{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:45:34Z","timestamp":1792806334888,"version":"4.5.0"},"reference-count":88,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2021,11,3]],"date-time":"2021-11-03T00:00:00Z","timestamp":1635897600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006601","name":"Ministero degli Affari Esteri e della Cooperazione Internazionale","doi-asserted-by":"publisher","award":["MAE00691612020-06-26"],"award-info":[{"award-number":["MAE00691612020-06-26"]}],"id":[{"id":"10.13039\/501100006601","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010767","name":"Innovative Medicines Initiative","doi-asserted-by":"publisher","award":["807015"],"award-info":[{"award-number":["807015"]}],"id":[{"id":"10.13039\/501100010767","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Creutzfeldt\u2013Jakob disease Foundation","award":["000"],"award-info":[{"award-number":["000"]}],"id":[{"id":"https:\/\/ror.org\/05d2ptg05","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nanomaterials"],"abstract":"<jats:p>Enzymes, as natural and potentially long-term treatment options, have become one of the most sought-after pharmaceutical molecules to be delivered with nanoparticles (NPs); however, their instability during formulation often leads to underwhelming results. Various molecules, including the Tween\u00ae polysorbate series, have demonstrated enzyme activity protection but are often used uncontrolled without optimization. Here, poly(lactic-co-glycolic) acid (PLGA) NPs loaded with \u03b2-glucosidase (\u03b2-Glu) solutions containing Tween\u00ae 20, 60, or 80 were compared. Mixing the enzyme with Tween\u00ae pre-formulation had no effect on particle size or physical characteristics, but increased the amount of enzyme loaded. More importantly, NPs made with Tween\u00ae 20:enzyme solutions maintained significantly higher enzyme activity. Therefore, Tween\u00ae 20:enzyme solutions ranging from 60:1 to 2419:1 mol:mol were further analyzed. Isothermal titration calorimetry analysis demonstrated low affinity and unquantifiable binding between Tween\u00ae 20 and \u03b2-Glu. Incorporating these solutions in NPs showed no effect on size, zeta potential, or morphology. The amount of enzyme and Tween\u00ae 20 in the NPs was constant for all samples, but a trend towards higher activity with higher molar rapports of Tween\u00ae 20:\u03b2-Glu was observed. Finally, a burst release from NPs in the first hour with Tween\u00ae:\u03b2-Glu solutions was the same as free enzyme, but the enzyme remained active longer in solution. These results highlight the importance of stabilizers during NP formulation and how optimizing their use to stabilize an enzyme can help researchers design more efficient and effective enzyme loaded NPs.<\/jats:p>","DOI":"10.3390\/nano11112946","type":"journal-article","created":{"date-parts":[[2021,11,3]],"date-time":"2021-11-03T21:57:49Z","timestamp":1635976669000},"page":"2946","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":31,"title":["Tween\u00ae Preserves Enzyme Activity and Stability in PLGA Nanoparticles"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2204-1981","authenticated-orcid":false,"given":"Jason Thomas","family":"Duskey","sequence":"first","affiliation":[{"name":"Te.Far.T.I.-Nanotech Lab, Department of Life Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2135-2422","authenticated-orcid":false,"given":"Ilaria","family":"Ottonelli","sequence":"additional","affiliation":[{"name":"Te.Far.T.I.-Nanotech Lab, Department of Life Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy"},{"name":"Clinical and Experimental Medicine PhD Program, Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arianna","family":"Rinaldi","sequence":"additional","affiliation":[{"name":"Te.Far.T.I.-Nanotech Lab, Department of Life Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy"},{"name":"Clinical and Experimental Medicine PhD Program, Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irene","family":"Parmeggiani","sequence":"additional","affiliation":[{"name":"Te.Far.T.I.