J Korean Orthop Res Soc.  2004 Oct;7(2):159-168.

Effect of Recombinant Fragment of Fibronectin on the Cellular Functions of Human Bone Marrow-Derived Mesenchymal Stem Cells

• Yeo BK
• Yoo JJ
• Lim JH
• Lee JE
• Jang JH
• Shin CS
• Chung MS
• Kim HJ

Abstract

PURPOSE
We investigated the effects of recombinant 9-10th type III repeat of fibronectin (rhFNIII9-10) on the adhesion, proliferation, and the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells(hMSCs).
MATERIALS AND METHODS
Adhesion and blocking assay for hMSCs were performed on the plates which had been coated with 100 microgram/ml rhFNIII9-10 or fibronectin. hMSCs seeded on the precoated plates were cultured in the osteogenic media for 3 weeks. MTS(Dimethylthiazole carboxymethoxyphenyl sulfophenyl tetrazolium compound) assay for the cell number, [Methyl-3H] thymidine incorporation study, alkaline phosphatase activity assay, calcium content assay and RT-PCR for alkaline phosphatase, osteopontin, cbfa-1, and type I collagen were performed during the osteogenic differentiation.
RESULTS
hMSCs showed significantly increased adhesion to rhFNIII9-10-coated plates and fibronectin-coated plates. A monoclonal antibody to the integrin alpha 5 beta 1 inhibited adhesion to rhFNIII9-10-coated plates and fibronectin-coated plates in dose-dependent manner. hMSCs seeded on the rhFNIII9-10-coated plates showed increased proliferation during the osteogenic differentiation. However, there was no significant difference in the alkaline phosphatase activity, calcium content and expression levels of mRNAs for alkaline phosphatase, osteopontin, cbfa-1, and type I collagen of hMSCs seeded on the rhFNIII9-10-coated plates.
CONCLUSION
rhFNIII9-10 stimulates hMSCs adhesion and increases hMSCs proliferation during the osteogenic differentiation. Although osteogenic differentiation is not promoted, adsorption of rhFNIII9-10 onto appropriate biomaterials can enhance integrin-mediated hMSCs adhesion and proliferation. This biomolecular engineering strategy represents a robust approach to increase biofunctional activity and integrin specificity of hMSCs.

Keyword

Fibronectin; Recombinant peptide; Integrin alpha 5 beta 1; Mesenchymal stem cell; Osteogenic differen-tiation; Bone marrow

MeSH Terms

Adsorption
Alkaline Phosphatase
Biocompatible Materials
Bone Marrow
Calcium
Cell Count
Collagen Type I
Fibronectins*
Humans*
Integrin alpha5beta1
Mesenchymal Stromal Cells*
Osteopontin
RNA, Messenger
Sensitivity and Specificity
Thymidine
Alkaline Phosphatase
Biocompatible Materials
Calcium
Collagen Type I
Fibronectins
Integrin alpha5beta1
Osteopontin
RNA, Messenger
Thymidine