Bone Conditioned Medium: Preparation and Bioassay

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:...

Full description

Saved in:
Bibliographic Details
Published in:Journal of visualized experiments 2015-07 (101), p.e52707-e52707
Main Authors: Caballé-Serrano, Jordi, Sawada, Kosaku, Schuldt Filho, Guenther, Bosshardt, Dieter D, Buser, Daniel, Gruber, Reinhard
Format: Article
Language:English
Subjects:
Animals
Biocompatible Materials - pharmacology
Biological Assay - methods
Bone and Bones - cytology
Bone Regeneration - physiology
Bone Transplantation - methods
Cell Culture Techniques - methods
Culture Media, Conditioned
Humans
Intercellular Signaling Peptides and Proteins - metabolism
Mesenchymal Stem Cells - cytology
Mesenchymal Stem Cells - metabolism
Molecular Biology
Swine
Transforming Growth Factor beta - biosynthesis
Transforming Growth Factor beta - genetics
Transplantation, Autologous
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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-β 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.
ISSN:1940-087X
1940-087X
DOI:10.3791/52707