Histological Evaluation of A unilateral Critical-Sized Mandibular Defect Reconstruction using human dental pulp stem cells by Light Microscope and Real-Time Quantitative PCR

Abstract

Objective: To evaluate the osteogenic differentiation potential of human dental pulp stem cells (DPSCs) isolated from the dental pulp of third molar teeth in vitro cultures, and to evaluate the bone regenerative capacity of human dental pulp stem cells (DPSCs) when transplanted into a unilateral critical-sized bone defect in the mandibular bone in vivo after receiving a hydroxyapatite matrix and polylactic-polyglycolic acid (HA/PLGA) scaffold. Material and methods: A total of 18 mandibular defects were made, and three groups (each n = 6) were created. The first group: the transplanted DPSCs implanted in the critical-sized bone defect after receiving (HA/PLGA) scaffold. The second group received only (HA/PLGA) scaffold. The third group, which served as the control, had a critical-sized defect left empty. After characterization, Von Kossa [VK] and Alizarin red staining were employed to identify differentiated osteoblasts at the 14th and 21st days, and histological analyses, as well as polymerase chain reactions (PCR), were also used. Results: It showed that DPSCs had high proliferation potential and typical fibroblastic shape. Additionally, osteogenic differentiation of DPSCs was validated by morphological alterations, histological examination, and the expression of lineage-specific genes confirmed osteogenic differentiation of DPSCs. Conclusion: High proliferation potential and the capacity to differentiate into osteoblasts are two characteristics of DPSCs taken from impacted third molars.