Periosteal-derived cells for bone bioengineering: a promising candidate
Periosteal-derived cells for bone bioengineering: a promising candidate
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DOI：
10.1111/j.1600-0501.2011.02287.x
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发表时间：
2012-10-01
影响因子：
4.3
通讯作者：
Granjeiro, Jose Mauro
中科院分区：
文献类型：
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作者：
Castro-Silva, Igor Iuco;Zambuzzi, Willian Fernando;Granjeiro, Jose Mauro
目的近年来，为了骨生物工程的成功，人们做了大量的努力来鉴定成骨细胞的天然来源。其中，骨膜组织已成为一个有趣的候选人。因此，我们决定通过描述从最初形态学变化到细胞外基质（ECM）矿化的一系列生物学事件来评估骨膜衍生细胞的成骨潜力。方法取成年大鼠颅骨骨膜来源细胞。贴壁细胞经原代培养和扩增后，分别于第7、14、21和28天在经典成骨培养基中培养，以评价这些细胞向成熟成骨细胞的分化。通过评估碱性磷酸酶（ALP）活性（成骨细胞分化的生物标志物）的时间线和随后的矿化结节（通过von Kossa染色和钙含量测量）来监测。结果相差显微镜下观察到的细胞形态变化主要为成纤维细胞（7天，指数生长时为半汇合培养）至多角体细胞（1428天，分化过程中为汇合培养）。ALP活性从1428天开始线性增加，与天数相似，而蛋白质量保持不变。有趣的是，我们的von Kossa染色数据显示矿化结节的发生率在28天时最高，与28天相似。结论骨膜源性细胞在体外具有向成熟成骨细胞分化的潜能，能通过向细胞外基质中加入钙离子促进矿化。从我们的观点来看，生物工程师可以探索这种成骨细胞的来源，以推进能够解决骨退行性病变的治疗方案。
Purpose Over the last years so many efforts have been made in order to indentify natural sources of osteogenic cells for the success of bone bioengineering. Among them, periosteum tissue has emerged as an interesting candidate. Thus, we decided to evaluate the osteogenic potential of periosteal-derived cells by describing a sequence of biological events since initial morphological changes to mineralization of extracellular matrix (ECM). Methods Periosteal-derived cells were obtained from calvarial of adult rats. After the primary culture and expansion, the adherent cells were cultured at 7, 14, 21 and 28 similar to days under a classical osteogenic culture medium in order to evaluate the differentiation of those cells in mature osteoblast. It was monitored by evaluating a time-line of alkaline phosphatase (ALP) activity (biomarker of osteoblast differentiation) and afterwards nodules of mineralization (measured by von Kossa staining and calcium content). Results Analysis from phase-contrast microscopy revealed mainly morphological changes ranging since fibroblast-shaped (7 similar to days, semi-confluent culture at exponential growth) to polyhedral-shaped cells (1428 similar to days, confluent culture during differentiation process). ALP activity was linearly increased since 1428 similar to days while amount of protein remained unchanged. Interesting, our data from von Kossa staining reveled a highest incidence of mineralization nodules at 28 similar to days. Conclusion Taken our results together, we can suggest that periosteal-derived cells present an interesting potential to differentiate in mature osteoblast able to promote mineralization in vitro by incorporating to ECM circulating calcium from extracellular compartment. From our point of view, this source of osteogenic cells can be explored by bioengineers in order to advance therapeutic protocols able to solve bone degenerative lesions.