TY - JOUR
T1 - Anti-cementoblastoma-derived protein antibody partially inhibits mineralization on a cementoblastic cell line
AU - Alvarez Pérez, Marco Antonio
AU - Pitaru, Sandu
AU - Alvarez Fregoso, Octavio
AU - Reyes Gasga, José
AU - Arzate, Higinio
N1 - Funding Information:
The authors wish to thank Dr. Larry W. Fisher for the gift of polyclonal antibodies against OPN (LF-123) and BSP (LF-100). Rogelio Fragoso from CINVESTAV-México, José Guzmán from IIM-UNAM, Luis Rendón, Pedro Mexía, Carlos Flores, and Roberto Hernández from IF-UNAM, for their technical assistance during the course of this study. Their support is greatly appreciated. This research was partially supported by DGAPA-UNAM (IN200599, IN200501) and CONACyT 30735-M.
PY - 2003/7/1
Y1 - 2003/7/1
N2 - The effect of human anti-cementoblastoma-derived protein antibody during cementogenesis in vitro was investigated by using human cementoblastoma-derived cells. Cultures treated with 5μg/ml of CP antibody from day 1 to day 15 revealed a significant decrease in alkaline phosphatase activity (ALP) 40% (p<0.005), 44% (p<0.001), 49% (p<0.1), and 45% (p<0.02) at 9, 11, 13, and 15 days, respectively. Immunoexpression of osteopontin revealed that in cultures treated with anti-CP antibody, the positive number of cementoblastoma cells was reduced by 87, 83, 69, and 52% at 5, 7, 9, and 11 days, respectively. Bone sialoprotein immunoexpression showed a decrease in positive cells of 82, 51, 60, 80, 83, and 87% at 5, 7, 9, 11, 13, and 15 days, respectively, as compared to controls. The Ca/P ratio of the mineral-like tissue deposited in vitro by cementoblastoma cells revealed that control cultures had a Ca/P ratio of 1.45 and 1.61 at 5 and 15 days, whereas experimental cultures revealed a Ca/P ratio of 0.50 and 0.79 at 5 and 15 days, respectively. Electron diffraction patterns showed inner double rings representing D-spacing that were consistent with those of hydroxyapatite in both control and experimental cultures. Examination of the crystallinity with high resolution transmission electron microscopy showed homogeneous and preferential spatial arrangement of hydroxyapatite crystallites in control and experimental cultures at 15 days. Atomic force microscopy images of control cultures at 5 and 15 days revealed small granular particles and grain agglomeration that favored the formation of crystalline plaques with a lamellar-like pattern of the mineral-like tissue. Experimental cultures at 5 and 15 days showed tiny and homogeneous granular morphology. The agglomerates maintained spherical morphology without organization of needle-like crystals to form plaque-like structures. Based on these findings, it is hypothesized that cementoblastoma-derived protein may be associated to crystal growth, compositional and morphological features during the mineralization process of cementum in vitro.
AB - The effect of human anti-cementoblastoma-derived protein antibody during cementogenesis in vitro was investigated by using human cementoblastoma-derived cells. Cultures treated with 5μg/ml of CP antibody from day 1 to day 15 revealed a significant decrease in alkaline phosphatase activity (ALP) 40% (p<0.005), 44% (p<0.001), 49% (p<0.1), and 45% (p<0.02) at 9, 11, 13, and 15 days, respectively. Immunoexpression of osteopontin revealed that in cultures treated with anti-CP antibody, the positive number of cementoblastoma cells was reduced by 87, 83, 69, and 52% at 5, 7, 9, and 11 days, respectively. Bone sialoprotein immunoexpression showed a decrease in positive cells of 82, 51, 60, 80, 83, and 87% at 5, 7, 9, 11, 13, and 15 days, respectively, as compared to controls. The Ca/P ratio of the mineral-like tissue deposited in vitro by cementoblastoma cells revealed that control cultures had a Ca/P ratio of 1.45 and 1.61 at 5 and 15 days, whereas experimental cultures revealed a Ca/P ratio of 0.50 and 0.79 at 5 and 15 days, respectively. Electron diffraction patterns showed inner double rings representing D-spacing that were consistent with those of hydroxyapatite in both control and experimental cultures. Examination of the crystallinity with high resolution transmission electron microscopy showed homogeneous and preferential spatial arrangement of hydroxyapatite crystallites in control and experimental cultures at 15 days. Atomic force microscopy images of control cultures at 5 and 15 days revealed small granular particles and grain agglomeration that favored the formation of crystalline plaques with a lamellar-like pattern of the mineral-like tissue. Experimental cultures at 5 and 15 days showed tiny and homogeneous granular morphology. The agglomerates maintained spherical morphology without organization of needle-like crystals to form plaque-like structures. Based on these findings, it is hypothesized that cementoblastoma-derived protein may be associated to crystal growth, compositional and morphological features during the mineralization process of cementum in vitro.
KW - Bone sialoprotein
KW - Cementoblastoma-derived protein
KW - Cementum
KW - Hydroxyapatite
KW - Mineralization
KW - Osteopontin
UR - http://www.scopus.com/inward/record.url?scp=0041666499&partnerID=8YFLogxK
U2 - 10.1016/S1047-8477(03)00116-3
DO - 10.1016/S1047-8477(03)00116-3
M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???
AN - SCOPUS:0041666499
SN - 1047-8477
VL - 143
SP - 1
EP - 13
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 1
ER -