{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:47:35Z","timestamp":1787028455420,"version":"build-2736575974"},"reference-count":32,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2021,7,23]],"date-time":"2021-07-23T00:00:00Z","timestamp":1626998400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Science Center, Poland","award":["2016\/21\/D\/ST8\/0185"],"award-info":[{"award-number":["2016\/21\/D\/ST8\/0185"]}]},{"name":"\u201dExcellence Initiative \u2013 Research University\u201d for the AGH University of Science and Technology,","award":["Krak\u00f3w, Poland"],"award-info":[{"award-number":["Krak\u00f3w, Poland"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Modular tissue engineering (MTE) is a novel \u201cbottom-up\u201d approach to create engineered biological tissues from microscale repeating units. Our aim was to obtain microtissue constructs, based on polymer microspheres (MSs) populated with cells, which can be further assembled into larger tissue blocks and used in bone MTE. Poly(L-lactide-co-glycolide) MS of 165 \u00b1 47 \u00b5m in diameter were produced by oil-in-water emulsification and treated with 0.1 M NaOH. To improve cell adhesion, MSs were coated with poly-L-lysine (PLL) or human recombinant collagen type I (COL). The presence of oxygenated functionalities and PLL\/COL coating on MS was confirmed by X-ray photoelectron spectroscopy (XPS). To assess the influence of medium composition on adhesion, proliferation, and osteogenic differentiation, preosteoblast MC3T3-E1 cells were cultured on MS in minimal essential medium (MEM) and osteogenic differentiation medium (OSG). Moreover, to assess the potential osteoblast\u2013osteoclast cross-talk phenomenon and the influence of signaling molecules released by osteoclasts on osteoblast cell culture, a medium obtained from osteoclast culture (OSC) was also used. To impel the cells to adhere and grow on the MS, anti-adhesive cell culture plates were utilized. The results show that MS coated with PLL and COL significantly favor the adhesion and growth of MC3T3-E1 cells on days 1 and 7, respectively, in all experimental conditions tested. On day 7, three-dimensional MS\/cell\/extracellular matrix constructs were created owing to auto-assembly. The cells grown in such constructs exhibited high activity of early osteogenic differentiation marker, namely, alkaline phosphatase. Superior cell growth on PLL- and COL-coated MS on day 14 was observed in the OSG medium. Interestingly, deposition of extracellular matrix and its mineralization was particularly enhanced on COL-coated MS in OSG medium on day 14. In our study, we developed a method of spontaneous formation of organoid-like MS-based cell\/ECM constructs with a few millimeters in size. Such constructs may be regarded as building blocks in bone MTE.<\/jats:p>","DOI":"10.3390\/ijms22157897","type":"journal-article","created":{"date-parts":[[2021,7,25]],"date-time":"2021-07-25T22:06:21Z","timestamp":1627250781000},"page":"7897","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Polymeric Microspheres\/Cells\/Extracellular Matrix Constructs Produced by Auto-Assembly for Bone Modular Tissue Engineering"],"prefix":"10.3390","volume":"22","author":[{"given":"Bartosz","family":"Mielan","sequence":"first","affiliation":[{"name":"Department of Biomaterials and Composites, Faculty of Science and Ceramics, AGH University of Science and Technology, Al. Mickiewicza 30, 30-059 Krak\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Sousa","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade (i3S), Rua Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"Instituto de Engenharia Biom\u00e9dica (INEB), Universidade do Porto, 4200-135 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9415-0282","authenticated-orcid":false,"given":"Ma\u0142gorzata","family":"Krok-Borkowicz","sequence":"additional","affiliation":[{"name":"Department of Biomaterials and Composites, Faculty of Science and Ceramics, AGH University of Science and Technology, Al. Mickiewicza 30, 30-059 Krak\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5370-1722","authenticated-orcid":false,"given":"Pierre","family":"Eloy","sequence":"additional","affiliation":[{"name":"Facult\u00e9 des Bioing\u00e9nieurs Institut de la Mati\u00e8re Condens\u00e9e et des Nanosciences, Universit\u00e9 Catholique de Louvain, Place L. Pasteur 1, 1348 Louvain-la-Neuve, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine","family":"Dupont","sequence":"additional","affiliation":[{"name":"Facult\u00e9 des Bioing\u00e9nieurs Institut de la Mati\u00e8re Condens\u00e9e et des Nanosciences, Universit\u00e9 Catholique de Louvain, Place L. Pasteur 1, 1348 Louvain-la-Neuve, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meriem","family":"Lamghari","sequence":"additional","affiliation":[{"name":"Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade (i3S), Rua Alfredo Allen 208, 4200-135 