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首页> 外文期刊>Acta biomaterialia >Cell viability, proliferation and extracellular matrix production of human annulus fibrosus cells cultured within PDLLA/Bioglass composite foam scaffolds in vitro.
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Cell viability, proliferation and extracellular matrix production of human annulus fibrosus cells cultured within PDLLA/Bioglass composite foam scaffolds in vitro.

机译：在PDLLA / Bioglass复合泡沫支架中体外培养的人环纤维细胞的细胞活力，增殖和细胞外基质产生。

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摘要

The objective of this study was to assess cell viability, attachment, morphology, proliferation, and collagen and sulphated glycosaminoglycan (s-GAG) production by human annulus fibrosus (HAF) cells cultured in vitro in poly(d,l-lactide) (PDLLA)/Bioglass composite foams. PDLLA foams with different percentages (0, 5 and 30wt.%) of Bioglass particles were prepared by thermally induced phase separation (TIPS) and characterized by scanning electron microscopy (SEM). HAF cell viability in the PDLLA/Bioglass foam was analysed using Live/Dead staining. HAF cell attachment was observed using SEM. An assessment of cell proliferation was conducted using the WST-1 assay. The level of s-GAG and collagen produced by HAF cells was quantified using the 1,9-dimethylmethylene blue (DMMB) assay and Sircoltrade mark assay after 4 weeks of culture. The presence of collagen types I and II within the PDLLA/Bioglass composite foams was analysed using immunohistochemistry. Live/dead staining showed that many viable HAF cells were present on the top surface of the foams as well as penetrating into the internal pore structure, suggesting that the PDLLA/Bioglass composite materials are non-toxic and that the presence of Bioglass particles within PDLLA scaffolds does not inhibit HAF cell growth. The SEM observations revealed that more clusters of HAF cells were attached to the pore walls of both the PDLLA/5BG foam and the PDLLA/30BG foam when compared with the PDLLA/0BG foam. WST-1 assay performed over a period of 4 weeks showed an increased tendency of HAF cells to proliferate within both the PDLLA/5BG foam and the PDLLA/30BG foam when compared with both the tissue culture plastic control and the PDLLA/0BG foam, indicating the presence of Bioglass in the foam has a positive effect on HAF cell proliferation. Sircoltrade mark and DMMB assays showed that HAF cells cultured within the PDLLA/30BG foam had a greater ability to deposit collagen and proteoglycan when compared with the control and the PDLLA/0BG foam after 4 weeks in culture,suggesting that the increase of Bioglass content may induce microenvironmental changes which promote the production of extracellular matrix containing abundant collagen and s-GAG. The immunohistochemical analysis of collagen production demonstrated that collagen produced in all cultures was predominantly of type I. These findings provide preliminary evidence for the use of PDLLA/Bioglass composite as cell-carrier materials for future treatments of the intervertebral disc with damaged AF region.
机译：这项研究的目的是评估在聚（d，l-丙交酯）（PDLLA）中体外培养的人环纤维（HAF）细胞产生的细胞活力，附着，形态，增殖以及胶原和硫酸化糖胺聚糖（s-GAG）的产生）/生物玻璃复合泡沫。通过热诱导相分离（TIPS）制备了具有不同百分比（0、5和30wt。％）生物玻璃颗粒的PDLLA泡沫，并通过扫描电子显微镜（SEM）进行了表征。使用活/死染色分析PDLLA /生物玻璃泡沫中的HAF细胞活力。使用SEM观察HAF细胞附着。使用WST-1分析进行细胞增殖评估。培养4周后，使用1,9-二甲基亚甲基蓝（DMMB）测定法和Sircoltrade mark测定法测定HAF细胞产生的s-GAG和胶原蛋白的水平。使用免疫组织化学分析了PDLLA / Bioglass复合泡沫中I型和II型胶原的存在。活/死染色表明，泡沫的顶表面上存在许多可行的HAF细胞，并且渗透到内部孔结构中，这表明PDLLA /生物玻璃复合材料无毒，并且PDLLA中存在生物玻璃颗粒支架不抑制HAF细胞生长。 SEM观察表明，与PDLLA / 0BG泡沫相比，更多簇的HAF细胞附着在PDLLA / 5BG泡沫和PDLLA / 30BG泡沫的孔壁上。与组织培养塑料对照和PDLLA / 0BG泡沫相比，在4周内进行的WST-1分析显示HAF细胞在PDLLA / 5BG泡沫和PDLLA / 30BG泡沫中增殖的趋势有所增加，这表明泡沫中存在生物玻璃对HAF细胞增殖具有积极作用。 Sircoltrade mark和DMMB分析表明，在培养4周后，与对照和PDLLA / 0BG泡沫相比，在PDLLA / 30BG泡沫中培养的HAF细胞具有更大的胶原蛋白和蛋白聚糖沉积能力，这表明生物玻璃含量可能增加。诱导微环境变化，从而促进含有丰富胶原蛋白和s-GAG的细胞外基质的产生。胶原蛋白产生的免疫组织化学分析表明，所有培养物中产生的胶原蛋白主要为I型。这些发现为PDLLA /生物玻璃复合材料作为细胞载体材料用于治疗AF区受损的椎间盘的初步证据。

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