{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,18]],"date-time":"2026-10-18T02:37:23Z","timestamp":1792291043303,"version":"4.4.0"},"reference-count":49,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2023,2,25]],"date-time":"2023-02-25T00:00:00Z","timestamp":1677283200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000200","name":"USAID","doi-asserted-by":"publisher","award":["7200AA20CA00019"],"award-info":[{"award-number":["7200AA20CA00019"]}],"id":[{"id":"10.13039\/100000200","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Bone regeneration using inorganic nanoparticles is a robust and safe approach. In this paper, copper nanoparticles (Cu NPs) loaded with calcium phosphate scaffolds were studied for their bone regeneration potential in vitro. The pneumatic extrusion method of 3D printing was employed to prepare calcium phosphate cement (CPC) and copper loaded CPC scaffolds with varying wt% of copper nanoparticles. A new aliphatic compound Kollisolv MCT 70 was used to ensure the uniform mixing of copper nanoparticles with CPC matrix. The printed scaffolds were studied for physico-chemical characterization for surface morphology, pore size, wettability, XRD, and FTIR. The copper ion release was studied in phosphate buffer saline at pH 7.4. The in vitro cell culture studies for the scaffolds were performed using human mesenchymal stem cells (hMSCs). The cell proliferation study in CPC-Cu scaffolds showed significant cell growth compared to CPC. The CPC-Cu scaffolds showed improved alkaline phosphatase activity and angiogenic potential compared to CPC. The CPC-Cu scaffolds showed significant concentration dependent antibacterial activity in Staphylococcus aureus. Overall, the CPC scaffolds loaded with 1 wt% Cu NPs showed improved activity compared to other CPC-Cu and CPC scaffolds. The results showed that copper has improved the osteogenic, angiogenic and antibacterial properties of CPC scaffolds, facilitating better bone regeneration in vitro.<\/jats:p>","DOI":"10.3390\/ph16030352","type":"journal-article","created":{"date-parts":[[2023,2,27]],"date-time":"2023-02-27T02:15:37Z","timestamp":1677464137000},"page":"352","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Multifunctional Three-Dimensional Printed Copper Loaded Calcium Phosphate Scaffolds for Bone Regeneration"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9900-5287","authenticated-orcid":false,"given":"Amit","family":"Pillai","sequence":"first","affiliation":[{"name":"PharmE3D Lab, Division of Molecular Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0039-0064","authenticated-orcid":false,"given":"Jaidev","family":"Chakka","sequence":"additional","affiliation":[{"name":"PharmE3D Lab, Division of Molecular Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4554-9780","authenticated-orcid":false,"given":"Niloofar","family":"Heshmathi","sequence":"additional","affiliation":[{"name":"PharmE3D Lab, Division of Molecular Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8925-2284","authenticated-orcid":false,"given":"Yu","family":"Zhang","sequence":"additional","affiliation":[{"name":"PharmE3D Lab, Division of Molecular Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faez","family":"Alkadi","sequence":"additional","affiliation":[{"name":"PharmE3D Lab, Division of Molecular Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3373-5586","authenticated-orcid":false,"given":"Mohammed","family":"Maniruzzaman","sequence":"additional","affiliation":[{"name":"PharmE3D Lab, Division of Molecular Pharmaceutics, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"682","DOI":"10.1002\/term.1563","article-title":"Fabrication of Porous Scaffolds by Three-Dimensional Plotting of a Pasty Calcium Phosphate Bone Cement under Mild Conditions: 3D Plotting of a Pasty Calcium Phosphate Bone Cement","volume":"8","author":"Lode","year":"2014","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"S3","DOI":"10.1016\/j.injury.2020.11.029","article-title":"Fracture Nonunion in Long Bones: A Literature Review of Risk Factors and Surgical Management","volume":"52","author":"Nicholson","year":"2021","journal-title":"Injury"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1186\/s13018-020-01904-2","article-title":"Prevalence and Influencing Factors of Nonunion in Patients with Tibial Fracture: Systematic Review and Meta-Analysis","volume":"15","author":"Tian","year":"2020","journal-title":"J. Orthop. Surg."