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Ribs were dissociated using collagenase P and strained through a 70μm nylon filter to remove ossified tissue, followed by 3 days in culture over 1.5% agarose in PBS to specifically select for chondrocytes [34].
Samples in both SIS and Blank groups mainly showed ossified tissue at all time points with partial fibrogenesis in a few samples.
Histological evaluation also revealed the presence of more mature bone in the new ossified tissue when the low dose of BMP-2 was delivered using the biomimetic supramolecular system.
Without staining of soft tissues, only ossified tissue is clearly visible in μCT scans.
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In this way the osteophyte volume measured by μCT may not be easily translatable to human OA, because mice have a greater propensity to ossify soft tissue structures that are not typically mineralized in humans.
We recently argued, based on MRI studies and especially on older pathological studies [ 45], that structural damage happens in two steps: first, inflammation causes erosive structural damage and these bony defects are then filled up with (fibrous) repair tissue; second, this repair tissue is subsequently ossified.
The tissue was extensively ossified at 14 and 28 days, and the newly formed bone exhibited normal bone histology.
In addition, unlike adult long bone fragments or embryonic calvarial explants often used in other models [ 33- 35], the intact femur retains the three-dimensional, multi-cell architecture including ossified bone and hematopoietic cells, stroma and matrix tissue.
Gross examination revealed a mass that was over 95% ossified, with the remainder being composed of lobulated adipose tissue.
In each case the ring is composed of fibrous tissue, part of which is converted into cartilage to form a female surface; the remainder may be ossified.
In addition to soft tissue windows, the bone windows (reconstructed with a bony algorithm) should be routinely evaluated if the tumour contacts ossified laryngeal cartilage.
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