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Mesenchymal stem cells (MSC) effects on tissue regeneration are mainly mediated by their secreted substances (secretome), inducing their paracrine activity. This Conditioned medium (CM), including soluble factors (proteins, nucleic acids, lipids) and extracellular vesicles is emerging as a potential alternative to cell therapy. However, the manufacturing of CM suffers from variable procedures and protocols leading to varying results between studies. Besides, there is no well-defined optimized procedure targeting specific applications in regenerative medicine.To focus on conditioned medium produced from dental MSC (DMSC-CM), we reviewed the current parameters and manufacturing protocols, in order to propose a standardization and optimization of these manufacturing procedures.We have selected all publications investigating the effects of dental MSC secretome inA total of 351 results were identified. And based on the inclusion criteria described above, 118 unique articles were included in the systematic review. DMSC-CM production was considered at three stages: before CM recovery (cell sources for CM), during CM production (culture conditions) and after production (CM treatment).No clear consensus could be recovered as evidence-based methods, but we were able to describe the most commonly used protocols: donors under 30 years of age, dental pulp stem cells and exfoliated deciduous tooth stem cells with cell passage between 1 and 5, at a confluence of 70% to 80%. CM were often collected during 48 h, and stored at -80 °C. It is important to point out that the preconditioning environment had a significant impact on DMSC-CM content and efficiency.

Hyaluronic acid (HA) is widely used in aesthetic medicine for its moisturizing and anti-aging action. This molecule, which is naturally present in the body, has an interesting response to aging, accentuated in totally edentulous patients. While its aesthetic benefits for facial rejuvenation are well-documented, there is a lack of description and investigation on its therapeutic usefulness for edentulous patients. The management of completely edentulous patients is a daily reality in dental practice and requires specific attention. The aesthetic and functional challenge is considerable. The displacement of the bone base, which is often marked, and lack of soft tissue support are sometimes difficult to correct with prosthetic reconstruction. This review aims to present the physiological processes appearing in completely edentulous patients and prosthetic solutions available to recreate oral functions and counteract facial aging. As prosthetic rehabilitations are not fully satisfying for counterbalancing the impression of excessive facial aging, we investigated the applications of HA injection in the perioral area, in order to improve edentulism treatment, and discussed the advantages and disadvantages, compared to other dermal fillers and rejuvenation therapies. Considering the specific situations of edentulous patients, dermal HA injections help to correct uncompensated bone losses and mucous volume losses and appear to be a therapeutically beneficial for treating completely edentulous patients, without the requirement to full rejuvenation therapy.

International audience; The aim of this work was the development of injectable radio-opaque and macroporous calcium phosphate cement (CPC) to be used as a bone substitute for the treatment of pathologic vertebral fractures. A CPC was first rendered radio-opaque by the incorporation of zirconium dioxide (ZrO2). In order to create macroporosity, poly lactic-co-glycolic acid (PLGA) microspheres around 100 μm were homogeneously incorporated into the CPC as observed by scanning electron microscopy. Physicochemical analyses by X-ray diffraction and Fourier transform infrared spectroscopy confirmed the brushite phase of the cement. The mechanical properties of the CPC/PLGA cement containing 30% PLGA (wt/wt) were characterized by a compressive strength of 2 MPa and a Young’s modulus of 1 GPa. The CPC/PLGA exhibited initial and final setting times of 7 and 12 min, respectively. Although the incorporation of PLGA microspheres increased the force necessary to inject the cement and decreased the percentage of injected mass as a function of time, the CPC/PLGA appeared fully injectable at 4 min. Moreover, in comparison with CPC, CPC/PLGA showed a full degradation in 6 weeks (with 100% mass loss), and this was associated with an acidification of the medium containing the CPC/PLGA sample (pH of 3.5 after 6 weeks). A cell viability test validated CPC/PLGA biocompatibility, and in vivo analyses using a bone defect assay in the caudal vertebrae of Wistar rats showed the good opacity of the CPC through the tail and a significant increased degradation of the CPC/PLGA cement a month after implantation. In conclusion, this injectable CPC scaffold appears to be an interesting material for bone substitution.

