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Vascular Adventitia Calcification and Its Underlying Mechanism.

Li N, Cheng W, Huang T, Yuan J, Wang X, Song M - PLoS ONE (2015)

Bottom Line: Explant cultured fibroblasts and smooth muscle cells (SMCs)were obtained from rat adventitia and media, respectively.Explant culture of fibroblasts, the primary cell type comprising the adventitia, was successfully induced for calcification after incubation with TGF-β1 (20 ng/ml) + mineralization media for 4 days, and the phenotype conversion vascular adventitia fibroblasts into myofibroblasts was identified.Culture of SMCs, which comprise only a small percentage of all cells in the adventitia, in calcifying medium for 14 days resulted in significant calcification.Vascular calcification can occur in the adventitia.

View Article: PubMed Central - PubMed

Affiliation: Department of Health Care, China-Japan Friendship Hospital, Ministry of Health, Beijing, China.

ABSTRACT
Previous research on vascular calcification has mainly focused on the vascular intima and media. However, we show here that vascular calcification may also occur in the adventitia. The purpose of this work is to help elucidate the pathogenic mechanisms underlying vascular calcification. The calcified lesions were examined by Von Kossa staining in ApoE-/- mice which were fed high fat diets (HFD) for 48 weeks and human subjects aged 60 years and older that had died of coronary heart disease, heart failure or acute renal failure. Explant cultured fibroblasts and smooth muscle cells (SMCs)were obtained from rat adventitia and media, respectively. After calcification induction, cells were collected for Alizarin Red S staining. Calcified lesions were observed in the aorta adventitia and coronary artery adventitia of ApoE-/-mice, as well as in the aorta adventitia of human subjects examined. Explant culture of fibroblasts, the primary cell type comprising the adventitia, was successfully induced for calcification after incubation with TGF-β1 (20 ng/ml) + mineralization media for 4 days, and the phenotype conversion vascular adventitia fibroblasts into myofibroblasts was identified. Culture of SMCs, which comprise only a small percentage of all cells in the adventitia, in calcifying medium for 14 days resulted in significant calcification.Vascular calcification can occur in the adventitia. Adventitia calcification may arise from the fibroblasts which were transformed into myofibroblasts or smooth muscle cells.

No MeSH data available.


Related in: MedlinePlus

Alizarin Red S staining was positive for myofibroblasts.Fibroblasts were incubated with control media for 4 days. Alizarin Red S staining of these cells was negative(Fig 5A). Calcification was not induced in the necrotic fibroblasts of mineralization group after incubation with mineralization media (DMEM containing 1% FBS, 50 ug/ml ascorbic acid, 5 mmol/L β-glycerophosphate) for 4 days. False positive was detected in fibroblasts by Alizarin Red S staining (Fig 5B).Calcification was detected in myofibroblasts of TGF-β1 (20 ng/ml) + mineralization group by Alizarin Red S staining (Fig 5C). Expression of α-actin in the myofibroblasts. Control group (Fig 6A), mineralization group (Fig 6B), TGF-β1 (20 ng/ml) + mineralization group (Fig 6C). Magnification *200
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pone.0132506.g005: Alizarin Red S staining was positive for myofibroblasts.Fibroblasts were incubated with control media for 4 days. Alizarin Red S staining of these cells was negative(Fig 5A). Calcification was not induced in the necrotic fibroblasts of mineralization group after incubation with mineralization media (DMEM containing 1% FBS, 50 ug/ml ascorbic acid, 5 mmol/L β-glycerophosphate) for 4 days. False positive was detected in fibroblasts by Alizarin Red S staining (Fig 5B).Calcification was detected in myofibroblasts of TGF-β1 (20 ng/ml) + mineralization group by Alizarin Red S staining (Fig 5C). Expression of α-actin in the myofibroblasts. Control group (Fig 6A), mineralization group (Fig 6B), TGF-β1 (20 ng/ml) + mineralization group (Fig 6C). Magnification *200

Mentions: Successful in vitro culture of fibroblasts was confirmed by Vimentin staining. Fibroblasts were then incubated with control media for 4 days. Alizarin Red S staining of these cells was negative (Fig 5A). In contrast to VSMCs, calcification was not induced in the necrotic fibroblasts of mineralization group after incubation with mineralization media (DMEM containing 1% FBS, 50 ug/ml ascorbic acid, 5 mmol/L β-glycerophosphate) for 4 days. False positive was detected in fibroblasts by Alizarin Red S staining (Fig 5B). Calcification was detected in myofibroblasts of TGF-β1 (20 ng/ml) + mineralization group by Alizarin Red S staining (Fig 5C).