-Nanotech Lab, Department of Life Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3876-0051","authenticated-orcid":false,"given":"Barbara","family":"Zambelli","sequence":"additional","affiliation":[{"name":"Laboratory of Bioinorganic Chemistry, Department of Pharmacy and Biotechnology, University of Bologna, 40127 Bologna, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leon Z.","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert K.","family":"Prud\u2019homme","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Angela","family":"Vandelli","sequence":"additional","affiliation":[{"name":"Te.Far.T.I.-Nanotech Lab, Department of Life Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8318-667X","authenticated-orcid":false,"given":"Giovanni","family":"Tosi","sequence":"additional","affiliation":[{"name":"Te.Far.T.I.-Nanotech Lab, Department of Life Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2400-9910","authenticated-orcid":false,"given":"Barbara","family":"Ruozi","sequence":"additional","affiliation":[{"name":"Te.Far.T.I.-Nanotech Lab, Department of Life Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/bs.irn.2017.08.006","article-title":"Current Strategies for the Delivery of Therapeutic Proteins and Enzymes to Treat Brain Disorders","volume":"137","author":"Duskey","year":"2017","journal-title":"Int. Rev. Neurobiol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Rigon, L., Salvalaio, M., Pederzoli, F., Legnini, E., Duskey, J.T., D\u2019Avanzo, F., De Filippis, C., Ruozi, B., Marin, O., and Vandelli, M.A. (2019). Targeting Brain Disease in MPSII: Preclinical Evaluation of IDS-Loaded PLGA Nanoparticles. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20082014"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"205","DOI":"10.2217\/nnm-2019-0367","article-title":"Drug Delivery across the Blood\u2013Brain Barrier: Recent Advances in the Use of Nanocarriers","volume":"15","author":"Mulvihill","year":"2020","journal-title":"Nanomedicine"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1080\/17425247.2020.1698544","article-title":"Nanoparticles as Carriers for Drug Delivery of Macromolecules across the Blood-Brain Barrier","volume":"17","author":"Tosi","year":"2020","journal-title":"Expert Opin. Drug Deliv."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e191","DOI":"10.3928\/19382359-20180424-01","article-title":"Enzyme Replacement Therapy: A Review and Its Role in Treating Lysosomal Storage Diseases","volume":"47","author":"Li","year":"2018","journal-title":"Pediatric Ann."},{"key":"ref_6","first-page":"64","article-title":"Long-Term Cognitive and Somatic Outcomes of Enzyme Replacement Therapy in Untransplanted Hurler Syndrome","volume":"13","author":"Eisengart","year":"2017","journal-title":"Mol. Genet. Metab. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s11011-017-0111-4","article-title":"How Close Are We to Therapies for Sanfilippo Disease?","volume":"33","author":"Gaffke","year":"2018","journal-title":"Metab. Brain Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.omtm.2017.01.004","article-title":"Chronic Enzyme Replacement to the Brain of a Late Infantile Neuronal Ceroid Lipofuscinosis Mouse Has Differential Effects on Phenotypes of Disease","volume":"4","author":"Wiseman","year":"2017","journal-title":"Mol. Ther.\u2014Methods Clin. Dev."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1663","DOI":"10.3389\/fphys.2018.01663","article-title":"New Approaches to Tay-Sachs Disease Therapy","volume":"9","author":"Solovyeva","year":"2018","journal-title":"Front. Physiol"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.colsurfb.2009.09.001","article-title":"Biodegradable Polymeric Nanoparticles Based Drug Delivery Systems","volume":"75","author":"Kumari","year":"2010","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1007\/s10545-006-0239-6","article-title":"Limitations of Enzyme Replacement Therapy: Current and Future","volume":"29","author":"Wraith","year":"2006","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1186\/s13052-018-0562-1","article-title":"Enzyme Replacement Therapy: Efficacy and Limitations","volume":"44","author":"Concolino","year":"2018","journal-title":"Ital. J. Pediatrics"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1007\/s10545-017-0032-8","article-title":"Enzyme Replacement Therapy and beyond\u2014in Memoriam Roscoe O. Brady, M.D. (1923\u20132016)","volume":"40","author":"Ries","year":"2017","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Nelemans, L.C., and Gurevich, L. (2020). Drug Delivery with Polymeric