scholarly journals The culture microenvironment of juvenile idiopathic arthritis synovial fibroblasts is favorable for endochondral bone formation through BMP4 and repressed by chondrocytes

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Megan M. Simonds ◽  
Amanda R. Schlefman ◽  
Suzanne M. McCahan ◽  
Kathleen E. Sullivan ◽  
Carlos D. Rose ◽  
...  
Keyword(s):  
Juvenile Idiopathic Arthritis ◽  
Bone Formation ◽  
Synovial Fibroblasts ◽  
Conditioned Media ◽  
Endochondral Bone Formation ◽  
Endochondral Bone ◽  
Member Gene ◽  
Bmp Antagonist ◽  
Fibroblast Like Synoviocytes ◽  
Tgfβ Superfamily

Abstract Background We examined influences of conditioned media from chondrocytes (Ch) on juvenile idiopathic arthritis synovial fibroblasts (JFLS) and potential for JFLS to undergo endochondral bone formation (EBF). Methods Primary cells from three control fibroblast-like synoviocytes (CFLS) and three JFLS were cultured in Ch-conditioned media and compared with untreated fibroblast-like synoviocytes (FLS). RNA was analyzed by ClariomS microarray. FLS cells cultured in conditioned media were exposed to either TGFBR1 inhibitor LY3200882 or exogenous BMP4 and compared with FLS cultured in conditioned media from Ch (JFLS-Ch). Media supernatants were analyzed by ELISA. Results In culture, JFLS downregulate BMP2 and its receptor BMPR1a while upregulating BMP antagonists (NOG and CHRD) and express genes (MMP9, PCNA, MMP12) and proteins (COL2, COLX, COMP) associated with chondrocytes. Important TGFβ superfamily member gene expression (TGFBI, MMP9, COL1A1, SOX6, and MMP2) is downregulated when JFLS are cultured in Ch-conditioned media. COL2, COLX and COMP protein expression decreases in JFLS-Ch. BMP antagonist protein (NOG, CHRD, GREM, and FST) secretion is significantly increased in JFLS-Ch. Protein phosphorylation increases in JFLS-Ch exposed to exogenous BMP4, and chondrocyte-like phenotype is restored in BMP4 presence, evidenced by increased secretion of COL2 and COLX. Inhibition of TGFBR1 in JFLS-Ch results in overexpression of COL2. Conclusions JFLS are chondrocyte-like, and Ch-conditioned media can abrogate this phenotype. The addition of exogenous BMP4 causes JFLS-Ch to restore this chondrocyte-like phenotype, suggesting that JFLS create a microenvironment favorable for endochondral bone formation, thereby contributing to joint growth disturbances in juvenile idiopathic arthritis.

scholarly journals Juvenile idiopathic arthritis fibroblast-like synoviocytes influence chondrocytes to alter BMP antagonist expression demonstrating an interaction between the two prominent cell types involved in endochondral bone formation

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Megan M. Simonds ◽  
Amanda R. Schlefman ◽  
Suzanne M. McCahan ◽  
Kathleen E. Sullivan ◽  
Carlos D. Rose ◽  
...  
Keyword(s):  
Juvenile Idiopathic Arthritis ◽  
Synovial Cell ◽  
Synovial Fibroblasts ◽  
Cell Types ◽  
Bmp Signaling ◽  
Conditioned Media ◽  
Endochondral Bone Formation ◽  
Direct Role ◽  
Endochondral Bone ◽  
Tgfβ Receptors

Abstract Background To examine critical interactions between juvenile idiopathic arthritis synovial fibroblasts (JFLS) and chondrocytes (Ch), and their role in bony overgrowth seen in patients with juvenile idiopathic arthritis (JIA). Methods Control (CFLS) and JFLS were cultured in synoviocyte media containing recombinant BMP4. Ch were cultured in either CFLS or JFLS conditioned-media without stimulation. Media supernatants were analyzed by ELISA. RNA from conditioned media experiment was analyzed by ClariomS microarray. Results As expected, genes expressed in untreated JFLS and CFLS cultured in synoviocyte media were similar to each other and this expression differed from untreated Ch cultured in chondrocyte media. JFLS favor BMP ligand gene expression while downregulating TGFβ receptors’ expression. Noggin and chordin, antagonists with high affinity for BMP4, are JFLS- but not Ch-preferred regulators of BMP signaling. Compared to Ch, JFLS overexpress collagen X (COLX), a marker of chondrocyte hypertrophy. Exogenous BMP4 causes JFLS to significantly decrease expression of noggin and collagen II (COL2), a marker of chondrocyte proliferation, and causes overexpression of COLX and alkaline-phosphatase (ALP). Chondrocytes cultured in JFLS-conditioned media (Ch-JFLS) express BMP genes and favor chordin protein expression over other antagonists. Ch-JFLS have significantly increased expression of COL2 and significantly decreased expression of COLX. Conclusions These data suggest JFLS, in the presence of BMP4, undergo hypertrophy and that JFLS-conditioned media influence chondrocytes to become highly proliferative. To the authors’ knowledge, no prior study has shown that JFLS and chondrocytes play a direct role in the bony overgrowth in joints of patients with JIA and that BMPs or regulation of these growth factors influence the interaction between two prominent synovial cell types.

