Toward cell therapy for vascular calcification: osteoclast-mediated demineralization of calcified elastin
Toward cell therapy for vascular calcification: osteoclast-mediated demineralization of calcified elastin
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DOI：
10.1016/j.carpath.2006.07.001
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发表时间：
2007-01-01
影响因子：
3.7
通讯作者：
Vyavahare, Narendra R.
中科院分区：
文献类型：
--
作者：
LaShan Simpson, Chartrisa;Lindley, Suzanne;Vyavahare, Narendra R.
背景：弹性蛋白导向的血管钙化是临床上的重要特征，它涉及异位骨样结构的形成。利用动脉钙化和骨调节之间的相似性，我们的假设是，限制血管钙化的治疗方法可以通过使用自体破骨细胞的位点特异性递送来开发。在本论文中，我们测试的假设，骨髓来源的破骨细胞有能力去矿化钙化的弹性蛋白，而没有显着改变弹性蛋白的完整性。研究方法：在维生素D-3和视黄酸存在下，通过大鼠骨髓来源的祖细胞体外成熟获得活性的多核破骨细胞。抗酒石酸酸性磷酸酶染色，羟基磷灰石包被的磁盘上形成的吸收坑，和RT-PCR鉴定组织蛋白酶K基因表达的细胞表型进行验证。将钙化的主动脉弹性蛋白与破骨细胞和钙一起接种，并监测凝胶和培养基中的磷水平以检测弹性蛋白的脱矿作用。还在培养基中监测可溶性弹性蛋白肽的弹性蛋白降解。在体内实验中，将纯主动脉弹性蛋白与同种异体破骨细胞共植入大鼠皮下，并使用矿物质分析和锁链素定量评价弹性蛋白钙化和降解的程度。结果：骨髓源性破骨细胞在体外可使钙化弹性蛋白的矿物质含量降低80%。此外，在体内植入同种异体破骨细胞附近的钙化弹性蛋白限制弹性蛋白矿化近50%，在没有可检测到的弹性蛋白降解。结论：破骨细胞具有使钙化弹性蛋白脱矿的能力，而弹性蛋白的完整性没有显著改变。(C)2007年爱思唯尔公司All right reserved.
Background: Elastin-oriented vascular calcification is a clinically significant feature, which involves formation of ectopic bone-like structures. Taking advantage of the similarities between arterial calcification and bone regulation, our hypothesis was that therapeutic approaches for limitation of vascular calcification could be developed using site-specific delivery of autologous osteoclasts. In the present paper, we tested the hypothesis that bone-marrow-derived osteoclasts have the ability to demineralize calcified elastin, without significant alterations in elastin integrity. Methods: Active, multinucleated osteoclasts were obtained by in vitro maturation of rat bone-marrow-derived progenitor cells in the presence of vitamin D-3 and retinoic acid. Cell phenotype was validated by staining for tartrate-resistant acid phosphatase, formation of resorption pits on hydroxyapatite-coated disks, and RT-PCR for identification of cathepsin K gene expression. Calcified aortic elastin was seeded with osteoclasts and calcium, and phosphorous levels were monitored in gels and culture media to detect demineralization of elastin. Soluble elastin peptides were also monitored in culture media for elastin degradation. For in vivo experiments, pure aortic elastin was coimplanted with allogenic osteoclasts subdermally into rats, and the degree of elastin calcification and degradation was evaluated using mineral analysis and desmosine quantitation. Results: Bone-marrow-derived osteoclasts reduced mineral content of calcified elastin in vitro by 80%. Moreover, in vivo implantation of allogenic osteoclasts in the vicinity of calcifying elastin limited elastin mineralization by almost 50%, in the absence of detectable elastin degradation. Conclusions: Osteoclasts have the ability to demineralize calcified elastin, without significant alterations in elastin integrity. (C) 2007 Elsevier Inc. All right reserved.