Ex Vivo Characterization of alphaSMA+ Periodontal Stem Cells and the Role of Bmp2 in Mus Musculus
Abstract
Background: Many approaches to regenerate lost periodontal tissues are studied with varying degrees of success, to include multiple sources of stem cells. The aim of this study was to examine the ex vivo differentiation of alpha smooth muscle actin positive periodontal stem cells (aSMA+ PSCs) and the effect of bone morphogenetic protein 2 (Bmp2) on differentiation capability. Methods: aSMA+ PSCs were isolated from Bmp2-floxed mice mandibles and implanted subcutaneously in mice using absorbable gelatin sponge and denuded molar after: no additional treatment (n=4), green fluorescent protein (GFP) labeling (n=6), or Bmp2 knockout (n=3). Mice were sacrificed at 4-8 weeks, and retrieved samples underwent selected staining and immunocytochemistry (H and E, Trichrome, Bsp-Ab, Postn-Ab, SMA-Ab, Bmp2-Ab, EdU ClickiT). Results: 8 of 13 transplants were successfully retrieved 5 were not present upon re-entry. Several teeth were lost from migration during implantation/healing. aSMA-Ab and GFP labeling confirmed implanted aSMA+ PSCs presence in retrieved samples. Strong bone sialoprotein signaling was observed in acellular cementum regions of several samples. Mineralized tissue consistent with bone was present and oriented adjacent to PDL-like fibroblasts in 6-week GFP sample. Periostin was seen in fibroblast cells adjacent to the root surface and throughout samples when a tooth was not present. Bmp2 was present in untreated and GFP samples. Initial observations showed Bmp2 knockout samples formed less bone and PDL fibroblasts. Conclusions: aSMA+ PSCs can differentiate ex vivo to produce bone, PDL fibroblasts, and possibly acellular cementum. The tooth acts as a matrix for differentiation. Knockout of Bmp2 disrupts this differentiation capacity.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 09, 2020
- Accession Number
- AD1182713
Entities
People
- Audrey Rakian
- Brian J. Alent
- Kristi N. Kennedy
- Obadah Austah
- Stephen E. Harris
Organizations
- Uniformed Services University of the Health Sciences
- University of Texas at Austin