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

SIMPLE SUMMARY: Tissue engineering of the dental pulp has been a goal of dental research for years. In this translational study, a chairside protocol is designed using endogenous dentin matrix proteins as signaling molecules for pulp regeneration. These bioactive molecules can be isolated from root canals by ultrasonic-activated irrigation, further processed chairside, and mixed with a hydrogel. The scaffold material is to be injected into the root canal and effect cell homing, i.e., allowing stem cells from the periapical space to migrate into the root canal. The aim of this innovative approach is the formation of an innervated and vascularized connective tissue that resembles the pulp in form and function. 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.