{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T10:02:29Z","timestamp":1784800949758,"version":"3.55.0"},"reference-count":51,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T00:00:00Z","timestamp":1784764800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Podophyllotoxin (PPT) inhibits tumors such as lung cancer and breast cancer. However, it has poor water solubility and causes gastrointestinal dysfunction and bone marrow suppression, which severely limit its clinical application. Based on the differential reactive oxygen species (ROS) levels between tumor microenvironments and normal tissues, we designed and constructed a ROS-responsive micelle delivery system, successfully fabricating blank micelles (M) and PPT-loaded micelles (M@PPT). Both micelles exhibited good particle size uniformity, colloidal stability, and biosafety. The ROS responsiveness experiment revealed that, after incubating blank micelles (M) with 10 mM H2O2, the particle size increased significantly, and the size distribution broadened. High-performance liquid chromatography (HPLC) confirmed the release of cinnamaldehyde from the micelles upon H2O2 exposure. Additionally, DCFH-DA assays demonstrated that treatment with blank micelles (M) enhanced intracellular ROS levels. In vitro release studies showed that drug-loaded micelles (M@PPT) achieved 77.76% cumulative PPT release within 24 h in a buffer containing 10 mM H2O2, significantly exceeding the release observed in the H2O2-free control group. These results collectively validate the ROS-responsive disintegration of micelles and the subsequent release of cinnamaldehyde. The liberated cinnamaldehyde further amplified intracellular ROS levels, establishing a positive feedback loop that accelerated drug release. Cellular assays revealed superior tumor growth inhibition by M@PPT over free PPT, mediated through apoptosis induction, G2\/M phase cell cycle arrest, downregulation of the anti-apoptotic protein Survivin, and upregulation of the p21 protein. In vivo studies further confirmed the enhanced antitumor efficacy and improved biosafety of M@PPT compared to free PPT. This ROS-responsive micellar system, by enabling tumor-targeted drug delivery and controlled release, provides a novel strategy to optimize the clinical utility of podophyllotoxin-based chemotherapeutics.<\/jats:p>","DOI":"10.3390\/ijms27156546","type":"journal-article","created":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T09:11:26Z","timestamp":1784797886000},"page":"6546","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["ROS-Responsive Micelles Loaded with Podophyllotoxin Inhibit Tumor Growth via ROS Self-Amplification and Regulation of Survivin and p21 Expression"],"prefix":"10.3390","volume":"27","author":[{"given":"Shuaiheng","family":"Song","sequence":"first","affiliation":[{"name":"Department of Organic Chemistry, College of Pharmacy, Harbin Medical University, Harbin 150081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiang","family":"Shao","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, College of Pharmacy, Harbin Medical University, Harbin 150081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siyi","family":"Liang","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, College of Pharmacy, Harbin Medical University, Harbin 150081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoyang","family":"Du","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, College of Pharmacy, Harbin Medical University, Harbin 150081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingnan","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, College of Pharmacy, Harbin Medical University, Harbin 150081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Guan","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, College of Pharmacy, Harbin Medical University, Harbin 150081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Harbin Medical University, Harbin 150081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, College of Pharmacy, Harbin Medical University, Harbin 150081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,23]]},"reference":[{"key":"ref_1","first-page":"47","article-title":"Cancer incidence and mortality in China, 2022","volume":"4","author":"Han","year":"2024","journal-title":"J. Natl. Cancer Cent."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e879","DOI":"10.1097\/MD.0000000000000879","article-title":"Is There a Survival Benefit in Patients with Stage IIIA (N2) Non-small Cell Lung Cancer Receiving Neoadjuvant Chemotherapy and\/or Radiotherapy Prior to Surgical Resection: A Systematic Review and Meta-analysis","volume":"94","author":"Xu","year":"2015","journal-title":"Medicine"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.jconrel.2021.10.027","article-title":"A modular ROS-responsive platform co-delivered by 10-hydroxycamptothecin and dexamethasone for cancer treatment","volume":"340","author":"Meng","year":"2021","journal-title":"J. Control Release"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1039\/C9BM01692A","article-title":"A bottlebrush-architectured dextran polyprodrug as an acidity-responsive vector for enhanced chemotherapy efficiency","volume":"8","author":"Zhang","year":"2020","journal-title":"Biomater. Sci."