Characterization of ex vivo-generated bovine and human cartilage by immunohistochemical, biochemical, and magnetic resonance imaging analyses.
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
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.
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
Tissue engineering. Part A
2010
2010-07
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1089/ten.TEA.2009.0717" target="_blank" rel="noreferrer noopener">10.1089/ten.TEA.2009.0717</a>
Development of bone and cartilage in tissue-engineered human middle phalanx models.
*Models; Aggrecans/genetics/metabolism; Animals; Biological; Bone Development/drug effects/*physiology; Calcium Phosphates/pharmacology; Cartilage/cytology/drug effects/*growth & development; Cattle; Chondrocytes/cytology/drug effects/metabolism; Collagen Type II/genetics/metabolism; Durapatite/pharmacology; Electron; Experimental; Finger Phalanges/cytology/diagnostic imaging/drug effects/*physiology; Gene Expression Profiling; Gene Expression Regulation/drug effects; Humans; Implants; Integrin-Binding Sialoprotein; Mice; Microscopy; Paraffin Embedding; Periosteum/cytology/drug effects; Polyesters/pharmacology; Radiography; Scanning; Sialoglycoproteins/genetics/metabolism; Tissue Engineering/*methods; Tissue Scaffolds/chemistry
Human middle phalanges were tissue-engineered with midshaft scaffolds of poly(L-lactide-epsilon-caprolactone) [P(LA-CL)], hydroxyapatite-P(LA-CL), or beta-tricalcium phosphate-P(LA-CL) and end plate scaffolds of bovine chondrocyte-seeded polyglycolic acid. Midshafts were either wrapped with bovine periosteum or left uncovered. Constructs implanted in nude mice for up to 20 weeks were examined for cartilage and bone development as well as gene expression and protein secretion, which are important in extracellular matrix (ECM) formation and mineralization. Harvested 10- and 20-week constructs without periosteum maintained end plate cartilage but no growth plate formation. They also consisted of chondrocytes secreting type II collagen and proteoglycan, and they were composed of midshaft regions devoid of bone. In all periosteum-wrapped constructs at like times, end plate scaffolds held chondrocytes elaborating type II collagen and proteoglycan and cartilage growth plates resembling normal tissue. Chondrocyte gene expression of type II collagen, aggrecan, and bone sialoprotein varied depending on midshaft composition, presence of periosteum, and length of implantation time. Periosteum produced additional cells, ECM, and mineral formation within the different midshaft scaffolds. Periosteum thus induces midshaft development and mediates chondrocyte gene expression and growth plate formation in cartilage regions of phalanges. This work is important for understanding developmental principles of tissue-engineered phalanges and by extension those of normal growth plate cartilage and bone.
Wada Yoshitaka; Enjo Mitsuhiro; Isogai Noritaka; Jacquet Robin; Lowder Elizabeth; Landis William J
Tissue engineering. Part A
2009
2009-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1089/ten.TEA.2009.0078" target="_blank" rel="noreferrer noopener">10.1089/ten.TEA.2009.0078</a>