Dataset Information

Profile of growth factors in calf costochondral junction – the site of initial bone formation

ABSTRACT: Endochondral bone formation is orchestrated by growth factors produced by chondrocytes and deposited in cartilage matrix. Some of these factors were identified, but their complete list and relationship remains unknown. In this work the growth factors were isolated from calcified cartilage of costochondral junctions. The isolation involved hyaluronidase digestion, guanidinium hydrochloride (GuHCl) extraction, HCl decalcification and GuHCl extraction of decalcified matrix. Growth factors were purified by heparin chromatography and estimated by enzyme-linked immunosorbent assay (ELISA) or used for protein sequence analysis. Bone morphogenetic protein-7 (BMP-7), Growth/differentiation factor 5 (GDF-5), and NEL-like protein 1 (NELL-1) quantitatively dominated in material obtained in all steps of isolation. During ossification perivascular cells and septoclasts enter the cartilage and release growth factors stimulating osteoclastogenesis. Osteoclasts dissolve calcified cartilage and transport released factors needed for stimulation of osteoprogenitor cells. Presence of BMP-7, GDF-5 and NELL-1 at the site of initial bone formation may suggest that their synergistic action favors osteogenesis.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Cartilage

SUBMITTER: Michał Kistowski

LAB HEAD: Michał Dadlez

PROVIDER: PXD021781 | Pride | 2021-05-03

REPOSITORIES: Pride

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Publications

Growth factor profile in calcified cartilage from the metaphysis of a calf costochondral junction, the site of initial bone formation.

Iwan Anna A Moskalewski Stanisław S Hyc Anna A

Biomedical reports 20210409 6

Endochondral bone formation is orchestrated by growth factors produced by chondrocytes and deposited in the cartilage matrix. Whilst some of these factors have been identified, the complete list and their relationship remains unknown. In the present study, the growth factors were isolated from non-calcified and calcified cartilage of costochondral junctions. Cartilage dissected from the ribs of 6-20-week-old calves was purchased from a local butcher within 24 h of the death of the animal. The is ...[more]

PMID: 33884197

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Project description:Successfully replacing damaged cartilage with tissue-engineered constructs requires integration with the host tissue and could benefit from leveraging the native tissue's intrinsic healing capacity; however, efforts are limited by a poor understanding of how cartilage repairs minor defects. Here, we investigated the conditions that foster natural cartilage tissue repair to identify strategies that might be exploited to enhance the integration of engineered/ grafted cartilage with host tissue. We damaged porcine articular cartilage explants and using a combination of pulsed SILAC-based proteomics, ultrastructural imaging, and catabolic enzyme blocking strategies reveal that integration of damaged cartilage surfaces is not driven by neo-matrix synthesis, but rather local depletion of proteoglycans. ADAMTS4 expression and activity are upregulated in injured cartilage explants, but integration could be reduced by inhibiting metalloproteinase activity with TIMP3. These observations suggest that catabolic enzyme-mediated proteoglycan depletion likely allows existing collagen fibrils to undergo cross-linking, fibrillogenesis, or entanglement, driving integration. Catabolic enzymes are often considered pathophysiological markers of osteoarthritis. Our findings suggest that damage-induced upregulation of metalloproteinase activity may be a part of a healing response that tips towards tissue destruction under pathological conditions and in osteoarthritis, but could also be harnessed in tissue engineering strategies to mediate repair. Statement of significance: Cartilage tissue engineering strategies require graft integration with the surrounding tissue; however, how the native tissue repairs minor injuries is poorly understood. We applied pulsed SILACbased proteomics, ultrastructural imaging, and catabolic enzyme blocking strategies to a porcine cartilage explant model and found that integration of damaged cartilage surfaces is driven by catabolic enzyme-mediated local depletion of proteoglycans. Although catabolic enzymes have been implicated in cartilage destruction in osteoarthritis, our findings suggest that damage-induced upregulation of metalloproteinase activity may be a part of a healing response that tips towards tissue destruction under pathological conditions. They also suggest that this natural cartilage tissue repair process could be harnessed in tissue engineering strategies to enhance the integration of engineered cartilage with host tissue.

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Dataset Information

Profile of growth factors in calf costochondral junction – the site of initial bone formation

Publications

Growth factor profile in calcified cartilage from the metaphysis of a calf costochondral junction, the site of initial bone formation.

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