International audience; Photobiomodulation (PBM) has been shown to improve cell proliferation and cell migration. Many cell types have been investigated, with most studies using deep penetrating red light irradiation. Considering the interest of surface biostimulation of oral mesenchymal cells after surgical wound, the present study aimed to assess green light irradiation effects on Dental Pulp Stem Cells’ (DPSC) proliferation and migration. To understand the mechanisms underlying these effects, we investigated cytoskeleton organization and subsequent cell shape and stiffness. A 532-nm wavelength Nd:YAG laser (30 mW) was applied between 30 and 600 s on DPSC in vitro. Cell proliferation was analyzed at 24, 48, and 72 h after irradiation, by cell counting and enzymatic activity quantification (paranitrophenylphosphate phosphatase (pNPP) test). A wound healing assay was used to study cell migration after irradiation. Effects of PBM on cytoskeleton organization and cell shape were assessed by actin filaments staining. Elasticity changes after irradiation were quantified in terms of Young’s modulus measured using Atomic Force Microscopy (AFM) force spectroscopy. Green light significantly improved DPSC proliferation with a maximal effect obtained after 300-s irradiation (energy fluence 5 J/cm2). This irradiation had a significant impact on cell migration, improving wound healing after 24 h. These results were concomitant with a decrease of cells’ Young’s modulus after irradiation. This cell softening was explained by actin cytoskeleton reorganization, with diminution of cell circularity and more abundant pseudopodia. This study highlights the interest of green laser PMB for the proliferation and migration of mesenchymal stem cells, with encouraging results for clinical application, especially for surgical wound healing procedures.

International audience; Extensive use of porous silicon (PSi) for tissue engineering is due to its convenient properties as it is both non-toxic and bioresorbable. Moreover, PSi surface modification is an important step to enhance cell adhesion and proliferation. In this work, a combination of optical and electrochemical studies is performed to elaborate a suitable PSi multilayer substrate for cell culture. For this study, we modified PSi surface by silanization and antibody grafting (APTES-anti STRO1), the 12-mer specific peptide to silicon p+type coating and the peptide modified with the antibody recognition sequence. Electrochemical characterization of PSi multilayers is performed to investigate its electrical behavior, determine the optimal measuring conditions and reveal the most stable PSi surfaces. Then, the behavior of dental pulp stem cells (DPSC) was investigated on various modified PSi surfaces. An electrochemical method was applied for the first time monitoring the electrical behavior of stem cell adhesion. The cells electrochemical behavior depends on the nature of the surface coating and the peptide-anti STRO1 improved adhesion and cell spreading onto the PSi surface compared to bare surface and the one coated with the peptide. Fluorescent microscopy revealed that all surface modi cation methods enhance cell adhesion compared to the bare PSi surface. An increased cell number is observed on APTES-anti STRO1, peptide and peptide-anti STRO1 coated PSi. The peptide-anti STRO1 provided the best cell proliferation results suggesting the improved accessibility of the recognition fragment of the antibody anti-STRO1.

The development of new diagnostic technologies based on the light scattering and autofluorescence properties of dental tissues is required to improve the diagnostic ability of initial caries lesions earlier than previously done and promoting the potential of treatment without surgical intervention. The aim of this study is to correlate fluorescence-based results provided by multiphoton microscopy (MPM) with confocal Raman microscopy records using phosphate level at 960 cm–1 and the organic matrix at ∼2,931 cm–1 in healthy and demineralized human enamel. Measurements on 14 teeth were made using two incident lights of different wavelengths, released by confocal Raman microscopy and MPM. Raman phosphate peak intensity at 960 cm–1 along with organic to mineral ratio at (2,931/430 cm–1) and nonlinear optical signals (second harmonic generation [SHG] and intrinsic two-photon excited fluorescence [I2PEF]) were recorded from the demineralized and healthy enamel sites. Raman spectral maps showed that the higher the organic/mineral ratio in the demineralized enamel, the lower the intensity of mineral component in the same zone. MPM revealed new optical indicators of carious lesion as shown by the presence of a red-shifted fluorescence peak in the 650- to 750-nm area of the fluorescence spectrum of demineralized enamel. Moreover, on sample regions with insignificant autofluorescence, the emergence of the SHG signal could be noted. By comparing I2PEF images with the structural motifs observed by the confocal Raman imaging system, the morphological similarity of the acquired images was quite evident. Any change in the I2PEF spectra reflects alterations in the chemical composition of enamel. These findings may provide an important basis for potentially valuable applications of photonic tools in the clinical diagnosis of tooth pathological conditions, besides exposing the fundamental role of organic matrix in enamel integrity and reparation.

