Department of Veterinary Surgery, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, 3-5 Manastur Street, 400372 Cluj-Napoca, Romania.
Raluca Ripan Institute for Research in Chemistry, Babeș-Bolyai University, 30 Fantanele Street, 400294 Cluj-Napoca, Romania.
Int J Mol Sci. 2023 Jun 30;24(13):10911. doi: 10.3390/ijms241310911.
Uniform filler distribution in composites is an important requirement. Therefore, BaO glass, nano hydroxyapatite and quartz filler distribution was realized through PCL microcapsules which progressively release filler during matrix polymerization. Two composites were realized based on a complex matrix containing BisGMA, UDMA, HEMA and PEG400 mixed with a previously described mineral filler: 33% for C1 and 31% for C2. The spreading efficiency was observed via SEM, revealing a complete disintegration of the microcapsules during C1 polymerization, while C2 preserved some microcapsule parts that were well embedded into the matrix beside BaO filler particles; this was confirmed by means of the EDS spectra. Mesenchymal stem cells of palatal origin were cultured on the composites for 1, 3, 5 and 7 days. The alkaline phosphatase (ALP) level was measured at each time interval and the cytotoxicity was tested after 3, 5 and 7 days of co-culture on the composite samples. The SEM investigation showed that both composites allowed for robust proliferation of the cells. The MSC cell pluripotency stage was observed from 1 to 3 days with an average level of ALP of 209.2 u/L for C1 and 193.0 u/L for C2 as well as a spindle cell morphology. Cell differentiation occurred after 5 and 7 days of culture, implied by morphological changes such as flattened, star and rounded shapes, observed via SEM, which were correlated with an increased ALP level (279.4 u/L for C1 and 284.3 u/L for C2). The EDX spectra after 7 days of co-culture revealed increasing amounts of P and Ca close to the hydroxyapatite stoichiometry, indicating the stimulation of the osteoinductive behavior of MSCs by C1 and C2. The MTT assay test showed a cell viability of 98.08% for C1 and 97.33% for C2 after 3 days, proving the increased biocompatibility of the composite samples. The cell viability slightly decreased at 5 and 7 days but the results were still excellent: 89.5% for C1 and 87.3% for C2. Thus, both C1 and C2 are suitable for further in vivo testing.
复合材料中均匀的填充物分布是一个重要的要求。因此，通过 PC L 微胶囊实现了 BaO 玻璃、纳米羟基磷灰石和石英填充物的分布，PC L 微胶囊在基质聚合过程中逐渐释放填充物。基于含有 BisGMA、UDMA、HEMA 和 PEG400 的复杂基质，实现了两种复合材料，与之前描述的矿物填充物混合：C1 为 33%，C2 为 31%。通过 SEM 观察到铺展效率，发现 C1 聚合过程中微胶囊完全解体，而 C2 则保留了一些微胶囊部分，这些部分很好地嵌入了基质和 BaO 填充物颗粒之间；这通过 EDS 光谱得到了证实。腭来源的间充质干细胞在复合材料上培养 1、3、5 和 7 天。在每个时间间隔测量碱性磷酸酶 (ALP) 水平，并在复合样品上共培养 3、5 和 7 天后测试细胞毒性。SEM 研究表明，两种复合材料都允许细胞大量增殖。从第 1 天到第 3 天观察到 MSC 细胞多能性阶段，C1 的平均 ALP 水平为 209.2u/L，C2 的平均 ALP 水平为 193.0u/L，细胞形态呈梭形。培养 5 和 7 天后发生细胞分化，通过 SEM 观察到细胞形态发生变化，如扁平、星形和圆形，这与 ALP 水平的升高（C1 为 279.4u/L，C2 为 284.3u/L）相关。共培养 7 天后的 EDX 光谱显示，靠近羟基磷灰石化学计量的 P 和 Ca 含量增加，表明 C1 和 C2 刺激 MSC 的成骨诱导行为。MTT 测定试验表明，C1 的细胞活力为 98.08%，C2 的细胞活力为 97.33%，3 天后，证明了复合样品的细胞相容性增加。5 和 7 天后，细胞活力略有下降，但结果仍然非常出色：C1 为 89.5%，C2 为 87.3%。因此，C1 和 C2 都适合进一步的体内测试。
