Osteocalcin Antibody - #DF12303

Fig. 5 Biocompatibility assessments. a CCK-8 assay about the proliferation of MC3T3-E1 preosteoblasts cultured on Ti-OH and Ti-PAA-NCl after cells were cultured for 1, 3 and 7 days (n = 3; P = 0.673, 0.639 and 0.145 for 1, 3 and 7 days compared with Ti-OH, respectively; Student’s t test). b Morphology of MC3T3-E1 preosteoblasts cultured on Ti-OH and Ti-PAA-NCl for 1 day (green for F-actin, blue for cell nucleus, scale bar = 20 μm). c ALP activity of MC3T3-E1 preosteoblasts cultured on Ti-OH and Ti-PAA-NCl for 7 and 14 days (n = 3; P = 0.132 and 0.954 for 7 and 14 days compared with Ti-OH; Student’s t test). d Alizarin Red S staining of MC3T3-E1 preosteoblasts cultured on Ti-OH and Ti-PAA-NCl for 21 days (red for calcium nodules, scale bar = 1 mm) and semi-quantitative measurement of calcium content (n = 3; P = 0.147 compared with Ti-OH; Student’s t test). e Western blot results of osteogenic-related proteins (OCN, OPN and RUNX2) expressed in MC3T3-E1 preosteoblasts cultured on Ti-OH and Ti-PAA-NCl for 3, 7 and 14 days after osteogenic induction. f RT-qPCR results of expression levels of osteogenic-related genes (OCN, OPN and RUNX2) in MC3T3-E1 preosteoblasts cultured on Ti-OH and Ti-PAA-NCl for 3, 7 and 14 days after osteogenic induction (n = 3; P > 0.05 for all time points of these three genes between groups; Student’s t test). g In vivo biocompatibility. HE staining and CD68 immunofluorescent staining of tissues around Ti-OH and Ti-PAA-NCl embedded in the backs of nude mice for 4 weeks (fluorescent green for CD68, fluorescent blue for cell nucleus, white scale bars in HE images = 100 μm, black scale bars in HE images = 25 μm, scale bars in CD68 immunofluorescent images = 100 μm). All error bars = s.d.

Figure 8 Representative sections of rat calvaria after implantation. (A) Masson trichrome staining of decalcified cranial sections at 8 weeks. (B) The results of Sirius red staining for new bone formation. (C) Typical images of alkaline phosphatase, osteocalcin and anti-CD31 staining observed by fluorescence microscopy; quantitative percentage of each fluorescent area for different groups; DAPI-DNA was stained in blue, Rhodamine-ALP, OCN and CD31 were stained in red. (***P<0.001)

Fig. 6 Representative images for immunohistochemical staining of COL-1 (A) and OCN (B). (scale bar at low magnification: 200 ​μm, scale bar at high magnification: 50 ​μm).