Roffi Alice, Filardo Giuseppe, Assirelli Elisa, Cavallo Carola, Cenacchi Annarita, Facchini Andrea, Grigolo Brunella, Kon Elizaveta, Mariani Erminia, Pratelli Loredana, Pulsatelli Lia, Marcacci Maurilio
Nano-Biotechnology Laboratory, Rizzoli Orthopaedic Institute, Via di Barbiano 1, 40136 Bologna, Italy.
II Clinic-Biomechanics Laboratory and Nano-Biotechnology Laboratory, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
Biomed Res Int. 2014;2014:692913. doi: 10.1155/2014/692913. Epub 2014 Jul 17.
PRP cryopreservation remains a controversial point. Our purpose was to investigate the effect of freezing/thawing on PRP molecule release, and its effects on the metabolism of chondrocytes and synoviocytes. PRP was prepared from 10 volunteers, and a half volume underwent one freezing/thawing cycle. IL-1β, HGF, PDGF AB/BB, TGF-β1, and VEGF were assayed 1 hour and 7 days after activation. Culture media of chondrocytes and synoviocytes were supplemented with fresh or frozen PRP, and, at 7 days, proliferation, gene expression, and secreted proteins levels were evaluated. Results showed that in the freeze-thawed PRP the immediate and delayed molecule releases were similar or slightly lower than those in fresh PRP. TGF-β1 and PDGF AB/BB concentrations were significantly reduced after freezing both at 1 hour and at 7 days, whereas HGF concentration was significantly lower in frozen PRP at 7 days. In fresh PRP IL-1β and HGF concentrations underwent a significant further increase after 7 days. Similar gene expression was found in chondrocytes cultured with both PRPs, whereas in synoviocytes HGF gene expression was higher in frozen PRP. PRP cryopreservation is a safe procedure, which sufficiently preserves PRP quality and its ability to induce proliferation and the production of ECM components in chondrocytes and synoviocytes.
富血小板血浆(PRP)的冷冻保存仍然是一个有争议的问题。我们的目的是研究冻融对PRP分子释放的影响,及其对软骨细胞和滑膜细胞代谢的影响。从10名志愿者中制备PRP,其中一半体积经历一个冻融循环。在激活后1小时和7天检测白细胞介素-1β(IL-1β)、肝细胞生长因子(HGF)、血小板衍生生长因子AB/BB(PDGF AB/BB)、转化生长因子-β1(TGF-β1)和血管内皮生长因子(VEGF)。软骨细胞和滑膜细胞的培养基补充新鲜或冷冻的PRP,并在7天时评估细胞增殖、基因表达和分泌蛋白水平。结果显示,冻融后的PRP中,即时和延迟分子释放与新鲜PRP相似或略低。在1小时和7天时,冷冻后TGF-β1和PDGF AB/BB浓度均显著降低,而冷冻PRP在7天时HGF浓度显著降低。在新鲜PRP中,IL-1β和HGF浓度在7天后进一步显著增加。用两种PRP培养的软骨细胞中发现相似的基因表达,而在滑膜细胞中,冷冻PRP中HGF基因表达较高。PRP冷冻保存是一种安全的方法,能充分保持PRP的质量及其诱导软骨细胞和滑膜细胞增殖和产生细胞外基质成分的能力。