Nanocarriers\u2014Cellular Uptake Mechanisms. Materials, 13.","DOI":"10.3390\/ma13020366"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Alven, S., and Aderibigbe, B.A. (2020). Efficacy of Polymer-Based Nanocarriers for Co-Delivery of Curcumin and Selected Anticancer Drugs. Nanomaterials, 10.","DOI":"10.3390\/nano10081556"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2939","DOI":"10.1002\/pat.5031","article-title":"Polymeric Nanocarriers in Targeted Drug Delivery Systems: A Review","volume":"31","author":"Abasian","year":"2020","journal-title":"Polym. Adv. Technol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1016\/j.jksus.2017.10.004","article-title":"Morphologies and Functionalities of Polymeric Nanocarriers as Chemical Tools for Drug Delivery: A Review","volume":"31","author":"Venditti","year":"2019","journal-title":"J. King Saud Univ.-Sci."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Avramovi\u0107, N., Mandi\u0107, B., Savi\u0107-Radojevi\u0107, A., and Simi\u0107, T. (2020). Polymeric Nanocarriers of Drug Delivery Systems in Cancer Therapy. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12040298"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Duskey, J.T., Baraldi, C., Gamberini, M.C., Ottonelli, I., Da Ros, F., Tosi, G., Forni, F., Vandelli, M.A., and Ruozi, B. (2020). Investigating Novel Syntheses of a Series of Unique Hybrid PLGA-Chitosan Polymers for Potential Therapeutic Delivery Applications. Polymers, 12.","DOI":"10.3390\/polym12040823"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Oddone, N., Boury, F., Garcion, E., Grabrucker, A.M., Martinez, M.C., Da Ros, F., Janaszewska, A., Forni, F., Vandelli, M.A., and Tosi, G. (2020). Synthesis, Characterization, and In Vitro Studies of an Reactive Oxygen Species (ROS)-Responsive Methoxy Polyethylene Glycol-Thioketal-Melphalan Prodrug for Glioblastoma Treatment. Front. Pharmacol., 11.","DOI":"10.3389\/fphar.2020.00574"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1016\/j.ejps.2017.07.020","article-title":"Anticancer Drug-Loaded Quantum Dots Engineered Polymeric Nanoparticles: Diagnosis\/Therapy Combined Approach","volume":"107","author":"Belletti","year":"2017","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3451","DOI":"10.1039\/C6PY00343E","article-title":"Engineered Non-Toxic Cationic Nanocarriers with Photo-Triggered Slow-Release Properties","volume":"7","author":"Dinu","year":"2016","journal-title":"Polym. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1016\/j.jconrel.2020.12.051","article-title":"Insights into Kinetics, Release, and Behavioral Effects of Brain-Targeted Hybrid Nanoparticles for Cholesterol Delivery in Huntington\u2019s Disease","volume":"330","author":"Birolini","year":"2021","journal-title":"J. Control. Release"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"102081","DOI":"10.1016\/j.jddst.2020.102081","article-title":"Development and Biocompatibility Assessments of Poly(3-Hydroxybutyrate-Co-\u03b5-Caprolactone) Microparticles for Diclofenac Sodium Delivery","volume":"60","author":"Musumeci","year":"2020","journal-title":"J. Drug Deliv. Sci. Technol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Pederzoli, F., Ruozi, B., Duskey, J., Hagmeyer, S., Sauer, A.K., Grabrucker, S., Coelho, R., Oddone, N., Ottonelli, I., and Daini, E. (2019). Nanomedicine against A\u03b2 Aggregation by \u03b2\u2013Sheet Breaker Peptide Delivery: In Vitro Evidence. Pharmaceutics, 11.","DOI":"10.3390\/pharmaceutics11110572"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2135","DOI":"10.1021\/acsbiomaterials.9b01794","article-title":"Encapsulation and Storage of Therapeutic Fibrin-Homing Peptides Using Conducting Polymer Nanoparticles for Programmed Release by Electrical Stimulation","volume":"6","year":"2020","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1097","DOI":"10.1021\/bm500169p","article-title":"Drug Carriers for the Delivery of Therapeutic Peptides","volume":"15","author":"Du","year":"2014","journal-title":"Biomacromolecules"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jconrel.2021.04.006","article-title":"Morphologic Design of Sugar-Based Polymer Nanoparticles for Delivery of Antidiabetic Peptides","volume":"334","author":"Elsabahy","year":"2021","journal-title":"J. Control. Release"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/bs.pbr.2019.03.001","article-title":"Nanomedicine