IADR Abstract Archives
POTENTIAL BIOACTIVE PROPERTIES OF PULP DERIVED EXTRACELLULAR MATRIX HYDROGELS
Objectives: The objective of this study was to prepare and characterize pulp derived Extracellular Matrix (P-ECM) hydrogels for regenerative endodontic applications.
Methods: Thirty Freshly extracted bovine molar teeth were used to prepare P-ECM; Bovine dental pulp tissues were harvested. For decellularization, pulp tissues were treated by trypsin/EDTA for 24 hours, washed by deionized water, treated with DNase, then washed with phosphate-buffered saline. Evaluation of the decellularization process was done using DNA quantification and histological examination compared to native tissues. This was followed by fabrication of extracellular matrix hydrogel scaffold and optimization of its concentration. The prepared scaffolds were evaluated for their protein content using BCA kit, protein release at 0,1 and 3 days, as well as MTT cell viability assay using Rabbit dental pulp stem cells (rDPSCs).
Results: Bovine pulp tissues were successfully decellularized. DNA was found to be below the cut-off point (50 ng/mg tissue) using Nanodrop spectrophotometer (32±2.4 ng/mg tissue) and using Quantifluor fluorescent dye (21.18±1.8 ng/mg tissue). Histological evaluation using H&E, Alcian Blue and Masson’s trichome revealed absence of nuclei, retention of glycosaminoglycans (GAGs) and collagen content, respectively. P-ECM hydrogels were fabricated with a final concentration of 3.00 mg/ml. Total protein content was found to be 493.12 µg/µl. Protein release was detected at time 0 showing burst release (109.6 µg/µl) followed by declined release at 24 hours (70.4 µg/µl) then increased release at 72 hours (102.7 µg/µl). P-ECM hydrogel maintained viability of rDPSCs as compared to cells cultured under controlled conditions.
Conclusions: The extracellular matrix pulp hydrogel scaffold retained its bioactive properties demonstrating a potential role as a scaffold for regenerative endodontic procedures.
Methods: Thirty Freshly extracted bovine molar teeth were used to prepare P-ECM; Bovine dental pulp tissues were harvested. For decellularization, pulp tissues were treated by trypsin/EDTA for 24 hours, washed by deionized water, treated with DNase, then washed with phosphate-buffered saline. Evaluation of the decellularization process was done using DNA quantification and histological examination compared to native tissues. This was followed by fabrication of extracellular matrix hydrogel scaffold and optimization of its concentration. The prepared scaffolds were evaluated for their protein content using BCA kit, protein release at 0,1 and 3 days, as well as MTT cell viability assay using Rabbit dental pulp stem cells (rDPSCs).
Results: Bovine pulp tissues were successfully decellularized. DNA was found to be below the cut-off point (50 ng/mg tissue) using Nanodrop spectrophotometer (32±2.4 ng/mg tissue) and using Quantifluor fluorescent dye (21.18±1.8 ng/mg tissue). Histological evaluation using H&E, Alcian Blue and Masson’s trichome revealed absence of nuclei, retention of glycosaminoglycans (GAGs) and collagen content, respectively. P-ECM hydrogels were fabricated with a final concentration of 3.00 mg/ml. Total protein content was found to be 493.12 µg/µl. Protein release was detected at time 0 showing burst release (109.6 µg/µl) followed by declined release at 24 hours (70.4 µg/µl) then increased release at 72 hours (102.7 µg/µl). P-ECM hydrogel maintained viability of rDPSCs as compared to cells cultured under controlled conditions.
Conclusions: The extracellular matrix pulp hydrogel scaffold retained its bioactive properties demonstrating a potential role as a scaffold for regenerative endodontic procedures.