猪骨髓来源富血小板血浆对骨髓间充质干细胞和内皮祖细胞的影响。
Effects of porcine bone marrow-derived platelet-rich plasma on bone marrow-derived mesenchymal stem cells and endothelial progenitor cells.
机构信息
Department of Pediatric Dentistry, Ninth People's Hospital, Shanghai Jiaotong University, School of Medicine, Shanghai, 200011, China; Visiting Scholar, Division of Orthodontics, College of Dentistry, The Ohio State University, Columbus, OH, USA.
Division of Biosciences, College of Dentistry, The Ohio State University, Columbus, OH, USA.
出版信息
Tissue Cell. 2021 Aug;71:101587. doi: 10.1016/j.tice.2021.101587. Epub 2021 Jun 29.
This study investigated the abundance of pro-regenerative growth factors in bone marrow-derived platelet-rich plasma (BM-PRP) and their effects on bone marrow-derived mesenchymal stem cells (BM-MSC) and bone marrow-derived endothelial progenitor cells (BM-EPC). Four 4-5 months-old domestic pigs were included, and each underwent bone marrow aspiration from its humerus bones and processed into bone marrow aspiration concentrate (BMAC) samples. The plasma and cellular portions of BMAC were subsequently separated and collected. The concentration of growth factors including BMP-2, PDGF-BB, TGF-β1 and VEGF in the plasma portion was measured and compared between BM-PRP and bone marrow-derived platelet-poor plasma (BM-PPP). It was found that platelet count was significantly higher in BM-PRP than in BM-PPP, but the concentration of above-mentioned growth factors was not significantly different between BM-PRP and BM-PPP. As most existing literature has indicated the regenerative potency of PRP, this study focused on assessing the effect of BM-PRP treatment on BM-MSC and BM-EPC proliferation, osteogenic differentiation and angiogenesis capacity by comparing samples with 2.5% BM-PRP treatment and samples without BM-PRP treatment (control). In response to BM-PRP treatment, the cellular doubling time increased with culturing time and was significantly shorter in the BM-PRP-treated samples than in control samples. For osteogenic differentiation, BM-PRP-treated BM-MSCs demonstrated a time-dependent increase in alkaline phosphatase (ALP) activity and expression levels of osteogenic differentiation markers. For the expression of angiogenic genes, none of the differences reached statistical significance despite a tendency of stronger expression at day 18 in BM-PRP-treated BM-EPCs. In conclusion, this in vitro study suggests that most BMP-2, PDGF-BB, TGF-β1 and VEGF-A contained in BM-PRP are not platelet-released and BM-PRP may have some stimulation (less than 1-fold) for MSC, EPC proliferation and MSC osteogenic differentiation.
本研究旨在探讨骨髓来源富血小板血浆（BM-PRP）中促再生生长因子的丰度及其对骨髓间充质干细胞（BM-MSC）和骨髓来源内皮祖细胞（BM-EPC）的影响。纳入 4 只 4-5 月龄的国产猪，分别从其肱骨骨髓抽吸并制成骨髓抽吸浓缩物（BMAC）样本。随后分离和收集 BMAC 的血浆和细胞部分。测量并比较血浆部分中包括 BMP-2、PDGF-BB、TGF-β1 和 VEGF 在内的生长因子浓度，以比较 BM-PRP 和骨髓来源血小板贫浆（BM-PPP）之间的差异。结果发现，BM-PRP 中的血小板计数明显高于 BM-PPP，但两种制剂中上述生长因子的浓度无显著差异。鉴于大多数现有文献均表明 PRP 具有再生潜能，因此本研究重点评估了 2.5%BM-PRP 处理对 BM-MSC 和 BM-EPC 增殖、成骨分化和血管生成能力的影响，并将接受 BM-PRP 处理的样本与未接受 BM-PRP 处理的样本（对照组）进行了比较。结果显示，随着培养时间的延长，细胞倍增时间增加，接受 BM-PRP 处理的样本明显短于对照组。对于成骨分化，接受 BM-PRP 处理的 BM-MSCs 碱性磷酸酶（ALP）活性和成骨分化标志物表达水平呈时间依赖性增加。对于血管生成基因的表达，尽管接受 BM-PRP 处理的 BM-EPCs 在第 18 天的表达趋势较强，但差异均无统计学意义。总之，本体外研究提示，BM-PRP 中大多数 BMP-2、PDGF-BB、TGF-β1 和 VEGF-A 并非血小板释放，且 BM-PRP 可能对 MSC、EPC 增殖和 MSC 成骨分化具有一定的刺激作用（<1 倍）。
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