Isolation of Endogenous TGF-β1 from Root Canals for Pulp Tissue Engineering: A Translational Study

Abstract

Cell homing for dental pulp tissue engineering has been advocated as a feasible approach to regenerate dental pulp in a clinical setting. In order to develop a translational protocol for clinical application, we wanted to determine the effects of disinfectants on the availability of growth factors from the root canal, the amount that can be obtained in this context, and whether they can be processed for use in tissue engineering procedures. The extraction of growth factors should also be confirmed in a clinical setting. Root canals were prepared in 36 extracted mature teeth, and the amount of TGF-β1 in solution was quantified after different irrigation protocols (sodium hypochlorite, chlorhexidine) and after intracanal medication (calcium hydroxide). Centrifugal filters with a cut-off of 10,000 Da and 3000 Da were used for efficient concentration, and volumes and amounts of retained TGF-β1 were measured at different time points. During conventional endodontic treatment, ethylenediaminotetraacetic acid (EDTA) solution was collected after ultrasonic activation from the root canals of mature teeth of 38 patients, and growth factor content was quantified via enzyme-linked immunosorbent assay (ELISA). Irrigation with sodium hypochlorite reduced TGF-β1 release into EDTA. This effect was partially reversed by canal enlargement after the use of sodium hypochlorite and by subsequent use of calcium hydroxide. A few minutes of centrifugation with a cut-off of 10,000 Da reduced the initial volume of the irrigant by 90% and led to a continuous increase in concentration to the same extent. Furthermore, TGF-β1 was obtained from root canals of mature teeth during endodontic treatment in quantities that have been shown to elicit desirable cellular responses in a subsequent clinical application. A mixture with a suitable scaffold material and injection into the root canal has the potential to promote dental pulp regeneration.

Effect of Calcium Silicate-Based Intracanal Medicament and Calcium Hydroxide on Growth Factor TGF-β1 Release from Root Canal Dentine

“…This scenario can be extremely detrimental in regenerative endodontic procedures, owing to the compromised viability of stem cells, which typically reside near the root apex and guide the root formation process jointly with Hertwig’s epithelial root sheath [ 11 , 12 ]. Unfortunately, younger patients with immature teeth, the primary receivers of regenerative endodontic procedures [ 13 ], are directly associated with a higher risk of inadvertent extrusion due to the limited patient cooperation and open root apices [ 14 ]. Moreover, the complete removal of calcium hydroxide paste from various root canal irregularities has also been reported to be difficult, resulting in the blockage of the dentinal tubules due to the remnants on the root canal surface [ 15 ].…”

Evaluation of immunohistochemical activity of dentin-pulp organs of dog’s teeth after stimulation with laser, MTA, and theracal therapies

“…(19)(20)(21) Additionally, it was stated that the different soluble proteins like TGF-β1can be extorted from the dentine matrix and can employ an orthodox biological effect on the stem cells of the pulp and encourage the repair mechanisms of the mineralized hard tissues. (22,23) Moreover, VEGF is a signal heparin-binding protein known as vascular permeability factor and it is released from the protein GFs, therefore, it indicates the amount of these protein GFs. (24) However, the quantity of TGF-β1 and VEGF that are liberated from the dentin matrix in response to the capping materials, has not been established.…”

Engineering the Future of Dental Health: Exploring Molecular Advancements in Dental Pulp Regeneration

“…In this context, stem cell-based or primarily cell-free approaches use specifically tailored scaffold materials and signaling molecules to achieve pulp regeneration, both in terms of tissue microarchitecture and functionality (Figure 1). Several of these approaches already take into account the requirements of potential clinical applications [9][10][11].…”