Abstract
Objective: Ex vivo and in vitro cell cultures require a basal medium with added supplements containing growth factors, proteins, and enzymes tosupport attachment, growth, and proliferation. Fetal bovine serum (FBS) is used to supplement cell culture media. However, human platelet lysate(hPL) represents an attractive alternative as it is nonxenogeneic.Methods: Human third molars were collected from six healthy donors (19–35 years old) with no history of regular alcohol consumption or smoking.Human dental pulp stem cells (hDPSCs) at the second passage were divided into two culture media groups, 10% FBS and 5% hPL, as well as a controlgroup after 24 h of serum starvation. A flow cytometry analysis was conducted to measure CD90, CD105, CD73, CD34, CD45, and Human LeukocyteAntigen-DR isotype (HLA-DR). Cellular proliferation was evaluated on days 1, 3, and 5.Results: The flow cytometry analysis revealed that the majority of the cells expressed positive mesenchymal stem cell surface markers, includingCD73 (98.5%), CD90 (98.3%), and CD105 (71.0%), and lacked CD34, CD45, and HLA-DR. There were significant differences among the 5% hPL, 10%FBS, and control groups on days 1, 3, and 5.Conclusion: For a nonxenogeneic culture, 5% hPL can be used as an alternative in culture media for hDPSC proliferation.
Highlights
Immature teeth are at risk for developing pulpal necrosis due to trauma, caries, and anatomical variations[1]
Flow cytometry revealed that the majority of cells expressed Human dental pulp stem cells (hDPSCs) positive surface markers, including CD73 (98.5%), CD90 (98.3%), and CD105 (71.0%), and were negative for CD34, CD45, and Human Leukocyte Antigen-DR isotype (HLA-DR) Fig. 1
The result of statistical analysis hDPSCs proliferation consists of three different growth media groups; the control group was cells after 24 h of serum starvation with 1% Fetal bovine serum (FBS), 10% FBS group, and 5% human platelet lysate (hPL) group
Summary
Immature teeth are at risk for developing pulpal necrosis due to trauma, caries, and anatomical variations[1]. Routine root canal treatment protocols to clean and obturate these teeth cannot be adequately performed on immature roots [2]. Regenerative endodontic procedures are intended to promote tooth survival and function [1]. Nakashima described the three essential components of tissue engineering: Stem cells, bioactive growth factors, and biomimetic three-dimensional scaffolds [3,4,5]. These three components play an essential roles in the restoration of previously damaged structures [4]
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