The Effect Of Hepatocyte Growth Factor (HGF) On The Biological Activity Of Human Degenerative Nucleus Pulposus Cells Cultured In Vitro

Background Disc degeneration (intervertebral disc degeneration, the IVD) is the leadingcause of spinal diseases such as disc herniation, spondylolisthesis, spinal stenosis. Studies showthat nucleus pulposus cells play a central role to maintain normal disc homeostasis and repairdegenerative disc change. Hepatocyte growth factor (HGF) is a multifunctional factor.It can acton a variety of tissues and cells, promote cell proliferation, angiogenesis, inhibit apoptosis andother effects.HGF plays a role in the ontogeny and tissue repair . This study investigated the roleof hepatocyte growth factor on nucleus pulposus cells proliferation activity and expression ofextracellular matrix, and to seek new ideas for the reversal of disc degeneration.Objective 1. To explore the effect of hepatocyte growth factor (HGF) on the biologicalactivity of the human degeneration nucleus pulposus cells cultured in vitro.2. To study the combined effects between HGF and IGF on the degeneration of thenucleus pulposus cells.Methods Collect the human nucleus pulposus, isolate and culture nucleus pulposus cells.Immunohistochemistry methods were used to detect the expression of type II collagen; Nucleuspulposus cells staining by HE. Observe HGF receptor expression in nucleus pulposus cells byImmunohistochemical staining. The experimental groups were cultured with 0、5、50、500ng/mlHGF, selects the optimal concentration by MTT. Select the 3rd generation of nucleus pulposuscells , the experiment group HGF、HGF+IGF、IGF and control group, reversetranscription-polymerase chain reaction (RT-PCR) was used to determine levels of type IIcollagen、proteoglycan and SOX9; flow cytometer are used to detect cell cycle.Results Nucleus pulposus cells express type II collagen; nuclei stained purple by HE staining.Nucleus pulposus cells express HGF receptor. HGF could significantly promote the proliferationof the human degeneration nucleus pulposus cells, which had a significant different comparedwith the control group. Various concentrations of HGF were effective to promote the nucleuspulposus cells proliferation and the optimal HGF concentration was 50ng/ml. The humandegeneration nucleus pulposus cells under HGF、IGF and HGF+IGF had shown to keep typeⅡcollagen、proteoglycan and SOX9 expression positive and increasing, and the proportion of cellsinto the proliferative phase increased compared with control group, which HGF+IGF is moreeffective.Conclusion Nucleus pulposus cells expressed HGF receptor. HGF can promote theproliferation of the human degeneration nucleus pulposus cells cultured in vitro, enhance theexpression of extracellular matrix. The combined effects of HGF and IGF are better than the two alone.