Vascular Adventitia Calcification and Its Underlying Mechanism.

Li N, Cheng W, Huang T, Yuan J, Wang X, Song M - PLoS ONE (2015)

Alizarin Red S staining was positive for myofibroblasts.Fibroblasts were incubated with control media for 4 days. Alizarin Red S staining of these cells was negative(Fig 5A). Calcification was not induced in the necrotic fibroblasts of mineralization group after incubation with mineralization media (DMEM containing 1% FBS, 50 ug/ml ascorbic acid, 5 mmol/L β-glycerophosphate) for 4 days. False positive was detected in fibroblasts by Alizarin Red S staining (Fig 5B).Calcification was detected in myofibroblasts of TGF-β1 (20 ng/ml) + mineralization group by Alizarin Red S staining (Fig 5C). Expression of α-actin in the myofibroblasts. Control group (Fig 6A), mineralization group (Fig 6B), TGF-β1 (20 ng/ml) + mineralization group (Fig 6C). Magnification *200
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492877&req=5

pone.0132506.g005: Alizarin Red S staining was positive for myofibroblasts.Fibroblasts were incubated with control media for 4 days. Alizarin Red S staining of these cells was negative(Fig 5A). Calcification was not induced in the necrotic fibroblasts of mineralization group after incubation with mineralization media (DMEM containing 1% FBS, 50 ug/ml ascorbic acid, 5 mmol/L β-glycerophosphate) for 4 days. False positive was detected in fibroblasts by Alizarin Red S staining (Fig 5B).Calcification was detected in myofibroblasts of TGF-β1 (20 ng/ml) + mineralization group by Alizarin Red S staining (Fig 5C). Expression of α-actin in the myofibroblasts. Control group (Fig 6A), mineralization group (Fig 6B), TGF-β1 (20 ng/ml) + mineralization group (Fig 6C). Magnification *200
Mentions: Successful in vitro culture of fibroblasts was confirmed by Vimentin staining. Fibroblasts were then incubated with control media for 4 days. Alizarin Red S staining of these cells was negative (Fig 5A). In contrast to VSMCs, calcification was not induced in the necrotic fibroblasts of mineralization group after incubation with mineralization media (DMEM containing 1% FBS, 50 ug/ml ascorbic acid, 5 mmol/L β-glycerophosphate) for 4 days. False positive was detected in fibroblasts by Alizarin Red S staining (Fig 5B). Calcification was detected in myofibroblasts of TGF-β1 (20 ng/ml) + mineralization group by Alizarin Red S staining (Fig 5C).

Bottom Line: Explant cultured fibroblasts and smooth muscle cells (SMCs)were obtained from rat adventitia and media, respectively.Explant culture of fibroblasts, the primary cell type comprising the adventitia, was successfully induced for calcification after incubation with TGF-β1 (20 ng/ml) + mineralization media for 4 days, and the phenotype conversion vascular adventitia fibroblasts into myofibroblasts was identified.Culture of SMCs, which comprise only a small percentage of all cells in the adventitia, in calcifying medium for 14 days resulted in significant calcification.Vascular calcification can occur in the adventitia.

View Article: PubMed Central - PubMed

Affiliation: Department of Health Care, China-Japan Friendship Hospital, Ministry of Health, Beijing, China.

ABSTRACT
Previous research on vascular calcification has mainly focused on the vascular intima and media. However, we show here that vascular calcification may also occur in the adventitia. The purpose of this work is to help elucidate the pathogenic mechanisms underlying vascular calcification. The calcified lesions were examined by Von Kossa staining in ApoE-/- mice which were fed high fat diets (HFD) for 48 weeks and human subjects aged 60 years and older that had died of coronary heart disease, heart failure or acute renal failure. Explant cultured fibroblasts and smooth muscle cells (SMCs)were obtained from rat adventitia and media, respectively. After calcification induction, cells were collected for Alizarin Red S staining. Calcified lesions were observed in the aorta adventitia and coronary artery adventitia of ApoE-/-mice, as well as in the aorta adventitia of human subjects examined. Explant culture of fibroblasts, the primary cell type comprising the adventitia, was successfully induced for calcification after incubation with TGF-β1 (20 ng/ml) + mineralization media for 4 days, and the phenotype conversion vascular adventitia fibroblasts into myofibroblasts was identified. Culture of SMCs, which comprise only a small percentage of all cells in the adventitia, in calcifying medium for 14 days resulted in significant calcification.Vascular calcification can occur in the adventitia. Adventitia calcification may arise from the fibroblasts which were transformed into myofibroblasts or smooth muscle cells.

No MeSH data available.


Related in: MedlinePlus