Synoviocyte Derived-Extracellular Matrix Enhances Human Articular Chondrocyte Proliferation and Maintains Re-Differentiation Capacity at Both Low and Atmospheric Oxygen Tensions
Synoviocyte Derived-Extracellular Matrix Enhances Human Articular Chondrocyte Proliferation and Maintains Re-Differentiation Capacity at Both Low and Atmospheric Oxygen Tensions
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DOI：
10.1371/journal.pone.0129961
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发表时间：
2015-06-15
期刊：
影响因子：
3.7
通讯作者：
Dennis, James E.
中科院分区：
文献类型：
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作者：
Kean, Thomas J.;Dennis, James E.
背景目前的组织工程方法不足以进行全关节表面置换，并且软骨细胞在组织培养塑料上扩增时经历去分化。去分化的软骨细胞在培养物中显示出较差的再分化，导致糖胺聚糖（GAG）和胶原基质积累减少。为了解决这个问题，猪滑膜细胞衍生的细胞外基质和低（5%）氧张力进行了评估，以提高人类关节软骨细胞的扩张和维持再分化potential.MethodsPorcine滑膜细胞基质失活使用3非洗涤剂的方法。将这些失活的滑膜细胞基质与组织培养塑料比较其支持人软骨细胞扩增的能力。在所有表面上的低氧张力（5%）和大气氧张力（20%）下进一步比较膨胀。然后，扩增的细胞在低氧和大气氧张力下在聚集体培养物中进行软骨形成再分化。聚集体进行了评估，他们的GAG和胶原蛋白含量生化和histologic.ResultsHuman软骨细胞扩大两倍快失活的滑膜细胞基质与组织培养塑料，细胞保留其再分化能力的两倍数量的人口doubling。低氧和大气氧分压下的生长速率无显著差异。与组织培养塑料相比，在滑膜细胞基质上扩增的细胞中I型胶原、II型胶原、聚集蛋白聚糖的表达显著减少，MMP 13的表达更多。由于氧张力对基因表达也有显著影响，其中与滑膜细胞基质相比，在组织培养塑料上有更高的I型胶原、II型胶原、SOX 9和更低的MMP 13表达。有一个显着增加GAG，但不是胶原蛋白，积累在软骨细胞聚集体重新分化在低氧张力在大气中的氧气conditions.ConclusionsSynoviocyte-derived矩阵支持增强的人类软骨细胞的扩张，使软骨细胞保持在一个状态，他们可以重新分化成一个软骨表型后，显着更多的人口doubling。此外，低氧张力支持GAG，但不支持胶原蛋白的积累。这些发现是朝着生产功能更强的组织工程软骨迈出的一步。
BackgroundCurrent tissue engineering methods are insufficient for total joint resurfacing, and chondrocytes undergo de-differentiation when expanded on tissue culture plastic. De-differentiated chondrocytes show poor re-differentiation in culture, giving reduced glycosaminoglycan (GAG) and collagen matrix accumulation. To address this, porcine synoviocyte-derived extracellular matrix and low (5%) oxygen tension were assessed for their ability to enhance human articular chondrocyte expansion and maintain re-differentiation potential.MethodsPorcine synoviocyte matrices were devitalized using 3 non-detergent methods. These devitalized synoviocyte matrices were compared against tissue culture plastic for their ability to support human chondrocyte expansion. Expansion was further compared at both low (5%), and atmospheric (20%) oxygen tension on all surfaces. Expanded cells then underwent chondrogenic re-differentiation in aggregate culture at both low and atmospheric oxygen tension. Aggregates were assessed for their GAG and collagen content both biochemically and histologically.ResultsHuman chondrocytes expanded twice as fast on devitalized synoviocyte matrix vs. tissue culture plastic, and cells retained their re-differentiation capacity for twice the number of population doublings. There was no significant difference in growth rate between low and atmospheric oxygen tension. There was significantly less collagen type I, collagen type II, aggrecan and more MMP13 expression in cells expanded on synoviocyte matrix vs. tissue culture plastic. There were also significant effects due to oxygen tension on gene expression, wherein there was greater collagen type I, collagen type II, SOX9 and less MMP13 expression on tissue culture plastic compared to synoviocyte matrix. There was a significant increase in GAG, but not collagen, accumulation in chondrocyte aggregates re-differentiated at low oxygen tension over that achieved in atmospheric oxygen conditions.ConclusionsSynoviocyte-derived matrix supports enhanced expansion of human chondrocytes such that the chondrocytes are maintained in a state from which they can re-differentiate into a cartilage phenotype after significantly more population doublings. Also, low oxygen tension supports GAG, but not collagen, accumulation. These findings are a step towards the production of a more functional, tissue engineered cartilage.