in Alzheimer\u2019s Disease: Amyloid Beta Targeting Strategy","volume":"245","author":"Tosi","year":"2019","journal-title":"Prog. Brain Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"12066","DOI":"10.1021\/acs.jpcb.5b06637","article-title":"PH-Triggered Reversible Multiple Protein\u2013Polymer Conjugation Based on Molecular Recognition","volume":"119","author":"Liu","year":"2015","journal-title":"J. Phys. Chem. B"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"430","DOI":"10.3109\/02652048.2011.576786","article-title":"Insulin-Loaded PLGA Microparticles: Flow Focusing versus Double Emulsion\/Solvent Evaporation","volume":"28","author":"Holgado","year":"2011","journal-title":"J. Microencapsul."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"935","DOI":"10.1021\/acs.biomac.5b01614","article-title":"Stimuli-Responsive Codelivery of Oligonucleotides and Drugs by Self-Assembled Peptide Nanoparticles","volume":"17","author":"Sigg","year":"2016","journal-title":"Biomacromolecules"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1600046","DOI":"10.1002\/advs.201600046","article-title":"Chaperonin\u2013Dendrimer Conjugates for SiRNA Delivery","volume":"3","author":"Nussbaumer","year":"2016","journal-title":"Adv. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1007\/s11671-009-9345-3","article-title":"Preparation and Characterization of Cationic PLA-PEG Nanoparticles for Delivery of Plasmid DNA","volume":"4","author":"Zou","year":"2009","journal-title":"Nanoscale Res. Lett."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1002\/wnan.73","article-title":"New Biotechnological and Nanomedicine Strategies for Treatment of Lysosomal Storage Disorders","volume":"2","author":"Muro","year":"2010","journal-title":"Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1016\/j.colsurfb.2017.06.033","article-title":"Development of Double Emulsion Nanoparticles for the Encapsulation of Bovine Serum Albumin","volume":"158","author":"Martinez","year":"2017","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_37","first-page":"38","article-title":"Polymeric Nanoparticles: Review of Synthesis Methods and Applications in Drug Delivery","volume":"1","author":"Jahangiri","year":"2018","journal-title":"J. Adv. Chem. Pharm. Mater. (JACPM)"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1481","DOI":"10.5958\/0974-360X.2019.00245.2","article-title":"A Comprehensive Review on PLGA-Based Nanoparticles Used for Rheumatoid Arthritis","volume":"12","author":"Mohanty","year":"2019","journal-title":"Rese. J. Pharm. Technol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1692","DOI":"10.1002\/jbm.b.33648","article-title":"Polymer Degradation and Drug Delivery in PLGA-Based Drug\u2013Polymer Applications: A Review of Experiments and Theories","volume":"105","author":"Xu","year":"2017","journal-title":"J. Biomed. Mater. Res. Part B Appl. Biomater."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"131","DOI":"10.15171\/apb.2017.017","article-title":"Preparation, Optimization and Activity Evaluation of PLGA\/Streptokinase Nanoparticles Using Electrospray","volume":"7","author":"Yaghoobi","year":"2017","journal-title":"Adv. Pharm. Bull."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"111427","DOI":"10.1016\/j.msec.2020.111427","article-title":"Use of MPEG-PLGA Nanoparticles to Improve Bioactivity and Hemocompatibility of Streptokinase: In-Vitro and in-Vivo Studies","volume":"118","author":"Hasanpour","year":"2021","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.1016\/j.msec.2017.12.036","article-title":"Recent Advances of PLGA Micro\/Nanoparticles for the Delivery of Biomacromolecular Therapeutics","volume":"92","author":"Ding","year":"2018","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"118710","DOI":"10.1016\/j.ijpharm.2019.118710","article-title":"Preparation, in Vitro and in Vivo Evaluation of PLGA\/Chitosan Based Nano-Complex as a Novel Insulin Delivery Formulation","volume":"572","author":"Mohammadpour","year":"2019","journal-title":"Int. J. Pharm."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1080\/09205063.2020.1760699","article-title":"Development of a Biocompatible PLGA Polymers Capable to Release Thrombolytic Enzyme Prourokinase","volume":"31","author":"Kaplan","year":"2020","journal-title":"J. Biomater. Sci. Polym. Ed."