},{"key":"ref_5","first-page":"70","article-title":"The side effects of chemotherapeutic agents","volume":"19","author":"Carr","year":"2008","journal-title":"Curr. Anaesth. Crit. Care"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"812","DOI":"10.1016\/j.drudis.2006.07.005","article-title":"Exploiting the enhanced permeability and retention effect for tumor targeting","volume":"11","author":"Iyer","year":"2006","journal-title":"Drug Discov. Today"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2130","DOI":"10.1080\/10717544.2022.2094498","article-title":"Nano-drug co-delivery system of natural active ingredients and chemotherapy drugs for cancer treatment: A review","volume":"29","author":"Li","year":"2022","journal-title":"Drug Deliv."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.addr.2020.07.002","article-title":"Liposomes: Advancements and innovation in the manufacturing process","volume":"154\u2013155","author":"Shah","year":"2020","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1715","DOI":"10.1016\/j.carbpol.2010.10.032","article-title":"Synthesis and anti-hepatitis B virus activity of acyclovir conjugated stearic acid-g-chitosan oligosaccharide micelle","volume":"83","author":"Huang","year":"2011","journal-title":"Carbohydr. Polym."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1186\/s12951-024-02376-w","article-title":"Glioma-targeted oxaliplatin\/ferritin clathrate reversing the immunosuppressive microenvironment through hijacking Fe2+ and boosting Fenton reaction","volume":"22","author":"Li","year":"2024","journal-title":"J. Nanobiotechnol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e1908","DOI":"10.1002\/wnan.1908","article-title":"Magnetite-based Janus nanoparticles, their synthesis and biomedical applications","volume":"15","author":"Madadi","year":"2023","journal-title":"Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1247","DOI":"10.1111\/j.1349-7006.2011.01941.x","article-title":"Nanomedicine in cancer therapy: Innovative trends and prospects","volume":"102","author":"Blanco","year":"2011","journal-title":"Cancer Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"566","DOI":"10.1038\/s41563-019-0566-2","article-title":"The entry of nanoparticles into solid tumours","volume":"19","author":"Sindhwani","year":"2020","journal-title":"Nat. Mater."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"6985","DOI":"10.1021\/acsnano.5b01324","article-title":"Positron Emission Tomography Based Elucidation of the Enhanced Permeability and Retention Effect in Dogs with Cancer Using Copper-64 Liposomes","volume":"9","author":"Hansen","year":"2015","journal-title":"ACS Nano"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5757","DOI":"10.1016\/j.biomaterials.2009.07.020","article-title":"Amphiphilic multi-arm-block copolymer conjugated with doxorubicin via pH-sensitive hydrazone bond for tumor-targeted drug delivery","volume":"30","author":"Prabaharan","year":"2009","journal-title":"Biomaterials"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3040","DOI":"10.1021\/acs.biomac.6b00916","article-title":"Specific Cancer Cytosolic Drug Delivery Triggered by Reactive Oxygen Species-Responsive Micelles","volume":"17","author":"Yu","year":"2016","journal-title":"Biomacromolecules"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/j.actbio.2016.07.020","article-title":"Tunable conjugation densities of camptothecin on hyaluronic acid for tumor targeting and reduction-triggered release","volume":"43","author":"Chen","year":"2016","journal-title":"Acta Biomater."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.actbio.2017.02.047","article-title":"Enzyme-responsive peptide dendrimer-gemcitabine conjugate as a controlled-release drug delivery vehicle with enhanced antitumor efficacy","volume":"55","author":"Zhang","year":"2017","journal-title":"Acta Biomater."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e2103836","DOI":"10.1002\/advs.202103836","article-title":"Tumor-Microenvironment-Responsive Nanomedicine for Enhanced Cancer Immunotherapy","volume":"9","author":"Peng","year":"2022","journal-title":"Adv. