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Publication:
The Chinese Journal of Dental Research

Year 2002
Volume 5 , Issue 2

Back
Pages: 59 - 63

Fibronectin in Adhesion, Spreading, and Proliferation of Mandibular Condylar Cartilage Cells in Cytodex-3 Microcarrier

Yantao Jiao, DDS, PhD/Xuchen Ma, DDS, PhD/Zhenkhang Zhang, DDS/Shifeng Yu, DDS, MS/Manjun Shao, PhD

Objective: To investigate the effect of exogenous fibronectin on mandibular condylar cartilage (MCC) cell attachment and growth on a DEAE-dextran microcarrier. Methods: MCC cells were harvested from newborn New Zealand white rabbits by sequential digestion with trypsin and collagenase. They were grown on the 20 mg/L fibronectin-coated and uncoated cytodex-3 microcarrier. Samples were collected at 1, 3, 5, 7, 9, and 11 days. The kinetics of adhesion and growth were observed with a phase contrast microscope and environmental scanning electronic microscope (ESEM) and quantified by the 0.1% crystal violet nucleus extrusion method. Results: MCC cells could rapidly attach to and spread on the fibronectin-coated cytodex-3; nearly 60% of cells attached to the microcarrier within 2 hours. Spreading cells showed flattening and low refraction when observed under a phase contrast microscope. The attachment and spreading of cells in the untreated group was slow: only 36% of cells attached to the uncoated microcarrier. However, most cells in both groups attached to the microcarrier after 24 hours. The morphology of MCC cells attached to the FN-coated cytodex-3 viewed through an environmental scanning electronic microscope showed them to be flattened and tightly adherent, with more processes (pseudopodia) extending from the cytoplasm. In contrast, cells on the untreated cytodex-3 remained regularly rounded even 24 hours after being plated. Meanwhile, the MCC cells on the fibronectin-treated cytodex-3 showed an accelerated growth rate. The cell density in the FN-treated group was higher than in the untreated group at the end of the culture. Conclusion: The results confirm that fibronectin, being a major glycoprotein of the extracellular matrix, plays an important role in cell attachment and spreading. A better understanding of the cell-extracellular matrix relationship will be helpful in selecting the optimal substrate for cell growth in vitro.

 

 

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