Effect of hyaluronic acid on proliferative activity, differentiation potential and expression of phenotypic traits of dental pulp stem cells
Abstract
Introduction: The aim of the study is to verify the dental pulp stem cells reaction to the presence of hyaluronic acid in the culture medium and to assess the possible influence of hyaluronic acid scaffolds on dental pulp stem cells. Methods: Two dental pulp stem cells lines were used in the experiment.
The cells were cultured in standard culture medium enriched with 0.1% hyaluronic acid at three different molar masses: medium 1:0.116 MDa, medium 2:0.54 MDa, medium 3:1.5 MDa. We used the Vi-Cell XR and Z2 Counter to determine the cell parameters and cell viability.
Results: We proved a phenotype typical for DPSC (high positivity for CD13, CD29, CD44, CD90 and OCT3/4), ability to exceed Hayflick limit and differentiation in osteogenic and chondrogenic tissues during long-term cultivation. During the experiment DPSC line 1 achieved in order control/ medium 1/medium 2/medium 3 in total 14.1/15.3/15.4/14.8 population doublings, the average doubling time with a standard deviation in the same order was 32.4+-3.0/29.1+-0.9/29.6+-1.8/30.9+-1.7 hours.
The viability of the DPSC obtained from the 7th passage was 92.3/93.1/91.8/92.6%. DPSC line 2 achieved in order control/ medium 1/ medium 2/ medium 3 in total 16.7/17.2/16.7/16.8 population doublings, the average doubling time with a standard deviation in the same order was 29,1+-4.1/28.0+-3.1/29.4+-5.5/29.0+-3.9 hours.
The viability of the dental pulp stem cells obtained from the 7th passage was 80.1/82.5/81.8/80.9%. Conclusion We verified that dental pulp stem cells in the presence of hyaluronic acid of molecular weights of 0.116, 0.54 and 1.5 MDa survive, proliferate and maintain the ability to differentiate.
We also verified the original assumption that the low molecular weight form of hyaluronic acid have a greater impact on the dental pulp stem cells phenotype than the high molecular weight form of hyaluronic acid. We demonstrated that hyaluronic acid is usable in tissue engineering as a material for scaffolding.