J Korean Orthop Res Soc.  2001 Oct;4(2):150-158.

The effect of proliferation and phenotype by ascorbic acid on human articular chondrocytes culture

• Lee HJ
• Woo JI
• Park SR
• Min BH

Abstract

For the successful autologous chondrocyte transplantation, it is important to maximize the number of chondrocyte and maintain its original morphology and phenotypic change of the chondrocyte in the culture. In this study, the effect of ascorbic acid and human serum which are known to promote cell proliferation and collagen synthesis was observed in the culture of human chondrocyte. Media were prepared with the conditions of fetal bovine Serum(FBS) treated group, FBS +ascorbic acid(asc) treated group, human serum(HuS) treated group, and HuS+asc treated group, respectively. Proliferation was measured by cell counting using trypan-blue staining method. We used to determine the degree of expression of aggrecan of mRNA and type II collagen using RT-PCR. Type II collagen in cultured cell and medium was measured by western blot analysis and proteoglycan synthesis by DMB (Dimethylene Blue) assay. Under all conditions, aggrecan on mRNA level was well expressed. On the other hand, expression of type II collagen was reduced on HuS treated group than FBS treated group, and ascorbic acid treated groups showed decreased expression of type II collagen. Western blot analysis showed increased expression of type II collagen on HuS treated group than FBS treated group, and ascorbic acid treated groups showed increased level. HuS+asc treated group showed the most significant effect than the other groups. The increased effects of ascorbic acid on the proliferation and collagen synthesis were more prominent in the culture with human serum. It might be due to the synergic effect with some growth factors which were present in human serum.

Keyword

Human articular chondrocyte; Ascorbic acid; Human serum; Proliferation; Type II collagen; proteoglycan

MeSH Terms

Aggrecans
Ascorbic Acid*
Blotting, Western
Cell Count
Cell Proliferation
Cells, Cultured
Chondrocytes*
Collagen
Collagen Type II
Hand
Humans*
Intercellular Signaling Peptides and Proteins
Phenotype*
Proteoglycans
RNA, Messenger
Aggrecans
Ascorbic Acid
Collagen
Collagen Type II
Intercellular Signaling Peptides and Proteins
Proteoglycans
RNA, Messenger