The effect of immobilization of heparin and bone morphogenic protein-2 to bovine bone substitute on osteoblast-like cell's function

Abstract

PURPOSEThis study was performed to investigate the ability of recombinant human-bone morphogenic protein-2 immobilized on a heparin-grafted bone substrate to enhance the osteoblastic functions.MATERIALS AND METHODSThe Bio-Oss®, not coated with any material, was used as a control group. In rhBMP-2-Bio-Oss® group, rhBMP-2 was coated with Bio-Oss® using only deep and dry methods (50 ng/mL, 24 h). In heparinized rhBMP-2-Bio-Oss® group, dopamine was anchored to the surface of Bio-Oss®, and coated with heparin. rhBMP-2 was immobilized onto the heparinized- Bio-Oss® surface. The release kinetics of the rhBMP-2-Bio-Oss® and heparinized rhBMP-2-Bio-Oss® were analyzed using an enzyme-linked immunosorbent assay. The biological activities of the MG63 cells on the three groups were investigated via cytotoxicity assay, cell proliferation assay, alkaline phosphatase (ALP) measurement, and calcium deposition determination. Statistical comparisons were carried out by one-way ANOVA test. Differences were considered statistically significant at *P<.05 and **P<.001.RESULTSThe heparinized rhBMP-2-Bio-Oss® showed more sustained release compared to the rhBMP-2-Bio-Oss® over an extended time. In the measurement of the ALP activity, the heparinized group showed a significantly higher ALP activity when compared with the non-heparinized groups (P<.05). The MG63 cells cultivated in the group with rhBMP-2 showed increased calcium deposition, and the MG63 cells from the heparinized group increased more than those that were cultivated in the non-heparinized groups.CONCLUSIONHeparin increased the rhBMP-2 release amount and made sustained release possible, and heparinized Bio-Oss® with rhBMP-2 successfully improved the osteoblastic functions.

Effects of BMP-2 Delivery in Calcium Phosphate Bone Graft Materials with Different Compositions on Bone Regeneration

“…To facilitate the sustained release of rhBMP-2, Huh et al [39] attempted to chemically immobilize rhBMP-2 to DOPA-heparin on xenogenous bone. However, the chemical approach is challenging clinically and potential chemical toxicity restricts its clinical application.…”

Improving Osteogenesis Activity on BMP-2-Immobilized PCL Fibers Modified by theγ-Ray Irradiation Technique

“…This result was in agreement with previous studies conducted by Kim et al [ 30 ] and Park et al [ 38 ]. However, in other previous studies, there was no significant increase in cell proliferation observed in substrates with BMP-2 as compared to the substrates without BMP-2 [ 27 – 29 , 39 ]. Thus, the effect of BMP-2 on cell proliferation is still not clear.…”

“…To demonstrate the osteogenic differentiation of MG-63 cells on PCL fibers and surface-modified PCL fibers, we assessed the ALP activity, calcium deposition, and the mRNA gene expressions of OCN and OPN. ALP activity is well known to be able to evaluate the early differentiation of osteoblast-like cells [ 30 , 39 ]. The ALP activity of MG-63 cells cultured on PCL fibers and surface-modified PCL fibers was increased throughout the 10 days of culture.…”

“…Moreover, the ALP activity of MG-63 cells on the BMP-2/AAc-PCL fibers surface on the 7th or 10th day was significantly higher than that on BMP-2/PCL fibers. The amount of calcium deposition was widely used to demonstrate the late differentiation of osteoblast-like cells [ 30 , 39 ]. From the ALP activity data, the amount of calcium deposition on MG-63 cells cultured on AAc-PCL fibers was found to be similar compared to the PCL fibers.…”

Fabrication of a BMP-2-immobilized porous microsphere modified by heparin for bone tissue engineering

“…It is well known that heparin is composed of many sulfate and carboxyl groups and has good binding affinities for BMP-2. Huh et al [41] and Kim et al [42] showed that BMP-2 from Ti and bovine bone (Bio-Oss) modified with heparin were released in a sustained fashion compared to BMP-2 from Ti and bovine bone (Bio-Oss). Consistent with the previous studies, this study showed the sustained release of BMP-2 from BMP-2/Hep-PMS.…”