International audience; Following the basis of tissue engineering (Cells-Scaffold-Bioactive molecules), regenerative endodontic has emerged as a new concept of dental treatment. Clinical procedures have been proposed by endodontic practitioners willing to promote regenerative therapy. Preserving pulp vitality was a first approach. Later procedures aimed to regenerate a vascularized pulp in necrotic root canals. However, there is still no protocol allowing an effective regeneration of necrotic pulp tissue either in immature or mature teeth. This review explores in vitro and preclinical concepts developed during the last decade, especially the potential use of stem cells, bioactive molecules, and scaffolds, and makes a comparison with the goals achieved so far in clinical practice. Regeneration of pulp-like tissue has been shown in various experimental conditions. However, the appropriate techniques are currently in a developmental stage. The ideal combination of scaffolds and growth factors to obtain a complete regeneration of the pulp-dentin complex is still unknown. The use of stem cells, especially from pulp origin, sounds promising for pulp regeneration therapy, but it has not been applied so far for clinical endodontics, in case of necrotic teeth. The gap observed between the hope raised from in vitro experiments and the reality of endodontic treatments suggests that clinical success may be achieved without external stem cell application. Therefore, procedures using the concept of cell homing, through evoked bleeding that permit to recreate a living tissue that mimics the original pulp has been proposed. Perspectives for pulp tissue engineering in the near future include a better control of clinical parameters and pragmatic approach of the experimental results (autologous stem cells from cell homing, controlled release of growth factors). In the coming years, this therapeutic strategy will probably become a clinical reality, even for mature necrotic teeth.

International audience; With their potent regenerative and protective capacities, stem cell-derived conditioned media emerged as an effective alternative to cell therapy, and have a prospect to be manufactured as pharmaceutical products for tissue regeneration applications. Our study investigates the neuroregenerative potential of human dental pulp cells (DPCs) conditioned medium (CM) and defines an optimization strategy of DPC-CM for enhanced neuronal outgrowth. Primary sensory neurons from mouse dorsal root ganglia were cultured with or without DPC-CM, and the lengths of βIII-tubulin positive neurites were measured. The impacts of several manufacturing features as the duration of cell conditioning, CM storage, and preconditioning of DPCs with some factors on CM functional activity were assessed on neurite length. We observed that DPC-CM significantly enhanced neurites outgrowth of sensory neurons in a concentration-dependent manner. The frozen storage of DPC-CM had no impact on experimental outcomes and 48 h of DPC conditioning is optimal for an effective activity of CM. To further understand the regenerative feature of DPC-CM, we studied DPC secretome by human growth factor antibody array analysis and revealed the presence of several factors involved in either neurogenesis, neuroprotection, angiogenesis, and osteogenesis. The conditioning of DPCs with the B-27 supplement enhanced significantly the neuroregenerative effect of their secretome by changing its composition in growth factors. Here, we show that DPC-CM significantly stimulate neurite outgrowth in primary sensory neurons. Moreover, we identified secreted protein candidates that can potentially promote this promising regenerative feature of DPC-CM.

International audience; Titanium dental implants are used routinely, with surgical procedure, to replace missing teeth. Even though they lead to satisfactory results, novel developments with implant materials can still improve implant treatment outcomes. The aim of this study was to investigate the efficiency of porous tantalum (Ta) dental implants for osseointegration, in comparison to classical titanium (Ti). Mesenchymal stem cells from the dental pulp (DPSC) were incubated on Ta, smooth titanium (STi), and rough titanium (RTi) to assess their adhesion, proliferation, osteodifferentiation, and mineralized matrix production. Cell proliferation was measured at 4 h, 24 h, 48 h with MTT test. Early osteogenic differentiation was followed after 4, 8, 12 days by alkaline phosphatase (ALP) quantification. Cells organization and matrix microstructure were studied with scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX). Collagen production and matrix mineralization were evaluated by immunostaining and histological staining. MTT test showed significantly higher proliferation of DPSC on Ta at 24 h and 48 h. However, APL quantification after 8 and 12 days was significantly lower for Ta, revealing a delayed differentiation, where cells were proliferating the more. After 3 weeks, collagen immunostaining showed an efficient production of collagen on all samples. However, Red Alizarin staining clearly revealed a higher calcification on Ta. The overall results tend to demonstrate that DPSC differentiation is delayed on Ta surface, due to a longer proliferation period until cells cover the 3D porous Ta structure. However, after 3 weeks, a more abundant mineralized matrix is produced on and inside Ta implants. Cell populations on porous Ta proliferate greater and faster, leading to the production of more calcium phosphate deposits than cells on roughened and smooth titanium surfaces, revealing a potential enhanced capacity for osseointegration

International audience; We investigated bone regeneration in vitro and in vivo, using porous silicon (pSi) microparticles coupled with nacre proteins. We prepared a composite powder formed by pSi particles loaded with phosphate, and nacre proteins carrying calcium. For in vitro experiments, we used primary culture of human Dental Pulp Stem Cells. Results showed efficiency for both cell adhesion, cell differentiation and CaPO4 formation. But the most interesting results were that this composite was highly efficient to regenerate bone in vivo, even without any additional stem cells grafting.

