In Vitro Proliferation and Osteoblastic Phenotype Expression of Cells Derived From Human Vertebral Lamina and Iliac Crest
In Vitro Proliferation and Osteoblastic Phenotype Expression of Cells Derived From Human Vertebral Lamina and Iliac Crest
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DOI：
10.1097/brs.0b013e3181a9c087
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发表时间：
2009-07-01
期刊：
影响因子：
3
通讯作者：
Rosa, Adalberto L.
中科院分区：
文献类型：
--
作者：
Defino, Helton L. A.;da Silva Herrero, Carlos F. P.;Rosa, Adalberto L.
研究设计。分离椎板和髂嵴来源的成骨细胞，并在相同条件（成骨培养基、pH、温度和CO2水平）下培养。 目的。比较源自椎板和髂骨移植的细胞的增殖和成骨细胞表型的表达。背景数据摘要。许多因素在脊柱融合中骨移植的成功中发挥着作用，其中包括成骨细胞群。两个常见的移植物来源是椎板和髂嵴，然而，尚未研究来自这些部位的细胞之间的增殖和成骨细胞表型表达的差异。方法。培养从椎板和髂嵴的松质骨中获得的细胞，通过直接细胞计数和台盼蓝检测活力来评估增殖情况。通过百里酚酞单磷酸释放的百里酚酞和茜素红 S 染色的基质矿化来评估碱性磷酸酶 (ALP) 活性。通过实时 PCR 分析 ALP、骨钙素、runt 相关转录因子 2、Msh 同源盒 2、骨形态发生蛋白 7、细胞间粘附分子 1 前体、骨保护素和 NF-kB 配体受体激活剂的基因表达。所有比较均与供体匹配。结果。与来自髂嵴的细胞相比，来自椎板的细胞在第7天和第10天的增殖更大，但细胞活力没有差异。第 7 天和第 10 天时，椎板细胞中的 ALP 活性高于髂嵴细胞。第 21 天时，椎板细胞中的矿化基质高于髂嵴细胞。第7天时，椎板来源的细胞中ALP、骨钙素、runt相关转录因子2、Msh同源盒2、骨形态发生蛋白7、细胞间粘附分子1前体、NF-kB配体受体激活剂和骨保护素的基因表达高于髂嵴细胞。结论。源自松质骨的细胞的细胞增殖和成骨细胞表型发育在椎板培养物中比在髂嵴培养物中更加旺盛。
Study Design. Osteoblastic cells derived from vertebral lamina and iliac crest were isolated and cultured under the same conditions (osteogenic medium, pH, temperature, and CO2 levels).Objective. To compare proliferation and expression of osteoblastic phenotype of cells derived from vertebral lamina and iliac grafting.Summary of Background Data. Many factors play a role in the success of bone graft in spinal fusion including osteoblastic cell population. Two common sources of graft are vertebral lamina and iliac crest, however, differences in proliferation and osteoblastic phenotype expression between cells from these sites have not been investigated.Methods. Cells obtained from cancellous bone of both vertebral lamina and iliac crest were cultured and proliferation was evaluated by direct cell counting and viability detected by Trypan blue. Alkaline phosphatase (ALP) activity was evaluated by thymolphthalein release from thymolphthalein monophosphate and matrix mineralization by staining with alizarin red S. Gene expression of ALP, osteocalcin, runt-related transcription factor 2, Msh homeobox 2, bone morphogenetic protein 7, intercellular adhesion molecule 1 precursor, osteoprotegerin, and receptor activator of NF-kB ligand was analyzed by real-time PCR. All comparisons were donor-matched.Results. Proliferation was greater at days 7 and 10 in cells from vertebral lamina compared with ones from iliac crest without difference in cell viability. ALP activity was higher in cells from vertebral lamina compared with cells from iliac crest at days 7 and 10. At 21 days, mineralized matrix was higher in cells derived from vertebral lamina than from iliac crest. At day 7, gene expression of ALP, osteocalcin, runt-related transcription factor 2, Msh homeobox 2, bone morphogenetic protein 7, intercellular adhesion molecule 1 precursor, receptor activator of NF-kB ligand, and osteoprotegerin was higher in cells derived from vertebral lamina compared with iliac crest.Conclusion. Cell proliferation and osteoblastic phenotype development in cells derived from cancellous bone were more exuberant in cultures of vertebral lamina than of iliac crest.