Jin Hexiu, Choung Han-Wool, Lim Ki-Taek, Jin Bin, Jin Chengbiao, Chung Jong-Hoon, Choung Pill-Hoon
1 Tooth Bioengineering Laboratory, Department of Oral and Maxillofacial Surgery, Dental Research Institute, School of Dentistry, Seoul National University , Seoul, Korea.
2 Program of Cell and Developmental Biology, Department of Oral Histology and Development Biology, Dental Research Institute, School of Dentistry, Seoul National University , Seoul, Korea.
Tissue Eng Part A. 2015 Dec;21(23-24):2817-28. doi: 10.1089/ten.TEA.2014.0399. Epub 2015 Aug 14.
The periodontium, consisting of gingiva, periodontal ligament (PDL), cementum, and alveolar bone, is necessary for the maintenance of tooth function. Specifically, the regenerative abilities of cementum with inserted PDL are important for the prevention of tooth loss. Periodontal ligament stem cells (PDLSCs), which are located in the connective tissue PDL between the cementum and alveolar bone, are an attractive candidate for hard tissue formation. We investigated the effects of recombinant human plasminogen activator inhibitor-1 (rhPAI-1) on cementogenic differentiation of human PDLSCs (hPDLSCs) in vitro and in vivo. Untreated and rhPAI-1-treated hPDLSCs mixed with hydroxyapatite/tricalcium phosphate (HA/TCP) and dentin matrix were transplanted subcutaneously into the dorsal surface of immunocompromised mice to assess their capacity for hard tissue formation at 8 and 10 weeks posttransplantation. rhPAI-1 accelerated mineral nodule formation and increased the mRNA expression of cementoblast-associated markers in hPDLSCs. We also observed that rhPAI-1 upregulated the levels of osterix (OSX) and cementum protein 1 (CEMP1) through Smad2/3 and p38 pathways, whereas specific inhibitors of Smad3 and p38 inhibited the enhancement of mineralization of hPDLSCs by rhPAI-1. Furthermore, transplantation of hPDLSCs with rhPAI-1 showed a great ability to promote cementogenic differentiation. Notably, rhPAI-1 induced hPDLSCs to regenerate cementum-like tissue with PDL fibers inserted into newly formed cementum-like tissue. These results suggest that rhPAI-1 may play a key role in cementogenic differentiation of hPDLSCs. rhPAI-1 with hPDLSCs may be a good candidate for future clinical applications in periodontal tissue regeneration and possibly in tooth root bioengineering.
牙周组织由牙龈、牙周韧带(PDL)、牙骨质和牙槽骨组成,对维持牙齿功能至关重要。具体而言,带有插入的PDL的牙骨质的再生能力对于预防牙齿脱落很重要。牙周韧带干细胞(PDLSCs)位于牙骨质和牙槽骨之间的结缔组织PDL中,是硬组织形成的有吸引力的候选细胞。我们研究了重组人纤溶酶原激活物抑制剂-1(rhPAI-1)在体外和体内对人PDLSCs(hPDLSCs)牙骨质形成分化的影响。将未处理的和经rhPAI-1处理的hPDLSCs与羟基磷灰石/磷酸三钙(HA/TCP)和牙本质基质混合,皮下移植到免疫受损小鼠的背部,以评估它们在移植后8周和10周形成硬组织的能力。rhPAI-1加速了矿物质结节的形成,并增加了hPDLSCs中与成牙骨质细胞相关标志物的mRNA表达。我们还观察到rhPAI-1通过Smad2/3和p38途径上调了osterix(OSX)和牙骨质蛋白1(CEMP1)的水平,而Smad3和p38的特异性抑制剂抑制了rhPAI-1对hPDLSCs矿化的增强作用。此外,用rhPAI-1移植hPDLSCs显示出极大的促进牙骨质形成分化的能力。值得注意的是,rhPAI-1诱导hPDLSCs再生出类似牙骨质的组织,PDL纤维插入新形成的类似牙骨质的组织中。这些结果表明,rhPAI-1可能在hPDLSCs的牙骨质形成分化中起关键作用。rhPAI-1与hPDLSCs可能是未来牙周组织再生以及可能的牙根生物工程临床应用的良好候选者。