Sui H X, Lv P J, Wang Y, Feng Y C
Department of Stomatology, Peking University First Hospital, Beijing 100034, China.
Center of Digital Dentistry, Department of Prosthodontics, Peking University School and Hospital of Stomatology & Research Center of Engineering and Technology for Digital Dentistry, Ministry of Health & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2018 Oct 18;50(5):868-875.
To explore the effects of low level laser irradiation (LLLI) on the osteogenic capacity of three-dimensional (3D) structure by 3D bio-printing construct used human adipose-derived stem cells (hASCs) as seed cells.
Using hASCs as seed cells, we prepared sodium alginate/gelatin/hASCs 3D bio-printing construct, and divided them into four groups: PM (proliferative medium), PM+LLLI, OM (osteogenic medium) and OM+LLLI, and the total doses of LLLI was 4 J/cm². Immunofluorescence microscopy was used to observe the viability of the cells, and analyze the expression of the osteogenesis-related protein Runt-related transcription factor 2 (Runx2) and osteocalcin (OCN).
The 3D constructs obtained by printing were examined by microscope. The sizes of these 3D constructs were 10 mm×10 mm×1.5 mm. The wall thickness of the printed gelatin mold was approximately 1 mm, and the pores were round and had a diameter of about 700 μm. The cell viability of sodium alginate/gelatin/hASCs 3D bio-printing construct was high, and the difference among the four groups was not significant. On day 7, the expression of OCN from high to low was group OM+LLLI, PM+LLLI, OM and PM. There were significant differences among these groups (P<0.01), but there was no significant difference between group PM+LLLI and OM. On day 14, the expression of OCN in each group was higher than that on day 7, and there was no significant difference between group OM+LLLI and OM. The expression of Runx2 in group OM+LLLI was more than 90%, significantly higher than that in group OM (P<0.01). But the expression of Runx2 in group PM+LLLI and OM+LLLI were significantly lower than that in the non-irradiated groups. The expression of osteogenesis-related protein Runx2 and OCN were higher in OM groups than in PM groups. Furthermore, the irradiated groups were significantly higher than the non-irradiated groups.
LLLI does not affect the cell viability of sodium alginate/gelatin/hASCs 3D bio-printing construct, and may promote the osteogenic differentiation of hASCs.
探讨低强度激光照射（LLLI）对以人脂肪来源干细胞（hASCs）为种子细胞的三维（3D）生物打印构建体的成骨能力的影响。
以hASCs为种子细胞，制备海藻酸钠/明胶/hASCs 3D生物打印构建体，并将其分为四组：增殖培养基（PM）组、PM+LLLI组、成骨培养基（OM）组和OM+LLLI组，LLLI的总剂量为4 J/cm²。采用免疫荧光显微镜观察细胞活力，并分析成骨相关蛋白Runx2相关转录因子2（Runx2）和骨钙素（OCN）的表达。
通过显微镜检查打印获得的3D构建体。这些3D构建体的尺寸为10 mm×10 mm×1.5 mm。打印的明胶模具壁厚约为1 mm，孔隙呈圆形，直径约为700μm。海藻酸钠/明胶/hASCs 3D生物打印构建体的细胞活力较高，四组之间差异不显著。第7天，OCN表达从高到低依次为OM+LLLI组、PM+LLLI组、OM组和PM组。这些组之间存在显著差异（P<0.01），但PM+LLLI组和OM组之间无显著差异。第14天，各组OCN表达均高于第7天，OM+LLLI组和OM组之间无显著差异。OM+LLLI组Runx2表达超过90%，显著高于OM组（P<0.01）。但PM+LLLI组和OM+LLLI组Runx2表达均显著低于未照射组。成骨相关蛋白Runx2和OCN在OM组中的表达高于PM组。此外，照射组显著高于未照射组。
LLLI不影响海藻酸钠/明胶/hASCs 3D生物打印构建体的细胞活力，并可能促进hASCs的成骨分化。
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