The Effects Of Scaffold Materials Of Epimedium/Natural Polymer/HA On The Proliferation Of Osteoblasts

Objective: Because the epimedium can invigorate the kidney and strong the bone, the epimedium was added to hydroxyapatite, silk fibroin and chitosan of bone matrix material. Through the technique of low temperature and cryodesiccate, HA/CS-SF-EP was prepared to study the law of effect on the adhesion, growth and proliferation of osteoblast, to provide theory basis for the repair of bone and also to provide reliable basis for clinical application.Method: The crania osteoblasts were obtained from newborn wistar rats within 24 hours. The crania of rats were disgested by the collagenase and primary cultured. In about seven days cells lined up in one layer, by differential velocity adherent technique separate osteoblasts and others.The cells were generated and their biological characteristic was examined by phase-contrast microscope, HE stain, histochemical staining of alkaline phosphatase (ALP) and mineralized nodules stain. The best cells were transplanted to HA/CS,HA-SF/CS and HA-SF/CS/EP at a density of 5×104/ml. Observe the biological characteristic of the osteoblasts by phase-contrast microscope and scanning electron microscope (SEM). Measure the cell proliferation by the assay of ALP activities and MTT.Results: 1. In the respect of morphology, we found the cultured rat crania cells possessed the morphological characteristic and biological behavior of osteoblasts. The shape of cells were triangle or polygon.2. In the respect of histology, black particles could be observed in the endochylema of the cultured cells with ALP stain by light microscope,in accordance with the standard of the osteoblasts.3. Under scanning electron microscape, few osteoblasts attached to HA/CS,HA-SF/CS and HA-SF/CS/EP on the second. Osteoblast could increase and extend fully on three materials on the9th day. The number of cells were most on HA-SF/CS/EP compared with others. 4. Through the testing of the cell's alksline phosphatase activity upon materials showed that the contents of ALP increased before the nine days. The contents of ALP of the different time was different (P< 0.05). Through MTT, experiment group was compared with control group (P< 0.05). The optical density of HA-SF/CS/EP was highest compared with the other two experiment groups and control group (P< 0.05). The resules showed that HA-SF/CS/EP has obviouseeffect on promoting the proliferation of the cell.Conclusion: 1. ALP and MTT showed that compared with HA/CS and HA-SF/CS, HA-SF/CS/EP has better effect on promoting proliferation of osteoblasts. SEM showed that HA-SF /CS and HA-SF/CS/EP possessed biocompatibility as HA/CS.2. Epimedium can promote the proliferation of the cell. HA-SF/CS/EP had actual feasibility acting as the scaffold materials of bone tissue engineering.