Characterization of ex vivo-generated bovine and human cartilage by immunohistochemical, biochemical, and magnetic resonance imaging analyses.

Title

Characterization of ex vivo-generated bovine and human cartilage by immunohistochemical, biochemical, and magnetic resonance imaging analyses.

Creator

Nugent Ashleigh E; Reiter David A; Fishbein Kenneth W; McBurney Denise L; Murray Travis; Bartusik Dorota; Ramaswamy Sharan; Spencer Richard G; Horton Walter E Jr

Publisher

Tissue engineering. Part A

Date

2010
2010-07

Description

Osteoarthritis (OA) is a prevalent age-associated disease involving altered chondrocyte homeostasis and cartilage degeneration. The avascular nature of cartilage and the altered chondrocyte phenotype characteristic of OA severely limit the capacity for in vivo tissue regeneration. Cell- and tissue-based repair has the potential to revolutionize treatment of OA, but those approaches have exhibited limited clinical success to date. In this study, we test the hypothesis that bovine and human chondrocytes in a collagen type I scaffold will form hyaline cartilage ex vivo with immunohistochemical, biochemical, and magnetic resonance (MR) endpoints similar to the original native cartilage. Chondrocytes were isolated from 1- to 3-week-old calf knee cartilage or from cartilage obtained from human total knee arthroplasties, suspended in 2.7 mg/mL collagen I, and plated as 300 microL spot cultures with 5 x 10(6) each. Medium formulations were varied, including the amount of serum, the presence or absence of ascorbate, and treatments with cytokines. Bovine chondrocytes generated metachromatic territorial and interstitial matrix and accumulated type II collagen over time. Type VI collagen was confined primarily to the pericellular region. The ex vivo-formed bovine cartilage contained more chondroitin sulfate per dry weight than native cartilage. Human chondrocytes remained viable and generated metachromatic territorial matrix, but were unable to support interstitial matrix accumulation. MR analysis of ex vivo-formed bovine cartilage revealed evidence of progressively maturing matrix, but MR-derived indices of tissue quality did not reach those of native cartilage. We conclude that the collagen-spot culture model supports formation and maturation of three-dimensional hyaline cartilage from active bovine chondrocytes. Future studies will focus on determining the capacity of human chondrocytes to show comparable tissue formation.

Subject

Aged; Animals; Articular/drug effects/growth & development/*metabolism; Cartilage; Cattle; Cell Proliferation/drug effects; Collagen Type II/metabolism; Culture Media/pharmacology; Cytokines/pharmacology; Glycosaminoglycans/metabolism; Humans; Immunohistochemistry; Magnetic Resonance Spectroscopy; Middle Aged; Tissue Engineering/*methods

Rights

Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).

Pages

2183–2196

Issue

7

Volume

16

Citation

Nugent Ashleigh E; Reiter David A; Fishbein Kenneth W; McBurney Denise L; Murray Travis; Bartusik Dorota; Ramaswamy Sharan; Spencer Richard G; Horton Walter E Jr, “Characterization of ex vivo-generated bovine and human cartilage by immunohistochemical, biochemical, and magnetic resonance imaging analyses.,” NEOMED Bibliography Database, accessed August 3, 2021, https://neomed.omeka.net/items/show/4167.

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