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1097\/00003086-200002000-00003","article-title":"Bone Graft Materials: An Overview of the Basic Science","volume":"371","author":"Bauer","year":"2000","journal-title":"Clin. Orthop. Relat. Res."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Wu, Y., Woodbine, L., Carr, A.M., Pillai, A.R., Nokhodchi, A., and Maniruzzaman, M. (2020). 3D Printed Calcium Phosphate Cement (CPC) Scaffolds for Anti-Cancer Drug Delivery. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12111077"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.actbio.2022.08.040","article-title":"\u03b1-TCP-Based Calcium Phosphate Cements: A Critical Review","volume":"151","author":"Tronco","year":"2022","journal-title":"Acta Biomater."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5002","DOI":"10.1016\/j.biomaterials.2005.01.015","article-title":"Premixed Rapid-Setting Calcium Phosphate Composites for Bone Repair","volume":"26","author":"Carey","year":"2005","journal-title":"Biomaterials"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"9558","DOI":"10.1016\/j.actbio.2013.08.009","article-title":"Ready-to-Use Injectable Calcium Phosphate Bone Cement Paste as Drug Carrier","volume":"9","author":"Vorndran","year":"2013","journal-title":"Acta Biomater."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"045002","DOI":"10.1088\/1758-5090\/aad36d","article-title":"Bioprinting of Mineralized Constructs Utilizing Multichannel Plotting of a Self-Setting Calcium Phosphate Cement and a Cell-Laden Bioink","volume":"10","author":"Ahlfeld","year":"2018","journal-title":"Biofabrication"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3801","DOI":"10.1016\/j.bioactmat.2021.03.028","article-title":"Hyaluronic Acid Facilitates Bone Repair Effects of Calcium Phosphate Cement by Accelerating Osteogenic Expression","volume":"6","author":"Cui","year":"2021","journal-title":"Bioact. Mater."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Hettich, G., Schierjott, R.A., Epple, M., Gbureck, U., Heinemann, S., Mozaffari-Jovein, H., and Grupp, T.M. (2019). Calcium Phosphate Bone Graft Substitutes with High Mechanical Load Capacity and High Degree of Interconnecting Porosity. Materials, 12.","DOI":"10.3390\/ma12213471"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1601","DOI":"10.1089\/ten.tea.2007.0370","article-title":"Angiogenesis in Calcium Phosphate Scaffolds by Inorganic Copper Ion Release","volume":"15","author":"Barralet","year":"2009","journal-title":"Tissue Eng. Part A"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1016\/j.actbio.2019.02.021","article-title":"Angiogenesis Involvement by Octacalcium Phosphate-Gelatin Composite-Driven Bone Regeneration in Rat Calvaria Critical-Sized Defect","volume":"88","author":"Kurobane","year":"2019","journal-title":"Acta Biomater."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"110819","DOI":"10.1016\/j.matdes.2022.110819","article-title":"Effect of Porosity and Phase Composition in 3D Printed Calcium Phosphate Scaffolds on Bone Tissue Regeneration in vivo","volume":"219","author":"Raja","year":"2022","journal-title":"Mater. Des."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1016\/j.addr.2021.04.019","article-title":"Emerging 3D Printing Technologies for Drug Delivery Devices: Current Status and Future Perspective","volume":"174","author":"Wang","year":"2021","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1002\/1439-2054(20001001)282:1<17::AID-MAME17>3.0.CO;2-8","article-title":"Desktop Manufacturing of Complex Objects, Prototypes and Biomedical Scaffolds by Means of Computer-Assisted Design Combined with Computer-Guided 3D Plotting of Polymers and Reactive Oligomers","volume":"282","author":"Landers","year":"2000","journal-title":"Macromol. Mater. Eng."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1016\/j.tibtech.2016.07.005","article-title":"Calcium Phosphates and Angiogenesis: Implications and Advances for Bone Regeneration","volume":"34","author":"Malhotra","year":"2016","journal-title":"Trends Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.actbio.2020.09.044","article-title":"Biological Properties of Copper-Doped Biomaterials for Orthopedic Applications: A Review of Antibacterial, Angiogenic and Osteogenic Aspects","volume":"117","author":"Jacobs","year":"2020","journal-title":"Acta Biomater."