Activation of mesenchymal stem cells promotes new bone formation within dentigerous cyst

Abstract

Background Dentigerous cyst (DC) is a bone destructive disease and remains a challenge for clinicians. Marsupialization enables the bone to regenerate with capsule maintaining, making it a preferred therapeutic means for DC adjacent to vital anatomical structures. Given that capsules of DC are derived from odontogenic epithelium remnants at the embryonic stage, we investigated whether there were mesenchymal stem cells (MSCs) located in DC capsules and the role that they played in the bone regeneration after marsupialization. Methods Samples obtained before and after marsupialization were used for histological detection and cell culture. The stemness of cells isolated from fresh tissues was analyzed by morphology, surface marker, and multi-differentiation assays. Comparison of proliferation ability between MSCs isolated from DC capsules before (Bm-DCSCs) and after (Am-DCSCs) marsupialization was evaluated by Cell Counting Kit-8 (CCK-8), fibroblast colony-forming units (CFU-F), and 5′-ethynyl-2′-deoxyuridine (EdU) assay. Their osteogenic capacity in vitro was detected by alkaline phosphatase (ALP) and Alizarin Red staining (ARS), combined with real-time polymerase chain reaction (RT-PCR) and immunofluorescence (IF) staining. Subcutaneous ectopic osteogenesis as well as cranial bone defect model in nude mice was performed to detect their bone regeneration and bone defect repairability. Results Bone tissue and strong ALP activity were detected in the capsule of DC after marsupialization. Two types of MSCs were isolated from fibrous capsules of DC both before (Bm-DCSCs) and after (Am-DCSCs) marsupialization. These fibroblast-like, colony-forming cells expressed MSC markers (CD44+, CD90+, CD31−, CD34−, CD45−), and they could differentiate into osteoblast-, adipocyte-, and chondrocyte-like cells under induction. Notably, Am-DCSCs performed better in cell proliferation and self-renewal. Moreover, Am-DCSCs showed a greater osteogenic capacity both in vitro and in vivo compared with Bm-DCSCs. Conclusions There are MSCs residing in capsules of DC, and the cell viability as well as the osteogenic capacity of them is largely enhanced after marsupialization. Our findings suggested that MSCs might play a crucial role in the healing process of DC after marsupialization, thus providing new insight into the treatment for DC by promoting the osteogenic differentiation of MSCs inside capsules.

miR-30a inhibits the osteogenic differentiation of the tibia-derived MSCs in congenital pseudarthrosis via targeting HOXD8

“…MSCs were identified by CD44, CD90, CD31, and CD34 detection, as described previously [ 19 ]. Briefly, third-generation periosteal MSCs were digested and resuspended with PBS.…”

Adult Mesenchymal Stem Cells from Oral Cavity and Surrounding Areas: Types and Biomedical Applications

“…Therefore, these previous findings supported the possibility of the presence of stem cells in the DC capsule, assigned the acronym DCCMC. In fact, Yu et al recently isolated two types of MSCs from the DC fibrous capsules of five patients, both before (Bm-DCSC) and after (Am-DCSC) marsupialization [ 242 ]. The authors reported positive expression for CD44 and CD90 cell surface antigens and negative expression for hematopoietic antigens (CD34– and CD45–).…”

“…Even better capacity to repair the bone defect was confirmed with greater capacity for ectopic bone regeneration. Following the results of this clinical trial, it stands out that MSCs reside in the DC capsule, and marsupialization improves osteogenic capacity both in vivo and in vitro [ 242 ]. This discovery provides a scientific basis for the treatment of DC after marsupialization and enables seamless subsequent orthodontic treatment as well as implant placement, paving the way for a multidisciplinary dental approach.…”