PLA-Based Mineral-Doped Scaffolds Seeded with Human Periapical Cyst-Derived MSCs: A Promising Tool for Regenerative Healing in Dentistry

Marco Tatullo; Gianrico Spagnuolo; Bruna Codispoti; Fausto Zamparini; Anqi Zhang; Micaela Degli Esposti; Conrado Aparicio; Carlo Rengo; Manuel Nuzzolese; Lucia Manzoli; Fabio Fava; Carlo Prati; Paola Fabbri; Maria Giovanna Gandolfi

doi:10.3390/ma12040597

PLA-Based Mineral-Doped Scaffolds Seeded with Human Periapical Cyst-Derived MSCs: A Promising Tool for Regenerative Healing in Dentistry

Materials (Basel). 2019 Feb 16;12(4):597. doi: 10.3390/ma12040597.

Authors

Affiliations

¹ Tecnologica Research Institute, Stem Cell Unit, 88900 Crotone, Italy. marco.tatullo@tecnologicasrl.com.
² Department of Neurosciences, Reproductive and Odontostomatological Sciences, University of Naples "Federico II", 80131 Napoli, Italy. gianrico.spagnuolo@gmail.com.
³ Tecnologica Research Institute, Stem Cell Unit, 88900 Crotone, Italy. bruna.codispoti@tecnologicasrl.com.
⁴ Laboratory of Biomaterials and Oral Pathology, School of Dentistry, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40125 Bologna, Italy. fausto.zamparini2@unibo.it.
⁵ MDRCBB-Minnesota Dental Research Center for Biomaterials and Biomechanics, School of Dentistry, University of Minnesota, Minneapolis, MN 55455, USA. azhang@umn.edu.
⁶ Department of Civil, Chemical, Environmental and Materials Engineering, University of Bologna, 40131 Bologna, Italy. micaela.degliesposti@unibo.it.
⁷ MDRCBB-Minnesota Dental Research Center for Biomaterials and Biomechanics, School of Dentistry, University of Minnesota, Minneapolis, MN 55455, USA. apari003@umn.edu.
⁸ Department of Prosthodontics and Dental Materials, School of Dental Medicine, University of Siena, 53100 Siena, Italy. carlo.rengo@alice.it.
⁹ University Hospitals Birmingham-NHS Foundation Trust, Birmingham B152GW, UK. manuelnuzzolese@libero.it.
¹⁰ Cellular Signalling Laboratory, Institute of Human Anatomy, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40126 Bologna, Italy. lucia.manzoli@unibo.it.
¹¹ Department of Civil, Chemical, Environmental and Materials Engineering, University of Bologna, 40131 Bologna, Italy. fabio.fava@unibo.it.
¹² Laboratory of Biomaterials and Oral Pathology, School of Dentistry, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40125 Bologna, Italy. carlo.prati@unibo.it.
¹³ Department of Civil, Chemical, Environmental and Materials Engineering, University of Bologna, 40131 Bologna, Italy. p.fabbri@unibo.it.
¹⁴ Laboratory of Biomaterials and Oral Pathology, School of Dentistry, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40125 Bologna, Italy. mgiovanna.gandolfi@unibo.it.

Abstract

Human periapical cyst mesenchymal stem cells (hPCy-MSCs) are a newly discovered cell population innovatively collected from inflammatory periapical cysts. The use of this biological waste guarantees a source of stem cells without any impact on the surrounding healthy tissues, presenting a valuable potential in tissue engineering and regenerative medicine applications. In the present study, hPCy-MSCs were collected, isolated, and seeded on three experimental mineral-doped porous scaffolds produced by the thermally-induced phase-separation (TIPS) technique. Mineral-doped scaffolds, composed of polylactic acid (PLA), dicalcium phosphate dihydrate (DCPD), and/or hydraulic calcium silicate (CaSi), were produced by TIPS (PLA-10CaSi, PLA-5CaSi-5DCPD, PLA-10CaSi-10DCPD). Micro-CT analysis evaluated scaffolds micromorphology. Collected hPCy-MSCs, characterized by cytofluorimetry, were seeded on the scaffolds and tested for cell proliferation, cells viability, and gene expression for osteogenic and odontogenic differentiation (DMP-1, OSC, RUNX-2, HPRT). Micro-CT revealed an interconnected highly porous structure for all the scaffolds, similar total porosity with 99% open pores. Pore wall thickness increased with the percentage of CaSi and DCPD. Cells seeded on mineral-doped scaffolds showed a superior proliferation compared to pure PLA scaffolds (control), particularly on PLA-10CaSi-10DCPD at day 12. A higher number of non-viable (red stained) cells was observable on PLA scaffolds at days 14 and 21. DMP-1 expression increased in hPCy-MSCs cultured on all mineral-doped scaffolds, in particular on PLA-5CaSi-5DCPD and PLA-10CaSi-10DCPD. In conclusion, the innovative combination of experimental scaffolds colonized with autologous stem cells from periapical cyst represent a promising strategy for regenerative healing of periapical and alveolar bone.

Keywords: bioactive scaffolds; calcium silicate-containing scaffolds; human periapical cyst mesenchymal stem cells (hPCy-MSCs); polylactic acid (PLA)-based scaffolds; porous scaffolds; regenerative dentistry; thermally induced phase separation technique (TIPS).