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Salvalaio, M., Rigon, L., Belletti, D., D\u2019Avanzo, F., Pederzoli, F., Ruozi, B., Marin, O., Vandelli, M.A., Forni, F., and Scarpa, M. (2016). Targeted Polymeric Nanoparticles for Brain Delivery of High Molecular Weight Molecules in Lysosomal Storage Disorders. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0156452"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1038\/jcbfm.2012.209","article-title":"Nanoparticles for Targeted Delivery of Antioxidant Enzymes to the Brain after Cerebral Ischemia and Reperfusion Injury","volume":"33","author":"Yun","year":"2013","journal-title":"J. Cereb Blood Flow Metab."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jconrel.2017.02.016","article-title":"Potential of Surfactant-Coated Nanoparticles to Improve Brain Delivery of Arylsulfatase A","volume":"253","author":"Schuster","year":"2017","journal-title":"J. Control. Release"},{"key":"ref_48","first-page":"301","article-title":"Recent Trends in Stabilizing Protein Structure upon Encapsulation and Release from Bioerodible Polymers","volume":"54","author":"Castellanos","year":"2002","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.ejpb.2016.12.003","article-title":"Influence of Different Stabilizers on the Encapsulation of Desmopressin Acetate into PLGA Nanoparticles","volume":"118","author":"Primavessy","year":"2017","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1023\/A:1026498209874","article-title":"Protein Instability in Poly(Lactic-Co-Glycolic Acid) Microparticles","volume":"17","author":"Hennink","year":"2000","journal-title":"Pharm. Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"3753","DOI":"10.1021\/bm5010383","article-title":"Stability Study Perspective of the Effect of Freeze-Drying Using Cryoprotectants on the Structure of Insulin Loaded into PLGA Nanoparticles","volume":"15","author":"Fonte","year":"2014","journal-title":"Biomacromolecules"},{"key":"ref_52","first-page":"407","article-title":"The Effects of Technical and Compositional Variables on the Size and Release Profile of Bovine Serum Albumin from PLGA Based Particulate Systems","volume":"9","author":"Taghipour","year":"2014","journal-title":"Res. Pharm. Sci."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1628","DOI":"10.1002\/jps.23988","article-title":"Characteristics of Sugar Surfactants in Stabilizing Proteins During Freeze\u2013Thawing and Freeze\u2013Drying","volume":"103","author":"Imamura","year":"2014","journal-title":"J. Pharm. Sci."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.ejpb.2010.03.005","article-title":"Effect of Various Additives and Polymers on Lysozyme Release from PLGA Microspheres Prepared by an s\/o\/w Emulsion Technique","volume":"75","author":"Tran","year":"2010","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/S0168-3659(00)00315-1","article-title":"Influence of the Co-Encapsulation of Different Non-Ionic Surfactants on the Properties of PLGA Insulin-Loaded Microspheres","volume":"69","author":"Rosa","year":"2000","journal-title":"J. Control. Release"},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Duskey, J.T., da Ros, F., Ottonelli, I., Zambelli, B., Vandelli, M.A., Tosi, G., and Ruozi, B. (2020). Enzyme Stability in Nanoparticle Preparations Part 1: Bovine Serum Albumin Improves Enzyme Function. Molecules, 25.","DOI":"10.3390\/molecules25204593"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1714","DOI":"10.1017\/S0007114508116270","article-title":"Prevention of Non-Enzymic Glycation of Proteins by Dietary Agents: Prospects for Alleviating Diabetic Complications","volume":"101","author":"Saraswat","year":"2008","journal-title":"Br. J. Nutr."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1186\/1477-3155-10-20","article-title":"Encapsulation of Alpha-1 Antitrypsin in PLGA Nanoparticles: In Vitro Characterization as an Effective Aerosol Formulation in Pulmonary Diseases","volume":"10","author":"Pirooznia","year":"2012","journal-title":"J. Nanobiotechnol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"3140","DOI":"10.1002\/jps.20847","article-title":"Effects of Various Cyclodextrins on the Stability of Freeze-Dried Lactate Dehydrogenase","volume":"96","author":"Iwai","year":"2007","journal-title":"J. Pharm. Sci."