Journal: International Journal of Molecular Sciences

Article Title: Preliminary Preclinical Evaluation of Innovative Bone Scaffolds Composed of Natural Sources–Whey Protein Isolate and Pearl Powder

doi: 10.3390/ijms26167939

Figure Lengend Snippet: Qualitative assessment of osteogenic differentiation of normal human osteoblasts cultured on control biomaterials–polystyrene and tested biomaterials composed of WPI without or with pearl powder (WPI/P0 and WPI/P2.5; WPI/P5; WPI/P7.5; WPI/P10, respectively). After 21 days of incubation, cells were incubated with primary anti-collagen I antibody or primary anti-osteocalcin antibody, followed by staining with specified secondary antibody-conjugated with Alexa Fluor 488 and additionally with Hoechst 33342. Then, the cells were observed using a confocal laser scanning microscope (CLSM). Cell nuclei = blue fluorescence; collagen or osteocalcin = green fluorescence. Magnification = 200× or 400×, bar scale = 70 or 30 μm.

Article Snippet: After incubation, the cells were stained with primary rabbit/IgG polyclonal anti-collagen I antibody (Invitrogen, ThermoFisher Scientific, Waltham, MA, USA), diluted 1:100 in 0.1% bovine serum albumin, BSA (Merck, Warsaw, Poland), or primary rabbit/IgG polyclonal anti-osteocalcin antibody (Bioss, ThermoFisher Scientific, Waltham, MA, USA), diluted 1:100 in 0.1% BSA (Merck, Warsaw, Poland), followed by staining with secondary goat polyclonal anti-rabbit IgG (H + L) antibody-conjugated with AlexaFluor ® 488 (Abcam, Cambridge, UK) , and diluted 1:1000 in PBS (Merck, Warsaw, Poland).

Techniques: Cell Culture, Control, Incubation, Staining, Laser-Scanning Microscopy, Fluorescence