Three-dimensional cartilage tissue engineering using adult stem cells from osteoarthritis patients.
OBJECTIVE
To determine whether it is possible to engineer 3-dimensional hyaline cartilage using mesenchymal stem cells derived from the bone marrow (BMSCs) of patients with osteoarthritis (OA).


METHODS
Expanded BMSCs derived from patients with hip OA were seeded onto polyglycolic acid scaffolds and differentiated using transforming growth factor beta3 in the presence or absence of parathyroid hormone-related protein (PTHrP) to regulate hypertrophy. Micromass pellet cultures were established using the same cells for comparison. At the end of culture, the constructs or pellets were processed for messenger RNA (mRNA) analysis by quantitative real-time reverse transcription-polymerase chain reaction. Matrix proteins were analyzed using specific assays.


RESULTS
Cartilage constructs engineered from BMSCs were at least 5 times the weight of equivalent pellet cultures. Histologic, mRNA, and biochemical analyses of the constructs showed extensive synthesis of proteoglycan and type II collagen but only low levels of type I collagen. The protein content was almost identical to that of cartilage engineered from bovine nasal chondrocytes. Analysis of type X collagen mRNA revealed a high level of mRNA in chondrogenic constructs compared with that in undifferentiated BMSCs, indicating an increased risk of hypertrophy in the tissue-engineered cells. However, the inclusion of PTHrP at a dose of 1 microM or 10 microM during the culture period resulted in significant suppression of type X collagen mRNA expression and a significant decrease in alkaline phosphatase activity, without any loss of the cartilage-specific matrix proteins.


CONCLUSIONS
Three-dimensional hyaline cartilage can be engineered using BMSCs from patients with OA. This method could thus be used for the repair of cartilage lesions.
Kafienah
Wael
W
Southmead Hospital, and University of Bristol, Bristol, UK.
Mistry
Sanjay
S
Dickinson
Sally C
SC
Sims
Trevor J
TJ
Learmonth
Ian
I
Hollander
Anthony P
AP
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D002417:Cattle; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D051457:Hyaline Cartilage / Q000166:cytology* / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D008875:Middle Aged; D015207:Osteoarthritis, Hip / Q000473:pathology*; D044162:Parathyroid Hormone-Related Protein / Q000494:pharmacology*; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D015291:Transforming Growth Factors / Q000494:pharmacology*
D024042:Collagen Type I; D024043:Collagen Type II; D004338:Drug Combinations; D044162:Parathyroid Hormone-Related Protein; D011509:Proteoglycans; D012333:RNA, Messenger; D015291:Transforming Growth Factors