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BACKGROUND: The commonly used culture methods for primary intervertebral disc cells are type Ⅱcol agenase alone digestion method, and type Ⅱ col agenase combined trypin digestion method. However, the acquired cells are few. OBJECTIVE: To acquire nucleus pulposus cells from human degenerative intervertebral disc using a systemic and simplified method. METHODS: Nucleus pulposus cells were isolated and cultured. The morphology of nucleus pulposus cells was observed under inverted phase contrast microscope every day. Primary and subcultured cellsuspension was applied for the determination of cel viability using trypan blue staining. The cel growth was detected with MTT assay. The cel morphology was observed under laser confocal microscopy. RESULTS AND CONCLUSION: Nucleus pulposus cells were successful y isolated from from human degenerative intervertebral disc using the improved method, and cells were subcultured to passage 3. Primary cells were fusiform shaped, while cells at passage 1 and 2 were triangle or polygonal, which were similar to fibroblasts. When cells almost reached the confluence, the cells showed slabstone-like appearance. Trypan blue staining showed that, the primary cel viability was 99%, and passage 3 cells had 93%-95% viability. The proliferation of nucleus pulposus cells gradual y decreased as the generation increased. Compared with passage 1 cells, passage 2 and 3 cells at logarithmic phase trended to be smoother. The cel morphology observed under laser confocal microscopy was similar to the results under phase contrast microscope. The improved simple method can successful y acquire a variety of cells from human degenerative intervertebral disc, and these cells show fine biological properties.

6.Application of free-form deformation algorithm in fast three-dimensional bone reconstruction.

Xiangsen ZENG ; Hai ZHOU ; Chengtao WANG ; Dongmei WANG ; Gong CHEN

Journal of Biomedical Engineering 2014;31(5):1121-1126

This paper presents a unit free-form deformation (FFD) method applied to rapid three-dimensioanl (3D) bone reconstruction, which was based on traditional FFD. With the femur as an example, we reconstructed a 3D model of femur from two X-ray images and a standardized model by taking advantage of unit FFD algorithm. The X-ray images and its parameters were taken by C-arm device. Those parameters and X-ray contour are contributed to 3D reconstruction. The out contours of X-ray image and standard model were connected by point matching algorithm. The unit-FFD lattice was built to reconstruct standard model and finally made the contour of X-ray image and standard model exactly the same. Experiments on shape accuracy, robustness and time consuming, carried out by 35 specimen from cadaver, showed that mean error of shape (0. 52 mm) and mean construction time (112 s) were lower than those using traditional method (0.7-2.6 mm, 8-20 min). The method proposed in this paper shows a good prospect in clinical application and related research.
Algorithms ; Femur ; anatomy & histology ; diagnostic imaging ; Humans ; Imaging, Three-Dimensional ; Models, Anatomic ; Radiography