Elisabeth Laemmel

Impact of serotonin on tumour growth

Several opposite effects of serotonin (5HT) on tumour growth have been reported. On one hand, 5HT is known as a growth factor for several types of nontumoural cells, and it has been proposed to take part in the autocrine loops of growth factors contributing to cell proliferation in aggressive tumours such as small cell lung carcinoma. Depending on the tumour type either 5HT2 or 5HT2 receptor antagonist have been found to inhibit the 5HT-induced increase in tumour growth. In contrast, several authors have also reported that 5HT and 5HT2 agonist can inhibit tumour growth. Most often this effect has been considered to be related with the specific vasoconstrictive effect of 5HT or 5HT2 agonists on the vessels irrigating the tumour, which has been evidenced by intravital microscopy. Intravital microscopy studies have also shown that vessels perfusing the tumour exhibit a specific vasconsmctive response to 5HT1 agonists. In addition, 5HT has been shown to be involved in the effects of several anticancer treatments associated with the reduction of tumour flow. Finally, the specific vasoconstrictive effect of 5HT or 5HT receptor subtype agonists might also be useful in inducing hypoxia in tumours, which could be exploited in a strategy using hypoxia-selective cytotoxins or hypoxia-selective gene therapy.

Characteristic features of in vivo skin microvascular reactivity in CADASIL

CADASIL (Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy) is caused by mutations in the Notch3 receptor expressed at the surface of vascular smooth muscle cells. The functional consequences of the disease at the peripheral microcirculation level are incompletely elucidated. In this study, we aimed to assess, in vivo, the endothelium-dependent and independent vasodilation of the skin microvasculature in CADASIL patients. Twenty-three affected subjects were compared with 23 gender and age-matched controls. The brachial artery endothelium-dependent and endothelium-independent vasodilation were assessed after forearm cuff occlusion and nitroglycerin administration. Skin vasoreactivity to transcutaneous administration of acetylcholine and sodium nitroprussiate, and after postocclusive hyperemia were measured by Laser Doppler flowmetry. The maximum changes in the diameter of the brachial artery after the cuff release or after nitroglycerin administration did not differ between patients and controls. With iontopheresis, only the peak value of the dose response was found decreased in normocholesterolemic patients after nitroprussiate administration. The postocclusive test revealed a large increase of the time to peak value and whole duration of the hyperemic response in CADASIL patients. The results of this study show that the skin vasoreactivity is altered in CADASIL. Particularly, the kinetics of reactive hyperemia after cuff occlusion is dramatically changed with a lengthened and delayed response. This characteristic pattern may be related to the specific ultrastructural modifications related to Notch3 gene mutations involving smooth muscle cells in the microvasculature.

Real-time and Spatial Quantification Using Contrast-enhanced Ultrasonography of Spinal Cord Perfusion During Experimental Spinal Cord Injury

Study Design. Experimental study in male Wistar rats. Objective. To quantify temporal and spatial changes simultaneously in spinal cord blood flow and hemorrhage during the first hour after spinal cord injury (SCI), using contrast-enhanced ultrasonography (CEU). Summary of Background Data. Post-traumatic ischemia and hemorrhage worsen the primary lesions induced by SCI. Previous studies did not simultaneously assess temporal and spatial changes in spinal cord blood flow. Methods. SCI was induced at Th10 in 12 animals, which were compared with 11 sham-operated controls. Spinal cord blood flow was measured in 7 adjacent regions of interest and in the sum of these 7 regions. Blood flow was quantified using CEU with intravenous microbubble injection. Spinal cord hemorrhage was measured on conventional B-mode sonogram slices. Results. CEU allowed us to measure the temporal and spatial changes in spinal cord blood flow in both groups. In the SCI group, spinal cord blood flow was significantly decreased in the global region of interest (P = 0.0016), at the impact site (epicenter), and in the 4 regions surrounding the epicenter, compared with the sham group. The blood flow decrease was maximum at the epicenter. No statistically significant differences between the sham groups were found for the most rostral and caudal regions of interest. Hemorrhage size increased significantly with time (P < 0.0001), from 30.3 mm2 (±2) after 5 minutes to 39.6 mm2 (±2.3) after 60 minutes. Conclusion. CEU seems reliable for quantifying temporal and spatial changes in spinal cord blood flow. After SCI, bleeding occurs in the spinal cord parenchyma and increases significantly throughout the first hour.

