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成骨细胞外泌体。一种用于评估骨诱导纳米材料的碱性磷酸酶无损定量方法。

Osteoblastic exosomes. A non-destructive quantitative approach of alkaline phosphatase to assess osteoconductive nanomaterials.

机构信息

Laboratorio de Medios e Interfases, Instituto Superior de Investigaciones Biológicas (INSIBIO), CONICET, Departamento de Bioingeniería, Facultad de Ciencias Exactas y Tecnología, UNT. Av. Independencia 1800, 4000 San Miguel de Tucumán, Argentina; Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain.

Laboratorio de Medios e Interfases, Instituto Superior de Investigaciones Biológicas (INSIBIO), CONICET, Departamento de Bioingeniería, Facultad de Ciencias Exactas y Tecnología, UNT. Av. Independencia 1800, 4000 San Miguel de Tucumán, Argentina; Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain.

出版信息

Mater Sci Eng C Mater Biol Appl. 2020 Oct;115:110931. doi: 10.1016/j.msec.2020.110931. Epub 2020 Apr 13.

Abstract

Alkaline phosphatase (ALP) is an essential biomarker of osteoblastic activity. Currently, ALP activity has been used to study bone mineralization mechanisms and osteoactive biomaterials among others. The ALP quantification is usually performed by destructive methods either on growing cells or cells lysate in which the osteoconductive biomaterial is being assessed. This work addresses the evaluation of a non-destructive colorimetric approach for the determination of ALP activity on osteoblast-derived exosomes from culture supernatants. The efficiency of the method was evaluated on osteoconductive electrospun scaffolds of PCL compounded with ZnO as a reference biomaterial. The results demonstrated that the osteoblast cell line mineralization induced by osteoconductive scaffolds can be monitorized over time by the non-destructive measurement of ALP activity on osteoblast derived exosomes. Consequently, this non-destructive approach suggested to be a reliable alternative technique for the quantification of biomaterials osteoconductivity or even evaluation of osteoblastic response at stem cells.

摘要

碱性磷酸酶(ALP)是成骨细胞活性的重要生物标志物。目前,ALP 活性已被用于研究骨矿化机制和骨活性生物材料等。ALP 的定量通常通过破坏性方法在生长细胞或细胞裂解物中进行,其中正在评估骨传导生物材料。本工作评估了一种用于测定培养上清液中骨细胞衍生的外体中 ALP 活性的非破坏性比色法。该方法的效率在与 ZnO 复合的 PCL 电纺支架等骨传导参考生物材料上进行了评估。结果表明,通过非破坏性测量骨细胞衍生的外体上的 ALP 活性,可以监测骨传导支架诱导的成骨细胞矿化随时间的变化。因此,这种非破坏性方法有望成为一种可靠的替代技术,用于定量生物材料的骨传导性,甚至评估干细胞中的成骨细胞反应。

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