载羟基磷灰石纳米颗粒的大孔透明质酸/壳聚糖聚电解质复合物水凝胶:制备、表征及体外评价

M. Drozdova, T. Demina, O. A. Dregval, A. Gaidar, E. Andreeva, A. Zelenetskii, T. Akopova, E. Markvicheva
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引用次数: 4

摘要

以透明质酸/壳聚糖(Hyal/Ch)多电解质复合物(PEC)负载均匀分布的羟基磷灰石纳米颗粒(nHAp)为基础,制备和表征复合大孔水凝胶,并在体外用小鼠成纤维细胞(L929)、成骨细胞样细胞(HOS)和人间充质基质细胞(hMSC)对其进行评价。利用扫描电镜(SEM)和共聚焦激光扫描显微镜(CLSM)研究了水凝胶形态与羟基磷灰石纳米颗粒含量的关系。Hyal/Ch凝胶的平均孔径为204±25 μm。nHAp (1 wt. %和5 wt. %)在Hyal/Ch水凝胶中的包埋导致平均孔径减小(相对为94±2和77±9 μm)。发现水凝胶在各种水介质中的溶胀率和失重率随着介质离子强度的增强而增加。用CLSM研究了水凝胶内的细胞形态和定位。细胞活力取决于nHAp含量,并在水凝胶中培养7天后用mtt法评估细胞活力。羟基磷灰石纳米颗粒负载在1-10 wt. %范围内的增加导致所有水凝胶细胞生长和增殖的增强。Hyal/Ch/nHAp-10样品的细胞存活率最高(10 wt. % nHAp),而Hyal/Ch/nHAp-1水凝胶的细胞存活率最低(1 wt. % nHAp)。因此,提出的简单的原始技术和PEC水凝胶的设计在组织工程,特别是骨组织修复方面具有前景。
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Macroporous Hyaluronic Acid/Chitosan Polyelectrolyte Complex-Based Hydrogels Loaded with Hydroxyapatite Nanoparticles: Preparation, Characterization and In Vitro Evaluation
The aim of the study was to fabricate and characterize composite macroporous hydrogels based on a hyaluronic acid/chitosan (Hyal/Ch) polyelectrolyte complex (PEC) loaded with homogeneously distributed hydroxyapatite nanoparticles (nHAp), and to evaluate them in vitro using mouse fibroblasts (L929), osteoblast-like cells (HOS) and human mesenchymal stromal cells (hMSC). Hydrogel morphology as a function of the hydroxyapatite nanoparticle content was studied using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The mean pore size in the Hyal/Ch hydrogel was 204 ± 25 μm. The entrapment of nHAp (1 and 5 wt. %) into the Hyal/Ch hydrogel led to a mean pore size decrease (94 ± 2 and 77 ± 9 μm, relatively). Swelling ratio and weight loss of the hydrogels in various aqueous media were found to increase with an enhancement of a medium ionic strength. Cell morphology and localization within the hydrogels was studied by CLSM. Cell viability depended upon the nHAp content and was evaluated by MTT-assay after 7 days of cultivation in the hydrogels. An increase of the hydroxyapatite nanoparticles loading in a range of 1–10 wt. % resulted in an enhancement of cell growth and proliferation for all hydrogels. Maximum cell viability was obtained in case of the Hyal/Ch/nHAp-10 sample (10 wt. % nHAp), while a minimal cell number was found for the Hyal/Ch/nHAp-1 hydrogel (1 wt. % nHAp). Thus, the proposed simple original technique and the design of PEC hydrogels could be promising for tissue engineering, in particular for bone tissue repair.
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