Specific response of mouse tumor-feeding arterioles to stimulation by 5-HT1 agonists

Using intravital microscopy, we compared the responses to 5-HT1 receptor stimulation by the host-modified arterioles feeding a Meth-A tumor implanted in the flank of female Balb/c mice with the responses of tumor-independent arterioles (TIA) and those of control arterioles from mice without tumor. Topical administration of 5 x 10(-5) M serotonin in the presence of 10(-4) M ketanserin (5-HT2 receptors inhibitor) induced arteriolar vasodilation in TIA (+13%) and in the control arterioles (+19%), but induced constriction (-14%) in the tumor-feeding arterioles (TFA). Topical administration of the general 5-HT1 agonist 5-carboxamidotryptamine maleate (10(-6) to 10(-4) M) or the 5-HT1A agonist buspirone (2 x 10(-6) to 2 x 10(-4) M) induced vasoconstriction that was dramatically higher in TFA than in TIA or control arterioles (p < 0.0001 in both cases). In addition, topical administration of the 5-HT1B agonist M-trifluoromethylphenylpiperazine (2 x 10(-6) to 2 x 10(-4) M) produced opposite responses, i.e., dose-dependent vasodilation in TIA and control arterioles, and dose-dependent constriction in TFA. Since we observed the same degree of vasodilation in response to 10(-4) M acetylcholine in all three groups of arterioles, the differences between the responses to 5-HT1 receptor stimulation were not due to the absence of endothelial-dependent dilatory mechanisms in the tumor-feeding arterioles. We conclude that 5-HT1 agonists are interesting pharmacologic tools for the modulation of tumoral blood flow, since they more dramatically constrict the microvasculature feeding the tumors than that feeding normal tissue.

Characterization of the specific response to serotonin of mouse tumour-feeding arterioles

PURPOSE To investigate the role of tumour versus non-tumour factors in the specific response to serotonin (5-HT) of tumour-feeding arterioles (TFA). MATERIALS AND METHODS Using mouse models of intra-vital microscopy, the response to topical administration of 5-HT was studied in arterioles feeding tumours: fibrosarcoma (Meth A), murine mammary adenocarcinoma (EMT6) and human colo-rectal carcinoma (HRT18) intra-cutaneously implanted. RESULTS For all types of tumour, 5-HT induced a far more pronounced constriction of TFA than of control arterioles. The presence of a tumour implanted in the connective tissue between the skin and the cremaster muscle also affected the reactivity of muscle arterioles. Conversely, the response to serotonin by neovessels grown after implantation of an exogenous element under the skin did not differ from that of control arterioles. CONCLUSIONS Changes in reactivity to serotonin were not dependent on the type of tumour implanted in the skin and were not present for a non-tumour implant. The presence of the tumour can alter the reactivity of vessels from tissue in contact with the tumour even if these vessels did not feed the tumour. This phenomenon is local and was not found in the vessels at a distance from the tumour.

Role of endothelial factors in the specific response of mouse tumour-feeding arterioles to stimulation of 5-HT1 receptors

PURPOSE To investigate the possible role of endothelial mediators on the increased vasoconstriction to 5-HT1 receptor stimulation by the host-modified arterioles feeding a Meth-A tumour implanted in the flank of female Balb/c mice. MATERIALS AND METHODS Using intravital microscopy, the response to the topical administration of the general 5-HT1 agonist 5-carboxamidotryptamine maleate (5-CT; 10(-6) M to 10(-4) M) by the tumour-feeding arterioles with the responses of tumour-independent arterioles and those of control arterioles from mice without tumour after antagonization or inhibition of the synthesis of endothelial mediators was compared. RESULTS The dramatically higher vasoconstriction to 5-CT observed in tumour-feeding arterioles than in tumour-independent or control arterioles still persisted when either nitric oxide synthase, cyclooxygenase, lipoxygenase, or phospholipase A2 were inhibited or when thromboxane A2 or endothelin were antagonized. CONCLUSIONS It was concluded that the higher reactivity to 5-HT1 stimulation by tumour-feeding arterioles is not due to changes in endothelial mediator release but probably due to changes affecting arteriolar smooth muscle.

