Thermal processing of bone: in vitro response of mesenchymal cells to bone-conditioned medium

Sawada, Kosaku; Caballé‐Serrano, Jordi; Filho, Guenther Schuldt; Bosshardt, Dieter D.; Schaller, Benoît; Buser, Daniel; Gruber, Reinhard

doi:10.1016/j.ijom.2015.03.012

Cited by 12 publications

(19 citation statements)

References 31 publications

(37 reference statements)

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“…Understanding how BCM affects mesenchymal cells can help to learn more about graft consolidation and properties of bone autografts. Based on this concept we have accumulated knowledge on the impact of BCM obtained from native 11,15 and processed bone 14,20 on gene expression of mesenchymal cells, but also on proliferation, migration, and differentiation into the three main lineages; osteoblasts, adipocytes and chondrocytes 11 . BCM was also examined for its capacity to target hematopoietic cells, for example with respect to the modulation of osteoclastogenesis 13 .…”

Section: Discussionmentioning

confidence: 99%

“…Among these genes are adrenomedullin (ADM), IL11, IL33, NADPH oxidase 4 (NOX4), proteoglycan 4 (PRG4, or lubricin) and pentraxin 3 (PTX3) 15 . BCM obtained from autoclaved bone chips failed to change the expression of the respective genes 14 . BCM from bone chips that underwent pasteurization and freezing was able to change gene expression 14 .…”

Section: Introductionmentioning

confidence: 98%

“…BCM obtained from autoclaved bone chips failed to change the expression of the respective genes 14 . BCM from bone chips that underwent pasteurization and freezing was able to change gene expression 14 . Also conditioned medium of demineralized bone matrix (DBM-CM) changes the expression of TGF-β-regulated genes 20 .…”

Section: Introductionmentioning

confidence: 98%

“…Third, autologous bone is osteoinductive as growth factors like bone morphogenetic proteins entombed in the matrix can initiate the process of endochondral or even intramembranous bone formation 9 . There is another aspect: freshly prepared bone chips hold a paracrine function based on the in vitro observations with "bone-conditioned medium" [10][11][12][13][14][15] . Also the impact of myelopoiesis should be mentioned 16 .…”

Section: Introductionmentioning

confidence: 99%

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Bone Conditioned Medium: Preparation and Bioassay

Caballé‐Serrano

Sawada

Filho

et al. 2015

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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. Video LinkThe video component of this article can be found at

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

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Bone Conditioned Medium: Preparation and Bioassay

Caballé‐Serrano

Sawada

Filho

et al. 2015

JoVE

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“…TGF-β targets regulated by bone-conditioned medium (BCM) include adrenomedullin (ADM), pentraxin 3 (PTX3), BTB/POZ domain-containing protein 11 (BTBD11), interleukin 11 (IL-11), proteoglycan 4 (PRG4), and NADPH oxidase 4 (NOX4) (21,22). This gene panel has recently provided a scientific basis for the development of a bioassay to determine paracrine-like activity in bone grafts that have undergone autoclaving, pasteurization, and freezing (23). A previous study assessed the response of oral fibroblasts exposed to supernatants of demineralized bone (24); however, it remains to be determined whether irradiation affects the paracrine-like activity of bone grafts.…”

Section: Introductionmentioning

confidence: 99%

High-dose irradiation of bone chips preserves the <i>in vitro</i> activity of bone-conditioned medium

et al. 2016

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Extracorporeal irradiation sterilizesresected tumor bone used as autografts in reconstruction surgery. Therapeutic irradiation is a standard technique in head and neck cancer therapy that aims to preserve organ function. Bone irradiation has a complex, mostly inhibitory, effect on remodeling and regeneration, although the underlying mechanisms are still not fully understood. It remains unclear if extracorporeal irradiation affects the paracrine-like activity of the corresponding autografts. We recently reported that bone-conditioned medium from autogenous bone chips contains a number of factors that might affect cell activity. In the present study, we investigated the effects of extracorporeal irradiation of porcine cortical bone chips on the activity of the corresponding bone-conditioned medium. The effects of bone-conditioned medium on the expressions of transforming growth factor-beta (TGF-β) target genes in oral fibroblasts were assessed. Bone-conditioned medium from bone chips exposed to a total radiation dose up to 120 Gy did not affect expressions of TGF-β target genes, including adrenomedullin, BTB/ POZ domain-containing protein 11, proteoglycan 4, NADPH oxidase 4, and interleukin 11, in oral fibroblasts. In conclusion, bone irradiation does not alter the capability of the corresponding bone-conditioned medium to provoke a robust fibroblastic cell response in vitro. (J Oral Sci 58, 325-331, 2016)

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