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/j.ijpharm.2011.02.013","article-title":"Enhanced Properties of Discrete Pulmonary Deoxyribonuclease I (DNaseI) Loaded PLGA Nanoparticles during Encapsulation and Activity Determination","volume":"408","author":"Osman","year":"2011","journal-title":"Int. J. Pharm."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1021\/acs.biomac.0c01663","article-title":"Single Enzyme Nanoparticles with Improved Biocatalytic Activity through Protein Entrapment in a Surfactant Shell","volume":"22","author":"Atkins","year":"2021","journal-title":"Biomacromolecules"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/S1359-0286(02)00117-1","article-title":"Biodegradable Nanoparticles for Drug Delivery and Targeting","volume":"6","author":"Hans","year":"2002","journal-title":"Curr. Opin. Solid State Mater. Sci."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1163\/156856206775997322","article-title":"Synthesis and Characterization of PLGA Nanoparticles","volume":"17","author":"Astete","year":"2006","journal-title":"J. Biomater. Sci. Polym. Ed."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"11390","DOI":"10.1021\/acs.jpcb.8b07368","article-title":"Effect of Surfactants on Surface-Induced Denaturation of Proteins: Evidence of an Orientation-Dependent Mechanism","volume":"122","author":"Arsiccio","year":"2018","journal-title":"J. Phys. Chem. B"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.cccn.2004.01.010","article-title":"Biochemical Study on \u03b2-Glucosidase in Individuals with Gaucher\u2019s Disease and Normal Subjects","volume":"343","author":"Michelin","year":"2004","journal-title":"Clin. Chim. Acta"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1016\/j.ijpharm.2018.03.061","article-title":"Hybrid Nanoparticles as a New Technological Approach to Enhance the Delivery of Cholesterol into the Brain","volume":"543","author":"Belletti","year":"2018","journal-title":"Int. J. Pharm."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1093\/chromsci\/bmr015","article-title":"Evaporative Light Scattering Detection Based HPLC Method for the Determination of Polysorbate 80 in Therapeutic Protein Formulations","volume":"50","author":"Nayak","year":"2012","journal-title":"J. Chromatogr. Sci."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.jfoodeng.2007.03.014","article-title":"Effects of Protein on Retention of ADH Enzyme Activity Encapsulated in Trehalose Matrices by Spray Drying","volume":"87","author":"Yoshii","year":"2008","journal-title":"J. Food Eng."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"8749","DOI":"10.2147\/IJN.S144545","article-title":"Development of Surface-Engineered PLGA Nanoparticulate-Delivery System of Tet1-Conjugated Nattokinase Enzyme for Inhibition of A\u03b240 Plaques in Alzheimer\u2019s Disease","volume":"12","author":"Bhatt","year":"2017","journal-title":"Int. J. Nanomed."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1007\/s12010-008-8232-1","article-title":"Superoxide Dismutase-Loaded PLGA Nanoparticles Protect Cultured Human Neurons Under Oxidative Stress","volume":"151","author":"Reddy","year":"2008","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"11","DOI":"10.3892\/ijmm.2012.1187","article-title":"New Strategies for the Treatment of Lysosomal Storage Diseases (Review)","volume":"31","author":"Parenti","year":"2013","journal-title":"Int. J. Mol. Med."},{"key":"ref_72","unstructured":"Ratko, T.A., Marbella, A., Godfrey, S., and Aronson, N. (2013). Enzyme-Replacement Therapies for Lysosomal Storage Diseases, Agency for Healthcare Research and Quality (US)."},{"key":"ref_73","unstructured":"Ullman, J.C., Arguello, A., Getz, J.A., Bhalla, A., Mahon, C.S., Wang, J., Giese, T., Bedard, C., Kim, D.J., and Blumenfeld, J.R. (2020). Brain Delivery and Activity of a Lysosomal Enzyme Using a Blood-Brain Barrier Transport Vehicle in Mice. Sci. Transl. Med., 12."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"6667","DOI":"10.1021\/nn402753y","article-title":"Direct Delivery of Functional Proteins and Enzymes to the Cytosol Using Nanoparticle-Stabilized Nanocapsules","volume":"7","author":"Tang","year":"2013","journal-title":"ACS Nano"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1007\/s11051-007-9332-5","article-title":"Novel Method for Immobilization of Enzymes to Magnetic Nanoparticles","volume":"10","author":"Johnson","year":"2008","journal-title":"J. Nanopart. Res."