De novo generation in an in vivo rat model and biomechanical characterization of autologous transplants for ligament and tendon reconstruction

Background: Surgical reconstruction of ligaments and tendons is frequently required in clinical practice. The commonly used autografts, allografts, or synthetic transplants present limitations in terms of availability, biocompatibility, cost, and mechanical properties that tissue bioengineering aims to overcome. It classically combines an exogenous extracellular matrix with cells, but this approach remains complex and expensive. Using a rat model, we tested a new bioengineering strategy for the in vivo and de novo generation of autologous grafts without the addition of extracellular matrix or cells, and analyzed their biomechanical and structural properties. Methods: A silicone perforated tubular implant (PTI) was designed and implanted in the spine of male Wistar rats to generate neo‐transplants. The tensile load to failure, stiffness, Young modulus, and ultrastructure of the generated tissue were determined at 6 and 12 weeks after surgery. The feasibility of using the transplant that was generated in the spine as an autograft for reconstruction of medial collateral ligaments (MCL) and Achilles tendons was also tested. Findings: Use of the PTI resulted in de novo transplant generation. Their median load to failure and Young modulus increased between 6 and 12 weeks (respectively 12 N vs 34 N and 48 MPa vs 178 MPa). At 12 weeks, the neo‐transplants exhibited collagen bundles (mainly type III) parallel to their longitudinal axis and elongated fibroblasts. Six weeks after their transfer to replace the MCL or the Achilles tendon, the transplants were still present, with their ends healed at their insertion point. Interpretation: This animal study is a first step in the design and validation of a new bioengineering strategy to develop autologous transplants for ligament and tendon reconstructions.

Effects of Adenosine Monophosphate Used in Combination with L-Arginine on Female Rabbit Corpus Cavernosum Tissue

Introduction Sexual dysfunction is significantly more prevalent in women than in men. However, to date, no satisfactory oral treatment is yet available. Aim The aim of this study was to study the effects of adenosine monophosphate (AMP) alone or its combination with L-Arginine on the relaxation of the female rabbit corpus cavernosum. Methods Cylinder strips from the corporal body of the excised clitoris from female New Zealand White rabbits were incubated in Krebs solution. Phenylephrine (PE) precontraction was achieved, then the drugs AMP and L-Arginine were administered either independently or in sequential combinations to the strips under precontracted conditions. Main Outcome Measures Contraction percentages were compared. Results When precontraction was induced by PE 8 μM or 20 μM, AMP was shown to induce relaxation up to 25% in a dose-dependent manner. The relaxation induced by L-Arginine reached 15.6% at 5.10−4 M vs. 16.5% at AMP 5.10−4 M under the same experimental conditions. Nitric oxide (NO) synthase inhibitor N-nitro-L-arginine strongly inhibited the relaxing effect provoked by AMP, suggesting that the action mechanism of this nucleotide is related to the NO pathway. The combination of L-Arginine at 5.10−4 M with AMP at different doses ranging from 5.10−4 M to 10−3 M significantly amplified the relaxing response up to 40.7% and 58%, respectively. Conclusions Our results demonstrate that AMP induces a relaxing effect on the female rabbit corpora. They also show that L-Arginine and AMP can potentiate each other and that a synergistic effect can be obtained by their combined use. Because only slight differences exist between both sexes in response to NO donors and/or nucleotide purines or in their use together, it is very likely that close biochemical mechanisms, although not to the same degree and not quite similar, are involved in the engorgement of the penis and the clitoris of New Zealand White rabbits. Stücker O, Pons C, Neuzillet Y, Laemmel E, and Lebret T. Original research-sexual medicine: Effects of adenosine monophosphate used in combination with L-Arginine on female rabbit corpus cavernosum tissue. Sex Med 2014;2:1–7.