The effect of surface morphology on endothelial and smooth muscle cells growth on blow-spun fibrous scaffolds

AbstractThis study aimed to analyze the growth of two types of blood vessel building cells: endothelial cells (ECs) and smooth muscle cells (SMCs) on surfaces with different morphology. Two types of materials, differing in morphology, were produced by the solution blow spinning technique. One-layer materials consisted of one fibrous layer with two fibrous surfaces. Bi-layer materials consisted of one fibrous-solid layer and one fibrous layer, resulting in two different surfaces. Additionally, materials with different average fiber diameters (about 200, 500, and 900 nm) were produced for each group. It has been shown that it is possible to obtain structures with a given morphology by changing the selected process parameters (working distance and polymer solution concentration). Both morphology (solid versus fibrous) and average fiber diameter (submicron fibers versus microfibers) of scaffolds influenced the growth of ECs. However, this effect was only visible after an extended period of culture (6 days). In the case of SMCs, it was proved that the best growth of SMCs is obtained for micron fibers (with an average diameter close to 900 nm) compared to the submicron fibers (with an average diameter below 900 nm).

Download Full-text

CYTOPLASMIC FILAMENTS IN DEVELOPING AND ADULT VERTEBRATE SMOOTH MUSCLE

The Journal of Cell Biology ◽

10.1083/jcb.50.2.484 ◽

1971 ◽

Vol 50 (2) ◽

pp. 484-497 ◽

Cited By ~ 64

Author(s):

Y. Uehara ◽

G. R. Campbell ◽

G. Burnstock

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Relative Number ◽

Muscle Cells ◽

Average Diameter ◽

Longitudinal Section ◽

Extensive Study ◽

Cytoplasmic Filaments ◽

Intimate Association ◽

Dark Body

An extensive study of adult and developing smooth muscle has revealed the widespread occurrence of a distinct filament with an average diameter of about 100 A (termed the 100 A filament). Unlike that of myofilaments, their appearance in longitudinal section is uniform, but in transverse section they have a round profile, occasionally exhibiting a less electron-opaque core. The 100 A filaments are almost invariably preserved under a variety of fixation procedures, whereas myofilaments, particularly the thicker filaments, are preserved inconsistently. The 100 A filaments appear to be randomly oriented throughout the cytoplasm, either singly or in small groups, although they are sometimes concentrated in the juxtanuclear region of the smooth muscle cells. The intimate association of 100 A filaments with dark bodies, in both developing and adult smooth muscle cells, may indicate that these filaments either play a role in dark body formation or, at least, constitute a part of the dark body. The 100 A filaments are conspicuous in developing smooth muscle cells and occasionally form networks or clusters; they appear to decrease in relative number as maturation proceeds, but considerable numbers are still present in adult tissue.

Download Full-text

A method for noninvasive longitudinal measurements of [Ca2+] in arterioles of hypertensive optical biosensor mice

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00182.2014 ◽

2014 ◽

Vol 307 (2) ◽

pp. H173-H181 ◽

Cited By ~ 7

Author(s):

Joseph R. H. Mauban ◽

Seth T. Fairfax ◽

Mark A. Rizzo ◽

Jin Zhang ◽

Withrow Gil Wier

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Vascular Function ◽

Resonance Energy ◽

Extended Period ◽

Muscle Cells ◽

Optical Biosensor ◽

Ang Ii ◽

Two Photon

We used two-photon (2-p) Förster resonance energy transfer (FRET) microscopy to provide serial, noninvasive measurements of [Ca2+] in arterioles of living “biosensor” mice. These express a genetically encoded Ca2+ indicator (GECI), either FRET-based exMLCK or intensity-based GCaMP2. The FRET ratios, Rmin and Rmax, required for in vivo Ca2+ calibration of exMLCK were obtained in isolated arteries. For in vivo experiments, mice were anesthetized (1.5% isoflurane), and arterioles within a depilated ear were visualized through the intact skin (i.e., noninvasively), by 2-p excitation of exMLCK (at 820 nm) or GCaMP2 (at 920 nm). Spontaneous or agonist-evoked [Ca2+] transients in arteriolar smooth muscle cells were imaged (at 2 Hz) with both exMLCK and GCaMP2. To examine changes in arteriolar [Ca2+] that might accompany hypertension, five exMLCK mice were implanted with telemetric blood pressure transducers and osmotic minipumps containing ANG II (350 ng·kg−1·min−1) and fed a high (6%)-salt diet for 9 days. [Ca2+] was measured every other day in five smooth muscle cells of two to three arterioles in each animal. Prior to ANG II/salt, [Ca2+] was 246 ± 42 nM. [Ca2+] increased transiently to 599 nM on day 2 after beginning ANG II/salt, then remained elevated at 331 ± 42 nM for 4 more days, before returning to 265 ± 47 nM 6 days after removal of ANG II/salt. In summary, two-photon excitation of exMLCK and GCaMP2 provides a method for noninvasive, longitudinal quantification of [Ca2+] dynamics and vascular structure in individual arterioles of a particular animal over an extended period of time, a capability that should enhance future studies of hypertension and vascular function.

