Department of Oral and Maxillofacial Surgery, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.
Department of Orthodontics, Seoul Saint Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.
Medicina (Kaunas). 2024 Apr 25;60(5):702. doi: 10.3390/medicina60050702.
: Tacrolimus is a macrolide lactone compound derived from the bacterium , widely known as an immunosuppressant. In basic research, the effects of tacrolimus on osteogenic differentiation have been tested using mesenchymal stem cells. In this study, tacrolimus's effects on the cellular survival and osteogenic differentiation of stem cell spheroids were investigated. : Concave microwells were used to form stem cell spheroids in the presence of tacrolimus at final concentrations of 0 μg/mL, 0.1 μg/mL, 1 μg/mL, 10 μg/mL, and 100 μg/mL. A microscope was used to test cellular vitality qualitatively, and an assay kit based on water-soluble tetrazolium salt was used to measure cellular viability quantitatively. Alkaline phosphatase activity and an anthraquinone dye test for measuring calcium deposits were used to assess osteogenic differentiation. To assess the expression of osteogenic differentiation, a quantitative polymerase chain reaction, Western blot, and RNA sequencing were performed. : Spheroids across all concentrations maintained a relatively uniform and spherical shape. Cell viability assay indicated that tacrolimus, up to a concentration of 100 μg/mL, did not significantly impair cell viability within spheroids cultured in osteogenic media. The increase in calcium deposition, particularly at lower concentrations of tacrolimus, points toward an enhancement in osteogenic differentiation. There was an increase in COL1A1 expression across all tacrolimus concentrations, as evidenced by the elevated mean and median values, which may indicate enhanced osteogenic activity. : This study showed that tacrolimus does not significantly impact the viability of stem cell spheroids in osteogenic media, even at high concentrations. It also suggests that tacrolimus may enhance osteogenic differentiation, as indicated by increased calcium deposition and COL1A1 expression. These findings advance our understanding of tacrolimus's potential roles in tissue repair, regeneration, and stem cell-based therapeutic applications.
他克莫司是一种大环内酯类化合物，来源于一种广泛用作免疫抑制剂的细菌。在基础研究中，已经使用间充质干细胞测试了他克莫司对成骨分化的影响。在这项研究中，研究了他克莫司对干细胞球状体细胞存活和成骨分化的影响。
使用凹微井在存在他克莫司的情况下形成干细胞球状体，他克莫司的终浓度分别为 0μg/mL、0.1μg/mL、1μg/mL、10μg/mL 和 100μg/mL。使用显微镜定性测试细胞活力，并使用基于水溶性四唑盐的测定试剂盒定量测量细胞活力。碱性磷酸酶活性和用于测量钙沉积的蒽醌染料试验用于评估成骨分化。为了评估成骨分化的表达，进行了定量聚合酶链反应、Western blot 和 RNA 测序。
所有浓度的球状体都保持相对均匀和球形的形状。细胞活力测定表明，在成骨培养基中培养的球状体中，他克莫司的浓度高达 100μg/mL，并不显著损害细胞活力。钙沉积的增加，特别是在较低浓度的他克莫司下，表明成骨分化增强。在所有他克莫司浓度下，COL1A1 的表达都增加，这表现为平均值和中位数的升高，这可能表明成骨活性增强。
这项研究表明，他克莫司在成骨培养基中对干细胞球状体的活力没有显著影响，即使在高浓度下也是如此。它还表明，他克莫司可能增强成骨分化，这表现为钙沉积和 COL1A1 表达的增加。这些发现增进了我们对他克莫司在组织修复、再生和基于干细胞的治疗应用中的潜在作用的理解。
