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Conditioned medium from fresh and demineralized bone enhances osteoclastogenesis in murine bone marrow cultures
Author(s) -
CaballéSerrano Jordi,
Schuldt Filho Guenther,
Bosshardt Dieter D.,
GargalloAlbiol Jordi,
Buser Daniel,
Gruber Reinhard
Publication year - 2016
Publication title -
clinical oral implants research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.407
H-Index - 161
eISSN - 1600-0501
pISSN - 0905-7161
DOI - 10.1111/clr.12573
Subject(s) - rankl , osteoclast , acid phosphatase , multinucleate , chemistry , bone resorption , bone marrow , microbiology and biotechnology , tartrate resistant acid phosphatase , stromal cell , paracrine signalling , osteolysis , in vitro , haematopoiesis , immunology , biology , stem cell , endocrinology , receptor , cancer research , biochemistry , medicine , activator (genetics) , dentistry , enzyme
Abstract Objectives Osteoclasts rapidly form on the surface of bone chips at augmentation sites. The underlying molecular mechanism, however, is unclear. Soluble factors released from bone chips in vitro have a robust impact on mesenchymal cell differentiation. Whether these soluble factors change the differentiation of hematopoietic cells into osteoclasts remains unknown. Methods Osteoclastogenesis, the formation of tartrate‐resistant acid phosphatase‐positive multinucleated cells, was studied with murine bone marrow cultures exposed to RANKL and M‐ CSF , and conditioned medium from fresh ( BCM ) and demineralized bone matrix ( DCM ). Histochemical staining, gene and protein expression, as well as viability assays were performed. Results This study shows that BCM had no impact on osteoclastogenesis. However, when BCM was heated to 85°C ( BCM h), the number of tartrate‐resistant acid phosphatase‐positive multinucleated cells that developed in the presence of RANKL and M‐ CSF approximately doubled. In line with the histochemical observations, there was a trend that BCM h increased expression of osteoclast marker genes, in particular the transcription factor c‐fos. The expression of c‐fos was significantly reduced by the TGF ‐β receptor I antagonist SB 431542. DCM significantly stimulated osteoclastogenesis, independent of thermal processing. Conclusions These data demonstrate that activated BCM by heat and DBM are able to stimulate osteoclastogenesis in vitro . These in vitro results support the notion that the resorption of autografts may be supported by as yet less defined paracrine mechanisms.