Porto, Portugal"},{"name":"Instituto de Engenharia Biom\u00e9dica (INEB), Universidade do Porto, 4200-135 Porto, Portugal"},{"name":"Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar (ICBAS), Universidade do Porto, 4050-313 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0464-6189","authenticated-orcid":false,"given":"El\u017cbieta","family":"Pamu\u0142a","sequence":"additional","affiliation":[{"name":"Department of Biomaterials and Composites, Faculty of Science and Ceramics, AGH University of Science and Technology, Al. Mickiewicza 30, 30-059 Krak\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,7,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1007\/s13233-014-2002-4","article-title":"In Vitro Micro-Mineralized Tissue Formation by the Combinatory Condition of Adipose-Derived Stem Cells, Macroporous PLGA Microspheres and a Bioreactor","volume":"22","author":"Jung","year":"2014","journal-title":"Macromol. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1007\/s10439-016-1609-3","article-title":"Modular Tissue Assembly Strategies for Biofabrication of Engineered Cartilage","volume":"45","author":"Schon","year":"2017","journal-title":"Ann. Biomed. Eng."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Ciucurel, E.C., Chamberlain, M.D., and Sefton, M.V. (2013). The Modular Approach, Elsevier Inc.. [1st ed.]. Chapter 7.","DOI":"10.1016\/B978-1-4557-2852-7.00007-X"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.colsurfb.2019.01.050","article-title":"Biomimetic Microspheres for 3D Mesenchymal Stem Cell Culture and Characterization","volume":"177","author":"Sempere","year":"2019","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1039\/b814285h","article-title":"Modular Tissue Engineering: Engineering Biological Tissues from the Bottom Up","volume":"5","author":"Nichol","year":"2010","journal-title":"Soft Matter"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1016\/j.jiec.2017.03.057","article-title":"Osteogenesis and New Bone Formation of Alendronate-Immobilized Porous PLGA Microspheres in a Rat Calvarial Defect Model","volume":"52","author":"Yong","year":"2017","journal-title":"J. Ind. Eng. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4227","DOI":"10.1016\/j.biomaterials.2009.04.025","article-title":"Biomaterials Cryopreservable and Tumorigenic Three-Dimensional Tumor Culture in Porous Poly (Lactic-Co-Glycolic Acid) Microsphere","volume":"30","author":"Kang","year":"2009","journal-title":"Biomaterials"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1002\/jbm.a.30217","article-title":"De Proliferation, Activity, and Osteogenic Differentiation of Bone Marrow Stromal Cells Cultured on Calcium Titanium Phosphate Microspheres","volume":"72","author":"Barrias","year":"2005","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1016\/j.tibtech.2015.09.003","article-title":"Cell-Laden Polymeric Microspheres for Biomedical Applications","volume":"33","author":"Leong","year":"2015","journal-title":"Trends Biotechnol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1016\/j.actbio.2013.09.002","article-title":"Poly (L -Glutamic Acid)\/Chitosan Polyelectrolyte Complex Porous Microspheres as Cell Microcarriers for Cartilage Regeneration","volume":"10","author":"Fang","year":"2014","journal-title":"Acta Biomater."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.colsurfb.2017.10.044","article-title":"Modification of Porous PLGA Microspheres by Poly- l -Lysine for Use as Tissue Engineering Scaffolds","volume":"161","author":"Yuan","year":"2018","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"457","DOI":"10.1016\/j.colsurfb.2014.06.057","article-title":"Effect of Lactoferrin-Impregnated Porous Poly (Lactide-Co-Glycolide) (PLGA) Microspheres on Osteogenic Differentiation of Rabbit Adipose-Derived Stem Cells (RADSCs)","volume":"122","author":"Eun","year":"2014","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1797","DOI":"10.1021\/bp0496981","article-title":"Biodegradable PLGA Microcarriers for Injectable Delivery of Chondrocytes: Effect of Surface Modification on Cell Attachment and Function","volume":"20","author":"Chun","year":"2004","journal-title":"Biotechnol. Prog."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.colsurfb.2008.06.014","article-title":"Chitosan Modified Poly (L-Lactide) Microspheres as Cell Microcarriers for Cartilage Tissue Engineering","volume":"66","author":"Lao","year":"2008","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/j.bioeng.2007.08.013","article-title":"Emulsion-Based Synthesis of PLGA-Microspheres for the in Vitro Expansion of Porcine Chondrocytes","volume":"24","author":"Gabler","year":"2007","journal-title":"Biomol. Eng."