Juvenile Idiopathic Arthritis Fibroblast-like Synoviocytes, through BMP signaling, play a central role in joint growth disturbances

2020 ◽  
Author(s):  
Megan Simonds ◽  
Amanda Schlefman ◽  
Suzanne McCahan ◽  
Kathleen Sullivan ◽  
Carlos Rose ◽  
...  
Keyword(s):  
Juvenile Idiopathic Arthritis ◽  
Synovial Cell ◽  
Synovial Fibroblasts ◽  
Cell Types ◽  
Bmp Signaling ◽  
Conditioned Media ◽  
Chondrocyte Proliferation ◽  
Direct Role ◽  
Tgfβ Receptors ◽  
Fibroblast Like Synoviocytes

Abstract Background: To examine critical interactions between juvenile idiopathic arthritis synovial fibroblasts (JFLS) and chondrocytes (Ch), and their role in bony overgrowth seen in patients with juvenile idiopathic arthritis (JIA). Methods: Control (CFLS) and JFLS were cultured in media containing recombinant BMP4. Ch were cultured in either CFLS or JFLS conditioned-media. Media supernatants were analyzed by ELISA. RNA was analyzed by ClariomS microarray. Results: As expected, genes expressed in JFLS and CFLS were similar to each other and this expression differed from Ch. JFLS favor BMP ligand gene expression while downregulating TGFβ receptors’ expression. Noggin and chordin, antagonists with high affinity for BMP4, are JFLS- but not Ch-preferred regulators of BMP signaling. Compared to Ch, JFLS overexpress collagen X (COLX), a marker of chondrocyte hypertrophy. Exogenous BMP4 causes JFLS to significantly decrease expression of noggin and collagen II (COL2), a marker of chondrocyte proliferation, and causes overexpression of COLX and alkaline-phosphatase (ALP). Chondrocytes cultured in JFLS-conditioned media (Ch-JFLS) express BMP genes and favor chordin protein expression over other antagonists. Ch-JFLS have significantly increased expression of COL2 and significantly decreased expression of COLX. Conclusions: These data suggest JFLS, in the presence of BMP4, undergo hypertrophy and that JFLS-conditioned media influence chondrocytes to become highly proliferative. To the authors’ knowledge, no prior study has shown that JFLS and chondrocytes play a direct role in the bony overgrowth in joints of patients with JIA and that BMPs or regulation of these growth factors influence the interaction between two prominent synovial cell types.

scholarly journals Intraflagellar transport is essential for endochondral bone formation

Development ◽  
2007 ◽  
Vol 134 (2) ◽  
pp. 307-316 ◽  
Author(s):  
C. J. Haycraft ◽  
Q. Zhang ◽  
B. Song ◽  
W. S. Jackson ◽  
P. J. Detloff ◽  
...  
Keyword(s):  
Bone Formation ◽  
Intraflagellar Transport ◽  
Endochondral Bone Formation ◽  
Endochondral Bone

scholarly journals Osteoblast‐Specific Loss of IGF1R Signaling Results in Impaired Endochondral Bone Formation During Fracture Healing

2015 ◽  
Vol 30 (9) ◽  
pp. 1572-1584 ◽  
Author(s):  
Tao Wang ◽  
Yongmei Wang ◽  
Alicia Menendez ◽  
Chak Fong ◽  
Muriel Babey ◽  
...  
Keyword(s):  
Bone Formation ◽  
Fracture Healing ◽  
Endochondral Bone Formation ◽  
Endochondral Bone ◽  
Specific Loss ◽  
Igf1r Signaling

scholarly journals Conditional Deletion of Prolyl Hydroxylase Domain-Containing Protein 2 (Phd2) Gene Reveals Its Essential Role in Chondrocyte Function and Endochondral Bone Formation

Endocrinology ◽  
2016 ◽  
Vol 157 (1) ◽  
pp. 127-140 ◽  
Author(s):  
Shaohong Cheng ◽  
Weirong Xing ◽  
Sheila Pourteymoor ◽  
Jan Schulte ◽  
Subburaman Mohan
Keyword(s):  
Bone Formation ◽  
Growth Plate ◽  
Prolyl Hydroxylase ◽  
Conditional Knockout ◽  
Hypertrophic Chondrocytes ◽  
Endochondral Bone Formation ◽  
Bone Surface ◽  
Endochondral Bone ◽  
Prolyl Hydroxylase Domain

