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Global Translational Medicine Gelatin-based cell carriers for tooth-germ organoids

material and structural properties in tooth-germ of growth factors, immune modulators, or cytokines. This
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organoids. This property is crucial, as material distribution dual-functionality carrier holds promise for applications in
during differentiation directly influences tooth germ other tissue regeneration contexts as well.
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development. Staining intensity was semi-quantitatively
analyzed using ImageJ software, examining over five 4. Conclusion
images per sample (>15 images per group) to assess Tooth organoids hold great promise for supporting tooth
levels of osteogenic differentiation (Figure 4B). Across all regeneration, creating a demand for cell carriers that are not
groups except GelMA HG, cells subjected to osteogenic only biocompatible and effective but also scalable for clinical
induction showed significantly stronger staining compared applications. In this study, we compared four different
to those in growth medium (+ versus -). Notably, induced gelatin-based cell carriers to evaluate their ability to support
cells on Gel MS (Gel MS+) exhibited stronger staining the growth and differentiation of hDPSCs. Our results
compared to those on GelMA MS or GelMA MP. The demonstrate that Gel MS, produced through a reproducible
higher cell density on Gel MS may contribute to these and scalable process, most effectively supported cell growth
observed differences in intensity. In addition, factors such and osteogenic differentiation of hDPSCs compared to
as sphere diameter, curvature, and size distribution have GelMA-based carriers (GelMA MS and GelMA MP). While
been shown to influence the osteogenic differentiation further research is needed to elucidate the mechanisms
of stem cells. Substrate stiffness is also known to affect underlying the differential effects of these carriers, Gel
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the osteogenic differentiation of stem cells, as reported MS has emerged as a strong candidate for tooth organoid
in several studies. 40-42 The Gel MS, GelMA MP, GelMA generation. Moving forward, pre-clinical studies using Gel
MS, and GelMA HG synthesized in this study have not MS will be crucial to determine its effectiveness in vivo and
been characterized for mechanical properties such as its potential to support tooth regeneration in clinical settings.
stiffness, which may differ due to the various crosslinking
methods used. Exploring the role of these features in Acknowledgments
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cell differentiation on cell carriers could provide valuable
insights into their impact on hDPSC differentiation and The authors would like to express their gratitude to
their potential as scaffolds. Dr. Pamela Yelick and Dr. Weibo Zhang at Tufts University
for providing hDPSCs. The authors also thank Dr. Derya
The mechanisms underlying the differential effects of Ozhava, Peter Winkler, and Krishi Patel for their technical
various cell carriers on hDPSCs require further investigation. assistance. Anisha Jackson acknowledges the support of the
Once these mechanisms are fully understood, it will be Aresty Research Center at Rutgers University. All illustrations
possible to develop cell carriers with optimized features that in this report were created using Biorender software.
facilitate tooth-germ organoid formation and harness the
advantages of multiple attributes. Using a combination of Funding
heterogeneous cell carriers to maximize the benefits of diverse None.
features could also be a viable approach. Future studies will
assess contributing factors such as base materials (gelatin Conflict of interest
versus GelMA) and particle sizes. In addition, to determine
whether these in vitro observations are translatable to in vivo The authors declare that they have no competing interests.
conditions and whether these organoids can support tooth Author contributions
regeneration, pre-clinical studies will be essential.
Conceptualization: All authors
Our results suggest that Gel MS not only provides
superior support for cell growth but also promotes Methodology: All authors
osteogenic differentiation in organoid-like aggregates of Investigation: Anisha Jackson, Yong Mao
Visualization: All authors
hDPSCs. Among the four cell carriers tested, Gel MS stands Writing – original draft: Anisha Jackson, Yong Mao
out as a promising candidate for dental organoid formation.
Notably, GelMA MS and GelMA MP showed similar Writing – review & editing: All authors
capabilities in supporting cell growth and differentiation. Ethics approval and consent to participate
However, given that the production of GelMA MP is simpler,
more reproducible, and scalable than that of GelMA MS, Not applicable.
GelMA MP may serve as a practical alternative. Importantly,
the photocrosslinking of GelMA MP does not compromise Consent for publication
protein integrity, making it suitable for the incorporation Not applicable.

Volume 4 Issue 1 (2025) 77 doi: 10.36922/gtm.5897