用于伤口愈合应用的经合减毒冻干人脐血血小板裂解液的体外再生效果。

In vitro regenerative effects of a pooled pathogen-reduced lyophilized human cord blood platelet lysate for wound healing applications.

机构信息

Institute of Clinical Physiology, CNR, Massa, Italy.

Transfusion Medicine and Transplant Biology, Pisa University Hospital, Pisa, Italy.

出版信息

Blood Transfus. 2024 Nov;22(6):514-524. doi: 10.2450/BloodTransfus.755. Epub 2024 Sep 16.

DOI:10.2450/BloodTransfus.755

PMID:39447186

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11576148/

Abstract

BACKGROUND

Cord blood platelets, easily obtained from blood units not suitable for hematopoietic stem cell transplantation, represent an abundant source of growth factors for use in wound healing. Although several protocols have been described for platelet lysate production, no standard manufacturing protocol is available. The use of pooled cord blood platelets could thus facilitate standardization. In this study, the effect of varying concentrations (up to 20%) of a pooled pathogen-reduced lyophilized cord blood platelet lysate (PRL-CBPL) was investigated in different cell types involved in the wound healing process. The effect of heparin addition was also evaluated. In parallel, a comparison was performed with a single donor cord blood platelet lysate (SD-CBPL).

MATERIALS AND METHODS

The effect of PRL-CBPL on the viability and proliferation of different cell lines (L929 mouse fibroblasts and HaCaT keratinocytes) and human primary cells (fibroblasts-NHDF, coronary artery smooth muscle cells-HCASMC and coronary artery endothelial cells-HCAEC), on HaCaT migration and the chemotactic effect on human monocytes (THP-1) was evaluated.

RESULTS

PRL-CBPL showed a lower PDGF-AB amount compared to SD-CBPL. Differing concentrations of both CBPL were necessary to influence cell viability and proliferation. 3% was the optimal concentration for L929 and HaCaT as well as for NHDF and HCASMC, while HCAEC required 10%. The effect of added heparin was more evident on SD-CBPL and in particular on NHDF and HCASMC proliferation. Keratinocyte scratch closure was obtained with 3 and 5% PRL-CBPL and SD-CBPL respectively. Both CBPLs caused an increase in the number of migrated THP-1 monocytes in a concentration-dependent manner up to 20% with a higher monocyte migration for SD-CBPL with respect to PRL-CBPL and in cells treated with heparin.

DISCUSSION

The data obtained suggest that PRL-CBPL is an effective standardized alternative to SD-CBPL.

摘要

背景

从不适合造血干细胞移植的血液单位中很容易获得脐带血血小板，这是一种丰富的生长因子来源，可用于伤口愈合。尽管已经描述了几种血小板裂解液（PL）的生产方案，但尚无标准的制造方案。因此，使用汇集的脐带血血小板可以促进标准化。在这项研究中，研究了浓度高达 20％的汇集的经病原体减少处理的冻干脐带血血小板裂解液（PRL-CBPL）对参与伤口愈合过程的不同细胞类型的影响。还评估了添加肝素的效果。同时，还与单个供体脐带血血小板裂解液（SD-CBPL）进行了比较。

材料和方法

评估了 PRL-CBPL 对不同细胞系（L929 小鼠成纤维细胞和 HaCaT 角质形成细胞）和人原代细胞（成纤维细胞-NHDF、冠状动脉平滑肌细胞-HCASMC 和冠状动脉内皮细胞-HCAEC）的活力和增殖、HaCaT 迁移以及对人单核细胞（THP-1）的趋化作用的影响。

结果

与 SD-CBPL 相比，PRL-CBPL 的 PDGF-AB 含量较低。两种 CBPL 的不同浓度都需要影响细胞活力和增殖。3％是 L929 和 HaCaT 以及 NHDF 和 HCASMC 的最佳浓度，而 HCAEC 需要 10％。添加肝素的效果在 SD-CBPL 上更为明显，尤其是在 NHDF 和 HCASMC 的增殖上。用 3％和 5％的 PRL-CBPL 和 SD-CBPL 分别获得了角质形成细胞划痕闭合。两种 CBPL 均以浓度依赖的方式导致迁移的 THP-1 单核细胞数量增加，在 20％时达到最大值，与 PRL-CBPL 相比，SD-CBPL 和用肝素处理的细胞中单核细胞的迁移更高。