研究目的
To investigate the regulation of osteoblast differentiation by osteocytes cultured on sclerostin antibody conjugated TiO2 nanotube array for bone fracture recovery, especially in osteoporotic conditions.
研究成果
The study demonstrates that sclerostin antibody-conjugated TiO2 nanotube arrays (TNTs-scl) promote the cyto-compatibility to MLO-Y4 cells and reduce the amount of sclerostin secreted by the MLO-Y4 cells. The lowered sclerostin secretion greatly stimulated Wnt signaling to promote the differentiation of osteoblasts, providing a simple and effective approach for surface engineering of titanium to improve the bone repair in osteoporotic fracture.
研究不足
The study focuses on the in vitro co-culture of osteoblasts with osteocytes on modified titanium surfaces. The in vivo effectiveness and long-term biocompatibility of the sclerostin antibody-conjugated TiO2 nanotube arrays need further investigation.
1：Experimental Design and Method Selection:
The study involved the fabrication of TiO2 nanotube arrays on titanium substrate to mimic the nano-scale structure of natural bone and then conjugated sclerostin antibody onto their surface via dopamine. Field emission scanning electron microscopy (SEM), contact angle measurement and confocal laser scanning microscope (CLSM) were employed to characterize the conjugation of sclerostin antibody onto the surface of TiO2 nanotube arrays.
2：Sample Selection and Data Sources:
Murine osteocyte-like cell line (MLO-Y4) and primer osteoblasts isolated from the calvaria of Kunming mice were used.
3：List of Experimental Equipment and Materials:
Titanium foils, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), alkaline phosphatase (ALP) and bicinchoninic acid (BCA) assay kit, dopamine hydrochloride, sclerostin antibody, etc.
4：Experimental Procedures and Operational Workflow:
The study included the preparation of TiO2 nanotube arrays, conjugation of sclerostin antibody, cell culture, evaluation of cell viability and morphology, detection of sclerostin levels in culture medium, and expression of sclerostin gene in MLO-Y4 cells.
5：Data Analysis Methods:
The statistical analysis was performed with OriginPro (version 9.0) via the Students’ t-test and one-way analysis of variance (ANOVA) at confidence levels of 95% and 99%.
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