Further development of biomaterials is expected as advanced therapeutic products must be compliant to good manufacturing practice regulations. A spraying method for building-up polyelectrolyte films followed by the deposition of dental pulp cells by spraying is presented. Physical treatments of UV irradiation and a drying/wetting process are applied to the system. Structural changes and elasticity modifications of the obtained coatings are revealed by atomic force microscopy and by Raman spectroscopy. This procedure results in thicker, rougher and stiffer film. The initially ordered structure composed of mainly α helices is transformed into random/β-structures. The treatment enhanced dental pulp cell adhesion and proliferation, suggesting that this system is promising for medical applications.

We report on the self-assembly based fabrication of fibrous polymers for tissue engineering applications.Directed self-assembly followed by polymerization of lysineappende ddiacetylenes generated a variety of polymers (P1−P5) with distinct chemical properties. The self-assembly along with the conjugated double and triple bonds and rigid geometry of diacetylene backbone imposed a nanofibrous morphology on the resulting polymers. Chemical properties including wettability of the polymers were tuned by using lysine (Lys) with orthogonal protecting groups (Boc andFmoc). These Lys-appended polydiacetylene scaffolds were compared in terms of their efficiency toward human mesenchymal stem cells adhesion and spreading. Interestingly, polymer P4 containing Lys Nα-NH2 and Lys Nε-Boc with balanced wettability supported cell adhesion better than the more hydrophobic polymer P2 with Nε-Boc and Nα-Fmoc or more hydrophilic polymer P5 containing free Nε and Nα amino groups. The molecular level control in the fabrication of nanofibrous polymers compared with other existing methods for the generation of fibrous polymers is the hallmark of this work. Refereed/Peer-reviewed

La découverte des cellules souches et de leurs potentialités a fait émerger de nouvelles formes de thérapies avec le développement de méthodes de bio-ingénierie cellulaire et tissulaire qui sous-tendent la médecine de demain. La disponibilité de cellules souches et leur conservation deviennent de ce fait un enjeu pour la santé de chacun. Parmi les différentes sources de cellules souches, celles de la pulpe dentaire présentent les avantages d’être pluripotentes, de pouvoir être cryopréservées et conservées durant de longues périodes sans perdre leur capacité de multiplication et de différenciation et enfin elles sont facilement accessibles. Les dents de sagesse ou les dents de lait extraites pour des raisons médicales sont une opportunité pour chacun de conserver des cellules souches pour un usage autologue. Des biobanques autorisées et spécialisées dans la préparation et la conservation des cellules souches pulpaires ouvrent l’accès à la médecine régénérative autologue de l’avenir.

International audience

International audience; The European Union (EU) directive 2010/63/EU on the protection of animals used for scientific purpose focused on reducing the number of animals and refinement of breeding. Animal studies are necessary to protect human health. Lots of animal models exist to study bone regeneration, but a reliable, well reproducible, and relatively inexpensive model with the possibility for multiple testing in the same animal is still missing. Rats may serve as good models for this due to the small animal size and good cost/benefit ratio. The present study aimed to develop a novel rat caudal vertebrae critical size defect model for bone regeneration and implant osseointegration studies The study was performed using Wistar rats with weight from 380 to 450 g. An incision was made on the dorsal side of the tail. After skin and muscles retractions, the vertebrae were exposed. Critical size defects for bone tissue engineering or implant placements for titanium body experiments were possible in each of the first four caudal vertebrae. Micro-computed tomography (CT) and histology were used to detect bone growth. There was no bone formation in the defects after 1 or 2 months of healing. When a calcium phosphate biomaterial was used (Bio-Oss®; Geistlich Pharma AG, Wolhusen, Switzerland), a good stability of the material in the defect was noted and bone growth was visible after 1 or 2 months. Results based on implant placement showed good primary stability after 3 months of healing. MicroCT showed integrated implant position through the vertebra. These results suggest that the rat caudal vertebrae may serve as a good new model for studying bone regeneration and implant osseointegration with the possibility of multiple testing within the same experimental animal and the potential to decrease number of experimental animals.

