Essential role of ADAM28 in regulating the proliferation and differentiation of human dental papilla mesenchymal cells (hDPMCs)

Abstract

Dental papilla mesenchymal cells (DPMCs) have been supposed to possess the relatively independent and critical role for tooth development and morphogenesis. Here, we characterized the role of ADAM28, a member of a disintegrin and metalloproteinase (ADAM) family, in the regulative mechanisms of odontogenic capability of hDPMCs. Immunofluorescence staining showed the ubiquitous expression of ADAM28 in multiple human dental mesenchymal and epithelial cells. After confirming the effect of eukaryotic expression plasmid containing ADAM28 coding region and ADAM28 antisense oligodeoxynucleotide (AS-ODN), we respectively transfected them into hDPMCs and observed the biological markers for proliferation and differentiation. Overexpression of ADAM28 favored the proliferation and lineage-specific differentiation of hDPMCs, while blockage of ADAM28 exerted the opposite effects and induced apoptosis. These results identified an unrecognized hypothesis that ADAM28 may function as positive regulator of growth and differentiation of hDPMCs and act as an important molecule mediating reciprocal epithelial-mesenchymal signaling during tooth organ development.

Extending the knowledge in histochemistry and cell biology

“…The role of a member of the disintegrin and metalloproteinase (ADAM) family, ADAM28, in diVerentiation of human dental papilla mesenchymal cells was the topic of a recent work by Zhao et al (2008). After conWrming ubiquitous expression of ADAM28 in multiple human dental mesenchymal and epithelial cells, the authors worked with overexpression and antisense constructs.…”

State-of-the-art technologies, current opinions and developments, and novel findings: news from the field of histochemistry and cell biology

“…Now, Zhao et al (2008) characterized the role of a member of the a disintegrin and metalloproteinase (ADAM) family of membrane-anchored and secreted proteins with proteolytic and/or adhesive properties, ADAM28, in regulating the proliferation and diVerentiation of human dental papilla mesenchymal cells (hDPMCs). Expression of ADAM28 was documented in multiple human dental cells of the developing and matured tooth.…”

ADAM28 Manipulates Proliferation, Differentiation, and Apoptosis of Human Dental Pulp Stem Cells

“…Eukaryotic expression plasmid pcDNA3.1(+)-ADAM28 was constructed and verified by PCR identification, restriction endonuclease digestion identification, and DNA sequencing as described previously (10). Meanwhile, glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 900 bp) was used as a control; specific primers were as follows: forward: 5'-AGCCGCATCTTCTTTTGCGTC-3' and reverse: 5'-TCATATTTGGCAGGTTTTTCT-3'.…”

“…Transient transfection was performed using the Lipofectamine 2000 system (Invitrogen, Carlsbad, CA) as described previously (10). After being transfected for 48 hours, cells on coverslips were washed with PBS, fixed in 4% paraformaldehyde for 2 hours, and then subjected to immunofluorescence staining as described previously (11).…”

“…The HDPSCs growth curve was determined by MTT (3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyl-2H-tetrazoliumbromide) method as described previously (10). Single-cell suspensions of the second-passage HDPCs and purified HDPSCs were seeded into a 96-bore plate with 2 Â 10 3 cell/bore, and each group contains 10 bores.…”