Mineral-Doped Poly(L-lactide) Acid Scaffolds Enriched with Exosomes Improve Osteogenic Commitment of Human Adipose-Derived Mesenchymal Stem Cells

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Abstract

Exosomes derived from mesenchymal stem cells are extracellular vesicles released to facilitate cell communication and function. Recently, polylactic acid (PLA), calcium silicates (CaSi), and dicalcium phosphate dihydrate (DCPD) have been used to produce bioresorbable functional mineral-doped porous scaffolds-through thermally induced phase separation technique, as materials for bone regeneration. The aim of this study was to investigate the effect of mineral-doped PLA-based porous scaffolds enriched with exosome vesicles (EVs) on osteogenic commitment of human adipose mesenchymal stem cells (hAD-MSCs). Two different mineral-doped scaffolds were produced: PLA-10CaSi-10DCPD and PLA-5CaSi-5DCPD. Scaffolds surface micromorphology was investigated by ESEM-EDX before and after 28 days immersion in simulated body fluid (HBSS). Exosomes were deposited on the surface of the scaffolds and the effect of exosome-enriched scaffolds on osteogenic commitment of hAD-MSCs cultured in proximity of the scaffolds has been evaluated by real time PCR. In addition, the biocompatibility was evaluated by direct-contact seeding hAD-MSCs on scaffolds surface-using MTT viability test. In both formulations, ESEM showed pores similar in shape (circular and elliptic) and size (from 10–30 µm diameter). The porosity of the scaffolds decreased after 28 days immersion in simulated body fluid. Mineral-doped scaffolds showed a dynamic surface and created a suitable bone-forming microenvironment. The presence of the mineral fillers increased the osteogenic commitment of hAD-MSCs. Exosomes were easily entrapped on the surface of the scaffolds and their presence improved gene expression of major markers of osteogenesis such as collagen type I, osteopontin, osteonectin, osteocalcin. The experimental scaffolds enriched with exosomes, in particular PLA-10CaSi-10DCPD, increased the osteogenic commitment of MSCs. In conclusion, the enrichment of bioresorbable functional scaffolds with exosomes is confirmed as a potential strategy to improve bone regeneration procedures.

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The biology of fracture healing is a complex biological process that follows specific regenerative patterns and involves changes in the expression of several thousand genes. Although there is still much to be learned to fully comprehend the pathways of bone regeneration, the over-all pathways of both the anatomical and biochemical events have been thoroughly investigated. These efforts have provided a general understanding of how fracture healing occurs. Following the initial trauma, bone heals by either direct intramembranous or indirect fracture healing, which consists of both intramembranous and endochondral bone formation. The most common pathway is indirect healing, since direct bone healing requires an anatomical reduction and rigidly stable conditions, commonly only obtained by open reduction and internal fixation. However, when such conditions are achieved, the direct healing cascade allows the bone structure to immediately regenerate anatomical lamellar bone and the Haversian systems without any remodelling steps necessary. In all other non-stable conditions, bone healing follows a specific biological pathway. It involves an acute inflammatory response including the production and release of several important molecules, and the recruitment of mesenchymal stem cells in order to generate a primary cartilaginous callus. This primary callus later undergoes revascularisation and calcification, and is finally remodelled to fully restore a normal bone structure. In this article we summarise the basic biology of fracture healing. Copyright © 2011 Elsevier Ltd. All rights reserved.

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Journal

Journal ID (nlm-ta): Nanomaterials (Basel)

Journal ID (iso-abbrev): Nanomaterials (Basel)

Journal ID (publisher-id): nanomaterials

Title: Nanomaterials

Publisher: MDPI

ISSN (Electronic): 2079-4991

Publication date (Electronic): 29 February 2020

Publication date Collection: March 2020

Volume: 10

Issue: 3

Electronic Location Identifier: 432

Affiliations

[1 ]Laboratory of Biomaterials and Oral Pathology, School of Dentistry, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40125 Bologna, Italy; fausto.zamparini2@ 123456unibo.it (F.Z.);

[2 ]Medical Sciences Department, University of Ferrara, 44100 Ferrara, Italy

[3 ]Department of Civil, Chemical, Environmental and Materials Engineering, University of Bologna, 40136 Bologna, Italy

[4 ]Department of Metallurgical and Materials Engineering, Middle East Technical University (METU), 06800 Ankara, Turkey

[5 ]Biomedical Engineering Program, METU, 06800 Ankara, Turkey

[6 ]BIOMATEN, Center of Excellence in Biomaterials and Tissue Engineering, METU, 06800 Ankara, Turkey

[7 ]Cellular Signaling Laboratory, Institute of Human Anatomy, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40126 Bologna, Italy

[8 ]Endodontic Clinical Section, School of Dentistry, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40125 Bologna, Italy

Author notes

[* ]Correspondence: mgiovanna.gandolfi@ 123456unibo.it (M.G.G.); barbara.zavan@ 123456unife.it (B.Z.)

Author information

Maria Giovanna Gandolfi https://orcid.org/0000-0001-7793-6227

Fausto Zamparini https://orcid.org/0000-0002-0071-4463

Micaela Degli Esposti https://orcid.org/0000-0002-4513-8527

Greta Parchi https://orcid.org/0000-0002-7764-4539

Barbara Zavan https://orcid.org/0000-0002-4779-4456

Article

Publisher ID: nanomaterials-10-00432

DOI: 10.3390/nano10030432

PMC ID: 7153699

PubMed ID: 32121340

SO-VID: f63ebcac-2975-4619-a3d4-13e703601b5e

License:

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

Mineral-Doped Poly(L-lactide) Acid Scaffolds Enriched with Exosomes Improve Osteogenic Commitment of Human Adipose-Derived Mesenchymal Stem Cells

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Photoluminescent Nanomaterials

Most cited references 71

The biology of fracture healing.

Clinical applications of mineral trioxide aggregate.

MicroRNA control of bone formation and homeostasis.

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