In Vitro Study of Composite Cements on Mesenchymal Stem Cells of Palatal Origin

Abstract

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.