The regulatory role of a disintegrin and metalloproteinase 28 on the biologic property of human periodontal ligament stem cells.
BACKGROUND
A disintegrin and metalloproteinase 28 (ADAM28) is considered to be the possible virulence gene for congenital hypoplasia of tooth root. Periodontal ligament stem cells (PDLSCs) are regarded as playing crucial roles in the developing process of the periodontium and are generally used in dental regenerative medicine. This study evaluates the influence of ADAM28 on the biologic property of human PDLSCs (HPDLSCs) and to postulate the possible mechanism of this influence.


METHODS
HPDLSCs were acquired by immunomagnetic bead selection and identified by immunofluorescence detection. After ADAM28 eukaryotic plasmid and antisense oligodeoxynucleotides (AS-ODNs) were constructed and respectively transfected into HPDLSCs by a transfection reagent, the expression differences of ADAM28 among various groups were assessed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide and cell-cycle assays were used to test the proliferation activity of HPDLSCs. Annexin V-fluorescein isothiocyanate/propidium iodide analysis was performed to detect the apoptotic level. Cell differentiation was tested by measuring the alkaline phosphatase level. Immunocytochemistry and Western blotting were carried out to determine the effects of ADAM28 AS-ODNs on HPDLSCs expressing cementum attachment protein (CAP), osteopontin, and osteocalcin.


RESULTS
The ADAM28 eukaryotic plasmid group showed the highest expression level in HPDLSCs, whereas the AS-ODN group displayed the lowest. Furthermore, the overexpression of ADAM28 enhanced the proliferation of HPDLSCs and inhibited the specific differentiation of HPDLSCs, whereas the inhibition of ADAM28 produced the opposite effects and induced apoptosis. ADAM28 AS-ODNs were able to significantly inhibit CAP expression, and ADAM28 had a positive correlation with CAP.


CONCLUSIONS
Our findings demonstrated that ADAM28 was able to effectively manipulate the proliferation, apoptosis, and differentiation of HPDLSCs.
Zhao
Zheng
Z
Institute of Stomatology, General Hospital of Chinese People's Liberation Army, Beijing, China.
Wang
Yi
Y
Wang
Dongsheng
D
Liu
Hongchen
H
D051722:ADAM Proteins / Q000235:genetics* / Q000502:physiology; D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology*; D017209:Apoptosis; D002453:Cell Cycle; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003771:Dental Papilla / Q000166:cytology / Q000201:enzymology*; D019483:Disintegrins / Q000235:genetics* / Q000502:physiology; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D006801:Humans; D008297:Male; D009805:Odontogenesis / Q000235:genetics*; D015675:Osteocalcin / Q000096:biosynthesis; D053495:Osteopontin / Q000096:biosynthesis; D010513:Periodontal Ligament / Q000002:abnormalities / Q000166:cytology* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D017027:Protein Tyrosine Phosphatases / Q000096:biosynthesis; D014071:Tooth Abnormalities / Q000235:genetics*; D014092:Tooth Root / Q000002:abnormalities*; D014162:Transfection; D055815:Young Adult
D019483:Disintegrins; D015675:Osteocalcin; D053495:Osteopontin; C404489:HACD1 protein, human; D017027:Protein Tyrosine Phosphatases; D051722:ADAM Proteins; C410224:ADAM28 protein, human