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1016\/j.msec.2019.02.042","article-title":"Concentration-Dependent Osteogenic and Angiogenic Biological Performances of Calcium Phosphate Cement Modified with Copper Ions","volume":"99","author":"Zhang","year":"2019","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2534","DOI":"10.1039\/C8TB03272F","article-title":"Copper-Nanoparticle-Embedded Hydrogel for Killing Bacteria and Promoting Wound Healing with Photothermal Therapy","volume":"7","author":"Tao","year":"2019","journal-title":"J. Mater. Chem. B"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Shimabukuro, M. (2020). Antibacterial Property and Biocompatibility of Silver, Copper, and Zinc in Titanium Dioxide Layers Incorporated by One-Step Micro-Arc Oxidation: A Review. Antibiotics, 9.","DOI":"10.3390\/antibiotics9100716"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"21159","DOI":"10.1021\/acsami.9b17815","article-title":"Antimicrobial Copper-Based Materials and Coatings: Potential Multifaceted Biomedical Applications","volume":"12","author":"Mitra","year":"2020","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"29770","DOI":"10.1016\/j.ceramint.2022.06.237","article-title":"Antimicrobial Properties of Co-Doped Tricalcium Phosphates Ca3\u22122x(M\u2032M\u2033)x(PO4)2 (M = Zn2+, Cu2+, Mn2+ and Sr2+)","volume":"48","author":"Deyneko","year":"2022","journal-title":"Ceram. Int."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"111032","DOI":"10.1016\/j.msec.2020.111032","article-title":"Improved Osteogenesis and Angiogenesis of a Novel Copper Ions Doped Calcium Phosphate Cement","volume":"114","author":"Lin","year":"2020","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_25","unstructured":"Bettinger, C., Borenstein, J.T., and Tao, S.L. (2013). Microfluidic Cell Culture Systems, William Andrew Publishing. Micro and Nano Technologies."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Pan, N., and Sun, G. (2011). Functional Textiles for Improved Performance, Protection and Health, Woodhead Publishing.","DOI":"10.1533\/9780857092878"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.matdes.2018.11.018","article-title":"Surface Functionalization of 3D Printed Polymer Scaffolds to Augment Stem Cell Response","volume":"161","author":"Jaidev","year":"2019","journal-title":"Mater. Des."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1023\/A:1018583706335","article-title":"Composition Effects on the PH of a Hydraulic Calcium Phosphate Cement","volume":"8","author":"Bohner","year":"1997","journal-title":"J. Mater. Sci. Mater. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"980545","DOI":"10.1155\/2014\/980545","article-title":"Synthesis of Copper Nanoparticles by Thermal Decomposition and Their Antimicrobial Properties","volume":"2014","year":"2014","journal-title":"J. Nanomater."},{"key":"ref_30","first-page":"100048","article-title":"Biosynthesis and Characterization of Eupatorium Adenophorum and Chitosan Mediated Copper Oxide Nanoparticles and Their Antibacterial Activity. Results Surf","volume":"6","author":"Maheo","year":"2022","journal-title":"Interfaces"},{"key":"ref_31","unstructured":"(2013). Hydroxyapatite, Powder X-Ray Diffraction, Crystal Structure Modelling. Am. J. Mater. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1002\/crat.201500143","article-title":"Preparation, Characterization and in Vitro Assessment of Ibuprofen Loaded Calcium Phosphate\/Gypsum Bone Cements","volume":"51","author":"Alexopoulou","year":"2016","journal-title":"Cryst. Res. Technol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1901407","DOI":"10.1002\/adfm.201901407","article-title":"3D Superelastic Scaffolds Constructed from Flexible Inorganic Nanofibers with Self-Fitting Capability and Tailorable Gradient for Bone Regeneration","volume":"29","author":"Wang","year":"2019","journal-title":"Adv. Funct. Mater."