Chiang Chih-Wei, Chen Wei-Chuan, Liu Hsia-Wei, Wang I-Chun, Chen Chih-Hwa
Bone and Joint Research Center, Department of Orthopedics and Traumatology, Taipei Medical University Hospital, School of Medicine, College of Medicine, Taipei Medical University, Taipei 110, Taiwan.
Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei 106, Taiwan.
Int J Mol Sci. 2015 Sep 28;16(10):23318-36. doi: 10.3390/ijms161023318.
Regenerative medicine is increasingly important in clinical practice. Ligamentum flava (LF) are typically removed during spine-related surgeries. LF may be a source of cells for spinal fusion that is conducted using tissue engineering techniques. In this investigation, LF cells of rabbits were isolated and then characterized by flow cytometry, morphological observation, and immunofluorescence staining. The LF cells were also cultivated in polyethylene (glycol) diacrylate (PEGDA) hydrogels that incorporated bone morphogenetic protein-2 (BMP-2) growth factor, to evaluate their proliferation and secretion of ECM and differentiation in vitro. The experimental results thus obtained that the proliferation, ECM secretion, and differentiation of the PEGDA-BMP-2 group exceeded those of the PEGDA group during the period of cultivation. The mineralization and histological staining results differed similarly. A nude mice model was utilized to prove that LF cells on hydrogels could undergo osteogenic differentiation in vivo. These experimental results also revealed that the PEGDA-BMP-2 group had better osteogenic effects than the PEGDA group following a 12 weeks after transplantation. According to all of these experimental results, LF cells are a source of cells for spinal fusion and PEGDA-BMP-2 hydrogel is a candidate biomaterial for spinal fusion by tissue engineering.
再生医学在临床实践中日益重要。在脊柱相关手术中,通常会切除黄韧带(LF)。黄韧带可能是利用组织工程技术进行脊柱融合的细胞来源。在本研究中,分离了兔的黄韧带细胞,然后通过流式细胞术、形态学观察和免疫荧光染色对其进行表征。还将黄韧带细胞培养在掺入骨形态发生蛋白-2(BMP-2)生长因子的聚(乙二醇)二丙烯酸酯(PEGDA)水凝胶中,以评估其增殖、细胞外基质(ECM)分泌及体外分化情况。由此获得的实验结果表明,在培养期间,PEGDA-BMP-2组的增殖、ECM分泌和分化均超过PEGDA组。矿化和组织学染色结果也有类似差异。利用裸鼠模型证明水凝胶上的黄韧带细胞在体内可发生成骨分化。这些实验结果还表明,移植12周后,PEGDA-BMP-2组的成骨效果优于PEGDA组。根据所有这些实验结果,黄韧带细胞是脊柱融合的细胞来源,PEGDA-BMP-2水凝胶是通过组织工程进行脊柱融合的候选生物材料。