Tissue Engineering Group, Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, North Campus Broad Lane, Sheffield, S3 7HQ, UK.
Tissue Eng Regen Med. 2020 Aug;17(4):445-458. doi: 10.1007/s13770-020-00263-7. Epub 2020 May 23.
Introduction of pro-angiogenic cells into tissue-engineered (TE) constructs (prevascularisation) is a promising approach to overcome delayed neovascularisation of such constructs post-implantation. Accordingly, in this study, we examined the contribution of human dermal microvascular endothelial cells (HDMECs) and human dermal fibroblasts (HDFs) alone and in combination on the formation of new blood vessels in ex-ovo chick chorioallantoic membrane (CAM) assay.
Poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV) and polycaprolactone (PCL) were first examined in terms of their physical, mechanical, and biological performances. The effect of gelatin coating and co-culture conditions on enhancing endothelial cell viability and growth was then investigated. Finally, the angiogenic potential of HDMECs and HDFs were assessed macroscopically and histologically after seeding on simple electrospun PHBV scaffolds either in isolation or in indirect co-culture using an ex-ovo CAM assay.
The results demonstrated that PHBV was slightly more favourable than PCL for HDMECs in terms of cell metabolic activity. The gelatin coating of PHBV scaffolds and co-culture of HDMECs with HDFs both showed a positive impact on HDMECs viability and growth. Both cell types induced angiogenesis over 7 days in the CAM assay either in isolation or in co-culture. The introduction of HDMECs to the scaffolds resulted in the production of more blood vessels in the area of implantation than the introduction of HDFs, but the co-culture of HDMECs and HDFs gave the most significant angiogenic activity.
Our findings showed that the in vitro prevascularisation of TE constructs with HDMECs and HDFs alone or in co-culture promotes angiogenesis in implantable TE constructs.
将促血管生成细胞引入组织工程(TE)构建体(血管预形成)是克服此类构建体植入后延迟新生血管形成的一种很有前途的方法。因此,在这项研究中,我们单独和联合检测了人真皮微血管内皮细胞(HDMEC)和人真皮成纤维细胞(HDF)在鸡胚绒毛尿囊膜(CAM)体外实验中形成新血管的作用。
首先研究了聚 3-羟基丁酸酯-共-3-羟基戊酸酯(PHBV)和聚己内酯(PCL)的物理、机械和生物学性能。然后研究了明胶涂层和共培养条件对增强内皮细胞活力和生长的影响。最后,通过鸡胚绒毛尿囊膜(CAM)体外实验,在简单静电纺 PHBV 支架上单独或间接共培养 HDMEC 和 HDF 后,宏观和组织学评估 HDMEC 和 HDF 的血管生成潜能。
结果表明,就细胞代谢活性而言,PHBV 对 HDMEC 比 PCL 稍有利。PHBV 支架的明胶涂层和 HDMEC 与 HDF 共培养均对 HDMEC 的活力和生长产生积极影响。两种细胞类型在 CAM 实验中均能在 7 天内诱导血管生成,无论是单独培养还是共培养。与单独引入 HDF 相比,将 HDMEC 引入支架可导致植入区域产生更多的血管,但 HDMEC 和 HDF 的共培养可产生最显著的血管生成活性。
我们的研究结果表明,单独或共培养 HDMEC 和 HDF 对 TE 构建体进行体外血管预形成可促进可植入 TE 构建体的血管生成。