Regenerative medicine brings promising applications for mesenchymal stem cells, such as dental pulp stem cells (DPSCs). Confocal Raman microscopy, a noninvasive technique, is used to study osteogenic differentiation of DPSCs. Integrated Raman intensities in the 2800 to 3000 cm− 1 region (C-H stretching) and the 960 cm− 1 peak ( ν 1 PO 4 3 − ) were collected (to image cells and phosphate, respectively), and the ratio of two peaks 1660 over 1690 cm− 1 (amide I bands) to measure the collagen cross-linking has been calculated. Raman spectra of DPSCs after 21 days differentiation reveal several phosphate peaks: ν 1 (first stretching mode) at 960 cm− 1 , ν 2 at 430 cm− 1 , and ν 4 at 585 cm− 1 and collagen cross-linking can also be calculated. Confocal Raman microscopy enables monitoring osteogenic differentiation in vitro and can be a credible tool for clinical stem cell based research.

The way cells explore their surrounding extracellular matrix (ECM) during development and migration is mediated by lamellipodia at their leading edge, acting as an actual motor pulling the cell forward. Lamellipodia are the primary area within the cell of actin microfilaments (filopodia) formation. In this work, we report on the use of porous silicon (pSi) scaffolds to mimic the ECM of mesenchymal stem cells from the dental pulp (DPSC) and breast cancer (MCF-7) cells. Our atomic force microscopy (AFM), fluorescence microscopy, and scanning electron microscopy (SEM) results show that pSi promoted the appearance of lateral filopodia protruding from the DPSC cell body and not only in the lamellipodia area. The formation of elongated lateral actin filaments suggests that pores provided the necessary anchorage points for protrusion growth. Although MCF-7 cells displayed a lower presence of organized actin network on both pSi and nonporous silicon, pSi stimulated the formation of extended cell protrusions.

Further development of biomaterials is expected as advanced therapeutic products must be compliant to good manufacturing practice regulations. A spraying method for building-up polyelectrolyte films followed by the deposition of dental pulp cells by spraying is presented. Physical treatments of UV irradiation and a drying/wetting process are applied to the system. Structural changes and elasticity modifications of the obtained coatings are revealed by atomic force microscopy and by Raman spectroscopy. This procedure results in thicker, rougher and stiffer film. The initially ordered structure composed of mainly α helices is transformed into random/β-structures. The treatment enhanced dental pulp cell adhesion and proliferation, suggesting that this system is promising for medical applications.

Regenerative medicine brings a huge application for Mesenchymal stem cells such as Dental Pulp Stem Cells (DPSCs). Confocal Raman microscopy, a non-invasive, label free , real time and high spatial resolution imaging technique is used to study osteogenic differentiation of DPSCs. Integrated Raman intensities in the 2800-3000 cm -1 region (C-H stretching) and 960 cm -1 peak (phosphate PO 4 3- ) were collected. In Dental Pulp Stem Cells 21 st day differentiated in buffer solution, phosphate peaks ν 1 PO 4 3- (first vibrational mode) at 960cm -1 and ν 2 PO 4 3- at 430cm –1 and ν 4 PO 4 3- at 585cm -1 are obviously present. Confocal Raman microscopy enables the detection of cell differentiation and it can be used to investigate clinical stem cell research.

Objective This study evaluated the hydrophobicity of dentin surfaces that were modified through chemical silanization with octadecyltrichlorosilane (OTS). Material and Methods An in vitro experimental study was performed using 40 human permanent incisors that were divided into the following two groups: non-silanized and silanized. The specimens were pretreated and chemically modified with OTS. After the chemical modification, the dentin hydrophobicity was examined using a water contact angle measurement (WCA). The effectiveness of the modification of hydrophobicity was verified by the fluid permeability test (FPT). Results and Conclusions Statistically significant differences were found in the values of WCA and FPT between the two groups. After silanization, the hydrophobic intraradicular dentin surface exhibited in vitro properties that limit fluid penetration into the sealed root canal. This chemical treatment is a new approach for improving the sealing of the root canal system.

