Abstract
Objective: Modification of the speed and time of centrifugation based on the low-speed centrifugation concept for platelet-rich fibrin (PRF) has resulted in a new type of PRF known as advanced PRF (A-PRF). A-PRF can release several types of growth factors (GFs) that participate in the process of differentiation, such as transforming GF-β1 (TGF-β1). The aim of this study was to analyze TGF-β1 expression in various concentrations of A-PRF in the differentiation process of human dental pulp stem cells (hDPSCs).Methods: hDPSC cultures were obtained from those of previous research (ethical approval form has been attached). These hDPSCs were in the 2nd–3rd passage, and serum starvation was done by reducing fetal bovine serum (FBS) levels in the hDPSC culture media. A-PRF was obtained using 10 ml blood collected from the cubital vein, which was centrifuged at 1500 rpm for 14 min and then divided into four concentration groups. TGF-β1 expression in 1%, 5%, and 25% A-PRF as well as in 10% FBS (control) was analyzed by ELISA on day 7.Results: Although no significant differences were observed in TGF-β1 expression between 1%, 5%, and 25% A-PRF, and 10% FBS, it was observed that the higher the concentration of A-PRF, the greater the TGF-β1 expression.Conclusion: The expression of TGF-β1 was consistent with the increase in A-PRF concentration. The highest TGF-β1 expression was detected in 25% A-PRF among all concentrations in the differentiation process of hDPSCs.
Highlights
In recent years, regenerative endodontics has presented new possibilities for the treatment of necrotic immature permanent teeth based on the combination and interplay of three key elements required for tissue regeneration, namely, stem cells, bioactive molecules, and scaffolds [1]
Human dental pulp stem cell cultures were obtained from those of previous research. These human dental pulp stem cells (hDPSCs) were in the 2nd–3rd passage before the serum starvation procedure, and serum starvation was done by reducing fetal bovine serum (FBS) levels till 1% concentration. hDPSCs were divided into four culture media groups; A-Platelet-rich fibrin (PRF) 1%, 5%, and 25%, and FBS 10% after 24 h of serum starvation, with each group containing three biological replicates
It was observed that cells after 24 h of starvation consisted of only a monolayer compared with other images after 7 days of advanced PRF (A-PRF) supplementation in the growth media, with the hDSPCs being developed into a multilayer
Summary
Regenerative endodontics has presented new possibilities for the treatment of necrotic immature permanent teeth based on the combination and interplay of three key elements required for tissue regeneration, namely, stem cells, bioactive molecules, and scaffolds [1]. The traditional triad of dental stem cells, bioactive signal molecules, and bioactive scaffolds as mentioned above fails to generate a healthy functional tissue. This is due to the fact that regenerated tissue does not completely match the normal healthy functional tissue. A modification of the preparation setting based on the previously described low-speed centrifugation concept (LSCC) is a first step in the reduction of the applied relative centrifugal force (RCF). This step is accompanied by a slight increase in centrifugation time, resulting in the so-called advanced PRF (A-PRF) [4]. A-PRF produces the maximum GF levels compared with PRP [5]
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