Abstract
Implementing the principles of tissue engineering within the clinical management of non-vital immature permanent teeth is of clinical interest. However, the ideal scaffold remains elusive. The aim of this work was to assess the feasibility of decellularising rat dental pulp tissue and evaluate the ability of such scaffold to support stem cell repopulation. Rat dental pulps were retrieved and divided into control and decellularised groups. The decellularisation protocol incorporated a low detergent concentration and hypotonic buffers. After decellularisation, the scaffolds were characterised histologically, immunohistochemistry and the residual DNA content quantified. Surface topography was also viewed under scanning electron microscopy. Biocompatibility was evaluated using cytotoxicity assays utilising L-929 cell line. Decellularised scaffolds were recellularised with human dental pulp stem cells up to 14 days in vitro. Cellular viability was assessed using LIVE/DEAD stain kit and the recellularised scaffolds were further assessed histologically and immunolabelled using makers for odontoblastic differentiation, cytoskeleton components and growth factors. Analysis of the decellularised scaffolds revealed an acellular matrix with histological preservation of structural components. Decellularised scaffolds were biocompatible and able to support stem cell survival following recellularisation. Immunolabelling of the recellularised scaffolds demonstrated positive cellular expression against the tested markers in culture. This study has demonstrated the feasibility of developing a biocompatible decellularised dental pulp scaffold, which is able to support dental pulp stem cell repopulation. Clinically, decellularised pulp tissue could possibly be a suitable scaffold for use within regenerative (reparative) endodontic techniques.
Highlights
Implementing the principles of tissue engineering within the clinical management of non-vital immature permanent teeth is of clinical interest
Immunolabelling with major histocompatibility complex class II antibodies revealed a positive immunoreactivity in the control pulp tissues, this positive staining was associated with specific cells scattered throughout the extracellular matrix (ECM) (Fig. 1a)
The ECM is regarded as a vital component of all tissues and, possibly, the ideal scaffold for tissue e ngineering[14]
Summary
Implementing the principles of tissue engineering within the clinical management of non-vital immature permanent teeth is of clinical interest. The aim of this work was to assess the feasibility of decellularising rat dental pulp tissue and evaluate the ability of such scaffold to support stem cell repopulation. This study has demonstrated the feasibility of developing a biocompatible decellularised dental pulp scaffold, which is able to support dental pulp stem cell repopulation. Careful consideration and incorporation of bioengineering principles (the use of stem cells, scaffold material and morphogenic signals) within such a treatment strategy is essential in the clinical application of r egenerative[5,6,7], or more accurately termed reparative[8], endodontics. The ECM is a vital and dynamic structure composed of tissue specific structural and functional proteins This complex integration between cells and the surrounding microenvironment (ECM proteins) regulates cellular activity and may provide a suitable environment for tissue regeneration[14]
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