Kantawong Fahsai, Saksiriwisitkul Chanidapa, Riyapa Chanakan, Limpakdee Suchalinee, Wanachantararak Phenphichar, Kuboki Thasaneeya
Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
The Dental Research Center, Faculty of Dentistry, Chiang Mai University, Chiang Mai 50200, Thailand.
Bioimpacts. 2018;8(2):129-138. doi: 10.15171/bi.2018.15. Epub 2018 Jan 10.
Induced neural stem cells (iNSCs) have the ability of differentiation into neurons, astrocytes and oligodendrocytes. iNSCs are very useful in terms of research and treatment. The present study offers an idea that biomaterials could be one of the tools that could modulate reprogramming process in the fibroblasts. Gelatin biomaterials were fabricated into 3 types, including (i) gelatin, (ii) gelatin with 1 mg/mL hydroxyapatite, and (iii) gelatin with hydroxyapatite and pig brain. NIH/3T3 fibroblasts were cultured on each type of biomaterial for 7, 9 and 14 days. RT-PCR was performed to investigate the gene expression of the fibroblasts on biomaterials compared to the fibroblasts on tissue culture plates. PI3K/Akt signaling was performed by flow cytometry after 24 hours seeding on the biomaterials. The biomaterials were also tested with the human APCs and PDL cells. The fibroblasts exhibited changes in the expression of the reprogramming factor; Klf4 and the neural transcription factors; NFIa, NFIb and Ptbp1 after 9 days culture. The cultivation of fibroblasts on the biomaterials for 7 days showed a higher expression of the transcription factor SOX9. The expression of epigenetic genes; Kat2a and HDAC3 were changed upon the cultivation on the biomaterials for 9 days. The fibroblasts cultured on the biomaterials showed an activation of PI3K/Akt signaling. The human APCs and human PDL cells developed mineralization process on biomaterials Changes in the expression of Klf4, NFIa, NFIb, Ptbp1 and SOX9 indicated that fibroblasts were differentiated into an astrocytic lineage. It is possible that the well-designed biomaterials could work as powerful tools in the reprogramming process of fibroblasts into iNSCs.
诱导神经干细胞（iNSCs）具有分化为神经元、星形胶质细胞和少突胶质细胞的能力。iNSCs在研究和治疗方面非常有用。本研究提出了一种观点，即生物材料可能是调节成纤维细胞重编程过程的工具之一。将明胶生物材料制成3种类型，包括（i）明胶，（ii）含1mg/mL羟基磷灰石的明胶，以及（iii）含羟基磷灰石和猪脑的明胶。将NIH/3T3成纤维细胞在每种生物材料上培养7天、9天和14天。进行RT-PCR以研究与组织培养板上的成纤维细胞相比，生物材料上的成纤维细胞的基因表达。在接种到生物材料上24小时后，通过流式细胞术检测PI3K/Akt信号通路。还对生物材料与人脂肪前体细胞（APCs）和牙周膜细胞（PDL细胞）进行了测试。培养9天后，成纤维细胞在重编程因子Klf4以及神经转录因子NFIa、NFIb和Ptbp1的表达上出现了变化。成纤维细胞在生物材料上培养7天显示转录因子SOX9的表达更高。在生物材料上培养9天后，表观遗传基因Kat2a和HDAC3的表达发生了变化。在生物材料上培养的成纤维细胞显示出PI3K/Akt信号通路的激活。人APCs和人PDL细胞在生物材料上形成了矿化过程。Klf4、NFIa、NFIb、Ptbp1和SOX9表达的变化表明成纤维细胞分化为星形胶质细胞谱系。精心设计的生物材料有可能在成纤维细胞重编程为iNSCs的过程中成为强大的工具。