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Cong, X., Chen, J., and Xu, R. (2022). Recent Progress in Bio-Responsive Drug Delivery Systems for Tumor Therapy. Front. Bioeng. Biotechnol., 10.","DOI":"10.3389\/fbioe.2022.916952"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"9052","DOI":"10.1021\/acs.jafc.0c03856","article-title":"ROS-Responsive Polymeric Micelle for Improving Pesticides Efficiency and Intelligent Release","volume":"68","author":"Li","year":"2020","journal-title":"J. Agric. Food Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"115787","DOI":"10.1016\/j.carbpol.2019.115787","article-title":"Novel dual ROS-sensitive and CD44 receptor targeting nanomicelles based on oligomeric hyaluronic acid for the efficient therapy of atherosclerosis","volume":"232","author":"Hou","year":"2020","journal-title":"Carbohydr. Polym."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"65","DOI":"10.2147\/IJN.S230237","article-title":"A pH-Responsive Charge-Reversal Drug Delivery System with Tumor-Specific Drug Release and ROS Generation for Cancer Therapy","volume":"15","author":"Xu","year":"2020","journal-title":"Int. J. Nanomed."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1039\/C8NH00417J","article-title":"DePEGylation strategies to increase cancer nanomedicine efficacy","volume":"4","author":"Kong","year":"2019","journal-title":"Nanoscale Horiz."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Shah, Z., Gohar, U.F., Jamshed, I., Mushtaq, A., Mukhtar, H., Zia-Ui-Haq, M., Toma, S.I., Manea, R., Moga, M., and Popovici, B. (2021). Podophyllotoxin: History, Recent Advances and Future Prospects. Biomolecules, 11.","DOI":"10.3390\/biom11040603"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2361","DOI":"10.1080\/10717544.2021.1995076","article-title":"ROS-Activated homodimeric podophyllotoxin nanomedicine with self-accelerating drug release for efficient cancer eradication","volume":"28","author":"Liang","year":"2021","journal-title":"Drug Deliv."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1080\/10717544.2021.1905750","article-title":"PH and ROS sequentially responsive podophyllotoxin prodrug micelles with surface charge-switchable and self-amplification drug release for combating multidrug resistance cancer","volume":"28","author":"Li","year":"2021","journal-title":"Drug Deliv."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Miranda-Vera, C., Hern\u00e1ndez, \u00c1.-P., Garc\u00eda-Garc\u00eda, P., D\u00edez, D., Garc\u00eda, P.A., and Castro, M.\u00c1. (2025). Bioconjugation of Podophyllotoxin and Nanosystems: Approaches for Boosting Its Biopharmaceutical and Antitumoral Profile. Pharmaceuticals, 18.","DOI":"10.3390\/ph18020169"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1021\/bm010042v","article-title":"Cytotoxicity of a unimolecular polymeric micelle and its degradation products","volume":"2","author":"Schmalenberg","year":"2001","journal-title":"Biomacromolecules"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.ejps.2015.12.031","article-title":"Recent advances in polymeric micelles for anti-cancer drug delivery","volume":"83","author":"Biswas","year":"2016","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"116730","DOI":"10.1016\/j.carbpol.2020.116730","article-title":"Inulin-pluronic-stearic acid based double folded nanomicelles for pH-responsive delivery of resveratrol","volume":"247","author":"Jangid","year":"2020","journal-title":"Carbohydr. Polym."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1517\/17425247.2012.689284","article-title":"Micelle-like nanoassemblies based on polymer-drug conjugates as an emerging platform for drug delivery","volume":"9","author":"Liu","year":"2012","journal-title":"Expert Opin. Drug Deliv."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"115072","DOI":"10.1016\/j.carbpol.2019.115072","article-title":"PH-sensitive amphiphilic chitosan-quercetin conjugate for intracellular delivery of doxorubicin enhancement","volume":"223","author":"Mu","year":"2019","journal-title":"Carbohydr. Polym."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3858","DOI":"10.1016\/j.biomaterials.2013.01.101","article-title":"Cellular uptake, intracellular trafficking, and antitumor efficacy of doxorubicin-loaded reduction-sensitive micelles","volume":"34","author":"Cui","year":"2013","journal-title":"Biomaterials"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e1595","DOI":"10.1002\/wsbm.1595","article-title":"Intracellular and microenvironmental regulation of mitochondrial membrane potential in cancer cells","volume":"15","author":"Begum","year":"2023","journal-title":"WIREs Mech. Dis."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"113935","DOI":"10.1016\/j.jconrel.2025.113935","article-title":"Structural stability and RNase resistance of mRNA Polyplex micelles for systemic delivery","volume":"384","author":"Dirisala","year":"2025","journal-title":"J. Control. Release"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"100271","DOI":"10.1016\/j.xcrp.2020.100271","article-title":"Mechanistic Investigation on Oxidative Degradation of ROS-Responsive Thioacetal\/Thioketal Moieties and Their Implications","volume":"1","author":"Liu","year":"2020","journal-title":"Cell Rep. Phys. