Induced Mesenchymal Stem Cells Generated from Periodontal Ligament Fibroblast for Regenerative Therapy

Overview

Journal Exp Biol Med (Maywood)

Specialty Biology

Date 2025 Feb 18

PMID 39963344

Authors

Hemanathan Vembuli

Sheeja Rajasingh

Patrick Nabholz

Jefferson Guenther

Brian R Morrow

Margaret M Taylor

Marziyeh Aghazadeh

Vinoth Sigamani

Johnson Rajasingh

Affiliations

Soon will be listed here.

Abstract

Bone fractures and bone loss represent significant global health challenges, with their incidence rising due to an aging population. Despite autologous bone grafts remain the gold standard for treatment, challenges such as limited bone availability, immune reactions, and the risk of infectious disease transmission have driven the search for alternative cell-based therapies for bone regeneration. Stem cells derived from oral tissues and umbilical cord mesenchymal stem cells (MSCs) have shown potential in both preclinical and clinical studies for bone tissue regeneration. However, their limited differentiation capacity and wound healing abilities necessitate the exploration of alternative cell sources. In this study, we generated induced pluripotent stem cells (iPSCs) using a safe, nonviral and mRNA-based approach from human periodontal ligament fibroblasts (PDLF), an easily accessible cell source. These iPSCs were subsequently differentiated into MSCs, referred to as induced MSCs (iMSCs). The resulting iMSCs were homogeneous, highly proliferative, and possessed anti-inflammatory properties, suggesting their potential as a superior alternative to traditional MSCs for regenerative therapy. These iMSCs demonstrated trilineage differentiation potential, giving rise to osteocytes, chondrocytes, and adipocytes. The iMSC-derived osteocytes (iOSTs) were homogeneous, patient-specific and showed excellent attachment and growth on commercial collagen-based membranes, highlighting their suitability for bone tissue regeneration applications. Given their promising characteristics compared to traditional MSCs, PDLF-derived iMSCs are strong candidates for future clinical studies in bone regeneration and other regenerative dental therapies.

References

Carnevale G, Riccio M, Pisciotta A, Beretti F, Maraldi T, Zavatti M . In vitro differentiation into insulin-producing β-cells of stem cells isolated from human amniotic fluid and dental pulp. Dig Liver Dis. 2013; 45(8):669-76. DOI: 10.1016/j.dld.2013.02.007. View

Iohara K, Zheng L, Ito M, Tomokiyo A, Matsushita K, Nakashima M . Side population cells isolated from porcine dental pulp tissue with self-renewal and multipotency for dentinogenesis, chondrogenesis, adipogenesis, and neurogenesis. Stem Cells. 2006; 24(11):2493-503. DOI: 10.1634/stemcells.2006-0161. View

Iwata T, Yamato M, Washio K, Yoshida T, Tsumanuma Y, Yamada A . Periodontal regeneration with autologous periodontal ligament-derived cell sheets - A safety and efficacy study in ten patients. Regen Ther. 2018; 9:38-44. PMC: 6222282. DOI: 10.1016/j.reth.2018.07.002. View

Holly D, Klein M, Mazreku M, Zamborsky R, Polak S, Danisovic L . Stem Cells and Their Derivatives-Implications for Alveolar Bone Regeneration: A Comprehensive Review. Int J Mol Sci. 2021; 22(21). PMC: 8583713. DOI: 10.3390/ijms222111746. View

Liang C, Park A, Guan J . In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc. 2007; 2(2):329-33. DOI: 10.1038/nprot.2007.30. View

Rajasingh S, Thangavel J, Czirok A, Samanta S, Roby K, Dawn B . Generation of Functional Cardiomyocytes from Efficiently Generated Human iPSCs and a Novel Method of Measuring Contractility. PLoS One. 2015; 10(8):e0134093. PMC: 4523188. DOI: 10.1371/journal.pone.0134093. View

Bose S, Roy M, Bandyopadhyay A . Recent advances in bone tissue engineering scaffolds. Trends Biotechnol. 2012; 30(10):546-54. PMC: 3448860. DOI: 10.1016/j.tibtech.2012.07.005. View

