[53] have also shown that the majority of DPSCs and SHED expressed several neural lineage markers, including nestin, Doublecortin (DCX; neuronal progenitor cells), βIII-tubulin (early neuronal cells), NeuN (mature neurons), GFAP (neural stem cells and astrocytes), S-100 (Schwann cells), A2B5, and CNPase (oligodendrocyte progenitor cells). A. Scheven, “Effects of glial cell line-derived neurotrophic factor on dental pulp cells,”, T. Yamamoto, Y. Osako, M. Ito et al., “Trophic effects of dental pulp stem cells on Schwann cells in peripheral nerve regeneration,”, T. Mita, Y. Furukawa-Hibi, H. Takeuchi et al., “Conditioned medium from the stem cells of human dental pulp improves cognitive function in a mouse model of Alzheimer's disease,”, T. Inoue, M. Sugiyama, H. Hattori, H. Wakita, T. Wakabayashi, and M. Ueda, “Stem cells from human exfoliated deciduous tooth-derived conditioned medium enhance recovery of focal cerebral ischemia in rats,”, K. Matsubara, Y. Matsushita, K. Sakai et al., “Secreted ectodomain of sialic acid-binding Ig-like lectin-9 and monocyte chemoattractant protein-1 promote recovery after rat spinal cord injury by altering macrophage polarity,”, Y. Sugimura-Wakayama, W. Katagiri, M. Osugi et al., “Peripheral nerve regeneration by secretomes of stem cells from human exfoliated deciduous teeth,”, A. Jarmalavičiūtė, V. Tunaitis, U. Pivoraitė, A. Venalis, and A. Pivoriūnas, “Exosomes from dental pulp stem cells rescue human dopaminergic neurons from 6-hydroxy-dopamine-induced apoptosis,”, M. P. de Miguel, S. Fuentes-Julián, A. Blázquez-Martínez et al., “Immunosuppressive properties of mesenchymal stem cells: advances and applications,”, Regulation (EC) No 2003/94/EC of the European Parliament and of the Council of 8 October 2003 laying down the principles and guidelines of good manufacturing practice in respect of medicinal products for human use and investigational medicinal products for human use, Regulation (EC) No 1394/2007 of the European parliament and of the council of 13 November 2007 on advanced therapy medicinal products and amending Directive 2001/83/EC and Regulation (EC) No 726/2004, S. Pacini, “Deterministic and stochastic approaches in the clinical application of mesenchymal stromal cells (MSCs),”, E. H. Javazon, K. J. Beggs, and A. W. Flake, “Mesenchymal stem cells: paradoxes of passaging,”, P. Hilkens, N. Meschi, P. Lambrechts, A. Bronckaers, and I. Lambrichts, “Dental stem cells in pulp regeneration: near future or long road ahead?”, P. G. Robey, S. A. Kuznetsov, J. Ren, H. G. Klein, M. Sabatino, and D. F. Stroncek, “Generation of clinical grade human bone marrow stromal cells for use in bone regeneration,”, A. Shahdadfar, K. Frønsdal, T. Haug, F. P. Reinholt, and J. E. Brinchmann, “In vitro expansion of human mesenchymal stem cells: choice of serum is a determinant of cell proliferation, differentiation, gene expression, and transcriptome stability,”, P. A. Sotiropoulou, S. A. Perez, M. Salagianni, C. N. Baxevanis, and M. Papamichail, “Characterization of the optimal culture conditions for clinical scale production of human mesenchymal stem cells,”, F. Mannello and G. A. Tonti, “Concise review: No breakthroughs for human mesenchymal and embryonic stem cell culture: Conditioned medium, feeder layer, or feeder-free; medium with fetal calf serum, human serum, or enriched plasma; serum-free, serum replacement nonconditioned medium, or ad hoc formula? Despite the constraints and limitations of current research approaches, it is safe to conclude that dental MSCs, including DPSCs, SHED, and SCAP, have been extensively studied in the past years by the dental research community using highly sophisticated in vitro and in vivo systems; this has led to a substantial understanding of their unique biological properties. Although SCAP have not been so closely investigated as DPSCs, several later reports provide significant insight into the particular molecular mechanisms responsible for SCAP biological responses to various microenvironments, providing data pivotally useful for the design of future regenerative strategies for targeted dental TE. It is significant to note that secretion of various soluble factors by MSCs may occur either via exocytosis or via release of extracellular vesicles (EVs). It has been recently shown that SHED can generate functional dental pulp when injected with scaffolds (Puramatrix or rhCollagen) into full-length root canals [85]. It has been further shown by means of genetic lineage tracing in rodent incisors that MSCs residing in the dental pulp may be of dual origin, consisting of not only NG2+ pericyte cells, whose presence is closely dependent on tissue vascularity, but also MSCs of nonpericyte origin, contributing to tissue growth and repair [17]. Regarding the neuroinductive culture media, most studies use either the Neurobasal A or conventional primarily Dulbecco’s Modified Eagle’s Medium (DMEM)/F12 media in their neural differentiation protocols. A very recent and interesting study mapping potential molecular differences between SHED and DPSCs identified several differentially regulated genes [96].

