School of Stomatology, Xuzhou Medical University, No. 209 Tongshan Road, Xuzhou, 221004, Jiangsu, China.
Affiliated Stomatological Hospital of Xuzhou Medical University, No.130 Huaihai West Road, Xuzhou, 221000, Jiangsu, China.
Clin Oral Investig. 2023 Sep;27(9):5153-5170. doi: 10.1007/s00784-023-05135-7. Epub 2023 Jul 10.
We aimed to explore the osteogenic potential of periodontal ligament stem cells (PDLSCs) in bioprinted methacrylate gelatine (GelMA) hydrogels in vitro and in vivo.
PDLSCs in GelMA hydrogels at various concentrations (3%, 5%, and 10%) were bioprinted. The mechanical properties (stiffness, nanostructure, swelling, and degradation properties) of bioprinted constructs and the biological properties (cell viability, proliferation, spreading, osteogenic differentiation, and cell survival in vivo) of PDLSCs in bioprinted constructs were evaluated. Then, the effect of bioprinted constructs on bone regeneration was investigated using a mouse cranial defect model.
Ten percent GelMA printed constructs had a higher compression modulus, smaller porosity, lower swelling rate, and lower degradation rate than 3% GelMA. PDLSCs in bioprinted 10% GelMA bioprinted constructs showed lower cell viability, less cell spreading, upregulated osteogenic differentiation in vitro, and lower cell survival in vivo. Moreover, upregulated expression of ephrinB2 and EphB4 protein and their phosphorylated forms were found in PDLSCs in 10% GelMA bioprinted constructs, and inhibition of eprhinB2/EphB4 signalling reversed the enhanced osteogenic differentiation of PDLSCs in 10% GelMA. The in vivo experiment showed that 10% GelMA bioprinted constructs with PDLSCs contributed to more new bone formation than 10% GelMA constructs without PDLSCs and constructs with lower GelMA concentrations.
Bioprinted PDLSCs with high-concentrated GelMA hydrogels exhibited enhanced osteogenic differentiation partially through upregulated ephrinB2/EphB4 signalling in vitro and promoted bone regeneration in vivo, which might be more appropriate for future bone regeneration applications.
Bone defects are a common clinical oral problem. Our results provide a promising strategy for bone regeneration through bioprinting PDLSCs in GelMA hydrogels.
本研究旨在探讨牙周膜干细胞（PDLSCs）在体外和体内生物打印甲基丙烯酰化明胶（GelMA）水凝胶中的成骨潜能。
将 PDLSCs 分别置于不同浓度（3%、5%和 10%）的 GelMA 水凝胶中进行生物打印。评估生物打印构建体的机械性能（硬度、纳米结构、溶胀和降解特性）以及生物打印构建体中 PDLSCs 的生物学特性（细胞活力、增殖、铺展、成骨分化和体内细胞存活）。然后，使用小鼠颅顶缺损模型研究生物打印构建体对骨再生的影响。
与 3% GelMA 相比，10% GelMA 打印构建体具有更高的压缩模量、更小的孔隙率、更低的溶胀率和更低的降解率。生物打印的 10% GelMA 构建体中 PDLSCs 的细胞活力较低，细胞铺展较少，体外成骨分化上调，体内细胞存活率较低。此外，在 10% GelMA 生物打印构建体中的 PDLSCs 中发现 EphrinB2 和 EphB4 蛋白及其磷酸化形式的表达上调，并且抑制 EphrinB2/EphB4 信号通路可逆转 10% GelMA 中 PDLSCs 增强的成骨分化。体内实验表明，与不含 PDLSCs 的 10% GelMA 构建体和浓度较低的 GelMA 构建体相比，含 PDLSCs 的 10% GelMA 生物打印构建体更有助于新骨形成。
体外高浓度 GelMA 水凝胶中生物打印的 PDLSCs 表现出增强的成骨分化，部分是通过 EphrinB2/EphB4 信号通路的上调，并且在体内促进了骨再生，这可能更适合未来的骨再生应用。
骨缺损是一种常见的临床口腔问题。我们的研究结果为通过生物打印 GelMA 水凝胶中的 PDLSCs 促进骨再生提供了一种有前景的策略。
