Study on the Role of Pressure in Regulating the Osteogenesis of Cranial Cells through the Exosomes of Immature Dural Cells
WANG Yu1,2,3#, HAN Yanjun1,2,3#, WANG Lei1, ZHANG Chunyang1,2,3, SUN Jianying1, ZHAO Zhijun1,2,3*, ZHANG Xiaolu1,2,3*
This study employed in vitro experiments to co-culture exosomes secreted by immature dural cells of SD rats under stress with cranial bone cells, simulating the growth and development process of rat cranial bones, to investigate the role of immature dura mater in cranial bone development. Primary dural cells were isolated and extracted from 3-day-old SD rats. In vitro experiments were conducted to subject the primary cultured dural cells to stress, with CCK-8 and colony formation assays used to assess dural cell proliferation, and TUNEL staining used to detect apoptosis. Cell supernatants were collected, and exosomes were extracted by differential centrifugation. Next-generation sequencing was employed to analyze the impact of stress on the osteogenic potential of immature dural cell exosomes. Co-cultures of cranial bone cells with exosomes derived from stress-treated immature dural cells were performed, and osteogenic activity was assessed by alizarin red and ALP staining, while real-time quantitative PCR was used to measure the expression of osteogenesis-related genes. Results demonstrated that stress stimulation enhanced the proliferative capacity of immature dural cells, and the exosomes produced contained higher levels of osteogenesis-regulating miRNAs (miR-21-5p, miR-132, miR-20a, miR-204). Immature dural cell exosomes promoted osteogenic activity in cranial bone cells, and co-culturing stress-treated exosomes with cranial bone cells further enhanced osteogenic capacity, accompanied by increased expression levels of Runx2, OPN, and OCN in cranial bone cells. The study found that the immature dura mater plays a crucial role in skull growth and development, and stimulation of the immature dura mater by pressure contributes to skull growth and development, providing a fundamental theoretical basis for selecting appropriate repair methods in clinical skull defect treatment.
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