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1016\/j.freeradbiomed.2008.10.025","article-title":"The cinnamon-derived Michael acceptor cinnamic aldehyde impairs melanoma cell proliferation, invasiveness, and tumor growth","volume":"46","author":"Cabello","year":"2009","journal-title":"Free Radic. Biol. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"121795","DOI":"10.1016\/j.biomaterials.2022.121795","article-title":"Cinnamaldehyde-based poly(thioacetal): A ROS-awakened self-amplifying degradable polymer for enhanced cancer immunotherapy","volume":"289","author":"Tu","year":"2022","journal-title":"Biomaterials"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1362","DOI":"10.1021\/acs.biomac.4c01731","article-title":"Cinnamaldehyde-Based ROS-Responsive Polymeric Gene Vectors for Efficient Gene Delivery and Tumor Cell Growth Inhibition","volume":"26","author":"Zhang","year":"2025","journal-title":"Biomacromolecules"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"5516","DOI":"10.1021\/acsapm.4c00779","article-title":"Self-Augmented Reactive Oxygen Species-Responsive Nanoformulation with Efficient Curcumin Delivery for Inhibiting Triple-Negative Breast Cancer Cell Growth and Migration","volume":"6","author":"Li","year":"2024","journal-title":"ACS Appl. Polym. Mater."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1007\/s11051-016-3583-y","article-title":"Factors affecting the stability of drug-loaded polymeric micelles and strategies for improvement","volume":"18","author":"Zhou","year":"2016","journal-title":"J. Nanopart Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"114622","DOI":"10.1016\/j.ejmech.2022.114622","article-title":"Intelligent triggering of nanomicelles based on a ROS-activated anticancer prodrug and photodynamic therapy (PDT)-synergistic therapy for lung cancers","volume":"241","author":"Chen","year":"2022","journal-title":"Eur. J. Med. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"115930","DOI":"10.1016\/j.carbpol.2020.115930","article-title":"In vitro\/vivo evaluation of novel mitochondrial targeting charge-reversal polysaccharide-based antitumor nanoparticle","volume":"234","author":"Fang","year":"2020","journal-title":"Carbohydr. Polym."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"6887","DOI":"10.1021\/acsnano.0c01018","article-title":"Nanoparticles Derived from the Natural Antioxidant Rosmarinic Acid Ameliorate Acute Inflammatory Bowel Disease","volume":"14","author":"Chung","year":"2020","journal-title":"ACS Nano"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/j.colsurfb.2016.08.042","article-title":"Influence of the graft density of hydrophobic groups on thermo-responsive nanoparticles for anti-cancer drugs delivery","volume":"148","author":"Wang","year":"2016","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e2304091","DOI":"10.1002\/adhm.202304091","article-title":"Fenton-like Reaction Inspired \u201c\u00b7OH Catalyzed\u201d Osteogenic Process for the Treatment of Osteoporosis","volume":"13","author":"Wang","year":"2024","journal-title":"Adv. Healthc. Mater."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.ijpharm.2014.05.010","article-title":"Polymeric micelles with citraconic amide as pH-sensitive bond in backbone for anticancer drug delivery","volume":"471","author":"Cao","year":"2014","journal-title":"Int. J. Pharm."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1016\/j.ijbiomac.2022.03.139","article-title":"Tumor-targeted hyaluronic acid-based oxidative stress nanoamplifier with ROS generation and GSH depletion for antitumor therapy","volume":"207","author":"Liu","year":"2022","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3109\/02652048.2011.621551","article-title":"In vitro antitumour activity of stearic acid-g-chitosan oligosaccharide polymeric micelles loading podophyllotoxin","volume":"29","author":"Huang","year":"2012","journal-title":"J. Microencapsul."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.ejmech.2017.03.011","article-title":"Synthesis and biological evaluation of novel podophyllotoxin-NSAIDs conjugates as multifunctional anti-MDR agents against resistant human hepatocellular carcinoma Bel-7402\/5-FU cells","volume":"131","author":"Zhang","year":"2017","journal-title":"Eur. J. Med. Chem."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/15\/6546\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T09:14:57Z","timestamp":1784798097000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/15\/6546"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,23]]},"references-count":51,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["ijms27156546"],"URL":"https:\/\/doi.org\/10.3390\/ijms27156546","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,23]]}}}