{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T14:07:27Z","timestamp":1790431647029,"version":"4.1.0"},"reference-count":36,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2021,1,15]],"date-time":"2021-01-15T00:00:00Z","timestamp":1610668800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["24593117 and 18K09570"],"award-info":[{"award-number":["24593117 and 18K09570"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Dentistry Journal"],"abstract":"<jats:p>Periodontal disease is a chronic inflammation of tooth-supporting tissues, and the destruction of these tissues results in tooth loss. Regeneration of periodontal tissues is the ultimate goal of periodontal treatment. We previously reported that transplantation of conditioned medium (CM) of periodontal ligament stem cells (PDLSCs) demonstrated the enhancement of periodontal tissue regeneration, compared to CM from fibroblasts (Fibroblast-CM). We hypothesized that the angiogenic effects of PDLSC-CM might participate in the enhanced wound healing of periodontal tissues. The aim of this study was to investigate the effect of PDLSC-CM on the functions of endothelial cells. PDLSCs were cultured from periodontal ligament tissues obtained from healthy volunteers. Human gingival epithelial cells, dermal fibroblasts, osteoblasts, and umbilical vein endothelial cells (HUVECs) were purchased from commercial sources. The functions of endothelial cells were examined using immunostaining of Ki67, observation of nuclear fragmentation and condensation (apoptosis), and network formation on Matrigel. Vascular endothelial cell growth factor (VEGF) level was measured using an ELISA kit. HUVECs demonstrated higher cell viability in PDLSC-CM when compared with those in Fibroblast-CM. HUVECs demonstrated a higher number of Ki67-positive cells and lower apoptosis cells in PDLSC-CM, compared to Fibroblast-CM. Additionally, HUVECs formed more capillary-like structures in PDLSC-CM than Fibroblast-CM. PDLSC-CM contained higher levels of angiogenic growth factor, VEGF, than Fibroblast-CM. Our results showed that PDLSC-CM increased cell viability, proliferation, and capillary formation of HUVECs compared to Fibroblast-CM, suggesting the angiogenic effects of PDLSC-CM, and the effect is a potential regenerative mechanism of periodontal tissues by PDLSC-CM.<\/jats:p>","DOI":"10.3390\/dj9010009","type":"journal-article","created":{"date-parts":[[2021,1,15]],"date-time":"2021-01-15T12:44:05Z","timestamp":1610714645000},"page":"9","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Angiogenic Effects of Secreted Factors from Periodontal Ligament Stem Cells"],"prefix":"10.3390","volume":"9","author":[{"given":"Kengo","family":"Iwasaki","sequence":"first","affiliation":[{"name":"Institute of Dental Research, Osaka Dental University, Osaka 573-1121, Japan"},{"name":"Department of Nanomedicine (DNP), Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo 113-8510, Japan"},{"name":"Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo 113-8510, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keiko","family":"Akazawa","sequence":"additional","affiliation":[{"name":"Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo 113-8510, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3874-2846","authenticated-orcid":false,"given":"Mizuki","family":"Nagata","sequence":"additional","affiliation":[{"name":"Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo 113-8510, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Motohiro","family":"Komaki","sequence":"additional","affiliation":[{"name":"Department of Nanomedicine (DNP), Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo 113-8510, Japan"},{"name":"Yokohama Clinic, Kanagawa Dental University, Yokohama Clinic, Kanagawa, Yokohama 221-0835, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yihao","family":"Peng","sequence":"additional","affiliation":[{"name":"Graduate School of Dentistry, Department of Periodontology, Osaka Dental University, Osaka 573-1121, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Makoto","family":"Umeda","sequence":"additional","affiliation":[{"name":"Department of Periodontology, Osaka Dental University, Osaka 573-1121, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5836-1309","authenticated-orcid":false,"given":"Tetsuro","family":"Watabe","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo 113-8510, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ikuo","family":"Morita","sequence":"additional","affiliation":[{"name":"Ochanomizu University, Tokyo 112-8610, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,1,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1111\/prd.12297","article-title":"The role of inflammation and genetics in periodontal disease","volume":"83","author":"Loos","year":"2020","journal-title":"Periodontol. 