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Guo, C., Niu, D., Liu, J., Bao, X., and Xu, G. (2021). Application of Biodegradable PLGA-PEG-PLGA\/CPC Composite Bone Cement in the Treatment of Osteoporosis. Coatings, 11.","DOI":"10.3390\/coatings11070827"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2675","DOI":"10.1016\/j.biomaterials.2009.01.022","article-title":"Mesenchymal Stem Cell Proliferation and Differentiation on an Injectable Calcium Phosphate\u2014Chitosan Composite Scaffold","volume":"30","author":"Moreau","year":"2009","journal-title":"Biomaterials"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1016\/S0140-6736(04)16727-5","article-title":"Osteomyelitis","volume":"364","author":"Lew","year":"2004","journal-title":"Lancet"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"417","DOI":"10.5435\/00124635-200510000-00006","article-title":"Chronic Posttraumatic Osteomyelitis and Infected Nonunion of the Tibia: Current Management Concepts","volume":"13","author":"Patzakis","year":"2005","journal-title":"JAAOS\u2014J. Am. Acad. Orthop. Surg."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1016\/S0195-5616(91)50085-1","article-title":"Morphology of Fracture Nonunion and Osteomyelitis","volume":"21","author":"Riser","year":"1991","journal-title":"Vet. Clin. N. Am. Small Anim. Pract."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1016\/j.biomaterials.2009.10.009","article-title":"The Stimulation of Angiogenesis and Collagen Deposition by Copper","volume":"31","author":"Bordeleau","year":"2010","journal-title":"Biomaterials"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1016\/j.actbio.2022.01.034","article-title":"Composites Consisting of Calcium Phosphate Cements and Mesoporous Bioactive Glasses as a 3D Plottable Drug Delivery System","volume":"156","author":"Richter","year":"2022","journal-title":"Acta Biomater."},{"key":"ref_41","unstructured":"Hellerbrand, K., Siedler, M., Schutz, A., Pompe, C., and Friess, W. (2009). In Situ Hardening Paste, Its Manufacturing and Use. (US20090048145A1), U.S. Patent."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1521","DOI":"10.1038\/s41596-018-0003-z","article-title":"Surface-Wetting Characterization Using Contact-Angle Measurements","volume":"13","author":"Tian","year":"2018","journal-title":"Nat. Protoc."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/S0927-7765(01)00200-4","article-title":"Surface Thermodynamics of Osteoblasts: Relation between Hydrophobicity and Bone Active Biomaterials","volume":"22","author":"Zanchetta","year":"2001","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"946","DOI":"10.1016\/j.protcy.2016.01.094","article-title":"Contact Angle Measurement on Medical Implant Titanium Based Biomaterials","volume":"22","author":"Strnad","year":"2016","journal-title":"Procedia Technol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1140\/epjp\/i2019-12375-x","article-title":"Effect of Copper Oxide Nanoparticles on Electrical Conductivity and Cell Viability of Calcium Phosphate Scaffolds with Improved Mechanical Strength for Bone Tissue Engineering","volume":"134","author":"Sahmani","year":"2019","journal-title":"Eur. Phys. J. Plus"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"8547","DOI":"10.1039\/C4TB01355G","article-title":"Evaluation of Borate Bioactive Glass Scaffolds as a Controlled Delivery System for Copper Ions in Stimulating Osteogenesis and Angiogenesis in Bone Healing","volume":"2","author":"Wang","year":"2014","journal-title":"J. Mater. Chem. B"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1111\/j.1939-165X.2007.tb00216.x","article-title":"Alkaline Phosphatase: Beyond the Liver","volume":"36","author":"Fernandez","year":"2007","journal-title":"Vet. Clin. Pathol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Deyneko, D., Zheng, Y., Barbaro, K., Lebedev, V.N., Aksenov, S.M., Borovikova, E.Y., Gafurov, M.R., Fadeeva, I.V., Antoniac, I., and Lazoryak, B. (2022). Dependence of Antimicrobial Properties on Site-Selective Arrangement and Concentration of Bioactive Cu2+ Ions in Tricalcium Phosphate. SSRN Electron. J.","DOI":"10.2139\/ssrn.4295801"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"025020","DOI":"10.1088\/1758-5090\/aa7077","article-title":"Fabrication of Biomimetic Bone Grafts with Multi-Material 3D Printing","volume":"9","author":"Sears","year":"2017","journal-title":"Biofabrication"}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/16\/3\/352\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:42:17Z","timestamp":1760121737000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/16\/3\/352"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,25]]},"references-count":49,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["ph16030352"],"URL":"https:\/\/doi.org\/10.3390\/ph16030352","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,2,25]]}}}