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.colsurfb.2014.09.026","article-title":"Enzyme\/PH Dual Sensitive Polymeric Nanoparticles for Targeted Drug Delivery to the Inflamed Colon","volume":"123","author":"Naeem","year":"2014","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"110512","DOI":"10.1016\/j.colsurfb.2019.110512","article-title":"Enzyme-Encapsulating Polymeric Nanoparticles: A Potential Adjunctive Therapy in Pseudomonas Aeruginosa Biofilm-Associated Infection Treatment","volume":"184","author":"Han","year":"2019","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"1751","DOI":"10.1021\/acssuschemeng.9b05240","article-title":"Enhanced Activity of the Cellulase Enzyme \u03b2-Glucosidase upon Addition of an Azobenzene-Based Surfactant","volume":"8","author":"Seidel","year":"2020","journal-title":"ACS Sustain. Chem. Eng."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"1879","DOI":"10.1002\/pro.3908","article-title":"Homodimerization of a Glycoside Hydrolase Family GH1 \u03b2-Glucosidase Suggests Distinct Activity of Enzyme Different States","volume":"29","author":"Otsuka","year":"2020","journal-title":"Protein Sci."},{"key":"ref_80","first-page":"137","article-title":"Dual Role of Imidazole as Activator\/Inhibitor of Sweet Almond (Prunus Dulcis) \u03b2-Glucosidase","volume":"10","author":"Caramia","year":"2017","journal-title":"Biochem. Biophys. Rep."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"102214","DOI":"10.1016\/j.jddst.2020.102214","article-title":"Brain Targeted Polysorbate-80 Coated PLGA Thymoquinone Nanoparticles for the Treatment of Alzheimer\u2019s Disease, with Biomechanistic Insights","volume":"61","author":"Yusuf","year":"2021","journal-title":"J. Drug Deliv. Sci. Technol."},{"key":"ref_82","first-page":"788","article-title":"The Effect of Tween Excipients on Expression and Activity of P-Glycoprotein in Caco-2 Cells","volume":"76","author":"Hodaei","year":"2014","journal-title":"Pharm. Ind."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/j.biopha.2011.10.007","article-title":"Reversal Effect of Tween-20 on Multidrug Resistance in Tumor Cells in Vitro","volume":"66","author":"Yang","year":"2012","journal-title":"Biomed. Pharmacother."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1211\/0022357001773805","article-title":"Effects of Some Non-Ionic Surfactants on Transepithelial Permeability in Caco-2 Cells","volume":"52","author":"Dimitrijevic","year":"2000","journal-title":"J. Pharm. Pharmacol."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.ijpharm.2011.03.053","article-title":"The MTT Assay as Tool to Evaluate and Compare Excipient Toxicity in Vitro on Respiratory Epithelial Cells","volume":"411","year":"2011","journal-title":"Int. J. Pharm."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1111\/j.1600-0773.1991.tb02057.x","article-title":"Polysorbate Toxicity in Neonatal Rats and Mice","volume":"68","author":"Farkas","year":"1991","journal-title":"Pharmacol. Toxicol."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"575","DOI":"10.2147\/IJN.S54750","article-title":"Tissue Inhibitor of Matrix Metalloproteinases-1 Loaded Poly(Lactic-Co-Glycolic Acid) Nanoparticles for Delivery across the Blood\u2013Brain Barrier","volume":"9","author":"Chaturvedi","year":"2014","journal-title":"Int J. Nanomed."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/j.ejpb.2009.09.003","article-title":"Drug Delivery to the Brain Using Surfactant-Coated Poly(Lactide-Co-Glycolide) Nanoparticles: Influence of the Formulation Parameters","volume":"74","author":"Gelperina","year":"2010","journal-title":"Eur. J. Pharm. Biopharm."}],"container-title":["Nanomaterials"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-4991\/11\/11\/2946\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:25:17Z","timestamp":1760167517000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-4991\/11\/11\/2946"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,3]]},"references-count":88,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["nano11112946"],"URL":"https:\/\/doi.org\/10.3390\/nano11112946","relation":{},"ISSN":["2079-4991"],"issn-type":[{"value":"2079-4991","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,3]]}}}