Osteogenic differentiation of human periodontal ligament stem cells expressing lentiviral NEL-like protein 1

Chen,Cai-Yun; Liu,Ya-Jing; Shi,Sheng-Gen; Chen,Fa-Ming; Cai,Chuan; Li,Bing; Wang,Jing; Shi,Liang; Li,Yan; Liu,Zhong-Yu; Niu,Zhong-Ying

doi:10.3892/ijmm.2012.1053

Osteogenic differentiation of human periodontal ligament stem cells expressing lentiviral NEL-like protein 1

Authors:
- Cai-Yun Chen
- Ya-Jing Liu
- Sheng-Gen Shi
- Fa-Ming Chen
- Chuan Cai
- Bing Li
- Jing Wang
- Liang Shi
- Yan Li
- Zhong-Yu Liu
- Zhong-Ying Niu
View Affiliations

Affiliations: Chinese PLA Postgraduate Medical School, Beijing, P.R. China, Institute of Stomatology, Chinese PLA General Hospital, Beijing, P.R. China, Center of Stomatology, The 306th Hospital of PLA, Beijing, P.R. China, Center Laboratory for Tissue Engineering, School of Stomatology, The Fourth Military Medical University, Xi'an, Shanxi, P.R. China, Department of Orthodontics, School of Stomatology, The Fourth Military Medical University, Xi'an, Shanxi, P.R. China, Department of Gynecology, Chinese PLA General Hospital, Beijing, P.R. China
Published online on: July 4, 2012 https://doi.org/10.3892/ijmm.2012.1053
Pages: 863-869

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

NEL-like protein 1 (NELL1) is a newly identified secreted protein involved in craniosynostosis and has been found to promote osteogenic differentiation of mesenchymal stem cells. The objective of this study was to investigate the effect of NELL1 on osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and the potential underlying mechanism. hPDLSCs underwent lentivirus-mediated NELL1 transfection (Lenti-NELL1) and markers of osteogenesis were assessed [alkaline phosphate (ALP), osteocalcin (OCN) and calcium deposition] to evaluate the effect of NELL1 on the differentiation of these cells. Quantitative polymerase chain reaction (qPCR) was employed to measure the mRNA expression of Msx2 and Runx2, and Lenti-enhanced green fluorescent protein (EGFP) served as a control. Western blot analysis and qPCR analyses confirmed that Lenti-NELL1-transfected hPDLSCs could express NELL1. Compared with the Lenti-EGFP group, ALP, OCN, calcium deposition and Msx2 mRNA expression were markedly increased (P<0.01), but there was no significant difference in Runx2 mRNA expression between the two groups (P>0.01). hPDLSCs can be transfected by Lenti-NELL1 and can stably express NELL1. NELL1 is able to promote the osteogenic differentiation of hPDLSCs, which may be related to the downregulation of Msx2 expression. Lenti-NELL1 transfection can be used during in vitro gene therapy for periodontal regeneration.

