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Research Biocompatibility of Osteoblasts with Nano Hydroxyapatite/Artificial Bone Collagen (NHAC) Adding Vascular Endothelial Growth Factor (VEGF)

Author: ZhaoHui
Tutor: KangYiJun
School: Central South University
Course: Surgery
Keywords: Nano- hydroxyapatite / collagen artificial bone (NHAC) Vascular endothelial growth factor (VEGF) Allogeneic bone Bone cement Osteoblasts Compatibility of bone cells
CLC: R318.08
Type: Master's thesis
Year: 2010
Downloads: 85
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Abstract

Objective: selection of MG-63 cell lines cultured osteoblasts with nano-hydroxyapatite / collagen bone substitute material (NHAC) combined with vascular endothelial growth factor (VEGF) co-cultured evaluation of the composite bone cells relative from the perspective of cytology capacitive, compared with allogeneic bone, bone cement and blank control group, provided the experimental basis for clinical application. : 106-generation MG-63 cell lines in vitro osteoblast cultured cell culture surface NHAC / VEGF, allogeneic bone, bone cement, three materials, the surface and the blank control group, four MTT Determination of trace enzyme reaction colorimetry (MTT) cell viability; using Coomassie brilliant blue, the determination of the total protein content of cultured cells with material composite; using reverse transcriptase polymerase chain reaction (RT-PCR) technology osteoblast culture ALP mRNA and osteocalcin hormone (OC) of the transcription of mRNA expression analysis. Quantitative analysis of osteoblasts secrete inorganic calcium material at the surface of various materials using calcium standard colorimetric analysis NHAC / VEGF osteoblasts compatibility, and allogeneic bone, NHAC / VEGF, bone cement the osteoblasts compatibility between the three groups whether the differences. Results: 1, the cell viability assessed by MTT assay results visible cells of each group of materials with the prolonged incubation time, the number has increased the cell absorbance values ??have statistically significant differences between groups of each time point ( P lt; 0.05). Osteoblasts total protein content in different materials, training the early allogeneic bone material group is higher than the other groups, NHAC / VEGF material group than the control group and the bone cement group, little difference in bone cement group and the control group . Than among the groups were statistically significant (P lt; 0.05). With the incubation time, the protein content has increased the trend train mid-protein content increased significantly, mature osteoblasts compared, the difference is not obvious. Osteoblast differentiation capacity determination: osteoblasts cultured on the three materials 7 days after ALPmRNA in each group were expressed, ALP mRNA relative expression levels of allogeneic bone material group gt; NHAC / of of VEGF materials group gt; bone cement group, the groups were compared in each group having a significant difference (P LT; 0.05); OC are not expressed. 14 days after the ALP mRNA relative expression level of allogeneic bone material group and the NHAC / VEGF materials group was no statistically significant difference (P gt; 0.05), but significantly higher than the bone cement group, comparison between groups was statistically difference (P lt; 0.05); OC mRNA in the bone cement group did not express, only a small group of allogeneic bone material and the NHAC / VEGF material group expression. 21 days after the ALP mRNA in each group were expressed, ALP mRNA relative expression level of bone cement group gt; NHAC / of of VEGF materials group gt; allogeneic bone material group, a statistically significant difference (P lt; 0.05) between groups; OC mRNA expression in the three groups, allogeneic bone material group and NHAC / of VEGF materials group was higher than the bone cement group, the relative expression levels between the two groups no statistically significant difference (P gt; 0.05), but statistically significant difference (P lt; 0.05) compared with the bone cement group. 4, into the determination of the inorganic calcium of the bone cells in the different materials: With the increase of the culture time, the cells in each group, has increased significantly in calcium content, white beginning to end allogeneic bone material group cells secrete inorganic calcium level higher than the other group, NHAC / VEGF material group the next best, both were no statistically significant differences (P gt; 0.05), but significantly better than bone cement group, with a statistically significant difference (P lt; 0.05) between groups. Conclusion: NHAC / VEGF materials, compatibility of bone cells significantly better than bone cement compared with allogeneic bone the NHAC the / of VEGF material of bone cells compatibility somewhat less. The 2, NHAC / VEGF materials have better bone Cytocompatibility, still regarded as a new composite artificial bone material for the repair of bone defects.

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