Adipose-derived stromal/stem cells are verified to be potential seed candidates for bio-root regeneration in three-dimensional culture

Abstract

Background Bio-root regeneration is a promising treatment for tooth loss. It has been reported that dental-derived stem cells are effective seed cells for bio-root construction, but further applications are limited by their few sources. Human adipose tissues have a wide range of sources and numerous studies have confirmed the ability of adipose-derived stromal/stem cells (ASCs) in regenerative medicine. In the current study, the odontogenic capacities of ASCs were compared with dental-derived stem cells including dental follicle cells (DFCs), and stem cells from human exfoliated deciduous teeth (SHEDs). Methods The biological characteristics of ASCs, DFCs, and SHEDs were explored in vitro. Two-dimensional (2D) and three-dimensional (3D) cultures were compared in vitro. Odontogenic characteristics of porcine-treated dentin matrix (pTDM) induced cells under a 3D microenvironment in vitro were compared. The complexes (cell/pTDM) were transplanted subcutaneously into nude mice to verify regenerative potential. RNA sequencing (RNA-seq) was used to explore molecular mechanisms of different seed cells in bio-root regeneration. Results 3D culture was more efficient in constructing bio-root complexes. ASCs exhibited good biological characteristics similar to dental-derived stem cells in vitro. Besides, pTDM induced ASCs presented odontogenic ability similar to dental-derived stem cells. Furthermore, 3D cultured ASCs/pTDM complex promoted regeneration of dentin-like, pulp-like, and periodontal fiber-like tissues in vivo. Analysis indicated that PI3K-Akt, VEGF signaling pathways may play key roles in the process of inducing ASCs odontogenic differentiation by pTDM. Conclusions ASCs are potential seed cells for pTDM-induced bio-root regeneration, providing a basis for further research and application. Graphical Abstract

Clinical application of a double-modified sulfated bacterial cellulose scaffold material loaded with FGFR2-modified adipose-derived stem cells in urethral reconstruction

“…The SMBC was cut into 2 cm diameter disks and stored at 4 °C. Subsequently, we digested and counted ADSCs that grew to 70–80% and added them to the scaffold material at a concentration of 1 × 10 6 cell/ml [ 18 , 19 ]. The medium was changed every 2–3 days for tissue-engineered urethra and the composite was used as a graft for urethral defect repair after 7 days.…”

Treated Dentin Matrix in Tissue Regeneration: Recent Advances

“…Extracted human teeth are the most common source for harvesting TDM raw material [ 43 , 46 , 49 , 50 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 ]. Experiments conducted with animal TDMs, such as bovine [ 43 , 48 ], canine [ 28 ], goat [ 49 ], monkey [ 28 ] and porcine TDMs [ 28 , 31 , 32 , 46 , 66 , 67 , 68 ], show that these are also feasible. It is worth noting that the porcine TDM possesses similar mineral phases, bioactive molecules and odontogenic induction ability to a human TDM [ 66 ].…”

“…TDM-based bio-root construction has not only achieved the histological goal of tissue regeneration regarding pulp, dentin, cementum, periodontal ligament and alveolar bone but has also realized the superior objective of bio-functioning [ 28 ], specifically load-bearing masticatory movement [ 54 , 81 ]. Dental pulp stem cells (DPSCs) [ 50 ], dental follicle cells (DFCs) [ 27 , 28 , 29 , 30 , 31 , 32 , 54 , 66 , 82 ], adipose-derived stromal/stem cells (ASCs) [ 67 ] and related products [ 68 ] were employed to explore the potential of constructing bio-root ( Figure 4 ) out of TDM or its derivatives. The porous structure of a treated dentin matrix has enabled it to become a promising bio-material as a drug carrier and delivery system.…”

Dental pulp stem cells-derived cannabidiol-pretreated micro-spheroids showed robust osteogenic potential via upregulation of WNT6

“…During stem cell-based bone tissue engineering, in vivo seeded MSCs should proliferate and survive long enough to differentiate in osteoblasts as well as to release the growth factors and chemokines to in uence the cells of surrounding tissues such as endothelial cells and macrophages (11,12). However, the grafted seed cells can survive only a few days in vivo fading effective bone regeneration (13,14).…”

“…3D-printed porous bone grafts are mainly designed with pore sizes 200-500 µm (22). Cell aggregates/spheroids produced by the conventional 3D culture techniques fail to control the size and cell numbers which causes cell necrosis and di culties in homogenous seeding of aggregates/spheroids throughout the micro-porous scaffolds (13). Therefore, the use of the micro-well technique that can produce functional and injectable cell microspheroids with < 100 µm size could allow the seeding of spheroids homogeneously in microporous bone grafts during the surgical procedure (23).…”