View Figures

View References

1.	JR ElterS OffenbacherJF TooleJD BeckRelationship of periodontal disease and edentulism to stroke/TIAJ Dent Res829981001200310.1177/15440591030820121214630902
2.	P DucyG KarsentyThe family of bone morphogenetic proteinsKidney Int5722072214200010.1046/j.1523-1755.2000.00081.x10844590
3.	J Rivera-FelicianoCJ TabinBmp2 instructs cardiac progenitors to form the heart-valve-inducing fieldDev Biol295580588200610.1016/j.ydbio.2006.03.04316730346
4.	M FranzA BerndtF WehrhanP SchleierJ ClementP HyckelEctopic bone formation as a complication of surgical rehabilitation in patients with Moebius’ syndromeJ Craniomaxillofac Surg35252257200717855104
5.	TE MrozJC WangR HashimotoDC NorvellComplications related to osteobiologics use in spine surgery: a systematic reviewSpine (Phila Pa 1976)35Suppl 9S86S104201010.1097/BRS.0b013e3181d81ef220407355
6.	K TingH VastardisJB MullikenHuman NELL-1 expressed in unilateral coronal synostosisJ Bone Miner Res148089199910.1359/jbmr.1999.14.1.809893069
7.	T AghalooX JiangC SooA study of the role of nell-1 gene modified goat bone marrow stromal cells in promoting new bone formationMol Ther1518721880200710.1038/sj.mt.630027017653100
8.	CM CowanS ChengK TingNell-1 induced bone formation within the distracted intermaxillary sutureBone384858200610.1016/j.bone.2005.06.02316243593
9.	T AghalooCM CowanYF ChouNell-1-induced bone regeneration in calvarial defectsAm J Pathol169903915200610.2353/ajpath.2006.05121016936265
10.	CM CowanX JiangT HsuSynergistic effects of Nell-1 and BMP-2 on the osteogenic differentiation of myoblastsJ Bone Miner Res22918930200710.1359/jbmr.07031217352654
11.	RK SiuSS LuW LiNell-1 protein promotes bone formation in a sheep spinal fusion modelTissue Eng Part A1711231135201110.1089/ten.tea.2010.048621128865
12.	J Louis-UgboHS KimSD BodenRetention of 125I-labeled recombinant human bone morphogenetic protein-2 by biphasic calcium phosphate or a composite sponge in a rabbit posterolateral spine arthrodesis modelJ Orthop Res2010501059200210.1016/S0736-0266(02)00011-6
13.	MD KofronCT LaurencinOrthopaedic applications of gene therapyCurr Gene Ther53761200510.2174/156652305299748815638710
14.	AW BaltzerJR LiebermanRegional gene therapy to enhance bone repairGene Ther11344350200410.1038/sj.gt.330219514724686
15.	SS LuX ZhangC SooThe osteoinductive properties of Nell-1 in a rat spinal fusion modelSpine J75060200710.1016/j.spinee.2006.04.02017197333
16.	BT FeeleyAH ConduahO SugiyamaL KrenekIS ChenJR LiebermanIn vivo molecular imaging of adenoviral versus lentiviral gene therapy in two bone formation modelsJ Orthop Res2417091721200610.1002/jor.2022916788987
17.	MS VirkA ConduahSH ParkInfluence of short-term adenoviral vector and prolonged lentiviral vector mediated bone morphogenetic protein-2 expression on the quality of bone repair in a rat femoral defect modelBone42921931200810.1016/j.bone.2007.12.21618295562
18.	BM SeoM MiuraS GronthosInvestigation of multipotent postnatal stem cells from human periodontal ligamentLancet364149155200410.1016/S0140-6736(04)16627-015246727
19.	Q TuP ValverdeJ ChenOsterix enhances proliferation and osteogenic potential of bone marrow stromal cellsBiochem Biophys Res Commun34112571265200610.1016/j.bbrc.2006.01.09216466699
20.	AW JamesA PanM ChiangA new function of Nell-1 protein in repressing adipogenic differentiationBiochem Biophys Res Commun411126131201110.1016/j.bbrc.2011.06.11121723263
21.	SJ LeeSW KangHJ DoEnhancement of bone regeneration by gene delivery of BMP2/Runx2 bicistronic vector into adipose-derived stromal cellsBiomaterials3156525659201010.1016/j.biomaterials.2010.03.01920413153
22.	W TechawattanawisalK NakahamaM KomakiM AbeY TakagiI MoritaIsolation of multipotent stem cells from adult rat periodontal ligament by neurosphere-forming culture systemBiochem Biophys Res Commun357917923200710.1016/j.bbrc.2007.04.03117459343
23.	T TruongX ZhangD PathmanathanC SooK TingCraniosynostosis-associated gene nell-1 is regulated by runx2J Bone Miner Res22718200710.1359/jbmr.06101217042739
24.	AO WilkieZ TangN ElankoFunctional haploinsufficiency of the human homeobox gene MSX2 causes defects in skull ossificationNat Genet24387390200010.1038/7422410742103
25.	D WideraWD GrimmJM MoebiusHighly efficient neural differentiation of human somatic stem cells, isolated by minimally invasive periodontal surgeryStem Cells Dev16447460200710.1089/scd.2006.006817610375
26.	GS CouraRC GarcezCB de AguiarM Alvarez-SilvaRS MaginiAG TrentinHuman periodontal ligament: a niche of neural crest stem cellsJ Periodontal Res43531536200810.1111/j.1600-0765.2007.01065.x18624954
27.	X ZhangD CarpenterN BokuiOverexpression of Nell-1, a craniosynostosis-associated gene, induces apoptosis in osteoblasts during craniofacial developmentJ Bone Miner Res1821262134200310.1359/jbmr.2003.18.12.212614672347
28.	X ZhangS KurodaD CarpenterCraniosynostosis in transgenic mice overexpressing Nell-1J Clin Invest110861870200210.1172/JCI1537512235118
29.	M MiyazakiO SugiyamaJ ZouComparison of lentiviral and adenoviral gene therapy for spinal fusion in ratsSpine (Phila Pa 1976)3314101417200810.1097/BRS.0b013e318176100318475244
30.	L NaldiniU BlomerP GallayIn vivo gene delivery and stable transduction of nondividing cells by a lentiviral vectorScience272263267199610.1126/science.272.5259.2638602510
31.	T KafriU BlomerDA PetersonFH GageIM VermaSustained expression of genes delivered directly into liver and muscle by lentiviral vectorsNat Genet17314317199710.1038/ng1197-3149354796
32.	CA YoshidaT FuruichiT FujitaCore-binding factor beta interacts with Runx2 and is required for skeletal developmentNat Genet32633638200210.1038/ng101512434152
33.	X ZhangK TingCM BessetteNell-1, a key functional mediator of Runx2, partially rescues calvarial defects in Runx2(+/−) miceJ Bone Miner Res26777791201120939017
34.	YX ZhouX XuL ChenC LiSG BrodieCX DengA Pro250Arg substitution in mouse Fgfr1 causes increased expression of Cbfa1 and premature fusion of calvarial suturesHum Mol Genet920012008200010.1093/hmg/9.13.200110942429
35.	K ShirakabeK TerasawaK MiyamaH ShibuyaE NishidaRegulation of the activity of the transcription factor Runx2 by two homeobox proteins, Msx2 and Dlx5Genes Cells6851856200110.1046/j.1365-2443.2001.00466.x11683913