Extracellular Matrix Derived From Dental Pulp Stem Cells Promotes Mineralization

Abstract

Background: Extracellular matrix (ECM) plays a pivotal role in many physiological processes. ECM macromolecules and associated factors differ according to tissues, impact cell differentiation, and tissue homeostasis. Dental pulp ECM may differ from other oral tissues and impact mineralization. Thus, the present study aimed to identify the matrisome of ECM proteins derived from human dental pulp stem cells (DPSCs) and its ability to regulate mineralization even in cells which do not respond to assaults by mineralization, the human gingival fibroblasts (GF).Methods: ECM were extracted from DPSCs cultured in normal growth medium supplemented with L-ascorbic acid (N-ECM) or in osteogenic induction medium (OM-ECM). ECM decellularization (dECM) was performed using 0.5% triton X-100 in 20 mM ammonium hydroxide after 21 days. Mass spectrometry and proteomic analysis identified and quantified matrisome proteins.Results: The dECM contained ECM proteins but lacked cellular components and mineralization. Interestingly, collagens (COL6A1, COL6A2, and COL6A3) and elastic fibers (FBN1, FBLN2, FN1, and HSPG2) were significantly represented in N-ECM, while annexins (ANXA1, ANXA4, ANXA5, ANXA6, ANXA7, and ANXA11) were significantly overdetected in OM-ECM. GF were reseeded on N-dECM and OM-dECM and cultured in normal or osteogenic medium. GF were able to attach and proliferate on N-dECM and OM-dECM. Both dECM enhanced mineralization of GF at day 14 compared to tissue culture plate (TCP). In addition, OM-dECM promoted higher mineralization of GF than N-dECM although cultured in growth medium.Conclusions: ECM derived from DPSCs proved to be osteoinductive, and this knowledge supported cell-derived ECM can be further utilized for tissue engineering of mineralized tissues.

Effects of decellularized extracellular matrix derived from Jagged1-treated human dental pulp stem cells on biological responses of stem cells isolated from apical papilla

“…The hFc-treated hDPSCs dECM did not show the remaining mineral content. This observation was consistent with a previous study showing that decellularized process could eliminate the mineral content in dECM of the hDPSCs culture in the OM medium ( Nowwarote et al, 2021 ). However, in the present study, we noted that decellularization could not eliminate mineral components in Jagged-1 treated hDPSCs dECM.…”

“…In this study, we generated and characterized the dECMs from immobilized Jagged1-treated hDPSCs and subsequently illustrated their osteoinductive activity on SCAPs. Previous reports showed that human gingival stem cells (hGSCs) could well attach, spread, and proliferate on hDPSC-derived dECMs ( Nowwarote et al, 2021 ). In addition, hDPSC-derived ECM markedly enhanced the osteogenic differentiation of hGSCs ( Nowwarote et al, 2021 ).…”

“…Previous reports showed that human gingival stem cells (hGSCs) could well attach, spread, and proliferate on hDPSC-derived dECMs ( Nowwarote et al, 2021 ). In addition, hDPSC-derived ECM markedly enhanced the osteogenic differentiation of hGSCs ( Nowwarote et al, 2021 ). Decellularization eliminates the genetic components while preserving proteins and ECM properties by chemical, enzymatic, physical, or combinative methods ( Gilpin and Yang 2017 ).…”

MatrisomeDB 2.0: 2023 updates to the ECM-protein knowledge database

“…MatrisomeDB 2.0 also includes two new studies on the ECM of hepatic metastasis from colorectal tumors ( 54 , 55 ) and an additional dataset on the ECM of aging lung ( 56 ). Last, we included three studies reporting the composition of the ECM produced in vitro by dental pulp stem cells ( 57 ), lung fibroblasts ( 58 ) and kidney organoids ( 59 ). Importantly, this release includes, for the first time, data on the ECM of aging tissues (skin, lung, intervertebral discs).…”

Potential of extracellular space for tissue regeneration in dentistry

“…For example, Melatonin-loaded hydrogels show favourable viscoelastic properties and promote accelerated and enhanced quality of bone formation in regeneration of furcation defect in dogs ( Abdelrasoul et al, 2022 ). Specifically, extracellular matrix derived from dental pulp stem cells can promote mineralization of human gingival fibroblasts ( Nowwarote et al, 2022 ). Dual ECM scaffolds derived from pulp and endothelial ECM promote attachments of mesenchymal stem cells and subsequently odontogenic differentiation.…”