2000"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1809","DOI":"10.1016\/S0140-6736(05)67728-8","article-title":"Periodontal diseases","volume":"366","author":"Pihlstrom","year":"2005","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1111\/prd.12331","article-title":"Current understanding of periodontal disease pathogenesis and targets for host-modulation therapy","volume":"81","author":"Hajishengallis","year":"2020","journal-title":"Periodontol. 2000"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1053","DOI":"10.1177\/0022034519846113","article-title":"Periodontal Medicine: 100 Years of Progress","volume":"98","author":"Beck","year":"2019","journal-title":"J. Dent. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1034\/j.1600-0757.2000.2230101.x","article-title":"Periodontal medicine: The emergence of a new branch of periodontology","volume":"23","author":"Williams","year":"2000","journal-title":"Periodontol. 2000"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1111\/j.1600-0757.2009.00317.x","article-title":"Does periodontal tissue regeneration really work?","volume":"51","author":"Bosshardt","year":"2009","journal-title":"Periodontol. 2000"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"S105","DOI":"10.1902\/jop.2015.140378","article-title":"Periodontal regeneration-intrabony defects: A consensus report from the AAP Regeneration Workshop","volume":"86","author":"Reynolds","year":"2015","journal-title":"J. Periodontol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Esposito, M., Grusovin, M.G., Papanikolaou, N., Coulthard, P., and Worthington, H.V. (2009). Enamel matrix derivative (Emdogain(R)) for periodontal tissue regeneration in intrabony defects. Cochrane Database Syst. Rev., CD003875.","DOI":"10.1002\/14651858.CD003875.pub3"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Needleman, I.G., Worthington, H.V., Giedrys-Leeper, E., and Tucker, R.J. (2006). Guided tissue regeneration for periodontal infra-bony defects. Cochrane Database Syst. Rev., CD001724.","DOI":"10.1002\/14651858.CD001724.pub2"},{"key":"ref_10","first-page":"393","article-title":"The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells","volume":"3","author":"Friedenstein","year":"1970","journal-title":"Cell Tissue Kinet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1126\/science.284.5411.143","article-title":"Multilineage potential of adult human mesenchymal stem cells","volume":"284","author":"Pittenger","year":"1999","journal-title":"Science"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1126\/science.276.5309.71","article-title":"Marrow stromal cells as stem cells for nonhematopoietic tissues","volume":"276","author":"Prockop","year":"1997","journal-title":"Science"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/S0140-6736(04)16627-0","article-title":"Investigation of multipotent postnatal stem cells from human periodontal ligament","volume":"364","author":"Seo","year":"2004","journal-title":"Lancet"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1111\/j.1600-0765.2005.00799.x","article-title":"Application of periodontal ligament cell sheet for periodontal regeneration: A pilot study in beagle dogs","volume":"40","author":"Akizuki","year":"2005","journal-title":"J. Periodontal Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1002\/jcp.25067","article-title":"Potential for Stem Cell-Based Periodontal Therapy","volume":"231","author":"Bassir","year":"2016","journal-title":"J. Cell. Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1111\/jre.12205","article-title":"Periodontal ligament-derived cells for periodontal regeneration in animal models: A systematic review","volume":"50","author":"Bright","year":"2015","journal-title":"J. Periodontal Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1186\/s13287-016-0288-1","article-title":"Treatment of periodontal intrabony defects using autologous periodontal ligament stem cells: A randomized clinical trial","volume":"7","author":"Chen","year":"2016","journal-title":"Stem Cell Res. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1089\/ten.2005.11.469","article-title":"Human periodontal ligament cell sheets can regenerate periodontal ligament tissue in an athymic rat model","volume":"11","author":"Hasegawa","year":"2005","journal-title":"Tissue Eng."},{"key":"ref_19","first-page":"693","article-title":"Periodontal regeneration using periodontal ligament stem cell-transferred amnion","volume":"20","author":"Iwasaki","year":"2014","journal-title":"Tissue Eng. Part A"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Iwasaki, K., Akazawa, K., Nagata, M., Komaki, M., Honda, I., Morioka, C., Yokoyama, N., Ayame, H., Yamaki, K., and Tanaka, Y. (2019). The Fate of Transplanted Periodontal Ligament Stem Cells in Surgically Created Periodontal Defects in Rats. