Rita Kandel MD Pathology and Laboratory Medicine, Sinai Health System, University of Toronto, Toronto ,ON Canada HISTOLOGY OF CARTILAGE REPAIR
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Rita Kandel MD
Pathology and Laboratory Medicine,
Sinai Health System,
University of Toronto,
Toronto ,ON Canada
HISTOLOGY OF CARTILAGE
REPAIR
AMERICAN SOCIETY FOR TESTING
AND MATERIALS (ASTM) GUIDELINES
- Gross evaluation - Histologic evaluation (validated scoring system)
- Biochemical evaluation (quantity and quality of collagen and proteoglycans)
- Biomechanical assessment
- Appropriate statistical analyses
GROSS ASSESSMENT
ICRS grading system
HISTOLOGICAL ANALYSIS OF
REPAIR
Implant
1) Conventional histology
Arzi B et al., Acta Biomater 2015; 1 (23):72-81
Synovium
HISTOLOGICAL APPEARANCE
OF JOINT SURFACE
Hyaline (non-
calcified)
Cartilage
Bone
Calcified
cartilage
SAMPLE PROCESSING FOR HISTOLOGY
1) EXCISE DEFECT
2) FIXATION: 10% buffered formalin duration dependent on tissue size
3) DECALCIFICATION: 0.5M EDTA duration dependent on tissue size at room temperature
4) EMBEDDING: paraffin vs plastic vs frozen section
5) SAMPLING OF DEFECT: standardize location
HISTOLOGICAL EVALUATION OF
REPAIR: tissue quality
Kandel RA et al. Biomaterials 2006
H&E
Toluidine blue
CPP
In vitro formed cartilage In vivo cartilage
Bone
1mm
Safranin O
EVALUATION OF REPAIR: interfaces
Hoemann C et al. Cartilage 2011
Safranin O
Native
Repair
200um
100um
Interface
LATERAL DEEP
one
one
Toluidine blue
1mm
Kandel RA et al. Biomaterials 2006
CLASSIFICATION SYSTEMS
Orth P et al Tiss Engineering Part C, 2012
100 μm
Hyaline cartilage
Calcified cartilage
ARTICULAR CARTILAGE: depth dependent
zonal architecture
10 -
20%
40 -
60%
30 -
40%
Co
llagen
Adapted from Klein TJ. et al., 2009
PRG4
Proteoglycan
Alkaline phosphatase
ALKALINE PHOSPHATASE
Bone
X X Xx
Hoemann C et al., Cartilage 2011
X X X x
HISTOLOGICAL ANALYSIS OF
REPAIR
1) Histology
2) Immunohistochemical staining
aggrecan
type II collagen type I collagen PRG4
PRG4
HISTOLOGICAL ANALYSIS OF
REPAIR
1) Histology 2) Immunohistochemical staining 3) Polarized light microscopy
REPAIR NORMAL
SAMPLING PITFALLS
Hoemann C et al. Cartilage 2011; 2:153 -172
Orth P et al. Progress in Histochemistry and Cytochemistry 2015
REPAIR
SAMPLING PITFALLS
HISTOLOGY PITFALLS
1) Fixation duration
2) Method of decalcification
3) Grading expertise
4) Poor quality images
European Cells and Materials 2016; 31:79-94
DOES HISTOLOGY PREDICT
FUNCTION?
HISTOLOGICAL APPEARANCE OF JOINT
SURFACE
Hyaline
Cartilage
Calcified
cartilage
Bone
Cells
Matrix
Normal
HISTOLOGICAL APPEARANCE autologous chondrocyte implantation at 1 year
Safranin O stain
Ref. Sancho-Tello M et al., Int J Artif Organs 2015; 38(4):210-223
Ref. Sancho-Tello M et al., Int J Artif Organs 2015; 38(4):210-223
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