Huang Jia-Ping, Wu Yan-Min, Liu Jia-Mei, Zhang Lan, Li Bo-Xiu, Chen Li-Li, Ding Pei-Hui, Tan Jing-Yi
Key Laboratory of Oral Biomedical Research of Zhejiang Province, School of Stomatology, The Affiliated Hospital of Stomatology, Zhejiang University School of Medicine, Hangzhou, China.
Department of Periodontology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
J Periodontal Res. 2021 Oct;56(5):929-939. doi: 10.1111/jre.12889. Epub 2021 Jun 26.
Recently, decellularized matrix (DCM) is considered as a new biomaterial for tissue regeneration. To explore the possible application of DCM in periodontal regeneration, the effect of DCM from three different cells on the proliferation and differentiation of human periodontal ligament stem cells (PDLSCs) was investigated.
DCM derived from human periodontal ligament cells (PDLCs), dental pulp cells (DPCs), and gingival fibroblasts (GFs) were fabricated using Triton X-100/NH OH combined with DNase I. Allogeneic PDLSCs were cultured on PDLC-DCM, DPC-DCM, and GF-DCM, respectively. The proliferative capacity of PDLSCs was evaluated by PicoGreen assay kit. The expression of alkaline phosphatase (ALP), runt-related transcription factor-2 (RUNX2), osteocalcin (OCN), collagen I (COL1), periostin (POSTN), and cementum protein 1 (CEMP1) were detected by qRT-PCR and western blotting.
PDLC-DCM, DPC-DCM, and GF-DCM had similar and integrated networks of extracellular matrix, as well as significantly decreased DNA content. Compared with control group in which PDLSCs were directly seeded in culture plates, PDLC-DCM, DPC-DCM, and GF-DCM promoted the proliferation of re-seeded PDLSCs. Additionally, PDLSCs on DCM exhibited higher mRNA and protein expression levels of ALP, RUNX2, OCN, and COL1. The expression of POSTN in PDLC-DCM group was significantly higher than control group at both mRNA and protein levels.
PDLC-DCM, DPC-DCM, and GF-DCM could enhance the proliferation of PDLSCs. PDLC-DCM facilitated osteogenic differentiation and periodontal ligament differentiation of PDLSCs, while DPC-DCM and GF-DCM promoted osteogenic differentiation.
近年来,脱细胞基质(DCM)被视为一种用于组织再生的新型生物材料。为探索DCM在牙周再生中的可能应用,研究了来自三种不同细胞的DCM对人牙周膜干细胞(PDLSCs)增殖和分化的影响。
使用Triton X - 100/NH OH联合DNase I制备源自人牙周膜细胞(PDLCs)、牙髓细胞(DPCs)和牙龈成纤维细胞(GFs)的DCM。将异体PDLSCs分别培养在PDLC - DCM、DPC - DCM和GF - DCM上。通过PicoGreen检测试剂盒评估PDLSCs的增殖能力。通过qRT - PCR和蛋白质印迹法检测碱性磷酸酶(ALP)、 runt相关转录因子2(RUNX2)、骨钙素(OCN)、胶原蛋白I(COL1)、骨膜蛋白(POSTN)和牙骨质蛋白1(CEMP1)的表达。
PDLC - DCM、DPC - DCM和GF - DCM具有相似且完整的细胞外基质网络,且DNA含量显著降低。与将PDLSCs直接接种于培养板的对照组相比,PDLC - DCM、DPC - DCM和GF - DCM促进了重新接种的PDLSCs的增殖。此外,DCM上的PDLSCs表现出更高的ALP、RUNX2、OCN和COL1的mRNA和蛋白质表达水平。PDLC - DCM组中POSTN在mRNA和蛋白质水平上的表达均显著高于对照组。
PDLC - DCM、DPC - DCM和GF - DCM可增强PDLSCs的增殖。PDLC - DCM促进PDLSCs的成骨分化和牙周膜分化,而DPC - DCM和GF - DCM促进成骨分化。