Triphasic 3D In Vitro Model of Bone-Tendon-Muscle Interfaces to Study Their Regeneration.
Triphasic 3D In Vitro Model of Bone-Tendon-Muscle Interfaces to Study Their Regeneration.
复制标题
骨-肌腱-肌肉界面三维三维体外模型研究其再生。
作者：
不同组织之间的过渡区域，称为组织界面，在损伤后再生能力有限，这可能导致不完全愈合。以前的研究集中在单一界面，最常见的是骨-肌腱和骨-软骨界面。在此，我们建立了一个三维体外模型来研究骨-肌腱-肌肉界面的再生。3D模型由胶原蛋白和琼脂糖制备，具有不同浓度的羟基磷灰石，以使组织从骨骼逐渐变为肌肉，从而产生刚度梯度。这种分级结构是使用间接3D打印制造的，以提供生物相关的表面形貌。发现MG-63、人真皮成纤维细胞和Sket.4U细胞分别适合用于骨骼、肌腱和肌肉的细胞模型。组成3D模型的两相和三相水凝胶显示出适合于细胞生长。将细胞在3D模型上共培养超过21天，然后评估细胞增殖、代谢活性、活力、细胞毒性、组织特异性标志物和基质沉积以确定界面形成。这些研究在新开发的生长室中进行，该生长室允许细胞通信，同时细胞培养基被区室化。3D模型在21天内促进了细胞活力、组织特异性标志物表达和新基质沉积，从而显示出开发新界面的前景。
The transition areas between different tissues, known as tissue interfaces, have limited ability to regenerate after damage, which can lead to incomplete healing. Previous studies focussed on single interfaces, most commonly bone-tendon and bone-cartilage interfaces. Herein, we develop a 3D in vitro model to study the regeneration of the bone-tendon-muscle interface. The 3D model was prepared from collagen and agarose, with different concentrations of hydroxyapatite to graduate the tissues from bones to muscles, resulting in a stiffness gradient. This graduated structure was fabricated using indirect 3D printing to provide biologically relevant surface topographies. MG-63, human dermal fibroblasts, and Sket.4U cells were found suitable cell models for bones, tendons, and muscles, respectively. The biphasic and triphasic hydrogels composing the 3D model were shown to be suitable for cell growth. Cells were co-cultured on the 3D model for over 21 days before assessing cell proliferation, metabolic activity, viability, cytotoxicity, tissue-specific markers, and matrix deposition to determine interface formations. The studies were conducted in a newly developed growth chamber that allowed cell communication while the cell culture media was compartmentalised. The 3D model promoted cell viability, tissue-specific marker expression, and new matrix deposition over 21 days, thereby showing promise for the development of new interfaces.
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