Abstract
Dental pulp is a major component of the dental body that serves to maintain the tooth life and function. The aim of the present work was to develop a system that functions as a growth-permissive microenvironment for dental pulp regeneration using a decellularized dental pulp (DDP) matrix, 5-Aza-2′-deoxycytidine (5-Aza), and Extracellular Vesicles (EVs) derived from human Dental Pulp Stem Cells (hDPSCs). Human dental pulps extracted from healthy teeth, scheduled to be removed for orthodontic purpose, were decellularized and then recellularized with hDPSCs. The hDPSCs were seeded on DDP and maintained under different culture conditions: basal medium (CTRL), EVs, 5-Aza, and EVs+-5-Aza. Immunofluorescence staining and Western blot analyses were performed to evaluate the proteins’ expression related to dentinogenesis, such as ALP, RUNX2, COL1A1, Vinculin, DMP1, and DSPP. Protein contents found in the DDP recellularized with hDPSCs were highly expressed in samples co-treated with EVs and 5-Aza compared to other culture conditions. This study developed a DDP matrix loaded by hDPSCs in co-treatment with EVs, which might enhance the dentinogenic differentiation with a high potentiality for endodontic regeneration.
Highlights
Published: 8 February 2022Teeth life and homeostasis are maintained by dental pulp, which can be damaged by caries, pulpitis, injury, and pulp necrosis
In the present study adn for the first time to our knowledge, we developed a system constituted by human decellularized dental pulp (DDP) and used it as an ECM 3D scaffold enriched with human Dental Pulp Stem Cells (hDPSCs), Extracellular Vesicles (EVs), and 5-Aza-2 -deoxycytidine (5-Aza) that lead to a progressive cellular differentiation in cells with an odontoblastic phenotype in order to promote dental pulp tissue regeneration
To explore the effect of EVs and 5-Aza on the odontogenic differentiation potential of hDPSCs seeded on DDP scaffold, we examined the expression of ALP, RUNX2, COL1A1, Vinculin, DMP1, and DSPP by means Western blot analysis
Summary
Teeth life and homeostasis are maintained by dental pulp, which can be damaged by caries, pulpitis, injury, and pulp necrosis. Often when dental pulp is affected, pulpal necrosis is an obvious consequence [1,2]. To ameliorate the quality of life, dental pulp damage must be restored in order to maintain the functionality of the pulp-dentin complex. When dental pulp is affected by trauma or caries, the most used treatment is root canal therapy, which leads to the loss of pulp tissue functions. After this type of treatment, teeth no longer survive. Tissue engineering could represent a valid alternative strategy to ‘save’
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