Paduano Francesco, Marrelli Massimo, Alom Noura, Amer Mahetab, White Lisa J, Shakesheff Kevin M, Tatullo Marco
a Tecnologica Research Institute, Biomedical Section , Crotone , Italy.
b Unit of Maxillofacial Surgery, Calabrodental , Crotone , Italy.
J Biomater Sci Polym Ed. 2017 Jun;28(8):730-748. doi: 10.1080/09205063.2017.1301770. Epub 2017 Mar 13.
Dental pulp tissue represents a source of mesenchymal stem cells that have a strong differentiation potential towards the osteogenic lineage. The objective of the current study was to examine in vitro osteogenic induction of dental pulp stem cells (DPSCs) cultured on hydrogel scaffolds derived from decellularized bone extracellular matrix (bECM) compared to collagen type I (Col-I), the major component of bone matrix. DPSCs in combination with bECM hydrogels were cultured under three different conditions: basal medium, osteogenic medium and medium supplemented with growth factors (GFs) and cell growth, mineral deposition, gene and protein expression were investigated. The DPSCs/bECM hydrogel constructs cultured in basal medium showed that cells were viable after three weeks and that the expression of runt-related transcription factor 2 (RUNX-2) and bone sialoprotein (BSP) were significantly upregulated in the absence of extra osteogenic inducers compared to Col-I hydrogel scaffolds. In addition, the protein expression levels of BSP and osteocalcin were higher on bECM with respect to Col-I hydrogel scaffolds. Furthermore, DPSCs/bECM hydrogels cultured with osteogenic or GFs supplemented medium displayed a higher upregulation of the osteo-specific markers compared to Col-I hydrogels in identical media. Collectively, our results demonstrate that bECM hydrogels might be considered as suitable scaffolds to support osteogenic differentiation of DPSCs.
牙髓组织是间充质干细胞的一个来源,这些干细胞对成骨谱系具有很强的分化潜力。本研究的目的是比较在源自脱细胞骨细胞外基质(bECM)的水凝胶支架上培养的牙髓干细胞(DPSC)与骨基质的主要成分I型胶原蛋白(Col-I)在体外的成骨诱导情况。将DPSC与bECM水凝胶组合在三种不同条件下培养:基础培养基、成骨培养基以及补充生长因子(GF)的培养基,并研究细胞生长、矿物质沉积、基因和蛋白质表达。在基础培养基中培养的DPSC/bECM水凝胶构建体显示,三周后细胞仍具有活力,并且与Col-I水凝胶支架相比,在没有额外成骨诱导剂的情况下, runt相关转录因子2(RUNX-2)和骨唾液蛋白(BSP)的表达显著上调。此外,相对于Col-I水凝胶支架,bECM上BSP和骨钙素的蛋白质表达水平更高。此外,与在相同培养基中的Col-I水凝胶相比,在补充有成骨或GF的培养基中培养的DPSC/bECM水凝胶显示出更高的骨特异性标志物上调。总的来说,我们的结果表明,bECM水凝胶可能被认为是支持DPSC成骨分化的合适支架。