Abstract The hypoxic growth plate cartilage requires hypoxia-inducible factor (HIF)-mediated pathways to maintain chondrocyte survival and differentiation. HIF proteins are tightly regulated by prolyl hydroxylase domain-containing protein 2 (Phd2)-mediated proteosomal degradation. We conditionally disrupted the Phd2 gene in chondrocytes by crossing Phd2 floxed mice with type 2 collagen-α1-Cre transgenic mice and found massive increases (>50%) in the trabecular bone mass of long bones and lumbar vertebra of the Phd2 conditional knockout (cKO) mice caused by significant increases in trabecular number and thickness and reductions in trabecular separation. Cortical thickness and tissue mineral density at the femoral middiaphysis of the cKO mice were also significantly increased. Dynamic histomorphometric analyses revealed increased longitudinal length and osteoid surface per bone surface in the primary spongiosa of the cKO mice, suggesting elevated conversion rate from hypertrophic chondrocytes to mineralized bone matrix as well as increased bone formation in the primary spongiosa. In the secondary spongiosa, bone formation measured by mineralizing surface per bone surface and mineral apposition rate were not changed, but resorption was slightly reduced. Increases in the mRNA levels of SRY (sex determining region Y)-box 9, osterix (Osx), type 2 collagen, aggrecan, alkaline phosphatase, bone sialoprotein, vascular endothelial growth factor, erythropoietin, and glycolytic enzymes in the growth plate of cKO mice were detected by quantitative RT-PCR. Immunohistochemistry revealed an increased HIF-1α protein level in the hypertrophic chondrocytes of cKO mice. Infection of chondrocytes isolated from Phd2 floxed mice with adenoviral Cre resulted in similar gene expression patterns as observed in the cKO growth plate chondrocytes. Our findings indicate that Phd2 suppresses endochondral bone formation, in part, via HIF-dependent mechanisms in mice.

Oxygen Tension Directs Chondrogenic Differentiation of Myelo-Monocytic Progenitors During Endochondral Bone Formation

2007 ◽  
Vol 13 (8) ◽  
pp. 2011-2019 ◽  
Author(s):  
Jessica Shafer ◽  
Alan R. Davis ◽  
Francis H. Gannon ◽  
Christine M. Fouletier-Dilling ◽  
Zawaunyka Lazard ◽  
...  
Keyword(s):  
Bone Formation ◽  
Oxygen Tension ◽  
Chondrogenic Differentiation ◽  
Endochondral Bone Formation ◽  
Endochondral Bone

Conditional inactivation of VEGF-A in areas of collagen2a1 expression results in embryonic lethality in the heterozygous state

Development ◽  
2000 ◽  
Vol 127 (7) ◽  
pp. 1445-1453 ◽  
Author(s):  
J.J. Haigh ◽  
H.P. Gerber ◽  
N. Ferrara ◽  
E.F. Wagner
Keyword(s):  
Bone Formation ◽  
Ischemic Cardiomyopathy ◽  
Embryonic Lethality ◽  
Limb Bud ◽  
Endochondral Bone Formation ◽  
Functional Importance ◽  
Mouse Development ◽  
Endochondral Bone ◽  
Myocardial Layers ◽  
Transgenic Lines

VEGF-A has been implicated in regulating the initial angiogenic invasion events that are essential for endochondral bone formation. VEGF-A mRNA expression was indeed found in the sclerotome of the developing somite and in the limb-bud mesenchyme at E10.5 in mouse development but declined during chondrogenesis and became upregulated in hypertrophic chondrocytes prior to angiogenic invasion. To determine the functional importance of VEGF-A expression in the developing chondrogenic tissues, VEGF-A was conditionally inactivated during early embryonic development using Collagen2a1-Cre transgenic lines. Deletion of a single VEGF-A allele in Collagen2a1-Cre-expressing cells results in embryonic lethality around E10.5. This lethality is characterized by aberrant development of the dorsal aorta and intersomitic blood vessels, along with defects in the developing endocardial and myocardial layers of the heart. A small percentage of VEGF(Flox)/+, Collagen2a1-Cre fetuses survive until E17.5, show aberrant endochondral bone formation and develop a heart phenotype resembling a dilated form of ischemic cardiomyopathy. These results provide insights into the function of VEGF-A in heart and endochondral bone formation and underscore the importance of tightly controlled levels of VEGF-A during development.

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