Safranin O reduces loss of glycosaminoglycans from bovine articular cartilage during histological specimen preparation

Király, Kari; Lammi, Mikko J.; Arokoski, Jari; Lapveteläinen, Tuomo; Tammi, Markku; Helminen, Heikki J.; Kiviranta, Ilkka

doi:10.1007/bf02331414

Cited by 59 publications

(39 citation statements)

References 24 publications

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“…After being washed in tap water for 5 minutes, they were decalcified in 15% EDTA for 1 week.. Serial sections of the TMJ were then performed with every 5 th section stained with H&E or Safranin O (13). …”

Section: Methodsmentioning

confidence: 99%

Analysis of microarchitectural changes in a mouse temporomandibular joint osteoarthritis model

Chen

Gupta

Barasz

et al. 2009

Archives of Oral Biology

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Objective-Little is known about the natural progression of the disease process of temporomandibular joint (TMJ) osteoarthritis (OA), which affects approximately 1 % of the US population. The goal of this study was to examine the early microarchitectural and molecular changes in the condylar cartilage and subchondral bone in biglycan/fibromodulin (Bgn/Fmod) doubledeficient mice, which develop TMJ-OA at 6 months.Methods-TMJs from 3 month old (n=44) and 9 month old (n=52) wild-type (WT n=46) and Bgn/ Fmod (n=50) double-deficient mice were evaluated. Micro-CT analysis of the subchondral bone (n=24), transmission electron microscopy for condylar cartilage fibril diameters (n=26), and real time PCR analysis for gene expression for bone and cartilage maturation markers (n=45) was performed.Results-A statistically significant increase in collagen fibril diameter of the condylar cartilage and a decrease in expression of Parathyroid related protein in the mandibular condylar head were observed in the 3 month Bgn/Fmod double-deficient mice compared to WT controls. The 9 month Bgn/Fmod double-deficient mouse demonstrated an increase in bone volume and total volume in subchondral bone, and an increase in the expression of Collagen Type X and Aggrecan in the mandibular condylar head compared to the WT controls.Conclusion-We found that changes in the microarchitecture of the condylar cartilage preceded changes in the subchondral bone during OA in the TMJ in Bgn/Fmod double-deficient mice.

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Section: Methodsmentioning

confidence: 99%

Analysis of microarchitectural changes in a mouse temporomandibular joint osteoarthritis model

Chen

Gupta

Barasz

et al. 2009

Archives of Oral Biology

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“…The specimen preparation protocol has been previously described by Király et al and Kiviranta et al32 33 All steps of tissue and section preparation were strictly standardised. Tissue sections were stained with safranin O (Fisher Scientific, Fair Lawn, NJ).…”

Section: Methodsmentioning

confidence: 99%

Articular cartilage superficial zone collagen birefringence reduced and cartilage thickness increased before surface fibrillation in experimental osteoarthritis

Panula

Hyttinen

Arokoski

et al. 1998

Annals of the Rheumatic Diseases

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127

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Objectives-To investigate articular cartilage collagen network, thickness of birefringent cartilage zones, and glycosaminoglycan concentration in macroscopically normal looking knee joint cartilage of young beagles subjected to experimental slowly progressive osteoarthritis (OA). Methods-OA was induced by a tibial 30°v algus osteotomy in 15 female beagles at the age of 3 months. Fifteen sisters were controls. Cartilage specimens were collected seven (Group 1) and 18 months (Group 2) postoperatively. Collagen induced optical path diVerence and cartilage zone thickness measurements were determined from histological sections of articular cartilage with smooth and intact surface by computer assisted quantitative polarised light microscopy. Volume density of cartilage collagen fibrils was determined by image analysis from transmission electron micrographs and content of glycosaminoglycans by quantitative digital densitometry from histological sections. Results-In the superficial zone of the lateral tibial and femoral cartilage, the collagen induced optical path diVerence (birefringence) decreased by 19 to 71% (p < 0.05) seven months postoperatively. This suggests that severe superficial collagen fibril network deterioration took place, as 18 months postoperatively, macroscopic and microscopic OA was present in many cartilage areas. Thickness of the uncalcified cartilage increased while the superficial zone became thinner in the same sites. In operated dogs, glycosaminoglycan content first increased (Group 1) in the lateral tibial condyle and then decreased (Group 2) (p < 0.05). Conclusion-In this OA model, derangement of the superficial zone collagen network was the probable reason for birefringence reduction. This change occurred well before macroscopic OA.

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“…Samples were fixed with 10% formalin, decalcified, dehydrated and embedded in paraffin as described earlier [Kiraly et al, 1997]. When processing samples for DD, safranin O stain (Fisher Scientific, Fair Lawn, N.J., Rieppo/Töyräs/Nieminen/Kovanen/ Hyttinen/Korhonen/Jurvelin/Helminen USA) was added to the fixative to minimize a possible loss of tissue PGs [Kiraly et al, 1996a].…”

Section: Microscopic Techniquesmentioning

confidence: 99%

Structure-Function Relationships in Enzymatically Modified Articular Cartilage

Rieppo¹,

Töyräs

Nieminen³

et al. 2003

Cells Tissues Organs

118

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The present study is aimed at revealing structure-function relationships of bovine patellar articular cartilage. Collagenase, chondroitinase ABC and elastase were used for controlled and selective enzymatic modifications of cartilage structure, composition and functional properties. The effects of the enzymatic degradations were quantitatively evaluated using quantitative polarized light microscopy, digital densitometry of safranin O-stained sections as well as with biochemical and biomechanical techniques. The parameters related to tissue composition and structure were correlated with the indentation stiffness of cartilage. In general, tissue alterations after enzymatic digestions were restricted to the superficial cartilage. All enzymatic degradations induced superficial proteoglycan (PG) depletion. Collagenase also induced detectable superficial collagen damage, though without causing cartilage fibrillation or tissue swelling. Quantitative microscopic techniques were more sensitive than biochemical methods in detecting these changes. The Young’s modulus of cartilage decreased after enzymatic treatments indicating significant softening of the tissue. The PG concentration of the superficial zone proved to be the major determinant of the Young’s modulus (r² = 0.767, n = 72, p < 0.001). Results of the present study indicate that specific enzymatic degradations of the tissue PGs and collagen can provide reproducible experimental models to clarify the structure-function relationships of cartilage. Effects of these models mimic the changes observed in early osteoarthrosis. Biomechanical testing and quantitative microscopic techniques proved to be powerful tools for detecting the superficial structural and compositional changes while the biochemical measurements on the whole uncalcified cartilage were less sensitive.

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Safranin O reduces loss of glycosaminoglycans from bovine articular cartilage during histological specimen preparation

Cited by 59 publications

References 24 publications

Analysis of microarchitectural changes in a mouse temporomandibular joint osteoarthritis model

Analysis of microarchitectural changes in a mouse temporomandibular joint osteoarthritis model

Articular cartilage superficial zone collagen birefringence reduced and cartilage thickness increased before surface fibrillation in experimental osteoarthritis

Structure-Function Relationships in Enzymatically Modified Articular Cartilage

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