ropn protein  (R&D Systems)

Journal: International Journal of Molecular Sciences

Article Title: Osteopontin on the Dental Implant Surface Promotes Direct Osteogenesis in Osseointegration

doi: 10.3390/ijms23031039

Figure Lengend Snippet: H&E-staining and OPN-immunoreactivity in the tissues surrounding the implants in the WT mice. ( a , g , m ) H&E-staining in the tissues surrounding the implants at day 5 after implant placement in the control, rOPN, and HABP-OPNpep groups. The infiltration of inflammatory cells and spindle-shaped or flattened cells is observed at the bone-implant interface. ( d , j , p ) There is a weak OPN positive immunoreaction at the bottom parts of threads and the cement lines of the pre-existing bone (arrowheads). ( b , c , h , i , n , o ) The formation of direct osteogenesis is clearly observed at week 1 (arrows) and week 2. ( e , f , k , l , q , r ) OPN-immunoreactivity gradually becomes intense (arrowheads) and elongates along the implant surface at week 2. B, bone. Scale bar = 50 μm.

Article Snippet: We soaked the HAB-implants [ ] in different OPN modified protein/peptides solution including rOPN protein (R&D Systems, Minneapolis, MN, USA; catalog no. 441-OP) (rOPN group; 20 μM in phosphate buffer saline [PBS]), Gly-Arg-Gly-Asp-Ser peptide (Peptide Institute Inc, Osaka, Japan; Fibronectin Active Fragment, #4189) (RGDS group; 3.1 mM in PBS), OPN mimic peptide (sequence: GRGDSLAYGLR [OPNpep group], theoretical molecular weight: 1164.27, purity: 97.95% [HPLC method], 3.1 mM in PBS), HABP-OPNpep (sequence: GGGLHAHKKPTQDIRGGGRGDSLAYGLR [HABP-OPNpep group], theoretical molecular weight: 3101.48, purity: 98.8% [HPLC method] 3.1 mM in PBS) (both peptides were provided from GenScript Japan [Tokyo, Japan] according to our order), and PBS (control) for 2 min in addition to filling the cavity with each solution before implant placement.

Techniques: Staining, Control

Journal: International Journal of Molecular Sciences

Article Title: Osteopontin on the Dental Implant Surface Promotes Direct Osteogenesis in Osseointegration

doi: 10.3390/ijms23031039

Figure Lengend Snippet: Azan-staining, OPN-immunoreactivity, and the rates of direct osteogenesis or OPN-positive perimeters in the WT mice. ( a – f ) Azan-staining in the tissues surrounding the implants at 2 weeks after implant placement in the rOPN, RGDS, and control groups. Partially, indirect osteogenesis progresses from the pre-existing bone in addition to the direct osteogenesis to achieve osseointegration at week 2. ( d – f ) are higher magnifications of the boxed areas in ( a – c ), respectively. ( g – i ) The OPN-immunoreactive lines coincide with the places where the direct osteogenesis occurs, although some areas lack the OPN reaction. ( j ) Quantification of the rates of direct osteogenesis and OPN-positive perimeters in rOPN ( n = 5), RGDS ( n = 5), OPNpep ( n = 5), HABP-OPNpep ( n = 5), and control ( n = 3) groups. The RGDS and HABP-OPNpep groups without following normal distribution were compared by Kruskal–Wallis test followed by the Bonferroni test for multiple comparisons for more than three groups or Mann–Whitney U test for two groups. In the rOPN group, the OPN-positive perimeter around the implant surface significantly increases compared with that in the RGDS and HABP-OPNpep groups at week 2 and shows the highest rate compared with other groups. The OPN-positive perimeter in the OPNpep group is higher than that in the RGDS group. Statistical analysis used a two-tailed Student’s t -test or Mann–Whitney U test. As to the percentage of the vertical axis, the numerator is direct osteogenesis or OPN-positive perimeter around implant surface and the denominator is the perimeter of external surface of implant body. The rates are the mean + SD. IS, implant space. Scale bars = ( a – c ) 500, ( d – i ) 50 μm.

Article Snippet: We soaked the HAB-implants [ ] in different OPN modified protein/peptides solution including rOPN protein (R&D Systems, Minneapolis, MN, USA; catalog no. 441-OP) (rOPN group; 20 μM in phosphate buffer saline [PBS]), Gly-Arg-Gly-Asp-Ser peptide (Peptide Institute Inc, Osaka, Japan; Fibronectin Active Fragment, #4189) (RGDS group; 3.1 mM in PBS), OPN mimic peptide (sequence: GRGDSLAYGLR [OPNpep group], theoretical molecular weight: 1164.27, purity: 97.95% [HPLC method], 3.1 mM in PBS), HABP-OPNpep (sequence: GGGLHAHKKPTQDIRGGGRGDSLAYGLR [HABP-OPNpep group], theoretical molecular weight: 3101.48, purity: 98.8% [HPLC method] 3.1 mM in PBS) (both peptides were provided from GenScript Japan [Tokyo, Japan] according to our order), and PBS (control) for 2 min in addition to filling the cavity with each solution before implant placement.

Techniques: Staining, Control, MANN-WHITNEY, Two Tailed Test