading system the results obtained when using the system should be compared with the results of a system already known to be valid. Unfortunately, a valid grading system capable of serving as "the gold standard" does not exist for OA articular cartilage at the histological level.Therefore, the pur...

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Section: Discussionmentioning

confidence: 74%

Validity of histopathological grading of articular cartilage from osteoarthritic knee joints

Østergaard

Andersen

Petersen

et al. 1999

Annals of the Rheumatic Diseases

102

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“…Specimens were formalin fixed and decalcified with 10% EDTA [19]. A 2mm coronal section of the tibial plateau centered on the collateral ligament attachments was prepared, paraffin embedded, and sectioned (5 µm) [19].…”

Section: Methodsmentioning

confidence: 99%

“…A 2mm coronal section of the tibial plateau centered on the collateral ligament attachments was prepared, paraffin embedded, and sectioned (5 µm) [19]. Three sections collected at 200µm intervals for each tibial plateau were processed for each stain.…”

Section: Methodsmentioning

confidence: 99%

Chronic in vivo load alteration induces degenerative changes in the rat tibiofemoral joint

Roemhildt

Beynnon

Gauthier

et al. 2013

Osteoarthritis and Cartilage

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Objective We investigated the relationship between the magnitude and duration of sustained compressive load alteration and the development of degenerative changes in the rat tibiofemoral joint. Methods A varus loading device was attached to the left hind limb of mature rats to apply increased compression to the medial compartment and decreased compression to the lateral compartment of the tibiofemoral joint of either 0% or 100% body weight for 0, 6 or 20 weeks. Compartment-specific assessment of the tibial plateaus included biomechanical measures (articular cartilage aggregate modulus, permeability and Poisson’s ratio, and subchondral bone modulus) and histological assessments (articular cartilage, calcified cartilage, and subchondral bone thicknesses, degenerative scoring parameters, and articular cartilage cellularity). Results Increased compression in the medial compartment produced significant degenerative changes consistent with the development of osteoarthritis including a progressive decrease in cartilage aggregate modulus (43% and 77% at 6 and 20 weeks), diminished cellularity (38% and 51% at 6 and 20 weeks), and increased histological degeneration. At 20 weeks, medial compartment articular cartilage thickness deceased 30% while subchondral bone thickness increased 32% and subchondral bone modulus increased 99%. Decreased compression in the lateral compartment increased calcified cartilage thickness, diminished region-specific subchondral bone thickness and revealed trends for reduced cellularity and decreased articular cartilage thickness at 20 weeks. Conclusions Altered chronic joint loading produced degenerative changes consistent with those observed clinically with the development of osteoarthritis and may replicate the slow development of non-traumatic osteoarthritis in which mechanical loads play a primary etiological role.

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“…The samples were first fixed in 4% formaldehyde in 0.07 M sodium phosphate buffer, pH of 7.0, for 48 h at 48C, and then decalcified for 14 days with 10% ethylenediaminetetraacetic acid and 4% formaldehyde in 0.1 M sodium phosphate buffer, pH of 7.4, at room temperature. 37 The samples were embedded in Tissue-Tek III embedding wax (Sakura Finetek Europe, Zoeterwoude, the Netherlands) and 5-mm-thick sections were prepared.…”

Section: Preparation Of Histologic Samplesmentioning

confidence: 99%

Poly-L-D-Lactic Acid Scaffold in the Repair of Porcine Knee Cartilage Lesions

et al. 2007

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Articular cartilage injuries cause a major clinical problem because of the negligible repair capacity of cartilage. Autologous chondrocyte transplantation is a surgical method developed to repair cartilage lesions. In the operation, cartilage defect is covered with a periosteal patch and the suspension of cultured autologous chondrocytes is injected into the lesion site. The method can form good repair tissue, but new techniques are needed to make the operation easier and to increase the postoperative biomechanical properties of the repair tissue. In this study, we investigated poly-L,D-lactic acid (PLDLA) scaffolds alone or seeded with autologous chondrocytes in the repair of circular 6-mm cartilage lesions in immature porcine knee joints. Spontaneous repair was used as a reference. Histologic evaluation of the repair tissue showed that spontaneous repair exhibited higher scores than either PLDLA scaffold group (with or without seeded chondrocytes). The scaffold material was most often seen embedded in the subchondral bone underneath the defect area, probably because of the hardness of the PLDLA material. However, some of the cell-seeded and nonseeded scaffolds contained cartilaginous tissue, suggesting that invasion of mesenchymal cells inside nonseeded scaffolds had occurred. Hyaluronan deposited in the scaffold had possibly acted as a chemoattractant for the cell recruitment. In conclusion, the PLDLA scaffold material used in this study was obviously mechanically too hard to be used for cartilage repair in immature animals.

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Fixation, decalcification, and tissue processing effects on articular cartilage proteoglycans

Cited by 13 publications

References 16 publications

Validity of histopathological grading of articular cartilage from osteoarthritic knee joints

Validity of histopathological grading of articular cartilage from osteoarthritic knee joints

Chronic in vivo load alteration induces degenerative changes in the rat tibiofemoral joint

Poly-L-D-Lactic Acid Scaffold in the Repair of Porcine Knee Cartilage Lesions

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