Objective To qualitatively analyze the expressions of key marker genes involved in osteogenic differentiation of bone marrow stromal cells (BMSCs) of mice at defined stages in vitro using real-time RT-PCR. Methods The BMSCs of mice were cultured in vitro and induced to osteoblasts for 7, 14, and 21 days. The total RNA was extracted from the cultured cells and the gene expression levels of collagenⅠ, bone alkaline phosphatase (APL), osteopontin (OPN) and bone sialoprotein (BSP) in the osteoblasts at the defined stages were determined by real-time RT-PCR. Results The ColⅠmRNA levels in 14 and 21 days were (0. 75?0. 04) and (0. 34?0. 03) times respectively of that in 7 days. ALP, OPN and BSP mRNA levels in 21 days were (19. 70?2. 36), (150. 12?9. 31) and (7. 73?0. 58) times of those in 7 days respectively. Conclusions ColⅠexpression levels tend to gradually decline while ALP, OPN and BSP levels gradually increase along with the culture time. Real-time RT-PCR is a reliable method for investigation of gene expressions in BMSCs cultured in vitro.

Background In previous study,peripheral retinal nerve fiber layer (RNFL) thickness is considered to be the earliest structural changes which can be detected.3D-OCT can measure the thickness of macular ganglion cell complex (mGCC),which makes the detection of primary glaucoma possible in the early stage.Objective This study was to measure the thickness of mGCC and disc-peripheral RNFL in early stage of primary glaucomous eyes by 3D-OCT and assess the anatomic basis of glaucoma-induced optical nerve damage.Methods 3D-OCT images from 10 patients with advanced stage primary glaucoma in one lateral eye and early stage glaucoma in fellow eye from December 2010 to December 2012 were prospectively analyzed in China-Japan Friendship Hospital.The patients were diagnosed based on the recommended standard of National glaucoma group (1987 version) and received routine eye examination.3D-OCT scanning was performed using 3D-macular mode,3D-macular Wide mode and 3D-disc mode with TOPCON 3D-OCT 2000 system,and the images at macular 6 mm×6 mm area were analyzed.The posterior pole area was divided into 5 concentric rings from fovea toward periphery and equally subdivided into 100 small checks,with the area of 0.6 mm×0.6 mm for each.The probable values in each check were calculated as the ratio of each figure and corresponding normal value.The probable values were expressed as red color (P＜ 1％),yellow color (P＜5％) and gray color (P≥ 5％).Then the disc-periphery RNFL thickness and disc cup were evaluated.Results No evident abnormality was found in the thicknesses of photoreceptors layer and bipolar cell layer in both advanced glaucomous eyes and the early stage of glaucomous eyes in the 10 patients.Serious damage of visual field was seen in the advanced glaucomous eyes and presented with red color in the parapapillary RNFL area,mGCC area and macular RNFL area,showing an evidently attenuation of the thicknesses of parapapillary RNFL,mGCC and RNFL.However,the visual field was close normal in the early stage glaucomous eyes,and mGCC and macular RNFL showed yellow color,while green or yellow color was exhibited in the parapapillary RNFL area,indicating mGCC and macular RNFL thickness was reduced,but parapapillar RNFL thickness was near normal.Conclusions The change of mGCC thickness is earlier than that of peripheral RNFL at optic disc in primary glaucomaous eyes,which may imply that the disappear of macular ganglion cell body is earlier than that of the axon.

Objective To explore the expression of transient receptor potential channel 6(TRPC6)in human breast cancer cells, and to clarify the correlation of TRPC6 with the invasion potential of breast cancer cells. Methods The human breast cancer cell strains MCF-7 (hypo-invasion group)and MDA-MB-231 (hyper-invasion group)were cultured.The expressions of TRPC6 mRNA and protein in in two groups were detected by RT-PCR and Western blotting methods.Then the MDA-MB-231 cells were divided into control group and SKF96365 group, the effects of SKF96365 on the invasion ability of MDA-MB-231 cells invitro were explored by wound healing assay and Transwell experiment.Results The results of Western blotting and RT-PCR showed that the expression levels of TRPC6 mRNA and protein in MDA-MB-231 cells were higher than that in MCF-7 cells(P<0.05).The wound healing assay showed the numbers of migrating cells in 5,25 and 40μmol·L-1 SKF96365 groups (76.24±7.54, 45.33±4.50,25.12±1.57)were lower than those in control group (130.48±9.55)(P<0.05).The Transwell experiment results indicated that the invasiveness of MDA-MB-231 cells were inhibited significantly by SKF96365 compared with control group (P<0.05).Conclusion The invasion ability of human breast cancer MDA-MB-231 cells is promoted by upregulating the TRPC6 expression, which indicates that the TRPC6 may play role in the metastasis of human breast cancer.

The application of bone tissue engineering regeneration technology is expected to repair maxillofacial bone tissue defects caused by tumors, trauma, etc. Surface patterning occupies an important position in bone tissue engineering. Microcontact printing is an emerging technology through which the elastic stamp contacts with the substance and materials used as ink can be transferred from stamp to substance to form patterns. The biggest characteristic of the technology is to fabricate high-throughput and high-accuracy patterned surface, making it widely applied. This review summarized the application and optimization of microcontact printing, and prospected its application in bone tissue engineering.

Bone defects caused by trauma, tumour resection, infection and congenital deformities, together with articular cartilage defects and cartilage-subchondral bone complex defects caused by trauma and degenerative diseases, remain great challenges for clinicians. Novel strategies utilising cell sheet technology to enhance bone and cartilage regeneration are being developed. The cell sheet technology has shown great clinical potential in regenerative medicine due to its effective preservation of cell-cell connections and extracellular matrix and its scaffold-free nature. This review will first introduce several widely used cell sheet preparation systems, including traditional approaches and recent improvements, as well as their advantages and shortcomings. Recent advances in utilising cell sheet technology to regenerate bone or cartilage defects and bone-cartilage complex defects will be reviewed. The key challenges and future research directions for the application of cell sheet technology in bone and cartilage regeneration will also be discussed.
Bone Regeneration ; Bone and Bones ; Cartilage, Articular ; Regeneration ; Tissue Engineering ; trends ; Tissue Scaffolds