Download Full-text

Evidence for a membrane site of action for 14,15-EET on expression of aromatase in vascular smooth muscle

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00321.2002 ◽

2002 ◽

Vol 283 (5) ◽

pp. H1936-H1942 ◽

Cited By ~ 35

Author(s):

Gary D. Snyder ◽

U. Murali Krishna ◽

J. R. Falck ◽

Arthur A. Spector

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Plasma Membrane ◽

Vascular Smooth Muscle ◽

Smooth Muscle Cells ◽

Muscle Cells ◽

Average Diameter ◽

Hydration Product ◽

Aromatase Activity ◽

Dibutyryl Camp

Epoxyeicosatrienoic acids (EETs) are synthesized in the endothelial cells of vascular tissues. They are released from the endothelial cells and produce relaxation of the smooth muscle cells by hyperpolarization. The present findings demonstrate that EETs also regulate aromatase activity in vascular smooth muscle cells. Exposure of cultured rat aortic smooth muscle cells to either 1 μM 14,15-EET or 1 μM 11,12-EET inhibits dibutyryl cAMP-induced aromatase activity by 80–100%. 11,12-Dihydroxyeicosatrienoic acid, the hydration product of 11,12-EET, has no effect on dibutyryl cAMP-induced vascular smooth muscle aromatase activity. In contrast to 14,15-EET, the N-methylsulfanilamide derivative of 14,15-EET (14,15-EET-SA) was neither metabolized nor incorporated into cell lipids, but it retained the ability to inhibit cAMP-induced aromatase activity. Furthermore, the 14,15-EET-SA inhibition of cAMP-induced aromatase activity persisted when the sulfanilamide derivative of 14,15-EET was covalently tethered to silica beads (average diameter, 0.5 μm), which restricted 14,15-EET-SA from entering the cell. These data are consistent with the presence of a receptor for EETs in the plasma membrane and support the hypothesis that the inhibition of aromatase by EETs is initiated by the interaction of EET with the putative plasma membrane receptor.

Download Full-text

Immunogold localization of HMB-45 antigen in pulmonary lymphangiomyomatosis

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100141366 ◽

1995 ◽

Vol 53 ◽

pp. 998-999

Author(s):

J.M. Minda ◽

E. Dessy ◽

G. G. Pietra

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Room Temperature ◽

Osmium Tetroxide ◽

Muscle Cells ◽

Ultrastructural Localization ◽

Reproductive Age ◽

Thin Sections ◽

Smooth Muscle Proliferation ◽

Hmb 45

Pulmonary lymphangiomyomatosis (PLAM) is a rare disease occurring exclusively in women of reproductive age. It involves the lungs, lymph nodes and lymphatic ducts. In the lungs, it is characterized by the proliferation of smooth muscle cells around lymphatics in the bronchovascular bundles, lobular septa and pleura The nature of smooth muscle proliferation in PLAM is still unclear. Recently, reactivity of the smooth muscle cells for HMB-45, a melanoma-related antigen has been reported by immunohistochemistry. The purpose of this study was the ultrastructural localization of HMB-45 immunoreactivity in these cells using gold-labeled antibodies.Lung tissue from three cases of PLAM, referred to our Institution for lung transplantation, was embedded in either Poly/Bed 812 post-fixed in 1% osmium tetroxide, or in LR White, without osmication. For the immunogold technique, thin sections were placed on Nickel grids and incubated with affinity purified, monoclonal anti-melanoma antibody HMB-45 (1:1) (Enzo Diag. Co) overnight at 4°C. After extensive washing with PBS, grids were treated with Goat-anti-mouse-IgG-Gold (5nm) (1:10) (Amersham Life Sci) for 1 hour, at room temperature.

Download Full-text