scholarly journals Effect of Activated Platelet-Rich Plasma on Chondrogenic Differentiation of Rabbit Bone Marrow-Derived Mesenchymal Stem Cells

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Zhen Wang ◽  
Zheng Wang ◽  
Bin Zhang ◽  
Qinghua Zhao ◽  
Yubao Liu ◽  
...  

We aimed to evaluate the effect of activated platelet-rich plasma (PRP) on proliferation and chondrogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). Six mature male rabbits were included in this study. PRP was obtained by two-step centrifugation from whole blood, and it was activated using CaCl2 solution. BMSCs were isolated and proliferated from bone marrow of rabbits and characterized by flow cytometry. Passage 3 BMSCs were cultured in high-glucose Dulbecco’s modified Eagle’s medium (HG-DMEM) with the four different compositions for consecutive 7 days, including 10% fetal bovine serum, 5% PRP, 10% PRP, and 15% PRP. Cell counting assays were performed to evaluate the cell proliferation of BMSCs. BMSCs ( 5 × 10 5 cells/well in 6-well plates) were induced in four conditions for 21 days to chondrogenic differentiation evaluation, including commercial chondrogenic medium (control), 5% PRP (HG-DMEM+5% PRP), 10% PRP (HG-DMEM+10% PRP), and 15% PRP (HG-DMEM+15% PRP). The gene expression levels of ACAN, COL2A1, and SOX9 in pellets were detected. Morphological and pathological assessments were performed by the blind observer. After purifying, the percentages of cells with CD105(+)/CD34(−) and CD44(+)/CD45(−) were 96.5% and 92.9%, respectively. The proliferation of BMSCs was enhanced in all groups, and 10% PRP revealed more significant outcome than the others from day 5. The levels of ACAN, COL2A1, and SOX9 were lower in the three PRP groups than control group, but the levels of ACAN and SOX9 were higher in 10% PRP group than 5% and 15% PRP groups. Histological examinations showed that 10% PRP-treated pellets had more regular appearance, larger size, and abundant extracellular matrix than 5% or 10% PRP groups, but still inferior to commercial chondrogenic medium. In conclusion, our results show that PRP may enhance the proliferation of rabbit BMSCs. However, PRP have limited effect on chondrogenic differentiation in comparison with commercial chondrogenic medium in pellets culture.

2021 ◽  
Author(s):  
Zhen Wang ◽  
Zheng Wang ◽  
Bin Zhang ◽  
Qinghua Zhao ◽  
Yubao Liu ◽  
...  

Abstract Background Platelet-rich plasma (PRP) has revealed benefits in tissue repair and regeneration, however, the effect of PRP on proliferation and chondrogenesis of mesenchymal stem cells remains controversial.This study is to evaluate the effect of different concentration PRP on proliferation and chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells(BMSCs). Methods PRP was obtained by centrifugation and activation, in which growth factors and cytokines were detected. BMSCs were isolated from rabbit bone marrow and characterized by flow cytometry. About 5 × 103 BMSCs were cultured in high glucose Dulbecco’s modified Eagle’s medium (HG-DMEM) with 4 different compositions: 10% fetal bovine serum, 5%PRP, 10%PRP, and 15%PRP for consecutive 7 days. Cell counting assays were performed on days 1, 3, 5, and 7 to evaluate the BMSCs proliferation. In chondrogenic differentiation, high-density cell pellets composed of 5 × 105 BMSCs were induced in 4 conditions: commercial chondrogenic medium (control), 5%PRP (HG-DMEM + 5%PRP), 10%PRP (HG-DMEM + 10%PRP), and 15%PRP (HG-DMEM + 15%PRP) for 21 days. The gene expression levels of aggrecan (ACAN), collagen type Ⅱ Alpha 1 (COL2A1), and SRY-Box Transcription Factor 9 (SOX9) in pellets were detected. Histological assessments were performed by morphologically observation and pathological stain. Independent-samples t-test and one-way analysis of variance were used in statistical analyses. Results The concentrations of growth factors and cytokines were elevated in PRP. BMSCs proliferation was enhanced in all groups, and 10% PRP revealed more obvious outcome than the others from day 5. In chondrogenic differentiation, the levels of ACAN, COL2A1, and SOX9 were lower in 3 PRP groups than control, but ACAN and SOX9 was higher in 10% PRP group than 5% and 15%. Histological examinations showed 10% PRP-treated pellets showed more regular appearance, larger size, and abundant extracellular matrix than 5% and 10% groups, but still inferior to commercial chondrogenic medium. Conclusions PRP may enhance the proliferation of rabbit BMSCs, while with limited effect on chondrogenic differentiation compared with commercial chondrogenic medium in pellets culture. Whether optimizing and modifying PRP components would lead to satisfying chondrogenesis of BMSCs, it is deserved furthermore study.


2021 ◽  
Vol 2021 ◽  
pp. 1-7
Author(s):  
Yi Dong ◽  
Long Chang ◽  
Long Hei ◽  
Sensen Yang ◽  
Wenxin Ma ◽  
...  