[30]. SCAP and SHED produced a more highly mineralized matrix in comparison with DPSCs but with lower crystallinity and carbonate substitution. In recent years, stem cell therapy has become a very promising and advanced scientific research topic. Specifically, DPSCs have demonstrated the capacity to differentiate into odontoblastic-like cells with characteristic cell polarity [67]. Sun et al., “Concise reviews: characteristics and potential applications of human dental tissue-derived mesenchymal stem cells,”, A. 1 Department of Fixed Prosthesis and Implant Prosthodontics, School of Dentistry, Aristotle University of Thessaloniki, Thessaloniki, Greece. Lee, G.-W. Huang, J.-N. Shiung et al., “Magnetic cryopreservation for dental pulp stem cells,”, T. K. Chatzistamatiou, A. C. Papassavas, E. Michalopoulos et al., “Optimizing isolation culture and freezing methods to preserve Wharton's jelly's mesenchymal stem cell (MSC) properties: an MSC banking protocol validation for the Hellenic Cord Blood Bank,”, S. Abbasalizadeh and H. Baharvand, “Technological progress and challenges towards cGMP manufacturing of human pluripotent stem cells based therapeutic products for allogeneic and autologous cell therapies,”, R. Martín-Ibáñez, A. M. Strömberg, O. Hovatta, and J. M. Canals, “UNIT 1C.8 cryopreservation of dissociated human embryonic stem cells in the presence of ROCK inhibitor,”, C. A. Lemos, V. E. de Souza Batista, D. A. Almeida, J. F. Santiago Júnior, F. R. Verri, and E. P. Pellizzer, “Evaluation of cement-retained versus screw-retained implant-supported restorations for marginal bone loss,”, S. A. Tarle, S. Shi, and D. Kaigler, “Development of a serum-free system to expand dental-derived stem cells: PDLSCs and SHEDs,”, E. J. Eubanks, S. A. Tarle, and D. Kaigler, “Tooth storage, dental pulp stem cell isolation, and clinical scale expansion without animal serum,”, N. F. Lizier, A. Kerkis, C. M. Gomes et al., “Scaling-up of dental pulp stem cells isolated from multiple niches,”, M. Ducret, H. Fabre, O. Degoul et al., “Manufacturing of dental pulp cell-based products from human third molars: current strategies and future investigations,”, A. Bakopoulou, G. Leyhausen, J. Volk et al., “Assessment of the impact of two different isolation methods on the osteo/odontogenic differentiation potential of human dental stem cells derived from deciduous teeth,”, P. Hilkens, P. Gervois, Y. Fanton et al., “Effect of isolation methodology on stem cell properties and multilineage differentiation potential of human dental pulp stem cells,”, M. Padial-Molina, F. O'Valle, A. Lanis et al., “Clinical application of mesenchymal stem cells and novel supportive therapies for oral bone regeneration,”, C. Jakobsen, J.
also detected several neurotrophic factors in SCAP secretome, including Midkine (MDK), Pleiotrophin (PTN), Mesencephalic Astrocyte-Derived Neurotrophic Factor (MANF), Neuroblast Differentiation-Associated Protein (AHNAK), and Neurophilin 2 (NRP2). Wan et al., “Autophagy in SDF-1, M. Seo, W. Sonoyama, T. Yamaza et al., “SHED repair critical-size calvarial defects in mice,”, A. Behnia, A. Haghighat, A. Talebi, N. Nourbakhsh, and F. Heidari, “Transplantation of stem cells from human exfoliated deciduous teeth for bone regeneration in the dog mandibular defect,”, A. Alkaisi, A. R. Ismail, S. S. Mutum, Z.

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