Self-assembly of differentiated dental pulp stem cells facilitates spheroid human dental organoid formation and prevascularization

Newswise — BACKGROUND

The self-assembly of solid organs from stem cells has the potential to greatly expand the applicability of regenerative medicine. Stem cells can self-organise into microsized organ units, partially modelling tissue function and regeneration. Dental pulp organoids have been used to recapitulate the processes of tooth development and related diseases. However, the lack of vasculature limits the utility of dental pulp organoids.

AIM

To improve survival and aid in recovery after stem cell transplantation, we demonstrated the three-dimensional (3D) self-assembly of adult stem cell-human dental pulp stem cells (hDPSCs) and endothelial cells (ECs) into a novel type of spheroid-shaped dental pulp organoid in vitro under hypoxia and conditioned medium (CM).

METHODS

During culture, primary hDPSCs were induced to differentiate into ECs by exposing them to a hypoxic environment and CM. The hypoxic pretreated hDPSCs were then mixed with ECs at specific ratios and conditioned in a 3D environment to produce prevascularized dental pulp organoids. The biological characteristics of the organoids were analysed, and the regulatory pathways associated with angiogenesis were studied.

RESULTS

The combination of these two agents resulted in prevascularized human dental pulp organoids (Vorganoids) that more closely resembled dental pulp tissue in terms of morphology and function. Single-cell RNA sequencing of dental pulp tissue and RNA sequencing of Vorganoids were integrated to analyse key regulatory pathways associated with angiogenesis. The biomarkers forkhead box protein O1 and fibroblast growth factor 2 were identified to be involved in the regulation of Vorganoids.

CONCLUSION

In this innovative study, we effectively established an in vitro model of Vorganoids and used it to elucidate new mechanisms of angiogenesis during regeneration, facilitating the development of clinical treatment strategies.

Core Tip: We demonstrated the three-dimensional self-assembly of adult stem cell-human dermal papilla cells and endothelial cells into a novel type of spheroid-shaped dental pulp organoid in vitro under hypoxia and conditioned medium. These organoids have been constructed to be morphologically and functionally closer to dental pulp tissue. Through the integration and analysis of single-cell RNA sequencing and RNA sequencing data, forkhead box protein O1 and fibroblast growth factor 2 were identified as crucial markers involved in the regulation of organoid angiogenesis. In this innovative study, we effectively established an in vitro model of prevascularized dental pulp organoids and used it to elucidate new mechanisms of angiogenesis during regeneration, facilitating the development of clinical treatment strategies.

• Citation: Liu F, Xiao J, Chen LH, Pan YY, Tian JZ, Zhang ZR, Bai XC. Self-assembly of differentiated dental pulp stem cells facilitates spheroid human dental organoid formation and prevascularization. World J Stem Cells 2024; 16(3): 287-304
• URL: https://www.wjgnet.com/1948-0210/full/v16/i3/287.htm
• DOI: https://dx.doi.org/10.4252/wjsc.v16.i3.287