生物活性玻璃离子诱导人脂肪干细胞在卡拉胶和 I 型胶原水凝胶中的有效成骨分化。
Bioactive glass ions induce efficient osteogenic differentiation of human adipose stem cells encapsulated in gellan gum and collagen type I hydrogels.
机构信息
Adult Stem Cell Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 100, FI-33014 Tampere, Finland; Research, Development and Innovation Centre, Tampere University Hospital, P.O. BOX 2000, FI-33521, Tampere, Finland.
Biomaterials and Tissue Engineering Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 527, FI-33101 Tampere, Finland; Heart Group, BioMediTech, Faculty of Medicine and Health Technology, Tampere University, P.O. BOX 100, FI-33014 Tampere, Finland.
出版信息
Mater Sci Eng C Mater Biol Appl. 2019 Jun;99:905-918. doi: 10.1016/j.msec.2019.02.035. Epub 2019 Feb 13.
BACKGROUND
Due to unmet need for bone augmentation, our aim was to promote osteogenic differentiation of human adipose stem cells (hASCs) encapsulated in gellan gum (GG) or collagen type I (COL) hydrogels with bioactive glass (experimental glass 2-06 of composition [wt-%]: NaO 12.1, KO 14.0, CaO 19.8, PO5 2.5, BO 1.6, SiO 50.0) extract based osteogenic medium (BaG OM) for bone construct development. GG hydrogels were crosslinked with spermidine (GG-SPD) or BaG extract (GG-BaG).
METHODS
Mechanical properties of cell-free GG-SPD, GG-BaG, and COL hydrogels were tested in osteogenic medium (OM) or BaG OM at 0, 14, and 21 d. Hydrogel embedded hASCs were cultured in OM or BaG OM for 3, 14, and 21 d, and analyzed for viability, cell number, osteogenic gene expression, osteocalcin production, and mineralization. Hydroxyapatite-stained GG-SPD samples were imaged with Optical Projection Tomography (OPT) and Selective Plane Illumination Microscopy (SPIM) in OM and BaG OM at 21 d. Furthermore, Raman spectroscopy was used to study the calcium phosphate (CaP) content of hASC-secreted ECM in GG-SPD, GG-BaG, and COL at 21 d in BaG OM.
RESULTS
The results showed viable rounded cells in GG whereas hASCs were elongated in COL. Importantly, BaG OM induced significantly higher cell number and higher osteogenic gene expression in COL. In both hydrogels, BaG OM induced strong mineralization confirmed as CaP by Raman spectroscopy and significantly improved mechanical properties. GG-BaG hydrogels rescued hASC mineralization in OM. OPT and SPIM showed homogeneous 3D cell distribution with strong mineralization in BaG OM. Also, strong osteocalcin production was visible in COL.
CONCLUSIONS
Overall, we showed efficacious osteogenesis of hASCs in 3D hydrogels with BaG OM with potential for bone-like grafts.
背景
由于对骨增强的需求未得到满足，我们的目的是促进包封在结冷胶（GG）或胶原 I 型（COL）水凝胶中的人脂肪干细胞（hASC）的成骨分化，并用基于生物活性玻璃（组成 [wt-%]：NaO 12.1、KO 14.0、CaO 19.8、PO5 2.5、BO 1.6、SiO 50.0）提取的成骨培养基（BaG OM）开发骨构建物。GG 水凝胶用亚精胺（GG-SPD）或 BaG 提取物（GG-BaG）交联。
方法
在成骨培养基（OM）或 BaG OM 中测试无细胞 GG-SPD、GG-BaG 和 COL 水凝胶的机械性能，时间分别为 0、14 和 21 天。将包埋 hASC 的水凝胶在 OM 或 BaG OM 中培养 3、14 和 21 天，并分析其活力、细胞数量、成骨基因表达、骨钙素产生和矿化。在 OM 和 BaG OM 中，用羟基磷灰石染色的 GG-SPD 样品在 21 天进行光学投影断层扫描（OPT）和选择性平面照明显微镜（SPIM）成像。此外，还使用拉曼光谱研究了在 BaG OM 中 21 天的 hASC 分泌 ECM 中的钙磷（CaP）含量在 GG-SPD、GG-BaG 和 COL 中的分布情况。
结果
结果表明 GG 中存在存活的圆形细胞，而 COL 中 hASC 呈长形。重要的是，BaG OM 在 COL 中诱导了更高的细胞数量和更高的成骨基因表达。在两种水凝胶中，BaG OM 诱导的矿化均强烈证实为 CaP 通过拉曼光谱，并且显著改善了机械性能。GG-BaG 水凝胶可在 OM 中挽救 hASC 矿化。OPT 和 SPIM 显示出在 BaG OM 中具有强矿化的均匀 3D 细胞分布。COL 中也可见强烈的骨钙素产生。
结论
总的来说，我们在 BaG OM 中显示了 hASC 在 3D 水凝胶中的有效成骨作用，具有潜在的骨样移植物。
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