Body weight, respiration and physiological lung parameters of IUGR rats.

<p>A: Body weight (g) and relative lung weight (lung weight/body weight) of rats after IUGR (white bars) and control rats (black bars) at days P1 and P70; n = 8–15 for each bar. B: Architectural changes in lung structure were evident in hematoxylin and eosin-stained lung sections from IUGR rats and control rats at day P70. Measurement of septal thickness (µm) and mean linear intercept (MLI; µm) in IUGR rats and control rats (CO) at day P70; n = 6–10 for each bar. C: Assessment of respiratory system compliance by whole body plethysmography in IUGR rats (white bars) and in the control group (CO; black bars) at P70. n = 15–17 for each bar. D: Expression pattern of genes encoding surfactant protein A (SP-A), SP-C, and SPD. IUGR rats (white bars) and control group (black bars). n = 6–15 for each bar. The significance for each bar is indicated by p values, IUGR vs. Co, n.s. = not significant; two-tailed Mann-Whitney test.</p

Effect of IUGR on the expression of TGF-β1 in lungs of neonatal and adult rats.

<p>A: Expression of genes encoding TGF-β1 and TGF-β-inducible plasminogen activator inhibitor-1 (PAI-1) during late lung development (day P1). The mRNA expression was assessed by quantitative real-time PCR. The control group was normalized to 1 as indicated by a scattered line; n = 15 for each group. The significance for each bar is indicated by p values, IUGR vs. Co; two-tailed Mann-Whitney test. B: representative immunoblots illustrating expression of TGF-β1 in lungs extracted at day P1 and P70 from rats with and without IUGR. β-actin served as loading control. Immunoblot data were quantified for both day P1 and P70; n = 6 for each bar. The significance for each bar is indicated by p values, IUGR vs. Co; two-tailed Mann-Whitney test.</p

Expression of the TGF-β signaling machinery in lungs of neonatal and adult rats.

<p>Changes in the expression of genes encoding components of the TGF-β signaling machinery as assessed by real-time RT-PCR at days P1 and P70; n = 8–15 for each bar. The significance for each bar is indicated by p values, IUGR vs. CO; two-tailed Mann-Whitney test. A: Expression of genes encoding the TGF-β receptors <i>tgfbr1</i>, <i>tgfbr2</i> and <i>tgfbr3</i> at P1 (white bar) and P70 (striped bar). B: Expression of genes encoding the regulatory <i>smad2</i>, <i>smad3</i> and <i>smad4</i> at days P1 (white bar) and P70 (striped bar). C: Expression of genes encoding the inhibitory <i>smad7</i>, <i>smurf2</i> and smad anchor for receptor activation (<i>sara</i>) at days P1 (white bar) and P70 (striped bar).</p

Primer pairs and TaqMan probes used.

<p>*TIMP-1, TIMP-2 and MMP-2 were detected by TaqMan realtime-PCR analysis.</p

Effect of IUGR on apoptosis in lungs of rats at days P1 and P70.

<p>Apoptosis is assessed by cleaved caspase-3 and cleaved fragment of Poly (ADP-ribose) polymerase (PARP). A: Representative immunoblots illustrating the expression of cleaved and total caspase-3, fragments of PARP and total PARP in lung homogenates of rats with IUGR and without IUGR (Co) at <i>day</i> P1 (A) and P70 (B). The β-actin served as loading control; n = 4–6 for each bar.</p

Effect of IUGR on the expression of extracellular matrix (ECM) proteins and modulators of the ECM in neonatal rat lungs.

