不同浓度透明质酸水凝胶的细胞相容性

Cytocompatibility of hyaluronic acid hydrogels with concentration variations

背景:水凝胶呈半凝固的胶体状态,可以实现细胞的三维立体培养,消除普通培养皿中出现的"接触抑制"现象,从而提高培养效率,透明质酸水凝胶作为三维培养基质在干细胞和组织工程研究领域的应用已有报道。目的:探讨透明质酸水凝胶浓度的改变对其基本性能以及对大鼠骨髓间充质干细胞三维培养形态和增殖的影响。方法:制备4,8,12g/L的透明质酸水凝胶,观察其凝胶时间、溶胀率、降解速率以及黏弹性;用这3种浓度透明质酸水凝胶包封骨髓间充质干细胞进行三维培养,并以普通培养皿的二维培养作对照。通过激光扫描共焦显微镜观察不同培养条件下骨髓间充质干细胞的形态,采用细胞计数法描绘骨髓间充质干细胞在不同浓度透明质酸水凝胶内的生长曲线。结果与结论:对于4-12g/L的透明质酸水凝胶,其浓度越大,凝胶时间越短,溶胀率越低,降解速率越慢,水凝胶弹性强度越大(P<0.05)。激光扫描共焦显微镜见骨髓间充质干细胞在透明质酸水凝胶内呈立体圆球形,与二维培养时呈现的梭形明显不同;同时,细胞计数结果显示骨髓间充质干细胞在透明质酸水凝胶内的增殖率高于二维培养,并以8g/L透明质酸水凝胶的促增殖作用最明显(P<0.05)。可见不同浓度的透明质酸水凝胶性能存在差异,对大鼠骨髓间充质干细胞的生长、增殖也有影响,8g/L的透明质酸水凝胶更适合大鼠骨髓间充质干细胞的三维培养。

BACKGROUND：Hydrogel, which is a kind of semi-solid materials, can be used for three-dimensional culture for mesenchymal stem cel s. So it would improve efficiency of cultured cel s by eliminating“contact inhibition”, which is common in two-dimensional culture. Hyaluronic acid hydrogel as a three-dimensional culture matrix has been reported in the field of stem cel s and tissue engineering research. OBJECTIVE：To investigate the morphology and proliferation of rat bone marrow mesenchymal stem cel s cultured in the three-dimensional hyaluronic acid hydrogels with different properties. METHODS：Three concentrations of hyaluronic acid hydrogels, 4, 8, and 12 g/L, were prepared and evaluated with their properties including gelation time, swel ing ratio, degradation rate and viscoelasticity. Rat bone marrow mesenchymal stem cel s were encapsulated and cultured in those hyaluronic acid hydrogels. Another rat bone marrow mesenchymal stem cel s cultured in the two-dimensional medium served as control. Morphology of the encapsulated bone marrow mesenchymal stem cel s was observed by laser scanning confocal microscope. Growth curves of bone marrow mesenchymal stem cel s cultured in the hyaluronic acid hydrogels with different concentrations were depicted using cel counting method. RESULTS AND CONCLUSION：The higher concentration of hyaluronic acid hydrogels leaded to the shorter gelation time, lower swel ing ratio, slower degradation rate and higher strength （P〈0.05）. Bone marrow mesenchymal stem cel s cultured in the three-dimensional hyaluronic acid hydrogels showed a spherical shape and higher proliferation compared with those in normal culture. Meanwhile, rat bone marrow mesenchymal stem cel s cultured in 8 g/L hyaluronic acid hydrogels showed higher proliferation than those in 4 and 12 g/L hyaluronic acid hydrogels （P〈0.05）. These findings indicate that concentration variations of hyaluronic acid hydrogels lead to changes in their mechanical properties. Hyaluronic acid hydrogel with a concentration of 8 g/L is more appropriate for three-dimensional culture of rat bone marrow mesenchymal stem cel s.