This study aims to evaluate the effect of peroxisome proliferator-activated receptor (PPAR) γ gene inhibition on the adipogenic differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs). Primary BMSCs were isolated from rabbit bone marrow, cultured, and the markers of BMSCs on cell’s surface were analyzed using flow cytometry. The experiment involved five groups, namely, control: untreated BMSCs; model: BMSCs treated with ethanol; empty siRNA: BMSCs treated with ethanol + empty siRNA; PPARγ: BMSCs treated with ethanol + PPARγ siRNA; and PPARγ inhibitor: BMSCs treated with ethanol + T0070907. RT-PCR and Western blotting were used to detect changes in the expression level of PPARγ, PETALA2 (AP2), lipoprotein lipase (LPL), fatty acid transport protein (FATP) 1, and fatty acid transporter (FAT). Adipocyte count and triacylglycerol content of the model and the empty siRNA groups were considerably greater than the control group ( P < 0.01 ). After the inhibition with PPARγ or T0070907, adipocyte count and triacylglycerol content of the PPARγ and T0070907 groups were significantly reduced ( P < 0.01 ), with no statistically significantly difference than the control group ( P > 0.05 ). The expression levels of PPARγ gene and protein in the model and empty siRNA groups were ominously enhanced than the control group ( P < 0.01 ), and after inhibition with PPARγ or T0070907, the PPARγ gene or protein expression level of PPARγ and T0070907 groups significantly reduced ( P < 0.01 ), with no statistically significance difference compared to the control group ( P > 0.05 ). The expression levels of Ap2, LPL, FATP1, and FAT genes in the model and empty siRNA groups were considerably greater compared to the control group ( P < 0.01 ). Inhibition with PPARγ or T0070907 in the PPARγ and T0070907 groups, respectively, lead to significantly reduced expression levels of adipogenic genes ( P < 0.01 ), with no statistically significance difference compared to the control ( P > 0.05 ). Inhibition of PPARγ gene downregulates the differentiation of BMSCs into adipocytes, indicating its putative role in the expression of adipogenic genes.


2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Basma Elsaadany ◽  
Samar El Kholy ◽  
Dalia El Rouby ◽  
Laila Rashed ◽  
Tarek Shouman

Normal tissue damage following radiotherapy is still a major problem in cancer treatment. Therefore, the current work aimed at exploring the possible role of systemically injected bone marrow derived mesenchymal stem cells (BM-MSCs) and/or locally injected platelet rich plasma (PRP) in ameliorating the side effects of ionizing radiation on the rat’s tongue. Twelve rats served as control group (N) and 48 rats received a single radiation dose of 13 Gy to the head and neck region; then, they were equally divided into 4 experimental groups: irradiated only (C), irradiated + MSCs (S), irradiated + (PRP) (P), and combined group (PS). Animal scarification occurred in 3 and 7 days after radiation. Then, tongues were dissected and examined histologically and for expression of bcl-2 by RT-PCR. Histological examination of the treated groups (S), (P), and (PS) revealed an obvious improvement in the histological structure of the tongue, compared to group (C), in addition to upregulated expression of bcl-2, indicating decreased apoptotic activity.Conclusion. BM-MSCs and PRP have shown positive effect in minimizing the epithelial atrophy of normal oral mucosa after regional radiotherapy, which was emphasized by decreasing apoptotic activity in these tissues. Nevertheless, combined use of BM-MSCs and PRP did not reveal the assumed synergetic effect in oral tissue protection.


2018 ◽  
Vol 24 (19-20) ◽  
pp. 1432-1443 ◽  
Author(s):  
Jr-Jiun Liou ◽  
Benjamin B. Rothrauff ◽  
Peter G. Alexander ◽  
Rocky S. Tuan

2021 ◽  
Vol 22 (13) ◽  
pp. 7058
Author(s):  
Thorsten Kirsch ◽  
Fenglin Zhang ◽  
Olivia Braender-Carr ◽  
Mary K. Cowman

Mesenchymal stem cells (MSCs) obtained from various sources, including bone marrow, have been proposed as a therapeutic strategy for the improvement of tissue repair/regeneration, including the repair of cartilage defects or lesions. Often the highly inflammatory environment after injury or during diseases, however, greatly diminishes the therapeutic and reparative effectiveness of MSCs. Therefore, the identification of novel factors that can protect MSCs against an inflammatory environment may enhance the effectiveness of these cells in repairing tissues, such as articular cartilage. In this study, we investigated whether a peptide (P15-1) that binds to hyaluronan (HA), a major component of the extracellular matrix of cartilage, protects bone-marrow-derived MSCs (BMSCs) in an inflammatory environment. The results showed that P15-1 reduced the mRNA levels of catabolic and inflammatory markers in interleukin-1beta (IL-1β)-treated human BMSCs. In addition, P15-1 enhanced the attachment of BMSCs to HA-coated tissue culture dishes and stimulated the chondrogenic differentiation of the multipotential murine C3H/10T1/2 MSC line in a micromass culture. In conclusion, our findings suggest that P15-1 may increase the capacity of BMSCs to repair cartilage via the protection of these cells in an inflammatory environment and the stimulation of their attachment to an HA-containing matrix and chondrogenic differentiation.


Sign in / Sign up

Export Citation Format

Share Document