Effect of Activated Platelet-Rich Plasma on Chondrogenic Differentiation of Rabbit Bone Marrow-Derived Mesenchymal Stem Cells.
Effect of Activated Platelet-Rich Plasma on Chondrogenic Differentiation of Rabbit Bone Marrow-Derived Mesenchymal Stem Cells.
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活化富血小板血浆对兔骨髓间充质干细胞软骨分化的影响
DOI：
10.1155/2021/9947187
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发表时间：
2021
影响因子：
4.3
通讯作者：
Qi W
中科院分区：
文献类型：
--
作者：
Wang Z;Wang Z;Zhang B;Zhao Q;Liu Y;Qi W
目的探讨活化富血小板血浆（PRP）对骨髓间充质干细胞（BMSCs）增殖及向软骨细胞分化的影响。本研究包括6只成年雄性家兔。从全血中通过两步离心获得PRP，并使用CaCl 2溶液对其进行活化。从兔骨髓中分离、扩增骨髓间充质干细胞，流式细胞仪检测其生物学特性。第3代骨髓基质细胞在高糖Dulbecco改良Eagle培养基（HG-DMEM）中连续培养7天，培养基中含有4种不同的成分，包括10%胎牛血清、5%PRP、10%PRP和15%PRP。采用细胞计数法检测BMSCs的增殖情况。BMSCs（5 × 105细胞/孔，6孔板）在四种条件下诱导21天以进行软骨分化评价，包括商业软骨形成培养基（对照）、5% PRP（HG-DMEM+5% PRP）、10% PRP（HG-DMEM+10% PRP）和15% PRP（HG-DMEM+15% PRP）。检测颗粒中ACAN、COL 2A 1和SOX 9的基因表达水平。由盲态观察者进行形态学和病理学评估。经纯化后，CD 105（+）/CD 34（-）和CD 44（+）/CD 45（-）的细胞百分比分别为96.5%和92.9%。BMSCs的增殖在所有组中均得到增强，从第5天开始，10%PRP显示出比其他组更显著的结果。ACAN、COL 2A 1和SOX 9在3个PRP组均低于对照组，但ACAN和SOX 9在10%PRP组高于5%PRP组和15%PRP组。组织学检查显示，10%PRP处理的颗粒比5%或10%PRP组具有更规则的外观，更大的尺寸和丰富的细胞外基质，但仍不如商业软骨形成培养基。总之，我们的结果表明PRP可以促进兔BMSCs的增殖。然而，与商业软骨形成培养基相比，PRP在颗粒培养中对软骨形成分化的影响有限。
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 × 105 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.
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影响因子：
2.8
作者：
Krueger, Jan Philipp;Hondke, Sylvia;Kaps, Christian
通讯作者：
Kaps, Christian
影响因子：
4
作者：
Park, Yong-Beom;Seo, Sinji;Ha, Chul-Won
通讯作者：
Ha, Chul-Won
影响因子：
4.8
作者：
de Almeida, Adriano Marques;Demange, Marco Kawamura;Hernandez, Arnaldo Jose
通讯作者：
Hernandez, Arnaldo Jose
影响因子：
7
作者：
Long DL;Ulici V;Chubinskaya S;Loeser RF
通讯作者：
Loeser RF
影响因子：
4.6
作者：
Ramezanifard R;Kabiri M;Hanaee Ahvaz H
通讯作者：
Hanaee Ahvaz H