<p>Expression of TGF-β-regulated genes encoding elastin (Eln), tenascin N (Ten), collagens (Coll) I, Coll III, and Fibrillin (A), and genes of ECM modulators including matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinases (TIMP)-1, and TIMP-2 (B) in lungs extracted at day P1 from neonatal rats after IUGR and control rats. The mRNA expression, illustrated as relative fold induction, was assessed by real-time PCR. The control group was normalized to 1 as indicated by a scattered line; n = 15 for each group. The significance for each bar is indicated by p values, IUGR vs. Co; two-tailed Mann-Whitney test.</p

Micro-CT vs. Whole Body Multirow Detector CT for Analysing Bone Regeneration in an Animal Model

<div><p>Objectives</p><p>Compared with multirow detector CT (MDCT), specimen (ex vivo) micro-CT (μCT) has a significantly higher (~ 30 x) spatial resolution and is considered the gold standard for assessing bone above the cellular level. However, it is expensive and time-consuming, and when applied in vivo, the radiation dose accumulates considerably. The aim of this study was to examine whether the lower resolution of the widely used MDCT is sufficient to qualitatively and quantitatively evaluate bone regeneration in rats.</p><p>Methods</p><p>Forty critical-size defects (5mm) were placed in the mandibular angle of rats and covered with coated bioactive titanium implants to promote bone healing. Five time points were selected (7, 14, 28, 56 and 112 days). μCT and MDCT were used to evaluate the defect region to determine the bone volume (BV), tissue mineral density (TMD) and bone mineral content (BMC).</p><p>Results</p><p>MDCT constantly achieved higher BV values than μCT (10.73±7.84 mm³ vs. 6.62±4.96 mm³, p<0.0001) and consistently lower TMD values (547.68±163.83 mm³ vs. 876.18±121.21 mm³, p<0.0001). No relevant difference was obtained for BMC (6.48±5.71 mm³ vs. 6.15±5.21 mm³, p = 0.40). BV and BMC showed very strong correlations between both methods, whereas TMD was only moderately correlated (r = 0.87, r = 0.90, r = 0.68, p < 0.0001).</p><p>Conclusions</p><p>Due to partial volume effects, MDCT overestimated BV and underestimated TMD but accurately determined BMC, even in small volumes, compared with μCT. Therefore, if bone quantity is a sufficient end point, a considerable number of animals and costs can be saved, and compared with in vivo μCT, the required dose of radiation can be reduced.</p></div

IUGR alters expression and phosphorylation of Smad proteins in rats.

<p>A: Representative immunoblots illustrating the expression of TGF-β-specific Smad2, Smad3, the co-Smad, Smad4, and the inhibitory Smad, Smad7, in lungs extracted at days P1 and P70 from rats with and without IUGR. β-actin served as loading control. Immunoblot data were quantified for Smad4 and Smad7 for both days P1 and P70 (Co as black bar, and IUGR as white bar); n = 4–6 for each bar. The significance for each bar is indicated by p values, IUGR vs. CO; two-tailed Mann-Whitney test. B: The expression of active TGF-β signaling components in lung homogenates of rats with and without IUGR was analyzed by immunoblotting of phosphorylated (p) and total Smad2 and Smad3. β-actin served as loading control. Immunoblot data were quantified for pSmad2 and pSmad3 for both days P1 and P70 (Co as black bar, and IUGR as white bar); n = 4–6 for each bar. The significance for each bar is indicated by p values, IUGR vs. CO; two-tailed Mann-Whitney test. C: Immunhistochemical localization and expression pattern of pSmad2 and pSmad3 in lungs of rats with IUGR (right column) and without IUGR (left column). A–F: representative fields illustrating the expression and localization of pSmad2 in bronchi (A–D) and in the alveoli (E–F) of lungs extracted on day P70. G–L: representative fields illustrating the expression and localization of pSmad3 in bronchi (G–J) and in the alveoli (K–L) of lungs extracted on day P70. M–N: negative control.</p

The role of transforming growth factor (TGF)-β signaling in lung disease subsequent to intrauterine growth restriction (IUGR).

<p>A proposed model depicting the effects of decreased TGF-β signaling during the development of IUGR-associated lung disease is shown. ECM - extracellular matrix.</p

Yellow rectangle representing the bone to be measured (left: μct, right: MDCT).

<p>Voxels that are filled to more than 50% are counted as bone (blue or red), while those that are filled to less than 50% are not counted (white). Because of the lower resolution of the larger voxels of the MDCT, it generates a higher BV than that of the real bone. In contrast, the μCt, with its high resolution and very small voxel size, generates a more precise volume.</p