Ideal Concentration of Advanced-Platelet Rich Fibrin (A-PRF) Conditioned Media for Human Dental Pulp Stem Cells Differentiation

Bagio, Dini Asrianti; Julianto, Indah; Suprastiwi, Endang; Margono, Anggraini

doi:10.4034/pboci.2019.191.109

Cited by 5 publications

(10 citation statements)

References 10 publications

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“…In this study, the highest expression of TGF-β1 was found in 25% A-PRF culture media supplement, whereas the lowest was observed at 1% concentration. This is consistent with the results of Bagio et al (2019) who reported that the highest expression of DSPP in the odontogenic differentiation of hDPSCs was obtained at 1% A-PRF and the lowest at a concentration of 25%. The presence of TGF-β1 in the culture media can stimulate the differentiation process, but if the amount is too high, it can be toxic and inhibit the process of differentiation itself [5,13,16,17].…”

Section: Resultssupporting

confidence: 93%

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TRANSFORMING GROWTH FACTOR-β1 EXPRESSION IN VARIOUS CONCENTRATIONS OF ADVANCED PLATELET-RICH FIBRIN MODULATING HUMAN DENTAL PULP STEM CELL DIFFERENTIATION

et al. 2020

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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.

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Section: Resultssupporting

confidence: 93%

“…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].…”

Section: Introductionmentioning

confidence: 99%

TRANSFORMING GROWTH FACTOR-β1 EXPRESSION IN VARIOUS CONCENTRATIONS OF ADVANCED PLATELET-RICH FIBRIN MODULATING HUMAN DENTAL PULP STEM CELL DIFFERENTIATION

et al. 2020

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“…Saaed et al also demonstrated that 10% PRF exudate (PRF-Ex) can replace FBS as CM for hDPSCs and increase their differentiation ability into osteogenic cells [23]. The potential of A-PRF as a culture medium for hDPSCs has also been reported by Bagio et al [24], who showed that 1%-5% A-PRF CM induces the dentin sialophosphoprotein (DSPP) expression of hDPSCs; furthermore, Alizarin red results showed the osteogenic differentiation of hDPSCs after 7 days of culture [24]. These results correlate with the osseointegration relationship in the ability of human MSCs (hMSCs) to express VEGF that exerts signaling with osteoblast differentiation [25].…”

Section: Introductionmentioning

confidence: 74%

Increased VEGF-A Expression of Human Dental Pulp Stem Cells (hDPSCs) Cultured with Advanced Platelet Rich Fibrin (A-PRF)

Bagio¹,

Julianto²,

Margono³

et al. 2021

TODENTJ

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Background: VEGF-A expression of human dental pulp stem cells (hDPSCs) can induce the angiogenesis process of dental pulp regeneration. This in vitro study aimed to analyze the effect of various concentrations of Advanced Platelet Rich Fibrin (A-PRF) conditioned media (CM) on the increased expression of vascular endothelial growth factor-A (VEGF-A) of hDPSCs. Methods: hDPSCs were collected from ten third molars extracted from nine healthy donors, cultured, and then harvested at the end of the third passage. The hDPSCs were seeded in four different CM (control group: hDPSCs + DMEM; 1% A-PRF CM group: hDPSCs + 1% A-PRF CM; 5% A-PRF CM group: hDPSCs + 5% A-PRF CM; 10% A-PRF CM group: hDPSCs + 10% A-PRF CM). All of the groups were cultured in biological triplicates (Triplo) and observed for 5, 12, and 24 hours. The VEGF-A protein expression of hDPSCs was measured using human VEGF-A ELISA at a wavelength of 405 nm. Data was analyzed with Kruskal Wallis and post hoc Mann Whitney test with p<0.05. Results: The VEGF-A expression rate of hDPSCs among all groups was statistically significantly different at 5, 12 and 24 hours of observations (p<0.05). Post hoc analysis test showed a statistically significant difference of hDPSCs’s VEGF-A expression between 5% A-PRF groups compared to other groups at 5 and 12 hours of observation (p<0.05). However, there were no statistically significant differences observed of hDPSCs’ VEGF-A expression at 24 hours of observation between 1%, 5% and 10% A-PRF groups (p>0.05). Conclusion: 5% A-PRF CM was superior in increasing VEGF-A expression of hDPSCs at 5, 12 and 24 hours of observations.

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“…Bagio et al proved that the ideal concentration of A-PRF, which have great potential ability of human DPCs' odontogenic differentiation, is at 1 to 5%. 24 From this result, various concentrations of A-PRF as well as L-PRF, as allogenic platelet concentrations, which are platelet-rich blood derivatives, have been widely used in stem-cell-based therapy as CM showed a great potential of stem cells' migration.…”

Section: Discussionmentioning

confidence: 99%

Changes in Migratory Speed Rate of Human Dental Pulp Stromal Cells Cultured in Advanced Platelet-Rich Fibrin

et al. 2022

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Objective Migratory speed rate evaluation of human dental pulp stromal cells (hDP-SCs) is one of the important steps in dental pulp regeneration. Therefore, the aim of the study is to analyze various concentrations of advanced platelet-rich fibrin (A-PRF) culture media toward hDP-SCs' migratory speed rate evaluations. Materials and Methods The hDP-SCs were divided into four groups: control: hDP-SCs in Dulbecco's modified Eagle medium + 10% fetal bovine serum group; hDP-SCs in 1% A-PRF group; hDP-SCs in 5% A-PRF group; and hDP-SCs in 10% A-PRF group, which were planted in 24-well (5 × 104 cell/well). The migratory speed rate of all groups was measured by using cell migration assay (scratch wound assay) after 24 hours. Cell characteristics were evaluated under microscope (Inverted microscope, Zeiss, Observer Z1, UK) that can be read through image-J interpretation. This image J represented the measurement of migratory speed rate (nm/h) data. Statistical analysis was conducted using one-way analysis of variance and post hoc Tamhane's test (p < 0.05) (IBM SPSS Statistics Software, version 22.0). Results There was a statistically significant difference in the migratory speed rates of hDP-SCs among various concentration groups of A-PRF (1, 5, and 10%) compared with the control group. Conclusion The increase in the migratory speed rate of hDP-SCs was highest in 10% A-PRF group.

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Ideal Concentration of Advanced-Platelet Rich Fibrin (A-PRF) Conditioned Media for Human Dental Pulp Stem Cells Differentiation

Cited by 5 publications

References 10 publications

TRANSFORMING GROWTH FACTOR-β1 EXPRESSION IN VARIOUS CONCENTRATIONS OF ADVANCED PLATELET-RICH FIBRIN MODULATING HUMAN DENTAL PULP STEM CELL DIFFERENTIATION

TRANSFORMING GROWTH FACTOR-β1 EXPRESSION IN VARIOUS CONCENTRATIONS OF ADVANCED PLATELET-RICH FIBRIN MODULATING HUMAN DENTAL PULP STEM CELL DIFFERENTIATION

Increased VEGF-A Expression of Human Dental Pulp Stem Cells (hDPSCs) Cultured with Advanced Platelet Rich Fibrin (A-PRF)

Changes in Migratory Speed Rate of Human Dental Pulp Stromal Cells Cultured in Advanced Platelet-Rich Fibrin

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