Nair Manitha B, Varma H K, John Annie
Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences & Technology, Trivandrum, Kerala, India.
Tissue Eng Part A. 2009 Jul;15(7):1619-31. doi: 10.1089/ten.tea.2008.0229.
New biotechnologies such as tissue engineering require functionally active cells within supportive matrices where the physical and chemical stimulus provided by the matrix is indispensable to determine the cellular behavior. This study has investigated the influence of platelet-rich plasma (PRP) and fibrin glue (FG) on the functional activity of goat bone marrow-derived mesenchymal stem cells (gBMSCs) that differentiated into the osteogenic lineage. To achieve this goal, PRP and FG were separately coated on bioactive ceramics like hydroxyapatite (HA) and silica-coated HA (HASi), on which gBMSCs were seeded and induced to differentiate into the osteogenic lineage for 28 days. The cells were then analyzed for viability (lactate dehydrogenase assay: acridine orange and ethidium bromide staining), morphology (scanning electron microscopy), proliferation (picogreen assay), cell cycle assay (propidium iodide staining), and differentiation (alkaline phosphatase [ALP] activity and real-time PCR analysis of ALP, osteocalcin, and osteopontin gene). It has been observed that PRP and FG have appreciably favored the viability, spreading, and proliferation of osteogenic-induced gBMSCs. The osteopontin and osteocalcin expression was significantly enhanced on PRP- and FG-coated HA and HASi, but PRP had effect on neither ALP expression nor ALP activity. The results of this study have depicted that FG-coated ceramics were better than PRP-coated and bare matrices. Among all, the excellent performance was shown by FG coated HASi, which may be attributed to the communal action of the stimulus emanated by Si in HASi and the temporary extracellular matrix provided by FG over HASi. Thus, we can conclude that PRP or FG in combination with bioactive ceramics could possibly enhance the functional activity of cells to a greater extent, promoting the hybrid composite as a promising candidate for bone tissue engineering applications.
组织工程等新生物技术需要在支持性基质中具有功能活性的细胞,其中基质提供的物理和化学刺激对于决定细胞行为是不可或缺的。本研究调查了富血小板血浆(PRP)和纤维蛋白胶(FG)对分化为成骨谱系的山羊骨髓间充质干细胞(gBMSCs)功能活性的影响。为实现这一目标,将PRP和FG分别涂覆在生物活性陶瓷如羟基磷灰石(HA)和二氧化硅涂层HA(HASi)上,在其上接种gBMSCs并诱导其分化为成骨谱系28天。然后对细胞进行活力分析(乳酸脱氢酶测定:吖啶橙和溴化乙锭染色)、形态分析(扫描电子显微镜)、增殖分析(PicoGreen测定)、细胞周期分析(碘化丙啶染色)以及分化分析(碱性磷酸酶[ALP]活性和ALP、骨钙素和骨桥蛋白基因的实时PCR分析)。据观察,PRP和FG明显有利于成骨诱导的gBMSCs的活力、铺展和增殖。在PRP和FG涂覆的HA和HASi上,骨桥蛋白和骨钙素的表达显著增强,但PRP对ALP表达和ALP活性均无影响。本研究结果表明,FG涂覆的陶瓷优于PRP涂覆的和裸露的基质。其中,FG涂覆的HASi表现出优异的性能,这可能归因于HASi中Si发出的刺激与FG在HASi上提供的临时细胞外基质的共同作用。因此,我们可以得出结论,PRP或FG与生物活性陶瓷结合可能在更大程度上增强细胞的功能活性,促进这种混合复合材料成为骨组织工程应用的有前途的候选材料。
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