Bone‐Conditioned Medium Inhibits Osteogenic and Adipogenic Differentiation of Mesenchymal Cells In Vitro

Peng, Jianbo; Nemec, Michael; Brolese, Eliane Lucia; Bosshardt, Dieter D.; Schaller, Benoît; Buser, Daniel; Gruber, Reinhard

doi:10.1111/cid.12200

Cited by 28 publications

(83 citation statements)

References 62 publications

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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%

“…For example, the TGF-β receptor I antagonist SB431542 blocked the effect of BCM on the expression of the gene panel ADM, IL-11, NOX4, PRG4, and PTX3 11,15 . Interestingly, alkaline phosphatase and IL33 were not reversed by SB431542 11,15 suggesting that other as yet unknown pathways are regulated by BCM. Another open question is what are the molecules in BCM being responsible for the cellular response?…”

Section: Discussionmentioning

confidence: 99%

“…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%

“…A similar term "demineralized bone matrix-conditioned medium" was already coined in 1996 and supports the overall concept of a paracrine function of bone, even when processed by demineralization 17 . For our purposes, BCM can be prepared from fresh pig mandibles 10,11 . Proteomic analyses of BCM revealed the complex composition, including growth factors and constituents of the extracellular matrix 10 , also extending existing knowledge on the proteasome of whole bone 18,19 .…”

Section: Introductionmentioning

confidence: 99%

“…In vitro, BCM reduces osteogenic and adipogenic differentiation, and provokes a strong increase of IL11 expression 11 . Genome wide microarray revealed more genes to be differentially expressed in mesenchymal cells in response to BCM.…”

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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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: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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The conditioned medium from osteo‐differentiating human mesenchymal stem cells affects the viability of triple negative MDA‐MB231 breast cancer cells

Librizzi

Tobiasch

Luparello

2015

Cell Biochemistry & Function

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This study aimed to investigate the effect of conditioned media (CM) from osteo-differentiating and adipo-differentiating human mesenchymal stem cells (MSCs) isolated from lipoaspirates of healthy female donors on the viability of triple-negative breast cancer cells MDA-MB231. The CM of undifferentiated and differentiating MSCs were collected after 7, 14, 21 and 28 days of culture. The effects of MSC CM on cell proliferation were assessed using an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay after 24 h. The effects of osteo-differentiating cell CM on apoptotic promotion, cell cycle impairment, mitochondrial transmembrane potential dissipation, production of reactive oxygen species and autophagosome accumulation were analysed by flow cytometry and Western blot. MTT assay showed that only CM collected from osteo-induced cells at day 28 (d28O-CM) reduced tumour cell viability. Treatment with d28O-CM restrained cell cycle progression through G2 phase, elicited a caspase-8-driven apoptotic effect already after 5 h of culture, and down-regulated autophagosome accumulation and beclin-1 expression. The finding that factor(s) secreted by osteo-differentiating MSCs shows properties of an apoptotic inducer and autophagy inhibitor on triple-negative breast cancer cells may have an important applicative potential that deserves further investigation.

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Acid bone lysates reduce bone regeneration in rat calvaria defects

Strauß

Küchler

Kobatake

et al. 2020

J Biomedical Materials Res

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Acid bone lysates (ABLs) represent the growth factors and other molecules released during autologous graft resorption. However, the impact of these bone‐derived growth factors on the healing of bone defects has not yet been investigated. The aim of the present study was, therefore, to examine the impact of ABLs adsorbed to collagen membranes on bone regeneration. To this end, in 16 female Sprague Dawley rats, a standardized 5‐mm‐diameter critical size defect on the calvarial bone was created. The defects were covered with collagen membranes that had been soaked either in serum‐free media or ABLs followed by lyophilization. After a healing period of 4 weeks, micro‐computed tomography (μCT) and histological analyses by means of undecalcified thin ground sections were performed. μCT analysis of the inner 4 mm of the calvaria defect showed a greater bone defect coverage in the control group when compared to ABL group, 29.8% (confidence interval [CI]: 17.7–50.3) versus 5.6% (CI: 1.0–29.8, p = .03), respectively. Moreover, we found significantly more absolute bone volume (BV) in the control group when compared to ABL group, 0.59 mm3 (CI: 0.27–1.25) versus 0.07 mm3 (CI: 0.06–0.59, p = .04), respectively. Histomorphometry confirmed these findings with a relative BV in the central compartment of 14.1% (CI: 8.4–20.6) versus 5.6% (CI: 3.4–7.9, p = .004), respectively. These findings indicate that bone‐derived growth factors contained in ABLs are able to attenuate bone regeneration within collagen membranes.

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Bone‐Conditioned Medium Inhibits Osteogenic and Adipogenic Differentiation of Mesenchymal Cells In Vitro

Abstract: The present work is a pioneer study on the paracrine activity of bone grafts. The findings suggest that cortical bone chips release soluble signals that can modulate differentiation of mesenchymal cells in vitro at least partially involving TGF-β signaling.

Cited by 28 publications

References 62 publications

Bone Conditioned Medium: Preparation and Bioassay

Bone Conditioned Medium: Preparation and Bioassay

The conditioned medium from osteo‐differentiating human mesenchymal stem cells affects the viability of triple negative MDA‐MB231 breast cancer cells

Acid bone lysates reduce bone regeneration in rat calvaria defects

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