Sakai K, Yamamoto A, Matsubara K, Nakamura S, Naruse M, Yamagata M . Human dental pulp-derived stem cells promote locomotor recovery after complete transection of the rat spinal cord by multiple neuro-regenerative mechanisms. J Clin Invest. 2011; 122(1):80-90. PMC: 3248299. DOI: 10.1172/JCI59251. View

Bakhtiar H, Mazidi S A, Mohammadi Asl S, Ellini M, Moshiri A, Nekoofar M . The role of stem cell therapy in regeneration of dentine-pulp complex: a systematic review. Prog Biomater. 2018; 7(4):249-268. PMC: 6304177. DOI: 10.1007/s40204-018-0100-7. View

10.

Secco M, Zucconi E, Vieira N, Fogaca L, Cerqueira A, Carvalho M . Mesenchymal stem cells from umbilical cord: do not discard the cord!. Neuromuscul Disord. 2007; 18(1):17-8. DOI: 10.1016/j.nmd.2007.11.003. View

11.

Oryan A, Alidadi S, Moshiri A, Maffulli N . Bone regenerative medicine: classic options, novel strategies, and future directions. J Orthop Surg Res. 2014; 9(1):18. PMC: 3995444. DOI: 10.1186/1749-799X-9-18. View

12.

Jin X, Lin T, Xu Y . Stem Cell Therapy and Immunological Rejection in Animal Models. Curr Mol Pharmacol. 2015; 9(4):284-288. DOI: 10.2174/1874467208666150928153511. View

13.

Kirankumar S, Gurusamy N, Rajasingh S, Sigamani V, Vasanthan J, Perales S . Modern approaches on stem cells and scaffolding technology for osteogenic differentiation and regeneration. Exp Biol Med (Maywood). 2021; 247(5):433-445. PMC: 8919323. DOI: 10.1177/15353702211052927. View

14.

Nakatsuka R, Nozaki T, Uemura Y, Matsuoka Y, Sasaki Y, Shinohara M . 5-Aza-2'-deoxycytidine treatment induces skeletal myogenic differentiation of mouse dental pulp stem cells. Arch Oral Biol. 2010; 55(5):350-7. DOI: 10.1016/j.archoralbio.2010.03.003. View

15.

Le Blanc K, Ringden O . Immunomodulation by mesenchymal stem cells and clinical experience. J Intern Med. 2007; 262(5):509-25. DOI: 10.1111/j.1365-2796.2007.01844.x. View

16.

Fang C, Yang Y, Wang Q, Yao Y, Zhang X, He X . Intraventricular injection of human dental pulp stem cells improves hypoxic-ischemic brain damage in neonatal rats. PLoS One. 2013; 8(6):e66748. PMC: 3682969. DOI: 10.1371/journal.pone.0066748. View

17.

Berner A, Reichert J, Muller M, Zellner J, Pfeifer C, Dienstknecht T . Treatment of long bone defects and non-unions: from research to clinical practice. Cell Tissue Res. 2011; 347(3):501-19. DOI: 10.1007/s00441-011-1184-8. View

18.

Liu M, Zeng X, Ma C, Yi H, Ali Z, Mou X . Injectable hydrogels for cartilage and bone tissue engineering. Bone Res. 2017; 5:17014. PMC: 5448314. DOI: 10.1038/boneres.2017.14. View

19.

Wada N, Wang B, Lin N, Laslett A, Gronthos S, Bartold P . Induced pluripotent stem cell lines derived from human gingival fibroblasts and periodontal ligament fibroblasts. J Periodontal Res. 2011; 46(4):438-47. DOI: 10.1111/j.1600-0765.2011.01358.x. View

20.

Song N, Scholtemeijer M, Shah K . Mesenchymal Stem Cell Immunomodulation: Mechanisms and Therapeutic Potential. Trends Pharmacol Sci. 2020; 41(9):653-664. PMC: 7751844. DOI: 10.1016/j.tips.2020.06.009. View