Stem cells discovery and their potential have led to the emergence of new forms of therapy with the development of bio-engineering cell and tissue methods underlying future medicine. The availability of stem cells and their preservation thus become an issue for everyone's health. Among the different sources of stem cells, those in the dental pulp have the advantage of being pluripotent, they can be cryopreserved and stored for long periods without losing their multiplication and differentiation capacities and finally they are easily accessible. The wisdom or natal teeth extracted for medical reasons are an opportunity for everyone to preserve stem cells for an autologous use. Biobanks authorized and specialized in the preparation and storage of pulp stem cells provide access to autologous regenerative medicine of tomorrow.

Porous silicon (pSi) is a promising biomaterial that is non-toxic and bioresorbable. Surface modifications can offer control over the degradation rate and can also impart properties that promote cell adhesion. Coupling the capacities of Dental Pulp Stem Cells (DPSC) with the pSi properties is a promising tool in regenerative medicine. P-type silicon wafers were etched at a constant current density of 30 mA/cm2 or 300 mA/cm2. The samples were oxidized or hydrosililzed. The topography of surface modified pSi samples was analysed by scanning electronic microscope (SEM) and water contact angle measurement. Dental pulp cells were collected from healthy adults and analyzed by flow cytometry. Cells were incubated on pSi samples for either 4 hours or 24 hours. Cellular morphology on pSi was evaluated with fluorescein diacetate (FDA) staining. Cell proliferation was measured through acid phosphatase activity. After oxidation or hydrosililation, at either 30 or 300 mA/cm2, pSi wafers became clearly hydrophilic. SEM revealed a highly porous surface, with a mean size of pore of 10nm±2 for 30mA/cm2, and 21nm±3 for 300mA/cm2. With flow cytometry, cells were 17% CD34+ and 77% CD146+. Acid phosphatase assay showed that samples etched with 300mA/cm2 tend to offer a better adhesion for cells; the same tendency was observed for hydrosililation treatment. Cells presented the same morphology on pSi as on culture plate. Surface modification, by turning pSi from hydrophobic to hydrophilic, allows cell adhesion. The two tested sizes of pore and the two tested surface treatments allowed adhesion of DPSC. Cell morphology on pSi was similar to culture plates. pSi resorption time is influenced by surface modification and DPSC adhesion is possible on both surface modifications, highlighting an interest for cell/tissue graft. And the tunable size of pore might permit to incorporate growth factors or nutriments inside the scaffold.

International audience

Mesenchymal stem cell secretome or conditioned medium (MSC-CM), is a combination of biomolecules and growth factors secreted by mesenchymal stem cells (MSCs) in the cell growth medium. MSC-CM emerge as an effective alternative to cell therapy for tissue regeneration applications. However, several issues such as manufacturing protocols must be addressed before the clinical application of these promising products. In this thesis, we focused on human dental pulp stem cells (DPSCs). After evaluating the impact of several manufacturing parameters on DPSC secretomes, we investigated DPSC-CM potentials for neuronal growth, bone regeneration, angiogenesis, and cancer therapy. Importantly, our work allowed the identification of standardized culture conditions providing factor-rich DPSC-CM and pointed towards promising avenues for the application of DPSC-CM to aide neuronal regeneration, and bone tissue repair. This thesis contributes to the qualitative and quantitative controls of DPSC-CM derived products necessary for their GMP-grade production, and their clinical translation in regenerative medicine.; Le sécrétome des cellules souches mésenchymateuses ou milieu conditionné (MSC-CM), est une combinaison de biomolécules et de facteurs de croissance sécrétés par les cellules souches mésenchymateuses (MSCs) dans un milieu de croissance cellulaire. Les MSC-CM apparaissent comme une alternative efficace à la thérapie cellulaire pour les applications de régénération tissulaire. Cependant, plusieurs questions telles que les protocoles de fabrication doivent être abordées avant l'application clinique de ces produits prometteurs. Dans cette thèse, nous nous sommes concentrés sur les cellules souches de la pulpe dentaire humaine (DPSCs). Après avoir évalué l'impact de plusieurs paramètres de fabrication sur les sécrétomes des DPSCs, nous avons étudié les potentiels des DPSC-CM pour la croissance neuronale, la régénération osseuse, l'angiogenèse et la thérapie contre le cancer. Ensemble, nos travaux ont permis d'identifier des conditions de culture standardisées fournissant des DPSC-CM riches en facteurs, et ont indiqué des pistes prometteuses pour l'application des DPSC-CM, afin de favoriser la régénération neuronale et la réparation des tissus osseux. Cette thèse contribue aux contrôles qualitatifs et quantitatifs des produits dérivés des DPSC-CM nécessaires à leur production selon les bonnes pratiques de fabrication, et à leur développement clinique en médecine régénérative.