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首页> 外文期刊>Osteoarthritis and cartilage >Tensile properties of engineered cartilage formed from chondrocyte- and MSC-laden hydrogels.
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Tensile properties of engineered cartilage formed from chondrocyte- and MSC-laden hydrogels.

机译：由软骨细胞和载有MSC的水凝胶形成的工程软骨的拉伸特性。

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OBJECTIVE: The objective of this study was to determine the capacity of chondrocyte- and mesenchymal stem cell (MSC)-laden hydrogel constructs to achieve native tissue tensile properties when cultured in a chemically defined medium supplemented with transforming growth factor-beta3 (TGF-beta3). DESIGN: Cell-laden agarose hydrogel constructs (seeded with bovine chondrocytes or MSCs) were formed as prismatic strips and cultured in a chemically defined serum-free medium in the presence or absence of TGF-beta3. The effects of seeding density (10 vs 30 million cells/mL) and cell type (chondrocyte vs MSC) were evaluated over a 56-day period. Biochemical content, collagenous matrix deposition and localization, and tensile properties (ramp modulus, ultimate strain, and toughness) were assessed biweekly. RESULTS: Results show that the tensile properties of cell-seeded agarose constructs increase with time in culture. However, tensile properties (modulus, ultimate strain, and toughness) achieved on day 56 were not dependent on either the initial seeding density or the cell type employed. When cultured in medium supplemented with TGF-beta3, tensile modulus increased and plateaued at a level of 300-400 kPa for each cell type and starting cell concentration. Ultimate strain and toughness also increased relative to starting values. Collagen deposition increased in constructs seeded with both cell types and at both seeding densities, with exposure to TGF-beta3 resulting in a clear shift toward type II collagen deposition as determined by immunohistochemical staining. CONCLUSIONS: These findings demonstrate that the tensile properties, an important and often overlooked metric of cartilage development, increase with time in culture in engineered hydrogel-based cartilage constructs. Under the free-swelling conditions employed in the present study, tensile moduli and toughness did not match that of the native tissue, though significant time-dependent increases were observed with the inclusion of TGF-beta3. Of note, MSC-seeded constructs achieved tensile properties that were comparable to chondrocyte-seeded constructs, confirming the utility of this alternative cell source in cartilage tissue engineering. Further work, including both modulation of the chemical and mechanical culture environment, is required to optimize the deposition of collagen and its remodeling to achieve tensile properties in engineered constructs matching the native tissue.
机译：目的：本研究的目的是确定载有软骨细胞和间充质干细胞（MSC）的水凝胶构建物在化学成分确定的培养基中培养的能力，从而获得天然组织拉伸特性，该培养基中添加了转化生长因子-β3（TGF-β3 ）。设计：充满细胞的琼脂糖水凝胶构建体（与牛软骨细胞或MSC一起播种）形成棱柱状条带，并在存在或不存在TGF-beta3的情况下，在化学成分明确的无血清培养基中培养。在56天的时间内评估了接种密度（10对3000万个细胞/ mL）和细胞类型（软骨细胞对MSC）的影响。每两周评估生化含量，胶原蛋白基质的沉积和定位以及拉伸性能（斜率模量，极限应变和韧性）。结果：结果表明，接种细胞的琼脂糖构建体的拉伸性能随培养时间的延长而增加。但是，在第56天达到的拉伸性能（模量，极限应变和韧性）不取决于初始播种密度或所使用的细胞类型。当在补充有TGF-β3的培养基中培养时，每种细胞类型和起始细胞浓度的拉伸模量均增加并稳定在300-400 kPa的水平。极限应变和韧性也相对于起始值增加。在两种细胞类型和两种接种密度下接种的构建体中，胶原蛋白沉积均增加，暴露于TGF-beta3导致免疫组织化学染色确定的向II型胶原蛋白沉积的明显转变。结论：这些发现表明，在基于工程水凝胶的软骨构建物中，抗张强度是重要的且经常被忽视的软骨发育指标，随着培养时间的增加而增加。在本研究中采用的自由溶胀条件下，拉伸模量和韧性与天然组织的拉伸模量和韧性不匹配，尽管在加入TGF-β3的情况下观察到了明显的时间依赖性增加。值得注意的是，播种了MSC的构建体获得了与软骨细胞播种的构建体相当的拉伸性能，从而证实了这种替代细胞来源在软骨组织工程中的实用性。需要进一步的工作，包括化学和机械培养环境的调节，以优化胶原蛋白的沉积及其重塑，以在与天然组织相匹配的工程构造中获得拉伸特性。

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