Bone Conditioned Medium: Preparation and Bioassay

作者：Caballe Serrano Jordi; Sawada Kosaku; Schuldt Filho Guenther; Bosshardt Dieter D; Buser Daniel; Gruber Reinhard^*

来源：Jove-Journal of Visualized Experiments, 2015, (101): e52707.

DOI：10.3791/52707

• Molecular Biology
• Issue 101
• Bone Conditioned Medium
• BCM
• bone autograft
• guided bone regeneration
• GBR
• dental implant
• membrane
• supernatant
• growth factors
• contour augmentation
• autologous bone

Autologous bone grafts are widely used in oral and maxillofacial surgery, orthopedics, and traumatology. Autologous bone grafts not only replace missing bone, they also support the complex process of bone regeneration. This favorable behavior of autografts is attributed to the three characteristics: osteoconductivity, osteogenicity, and osteoinductivity. However, there is another aspect: Bone grafts release a myriad of molecules, including growth factors, which can target mesenchymal cells involved in bone regeneration. The paracrine properties of bone grafts can be studied in vitro by the use of bone-conditioned medium (BCM). Here we present a protocol on how to prepare bone-conditioned medium from native pig cortical bone, and bone that underwent thermal processing or demineralization. Cells can be directly exposed to BCM or seeded onto biomaterials, such as collagen membranes, previously soaked with BCM. We give examples for in vitro bioassays with mesenchymal cells on the expression of TGF-beta regulated genes. The presented protocols should encourage to further reveal the paracrine effects of bone grafts during bone regeneration and open a path for translational research in the broad field of reconstructive surgery.

• 出版日期2015-7