Arthritis & Rheumatism, Volume 62,
November 2010 Abstract Supplement

Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Atlanta, Georgia November 6-11, 2010.


Chondrogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Grown on Type I Collagen and Heparan Sulfate Scaffolds.

Diaz-Prado4,  Silvia, Muinos3,  Emma, Trejo-Iriarte1,  C. G., Lozano2,  D., Garcia-Honduvilla1,  N., Fuentes3,  Isaac, De Toro3,  Francisco J.

Department of Medical Specialties. University of Alcalá de Henares, Madrid, Spain
Laboratory of Bone and Mineral Metabolism. Fundación Jiménez Díaz (Capio Group). Madrid, Spain
Osteoarticular and Aging Res. Lab. CIBER-BBN, Rheumatology Div, INIBIC-Complejo Hosp, Univ. A Coruña, A Coruña, Spain
Osteoarticular and Aging Res. Lab. CIBER-BBN. Rheumatology Div. INIBIC-Complejo Hosp. Univ. A Coruña, Coruña, Spain

Background:

Articular cartilage lesions are not able to repair. To avoid the need for prosthetic replacement different cell treatments were developed with the aim of generating a repaired tissue. The transplantation of mesenchymal stem cells (MSCs) has been suggested as an alternative therapeutic approach for treatment of cartilage defects.

Aim:

Evaluate the chondrogenic potential of bone marrow mesenchymal stem cells (BM-MSCs) grown on type I collagen and different concentrations of heparan sulfate (HS) scaffolds.

Materials and Methods:

BM-MSCs and chondrocytes were cultured on type I collagen and different concentrations of HS scaffolds for 16 and 30 days. BM-MSCs were cultured in chondrogenic medium or DMEM with 20% FBS plus 100 nM PTHrP. Chondrocytes were grown in DMEM with 10% FBS plus 100 nM PTHrP. Chondrogenic differentiation was confirmed by histochemical and immunohistochemical analysis.

Results:

BM-MSCs and chondrocytes were able to proliferate on type I collagen and various concentrations of HS scaffolds, since cells showed high percentages of positivity for PCNA proliferation marker. Increased cell proliferation has been associated with a high rate of scaffold degradation, although the thicker collagen fibers take longer to be degraded. The results indicated that BM-MSCs proliferated better in chondrogenic medium than in the usual growth medium (DMEM 20%). The study groups with BM-MSCs grown in chondrogenic medium +100 nM PTHrP, regardless the HS concentration, showed high percentage of cells regarding the scaffold area (more than 80% after 16 days in culture and more than 90% after 30 days in culture). Moreover, they also showed high percentages of positivity for safranin O (Saf O), aggrecan Agg) and type II collagen (Col II) (Table 1 and Figure 1). Masson's trichrome (Mt) staining showed that cells formed aggregates and produced extracellular matrix. Von Kossa (VKos) stainings, performed in BM-MSCs grown for 30 days in chondrogenic medium, exhibited the presence of calcifications.

Figure 1. Histochemical and immunohistochemistry analysis of BM-MSCs cultured over type 1 collagen and 3% HS scaffolds on chondrogenic medium after 30 days in culture.

Conclusions:

Our data demonstrated that type I collagen and heparan sulfate scaffolds were optimal for BM-MSCs and chondrocyte growth and that BM-MSCs cultured over these scaffolds on chondrogenic medium were able to differentiate to chondrocyte-like cells.

Acknowledgements:

B. Parma; CIBER BBN CB06-01-0040; SDP is beneficiary of IPP contract and stays grant (IN809 A 2007/193, 2008/237and 2009/23) from XUGA, Spain.

To cite this abstract, please use the following information:
Diaz-Prado, Silvia, Muinos, Emma, Trejo-Iriarte, C. G., Lozano, D., Garcia-Honduvilla, N., Fuentes, Isaac, et al; Chondrogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Grown on Type I Collagen and Heparan Sulfate Scaffolds. [abstract]. Arthritis Rheum 2010;62 Suppl 10 :1475
DOI: 10.1002/art.29241

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