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2475","DOI":"10.1016\/S0142-9612(00)00115-0","article-title":"The Manufacturing Techniques of Various Drug Loaded Biodegradable Poly (Lactide- Co -Glycolide) (PLGA) Devices","volume":"21","author":"Jain","year":"2000","journal-title":"Biomaterials"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1007\/s10118-014-1507-9","article-title":"Preparation of PLGA Microspheres with Different Porous Morphologies","volume":"33","author":"Wanh","year":"2015","journal-title":"Chin. J. Polym. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.colsurfa.2016.04.028","article-title":"Physicochemical and Engineering Aspects Direct and Controllable Preparation of Uniform PLGA Particles with Various Shapes and Surface Morphologies","volume":"500","author":"Fan","year":"2016","journal-title":"Colloids Surf. A Physicochem. Eng. Asp."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6034","DOI":"10.1016\/j.biomaterials.2011.04.048","article-title":"Biomaterials Biodegradation and in Vivo Biocompatibility of a Degradable, Polar\/Hydrophobic\/Ionic Polyurethane for Tissue Engineering Applications","volume":"32","author":"Mcbane","year":"2011","journal-title":"Biomaterials"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.matlet.2015.04.064","article-title":"Optimization and Characterization of Chemically Modified Polymer Microspheres and Their Effect on Cell Behavior","volume":"154","author":"Li","year":"2015","journal-title":"Mater. Lett."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/S0168-3659(99)00076-0","article-title":"Influence of formulation parameters on the characteristics of poly (D, L-lactide-co-glycolide) microspheres containing poly (L-lysine) complexed plasmid DNA","volume":"60","author":"Capan","year":"1999","journal-title":"J. Control. Release"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4453","DOI":"10.1016\/j.biomaterials.2006.04.011","article-title":"The Use of Poly(l-Lactide) and RGD Modified Microspheres as Cell Carriers in a Flow Intermittency Bioreactor for Tissue Engineering Cartilage","volume":"27","author":"Chen","year":"2006","journal-title":"Biomaterials"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"970","DOI":"10.2144\/97225rr06","article-title":"Collagen-Coated Ba2+-Alginate Microcarriers for the Culture of Anchorage-Dependent Mammalian Cells","volume":"22","author":"Zimmermann","year":"1997","journal-title":"Biotechniques"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.matbio.2012.11.001","article-title":"Collagen XVI in Health and Disease","volume":"32","author":"Bauer","year":"2013","journal-title":"Matrix Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/j.cis.2019.05.005","article-title":"A Practical Review on the Measurement Tools for Cellular Adhesion Force","volume":"269","author":"Peter","year":"2019","journal-title":"Adv. Colloid Interface Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.jcis.2012.08.008","article-title":"Influence of PH on the Build-up of Poly-L-Lysine\/Heparin Multilayers","volume":"388","author":"Barrantes","year":"2012","journal-title":"J. Colloid Interface Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1021\/acs.biomac.5b01434","article-title":"Layer-by-Layer Assembly of Supported Lipid Bilayer Poly-L-Lysine Multilayers","volume":"17","author":"Heath","year":"2016","journal-title":"Biomacromolecules"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1016\/j.actbio.2019.01.006","article-title":"Exosome-Integrated Titanium Oxide Nanotubes for Targeted Bone Regeneration","volume":"86","author":"Wei","year":"2019","journal-title":"Acta Biomater."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"116861","DOI":"10.1016\/j.lfs.2019.116861","article-title":"The Potential of Exosomes in the Therapy of the Cartilage and Bone Complications; Emphasis on Osteoarthritis","volume":"236","author":"Pourakbari","year":"2019","journal-title":"Life Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"36652","DOI":"10.1021\/acsami.8b10992","article-title":"Combinatorial Surface Roughness Effects on Osteoclastogenesis and Osteogenesis","volume":"10","author":"Zhang","year":"2018","journal-title":"Appl. Mater. Interfaces"},{"key":"ref_31","first-page":"2","article-title":"Optimizing Manufacturing Conditions of Polymer Microspheres as Cell Carriers for Modular Tissue Engineering","volume":"156","author":"Mielan","year":"2020","journal-title":"Eng. Biomater."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/srep33470","article-title":"Ablation of Y 1 Receptor Impairs Osteoclast Bone-Resorbing Activity","volume":"6","author":"Sousa","year":"2016","journal-title":"Sci. Rep."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/15\/7897\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:34:17Z","timestamp":1760164457000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/15\/7897"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,23]]},"references-count":32,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2021,8]]}},"alternative-id":["ijms22157897"],"URL":"https:\/\/doi.org\/10.3390\/ijms22157897","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,23]]}}}