Innovative Research Laboratory for Wound Healing, Health Sciences Department, University Eastern Piedmont, Novara, Italy.
Eur Rev Med Pharmacol Sci. 2020 Sep;24(18):9658-9664. doi: 10.26355/eurrev_202009_23055.
Platelet rich plasma (PRP) is a haemoderivative used in clinical practice for the treatment of hard-to-heal wounds. Platelet (PLT) activation is a key factor in the wound healing process leading to the production of extracellular vesicles. We obtained PRP and PRP-derived microvesicles (PLT-MVs) from healthy donors and compared their pro-healing efficacy in an in vitro wound model using human keratinocytes.
We evaluated PLT-MVs' direct effect on an in vitro model of wound healing. PRP, PRP activated using calcimycin, and PLT-MVs separated by high speed centrifugation were added to scratched keratinocyte monolayers. Fluorescein diacetate was used in flow cytometry to distinguish PLTs and PLT-MVs from debris, and then, PLT-MVs were quantified on the basis of relative dimensions (Forward Scatter signals).
Wound areas were measured at time 0 and after 24 hours and they were healed by 24.80 ± 4.28% in control conditions, while PRP, activated PRP, and PLT-MVs increased closure of 62.94 ± 0.96%, 52.69 ± 17.20% and 52.76 ± 9.44%, respectively.
Our results indicate that PRP pro-healing effects were fully replicable by PLT-MVs, suggesting a key role of microvesicles in the healing process and a possible clinical use as an alternative to PRP.
富含血小板的血浆（PRP）是一种血液衍生品，在临床实践中用于治疗难以愈合的伤口。血小板（PLT）的激活是导致细胞外囊泡产生的伤口愈合过程中的关键因素。我们从健康供体中获得 PRP 和 PRP 衍生的微泡（PLT-MVs），并在使用人角质形成细胞的体外伤口模型中比较它们的促愈合功效。
我们评估了 PLT-MVs 对体外伤口愈合模型的直接影响。将 PRP、使用钙调蛋白激活的 PRP 和通过高速离心分离的 PLT-MVs 添加到划痕的角质形成细胞单层中。使用荧光二乙酸酯在流式细胞术中区分 PLTs 和 PLT-MVs 与碎片，然后根据相对尺寸（前向散射信号）定量 PLT-MVs。
在 0 时间和 24 小时后测量伤口面积，在对照条件下愈合率为 24.80 ± 4.28%，而 PRP、激活的 PRP 和 PLT-MVs 分别增加了 62.94 ± 0.96%、52.69 ± 17.20%和 52.76 ± 9.44%的闭合。
我们的结果表明，PRP 的促愈合作用可以被 PLT-MVs 完全复制，这表明微泡在愈合过程中起着关键作用，并可能作为 PRP 的替代物在临床上使用。
