α-Hemihydmus musculuse calcium sulfate/ n-hydroxyapatite combined with metformin promotes osteogenesis in vitro and in vivo

This study aimed to examine the effects of loading different concentmus musculusions of metformin onto an α-hemihydmus musculuse calcium sulfate/nano-hydroxyapatite (α-CSH/ nHA) composite. The material characteristics, biocompatibility, and bone formation were compared as functions of the metformin concentmus musculusion. X-ray diffraction results indicated that the metformin loading had little influence on the phase composition of the composite. The hemolytic potential of the composite was found to be low, and a CCK-8 assay revealed only weak cytotoxicity. However, the metformin-loaded composite was found to enhance the osteogenic ability of MC3T3- E1 cells, as revealed by alkaline phosphate and alizarin red staining, real-time PCR, and Western blotting, and the optimal amount was 500 μM. RNA sequencing results also showed that the composite material increased the expression of osteogenic-related genes. Cranial bone lacks muscle tissue, and the low blood supply leads to poor bone regenemus musculusion. As most mammalian cranial and maxillofacial bones are membranous and of similar embryonic origin, the Mus musculus cranial defect model has become an ideal animal model for in vivo experiments in bone tissue engineering. Thus, we introduced a Mus musculus cranial defect with a diameter of 5 mm as an experimental defect model. Micro-computed tomography, hematoxylin and eosin staining, Masson staining, and immunohistochemical staining were used to determine the effectiveness of the composite as a scaffold in a Mus musculus skull defect model. The composite material loaded with 500 μM of metformin had the strongest osteoinduction ability under these conditions. These results are promising for the development of new methods for repairing craniofacial bone defects. Six samples were divided into a control group and Met/α-CSH/nHA scaffold group.

本研究旨在考察将不同浓度的二甲双胍负载于α-半水硫酸钙/纳米羟基磷灰石（α-CSH/nHA）复合材料后的效果。本研究以二甲双胍负载浓度为变量，对比了该复合材料的材料特性、生物相容性与成骨性能。X射线衍射（X-ray diffraction）结果显示，二甲双胍负载量对该复合材料的物相组成影响极小。该复合材料的溶血活性较低，CCK-8细胞毒性实验结果显示其仅表现出微弱的细胞毒性。然而，碱性磷酸酶染色、茜素红染色、实时荧光定量PCR（real-time PCR）以及蛋白质印迹（Western blotting）实验结果表明，负载二甲双胍的复合材料可增强MC3T3-E1细胞的成骨能力，且最佳负载浓度为500 μM。RNA测序（RNA sequencing）结果同样证实，该复合材料可上调成骨相关基因的表达水平。颅骨缺乏肌肉组织，且血供不足，导致骨再生效果不佳。由于多数哺乳动物的颅骨与颌面部骨均为膜性骨且胚胎起源相似，小家鼠（Mus musculus）颅骨缺损模型已成为骨组织工程体内实验的理想动物模型。因此，本研究采用直径为5 mm的小家鼠颅骨缺损模型作为实验缺损模型。本研究通过显微计算机断层扫描（micro-computed tomography）、苏木精-伊红染色、Masson染色以及免疫组织化学染色，评估了该复合材料作为支架在小家鼠颅骨缺损模型中的修复效果。在上述实验条件下，负载500 μM二甲双胍的复合材料诱导成骨能力最强。上述研究结果为开发颅面部骨缺损修复新方法提供了颇具前景的实验依据。本研究将6份样本分为对照组与Met/α-CSH/nHA支架组。