Collado-González M, Pecci-Lloret M P, García-Bernal D, Aznar-Cervantes S, Oñate-Sánchez R E, Moraleda J M, Cenis J L, Rodríguez-Lozano F J
Cellular Therapy and Hematopoietic Transplant Unit, Haematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain.
School of Dentistry, Faculty of Medicine, University of Murcia, Murcia, Spain.
Odontology. 2018 Apr;106(2):125-134. doi: 10.1007/s10266-017-0310-9. Epub 2017 Jun 14.
The aim is to investigate in vitro biological effects of silk fibroin 3D scaffolds on stem cells from human exfoliated deciduous teeth (SHEDs) in terms of proliferation, morphological appearance, cell viability, and expression of mesenchymal stem cell markers. Silk fibroin 3D scaffolding materials may represent promising suitable scaffolds for their application in regenerative endodontic therapy approaches. SHEDs were cultured in silk fibroin 3D scaffolds. Then, cell numbers were counted and the Alamar blue colorimetric assay was used to analyse cell proliferation after 24, 48, 72, and 168 h of culture. The morphological features of SHEDs cultured on silk fibroin scaffolds were evaluated by scanning electron microscopy (SEM). Finally, cell viability and the expression of mesenchymal stem cell markers were analysed by flow cytometry. One-way analysis of variance (ANOVA) followed by a Bonferroni post-test was performed (P < 0.05). At 24 and 48 h of culture, SHED proliferation on scaffolds was modest compared to the control although still significant (p < 0.05). However, cell proliferation progressively increased from 72 to 168 h compared with the control (p < 0.001; p < 0.01). In addition, flow cytometry analysis showed that the culture of SHEDs on silk fibroin scaffolds did not significantly alter the level of expression of the mesenchymal markers CD73, CD90, or CD105 up to 168 h; in addition, cell viability in silk fibroin was similar to than obtained in plastic. Moreover, SEM studies revealed a suitable degree of proliferation, cell spreading, and attachment, especially after 168 h of culture. The findings from the current study suggest that silk fibroin 3D scaffolds had a favourable effect on the biological responses of SHEDs. Further in vivo investigations are required to confirm these results.
目的是研究丝素蛋白三维支架对人乳牙脱落干细胞(SHEDs)在增殖、形态外观、细胞活力和间充质干细胞标志物表达方面的体外生物学效应。丝素蛋白三维支架材料可能是再生牙髓治疗方法中很有前景的合适支架。将SHEDs培养在丝素蛋白三维支架中。然后,计数细胞数量,并使用阿拉玛蓝比色法分析培养24、48、72和168小时后的细胞增殖情况。通过扫描电子显微镜(SEM)评估在丝素蛋白支架上培养的SHEDs的形态特征。最后,通过流式细胞术分析细胞活力和间充质干细胞标志物的表达。进行单因素方差分析(ANOVA),然后进行Bonferroni事后检验(P < 0.05)。在培养24和48小时时,与对照组相比,支架上SHEDs的增殖适度,尽管仍有显著差异(p < 0.05)。然而,与对照组相比,从72小时到168小时细胞增殖逐渐增加(p < 0.001;p < 0.01)。此外,流式细胞术分析表明,在丝素蛋白支架上培养SHEDs长达168小时,间充质标志物CD73、CD90或CD105的表达水平没有显著改变;此外,丝素蛋白中的细胞活力与在塑料中获得的相似。而且,SEM研究显示增殖、细胞铺展和附着程度合适,尤其是在培养168小时后。本研究结果表明,丝素蛋白三维支架对SHEDs的生物学反应有有利影响。需要进一步的体内研究来证实这些结果。