Sartoneva Reetta, Kuismanen Kirsi, Juntunen Miia, Karjalainen Sanna, Hannula Markus, Kyllönen Laura, Hyttinen Jari, Huhtala Heini, Paakinaho Kaarlo, Miettinen Susanna
Adult Stem Cell Research Group, BioMediTech, Faculty of Medicine and Life Sciences, University of Tampere, Arvo Ylpönkatu 34, 4th Floor, 33520 Tampere, Finland.
Science Centre, Tampere University Hospital, Tampere, Finland.
R Soc Open Sci. 2018 Aug 15;5(8):180811. doi: 10.1098/rsos.180811. eCollection 2018 Aug.
The surgical reconstruction of functional neovagina is challenging and susceptible to complications. Therefore, developing tissue engineering-based treatment methods for vaginal defects is important. Our aim was to develop and test a novel supercritical carbon dioxide foamed poly-l-lactide-co-ɛ-caprolactone (scPLCL) scaffold for vaginal reconstruction. The scaffolds were manufactured and characterized for porosity (65 ± 4%), pore size (350 ± 150 µm) and elastic modulus (2.8 ± 0.4 MPa). Vaginal epithelial (EC) and stromal cells (SC) were isolated, expanded and characterized with flow cytometry. Finally, cells were cultured with scPLCL scaffolds in separate and/or co-cultures. Their attachment, viability, proliferation and phenotype were analysed. Both cell types strongly expressed cell surface markers CD44, CD73 and CD166. Strong expression of CD326 was detected with ECs and CD90 and CD105 with SCs. Both ECs and SCs attached and maintained viability on scPLCL. Further, scPLCL supported the proliferation of especially ECs, which also maintained epithelial phenotype (cytokeratin expression) during 14-day assessment period. Interestingly, ECs expressed uroplakin (UP) Ia, UPIb and UPIII markers; further, UPIa and UPIII expression was significantly higher on ECs cultured on scPLCL than on cell culture plastic. In conclusion, the scPLCL is potential scaffold for vaginal tissue engineering and the results of this study further illustrate the excellent biocompatibility of PLCL.
功能性新阴道的手术重建具有挑战性且易发生并发症。因此，开发基于组织工程的阴道缺损治疗方法很重要。我们的目标是开发并测试一种用于阴道重建的新型超临界二氧化碳发泡聚左旋乳酸-共-ε-己内酯（scPLCL）支架。对支架进行制造，并对其孔隙率（65±4%）、孔径（350±150µm）和弹性模量（2.8±0.4MPa）进行表征。分离、扩增阴道上皮细胞（EC）和基质细胞（SC），并用流式细胞术进行表征。最后，将细胞与scPLCL支架进行单独和/或共培养。分析它们的附着、活力、增殖和表型。两种细胞类型均强烈表达细胞表面标志物CD44、CD73和CD166。在EC中检测到CD326的强表达，在SC中检测到CD90和CD105的强表达。EC和SC均可附着在scPLCL上并保持活力。此外，scPLCL尤其支持EC的增殖，在14天的评估期内，EC也保持上皮表型（细胞角蛋白表达）。有趣的是，EC表达uroplakin（UP）Ia、UPIb和UPIII标志物；此外，在scPLCL上培养的EC上UPIa和UPIII的表达明显高于在细胞培养塑料上培养的EC。总之，scPLCL是阴道组织工程的潜在支架，本研究结果进一步说明了PLCL具有优异的生物相容性。
