Department of Cranio-Maxillofacial Surgery, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland.
Department of Oral and Maxillofacial Surgery, School of Life Dentistry at Tokyo, The Nippon Dental University, Tokyo, Japan.
J Biomed Mater Res A. 2022 Aug;110(8):1524-1532. doi: 10.1002/jbm.a.37392. Epub 2022 Apr 15.
The present study investigated the osteoclast differentiation potential and paracrine effects of osteoclasts on osteoblast differentiation when the cells were cultured directly on two bone substitutes (BSs): deproteinized bovine bone mineral (DBBM) and carbonate apatite (CO Ap). Human primary osteoclasts cultured on the BSs were assessed by tartrate-resistant acid phosphatase (TRAP) and actin ring staining. Thereafter, the mRNA levels of osteoclastic differentiation markers were quantified by real-time PCR. Osteoblast behaviors in response to conditioned media collected from osteoclast cultures were investigated. Interestingly, mature osteoclasts were occasionally observed on the surface of the CO Ap granules, whereas very few and small osteoclasts were observed on DBBM. Similarly, real-time PCR analysis showed higher mRNA levels of osteoclast markers, including cathepsin K and TRAP, in the cells cultured on CO Ap than in those cultured on DBBM. Furthermore, compared to DBBM, CO Ap promoted osteoblast differentiation in human primary osteoblasts, whereas few paracrine effects of osteoclasts cultured with either BS were observed on the osteoblast differentiation potential. These limited results showed that CO Ap provided a favorable surface for osteoclast differentiation, as well as osteoblasts, compared to DBBM in vitro.
本研究探讨了将细胞直接培养在两种骨替代物（BS）：脱蛋白牛骨矿物质（DBBM）和碳酸磷灰石（CO Ap）上时，破骨细胞的破骨细胞分化潜能和旁分泌效应对成骨细胞分化的影响。通过抗酒石酸酸性磷酸酶（TRAP）和肌动蛋白环染色评估培养在 BS 上的人原代破骨细胞。此后，通过实时 PCR 定量测定破骨细胞分化标志物的 mRNA 水平。研究了破骨细胞培养物收集的条件培养基对成骨细胞行为的影响。有趣的是，偶尔会在 CO Ap 颗粒表面观察到成熟的破骨细胞，而在 DBBM 上则观察到很少且很小的破骨细胞。同样，实时 PCR 分析显示，在 CO Ap 上培养的细胞中破骨细胞标志物（包括组织蛋白酶 K 和 TRAP）的 mRNA 水平高于在 DBBM 上培养的细胞。此外，与 DBBM 相比，CO Ap 促进了人原代成骨细胞的分化，而在成骨细胞分化潜能方面，很少观察到两种 BS 培养的破骨细胞的旁分泌作用。这些有限的结果表明，与 DBBM 相比，CO Ap 为体外破骨细胞分化以及成骨细胞提供了有利的表面。
