Comparative study of periodontal differentiation propensity of induced pluripotent stem cells from different tissue...

Despite being almost identical to embryonic stem cells, induced pluripotent stem cells (iPSCs) have been shown to possess a residual somatic memory that favors their differentiation propensity into donor tissue. To further confirm this assumption, we compare for the first time the periodontal differentiation tendency of human gingival fibroblast-derived iPSCs (G-iPSCs) and human neonatal skin fibroblast-derived iPSCs (S-iPSCs) to assess whether G-iPSCs could be more efficiently induced toward periodontal cells.

We induced G- and S-iPSCs under the treatment of growth/differentiation factor-5 and connective tissue growth factor, respectively, for 14 days. Immunofluorescence staining and real-time polymerase chain reaction were used to compare their expression levels of related markers. Furthermore, a hydrogel carrier was developed to seed these periodontal progenitors for subcutaneous implantation in non-obese diabetic-severe combined immunodeficiency disease mice. Their differentiated periodontal phenotype maintenance was further assayed by HE observation, immunohistochemical staining and immunofluorescence co-localization with pre-labeled PKH67.

As expected, both iPSCs were inclined to differentiate back into their original lineage by expressing higher markers at both gene and protein levels in vitro. HE observation of G-iPSCs-seeded hydrogel constructs present more mineralized structure formation than S-iPSCs-seeded ones. Immunohistochemical staining and immunofluorescence analysis also showed stronger positive staining for periodontal related markers in G-iPSCs-seeded hydrogel constructs.

Our results preliminarily confirmed that both G- and S-iPSCs were inclined to differentiate back into their original tissue in vitro. Animal study further confirmed the phenotype maintenance of periodontal differentiated G-iPSCs, which highlighted their significant implications for therapeutic use in periodontal regeneration.