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Phosphorylated oligosaccharides of calcium (POs\u2010Ca), a novel calcium salt derived from potato starch, has recently attracted attention for its remineralization capabilities and potential to promote stem cell differentiation. Here, we investigated the impact of POs\u2010Ca on the osteogenic differentiation of hDPSCs and its underlying mechanism. Isolated hDPSCs were characterized via flow cytometry based on mesenchymal surface markers. Biocompatibility and osteogenic differentiation were assessed via Cell Counting Kit\u20108 (CCK\u20108) assay, alkaline phosphatase (ALP) activity, Alizarin Red S staining, and protein levels of osteogenic (Collagen I, DSPP, DMP1, and RUNX2). Intracellular Ca\n                    <jats:sup>2+<\/jats:sup>\n                    flux was monitored using Fluo\u20104 AM, while AMP\u2010activated protein kinase (AMPK) signaling and autophagic flux were analyzed by western blot (p\u2010AMPK, p\u2010ULK1, and light chain 3 (LC3)\u2010II), TEM, and LC3\u2010GFP imaging. Mechanistic studies employed verapamil (Ca\n                    <jats:sup>2+<\/jats:sup>\n                    channel blocker), Compound C (CC;AMPK inhibitor), and chloroquine (CQ;autophagy inhibitor). POs\u2010Ca (5\u2009mg\/mL) exhibited excellent biocompatibility and significantly promoted osteogenic differentiation, as evidenced by a 3.22\u2010fold increase in ALP activity and markedly enhanced mineralized nodule formation as shown by Alizarin Red S staining. Mechanistic studies revealed that POs\u2010Ca triggers rapid intracellular Ca\n                    <jats:sup>2+<\/jats:sup>\n                    influx, activating the AMPK pathway and inducing autophagic flux. Pharmacological inhibition established the essential causality of this cascade: verapamil abolished osteogenic enhancement, while CC and CQ suppressed ALP activity, mineralization, and osteogenic marker expression. Notably, CQ reciprocally attenuated POs\u2010Ca\u2010induced Ca\n                    <jats:sup>2+<\/jats:sup>\n                    influx, revealing novel bidirectional Ca\n                    <jats:sup>2+<\/jats:sup>\n                    \u2010autophagy crosstalk. In conclusion, POs\u2010Ca might promote hDPSCs osteogenesis via a calcium influx\u2010driven AMPK\u2010autophagy axis, providing a foundation for novel biomaterials that exploit physiological calcium signaling. These findings offer immediate translational potential for developing minimally invasive, cost\u2010effective strategies in dental pulp regeneration and bone defect repair.\n                  <\/jats:p>","DOI":"10.1155\/sci\/6150093","type":"journal-article","created":{"date-parts":[[2025,12,29]],"date-time":"2025-12-29T09:17:45Z","timestamp":1766999865000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["POs\u2010Ca Drives Osteogenic Differentiation of Human Dental Pulp Stem Cells Via AMPK\u2010Dependent Autophagy Activation and Reciprocal Calcium\u2010Autophagy Crosstalk"],"prefix":"10.1155","volume":"2025","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-2874-8697","authenticated-orcid":false,"given":"Jiayuan","family":"Zhang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2296-9205","authenticated-orcid":false,"given":"Yunqing","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuhei","family":"Hoshika","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chiharu","family":"Kawamoto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hidehiko","family":"Sano","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1638-4008","authenticated-orcid":false,"given":"Atsushi","family":"Tomokiyo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-3056-3265","authenticated-orcid":false,"given":"Jie","family":"Gao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,12,29]]},"reference":[{"key":"e_1_2_12_1_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21176172"},{"key":"e_1_2_12_2_2","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2021.654559"},{"key":"e_1_2_12_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s13577-020-00327-9"},{"key":"e_1_2_12_4_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms222112018"},{"key":"e_1_2_12_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.carbpol.2024.122401"},{"key":"e_1_2_12_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.colsurfb.2024.113775"},{"key":"e_1_2_12_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-96484-4"},{"key":"e_1_2_12_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-025-99979-6"},{"key":"e_1_2_12_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/jbm.a.36897"},{"key":"e_1_2_12_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.msec.2021.112466"},{"key":"e_1_2_12_11_2","doi-asserted-by":"publisher","DOI":"10.1002\/bab.2126"},{"key":"e_1_2_12_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cellsig.2020.109740"},{"key":"e_1_2_12_13_2","doi-asserted-by":"publisher","DOI":"10.3390\/ijms221910459"},{"key":"e_1_2_12_14_2","doi-asserted-by":"publisher","DOI":"10.3390\/molecules25215103"},{"key":"e_1_2_12_15_2","doi-asserted-by":"publisher","DOI":"10.5458\/jag.jag.JAG-2020_0001"},{"key":"e_1_2_12_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.archoralbio.2019.03.018"},{"key":"e_1_2_12_17_2","doi-asserted-by":"crossref","DOI":"10.2903\/j.efsa.2016.4488","article-title":"Nutrient Sources Added to F. 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