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20010192"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1089\/ten.tea.2016.0274","article-title":"Conditioned Medium from Periodontal Ligament Stem Cells Enhances Periodontal Regeneration","volume":"23","author":"Nagata","year":"2017","journal-title":"Tissue Eng. Part A"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"28126","DOI":"10.1038\/srep28126","article-title":"The use of platelet-rich fibrin combined with periodontal ligament and jaw bone mesenchymal stem cell sheets for periodontal tissue engineering","volume":"6","author":"Wang","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1186\/s13287-017-0731-y","article-title":"Adult human periodontal ligament-derived stem cells delay retinal degeneration and maintain retinal function in RCS rats","volume":"8","author":"Huang","year":"2017","journal-title":"Stem Cell Res. Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"585530","DOI":"10.3389\/fimmu.2020.585530","article-title":"Maresin-1 and Resolvin E1 Promote Regenerative Properties of Periodontal Ligament Stem Cells Under Inflammatory Conditions","volume":"11","author":"Schulte","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1016\/j.archoralbio.2010.10.013","article-title":"Comparison of characteristics of periodontal ligament cells obtained from outgrowth and enzyme-digested culture methods","volume":"56","author":"Tanaka","year":"2011","journal-title":"Arch. Oral Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1002\/term.366","article-title":"Cell-printing and transfer technology applications for bone defects in mice","volume":"5","author":"Tsugawa","year":"2011","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2239","DOI":"10.21873\/anticanres.11560","article-title":"Detection and Quantification of Nuclear Morphology Changes in Apoptotic Cells by Fluorescence Microscopy and Subsequent Analysis of Visualized Fluorescent Signals","volume":"37","author":"Mandelkow","year":"2017","journal-title":"Anticancer Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"H1106","DOI":"10.1152\/ajpheart.00265.2005","article-title":"Endothelial microparticles affect angiogenesis in vitro: Role of oxidative stress","volume":"289","author":"Mezentsev","year":"2005","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"990","DOI":"10.1681\/ASN.2015030286","article-title":"Acute Dialysis Quality Initiative (ADQI) XIII Work Group Targeting Endogenous Repair Pathways after AKI","volume":"27","author":"Humphreys","year":"2016","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"602597","DOI":"10.1155\/2015\/602597","article-title":"Mesenchymal Stromal Cell Therapy in Ischemia\/Reperfusion Injury","volume":"2015","author":"Rowart","year":"2015","journal-title":"J. Immunol. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1425","DOI":"10.1902\/jop.2012.120547","article-title":"Periodontal ligament stem cells possess the characteristics of pericytes","volume":"84","author":"Iwasaki","year":"2013","journal-title":"J. Periodontol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"972313","DOI":"10.1155\/2015\/972313","article-title":"Periodontal ligament stem cells: Current status, concerns, and future prospects","volume":"2015","author":"Zhu","year":"2015","journal-title":"Stem Cells Int."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1248","DOI":"10.1016\/j.cell.2019.01.021","article-title":"VEGF in Signaling and Disease: Beyond Discovery and Development","volume":"176","author":"Apte","year":"2019","journal-title":"Cell"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Karaman, S., Lepp\u00e4nen, V.-M., and Alitalo, K. (2018). Vascular endothelial growth factor signaling in development and disease. Development, 145.","DOI":"10.1242\/dev.151019"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"386","DOI":"10.14348\/molcells.2017.0004","article-title":"The Significance of SDF-1\u03b1-CXCR4 Axis in in vivo Angiogenic Ability of Human Periodontal Ligament Stem Cells","volume":"40","author":"Bae","year":"2017","journal-title":"Mol. Cells"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1188","DOI":"10.1089\/ten.tea.2013.0512","article-title":"Stem cells derived from tooth periodontal ligament enhance functional angiogenesis by endothelial cells","volume":"20","author":"Yeasmin","year":"2014","journal-title":"Tissue Eng. Part A"}],"container-title":["Dentistry Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2304-6767\/9\/1\/9\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:11:32Z","timestamp":1760159492000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2304-6767\/9\/1\/9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,15]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,1]]}},"alternative-id":["dj9010009"],"URL":"https:\/\/doi.org\/10.3390\/dj9010009","relation":{},"ISSN":["2304-6767"],"issn-type":[{"value":"2304-6767","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,1,15]]}}}