Decellularized swine dental pulp extracellular matrix: a comparison between formed and developing teeth

Abstract

Objective: To decellularize and characterize, histologically and biochemically, the extracellular matrix (ECM) of dental pulp from fully formed (M1) and developing (M2) swine molars, comparing native and decellularized tissues. Methods: Experimental in vitro study using coronal pulps from molars of commercially slaughtered swine, divided into M1 (erupted, with a complete root) and M2 (intraosseous, developing). An enzyme-free, Triton X-100-free protocol based on EDTA and SDS was applied. DNA and collagen were quantified, and histology (H&E, Masson's trichrome and DAPI) and scanning electron microscopy were performed; data were analysed by one-way ANOVA with Tukey's post-hoc test (p<0.05). Results: Decellularization removed the cellular content of both pulps, with absence of nuclei and DNA reduction of 92.7% in M1 and 96.8% in M2 (p<0.0001), with no residual difference between groups (p=0.60). M2 pulps, collected from the maxilla only, showed a greater yield per tooth (1.173 g versus 0.377 g) and maintained stable collagen. A porous extracellular matrix scaffold was observed in both groups, with mineralized deposits suggestive of intrinsic mineralization, exclusive to decellularized M2. Conclusion: The conservative protocol effectively decellularized both pulps. Teeth at distinct developmental stages provide ECM with particular characteristics, which is relevant to scaffold selection in regenerative endodontics.