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聚(羟基丁酸酯-羟基戊酸酯)、聚(ε-己内酯)支架上培养的大鼠骨髓间充质干细胞的成骨分化。

Osteogenic differentiation of rat bone mesenchymal stem cells cultured on poly (hydroxybutyrate-co-hydroxyvalerate), poly (ε-caprolactone) scaffolds.

机构信息

Laboratório de Biomateriais em Ortopedia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Campinas, SP, Brazil.

Centro de Engenharia, Modelagem e Ciências Sociais Aplicadas (CECS), Universidade Federal do ABC, São Bernardo do Campo, SP, Brazil.

出版信息

J Mater Sci Mater Med. 2021 Oct 30;32(11):138. doi: 10.1007/s10856-021-06615-6.

Abstract

Bioresorbable biomaterials can fill bone defects and act as temporary scaffold to recruit MSCs to stimulate their differentiation. Among the different bioresorbable polymers studied, this work focuses on poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL). Were prepared blends of PHBV and PCL to obtain PHBV based biomaterials with good tenacity, important for bone tissue repair, associated with biocompatible properties of PCL. This study assesses the viability of Vero cells on scaffolds of PHBV, PCL, and their blends and the osteogenic differentiation of mesenchymal stem cells (MSCs). Materials were characterized in surface morphology, DSC and Impact Strength (IS). Vero cells and MSCs were assessed by MTT assay, cytochemical and SEM analysis. MSC osteogenic differentiation was evaluated through alizarin red staining and ALP activity. We found some roughness onto surface materials. DSC showed that the blends presented two distinct melting peaks, characteristic of immiscible blends. IS test confirmed that PHBV-PCL blends is an alternative for increase the tenacity of PHBV. MTT assay showed cells with high metabolic activities on extract toxicity test, but with low activity in the direct contact test. SEM analysis showed spreading cells with irregular and flattened morphology on different substrates. Cytochemical study revealed that MSCs maintained their morphology, although in smaller number for MSCs. The development of nodules of mineralized organic matrix in MSC cultures was identified by alizarin red staining and osteogenic differentiation was confirmed by the quantification of ALP activity. Thus, our scaffolds did not interfere on viability of Vero cells or the osteogenic differentiation of MSCs.

摘要

可生物吸收的生物材料可以填充骨缺损并作为临时支架招募间充质干细胞,以刺激其分化。在研究的不同可生物吸收聚合物中,这项工作集中在聚(羟基丁酸酯-co-羟基戊酸酯)(PHBV)和聚(ε-己内酯)(PCL)上。制备了 PHBV 和 PCL 的共混物,以获得具有良好韧性的基于 PHBV 的生物材料,这对于骨组织修复很重要,同时具有 PCL 的生物相容性。本研究评估了 Vero 细胞在 PHBV、PCL 及其共混物支架上的活力以及间充质干细胞(MSCs)的成骨分化。对材料的表面形态、DSC 和冲击强度(IS)进行了表征。通过 MTT 测定、细胞化学和 SEM 分析评估了 Vero 细胞和 MSCs。通过茜素红染色和 ALP 活性评估 MSC 成骨分化。我们发现表面材料有些粗糙。DSC 表明共混物呈现出两个独特的熔融峰,这是不混溶共混物的特征。IS 测试证实 PHBV-PCL 共混物是提高 PHBV 韧性的一种替代方法。MTT 测定显示,在提取物毒性测试中细胞具有高代谢活性,但在直接接触测试中活性较低。SEM 分析表明,细胞在不同基质上呈不规则和平坦形态的扩展。细胞化学研究表明,MSC 保持其形态,尽管 MSC 的数量较少。茜素红染色鉴定 MSC 培养物中矿化有机基质结节的形成,并通过 ALP 活性的定量证实成骨分化。因此,我们的支架不会干扰 Vero 细胞的活力或 MSC 的成骨分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7a8f/8557177/e6b4c6a759ec/10856_2021_6615_Fig1_HTML.jpg

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