ABSTRACT
Objective:
The
biomimetic coating on
titanium surface
affects the adhesion, proliferation, and differentiation of
bone cells on the surface of implants. Naringin-7-O-Neohesperidoside (NRG) positively
affects the proliferation and differentiation of
bone cells, while inhibiting the formation of
osteoclasts, thereby affecting the osteogenic effect. This study aimed to construct
biomimetic coatings on pure
titanium surfaces using layer by layer (LBL)
self-assembly of NRGat different concentrations. The effects of the assembled NRG
biomimetic coatings on the proliferation and differentiation of
mouse preosteoblast
cells (MC3T3-E1)
in vitro were investigated. The influence of NRG concentration and
culture time on MC3T3-E1
cells was also explored.
Methods:
LBL is a
technology that allows for the creation of thin
membranes made of
polyelectrolytes through
electrostatic attraction between polyanions and polycations, which effectively incorporates charged
polyelectrolytes onto solid surfaces while preserving their
biological activity.
Alkaline phosphatase (ALP)
plays a crucial
role in
biomineralization, and its activity is considered as a marker for
osteoblast differentiation. Real-
time quantitative
PCR accurately and quantitatively
measures gene expression levels, which reflect the transcriptional activity of
genes and thus reflect the proliferation and differentiation of
osteoblasts. The
research different concentrations of NRG
biomimetic coatings (1×10-4 mol/L, 1×10-5 mol/L, 1×10-6 mol/L, and 1×10-7 mol/L) were constructed on
titanium surfaces using the LBL
self-assembly
technique. The
control groups included the blank group and the group without
drugs. The effects of the coatings on the proliferation of MC3T3-E1
cells were evaluated by ALP activity assay. The differentiation of MC3T3-E1
cells was evaluated by ALP activity assay. Real-
time quantitative
PCR was performed to detect the
gene expressions of OC
mRNA, Runx2
mRNA, and Col1a1
mRNA in MC3T3-E1
cells grown on the
titanium samples of different experimental groups.
Results:
The proliferation indices of all NRG concentration groups were higher than those of the groups without
drug and blank groups. The highest ALP value was detected at a concentration of 10-4 mol/L. All NRG concentrations upregulated the expression of Col1al
mRNA compared to the group without the
drug, and the concentrations of 10-5 mol/L and 10-6 mol/L showed statistically significant differences (P < .01). NRG at a concentration of 10-6 mol/L significantly upregulated the expression of Runx2
mRNA (P < .05), while all NRG concentration groups upregulated the expression of OC
mRNA. NRG at a concentration of 10-6 mol/L demonstrated a 4 times increase in Runx2
mRNA expression, indicating a significant impact on osteogenic differentiation.
Conclusions:
NRG
biomimetic coatings on
titanium surfaces were successfully constructed using the LBL
technique. NRG at different concentrations had stronger effects on the proliferation and differentiation of MC3T3-E1
cells compared to the groups without
drug and blank groups, with the concentration of 10-6 mol/L demonstrating the best effect. These findings suggest that NRG-loaded
biomimetic coatings may enhance the
osseointegration of
titanium implants, offering promising prospects for dental and orthopedic applications.