Decellularized Human Dental Pulp as a Scaffold for Regenerative Endodontics

Abstract

Teeth undergo postnatal organogenesis relatively late in life and only complete full maturation a few years after the crown first erupts in the oral cavity. At this stage, development can be arrested if the tooth organ is damaged by either trauma or caries. Regenerative endodontic procedures (REPs) are a treatment alternative to conventional root canal treatment for immature teeth. These procedures rely on the transfer of apically positioned stem cells, including stem cells of the apical papilla (SCAP), into the root canal system. Although clinical success has been reported for these procedures, the predictability of expected outcomes and the organization of the newly formed tissues are affected by the lack of an available suitable scaffold that mimics the complexity of the dental pulp extracellular matrix (ECM). In this study, we evaluated 3 methods of decellularization of human dental pulp to be used as a potential autograft scaffold. Tooth slices of human healthy extracted third molars were decellularized by 3 different methods. One of the methods generated the maximum observed decellularization with minimal impact on the ECM composition and organization. Furthermore, recellularization of the scaffold supported the proliferation of SCAP throughout the scaffold with differentiation into odontoblast-like cells near the dentinal walls. Thus, this study reports that human dental pulp from healthy extracted teeth can be successfully decellularized, and the resulting scaffold supports the proliferation and differentiation of SCAP. The future application of this form of an autograft in REPs can fulfill a yet unmet need for a suitable scaffold, potentially improving clinical outcomes and ultimately promoting the survival and function of teeth with otherwise poor prognosis.

A biocompatible decellularized pulp scaffold for regenerative endodontics

“…Recently, decellularization of the pulp, using human tooth slices, which were essentially pulp–dentine discs 1.5 mm in thickness, has been reported (Song et al . ). In contrast, the present study reports decellularization of the whole pulp retrieved from extracted teeth.…”

“…Song et al . () evaluated three different protocols for decellularization, the most effective protocol being the one which incorporated three cycles of 1% SDS and 1 cycle of 1% Triton X‐100. Similar detergents were also used by Chen et al .…”

“…The DNA removal efficiency (approximately 98% of DNA content) in this study was comparable to that reported by Song et al . () in spite of using a single cycle of SDS at 33 times lower concentration and without the use of Triton X‐100. The sufficient removal of cellular materials is an important step for future clinical usage as remaining materials could act as foreign body triggering a host immune reaction (Wilshaw et al .…”

“…() and Song et al . () reported successful decellularization of dental tissues utilizing 1% sodium dodecyl sulphate (SDS) and 1% Triton X‐100. Despite these results, the use of decellularization protocols with lower concentrations of toxic detergents is advantageous for future tissue regeneration.…”

“…Within the dental field attempts to decellularize porcine tooth buds (Traphagen et al 2012, Zhang et al 2017, pulp tissues obtained from miniature swine teeth (Chen et al 2015) and human pulp tissues within root slices (Song et al 2017) have been reported. Chen et al (2015) and Song et al (2017) reported successful decellularization of dental tissues utilizing 1% sodium dodecyl sulphate (SDS) and 1% Triton X-100. Despite these results, the use of decellularization protocols with lower concentrations of toxic detergents is advantageous for future tissue regeneration.…”