Characterization and Safety Profile of a New Combined Advanced Therapeutic Medical Product Platelet Lysate-Based Fibrin Hydrogel for Mesenchymal Stromal Cell Local Delivery in Regenerative Medicine.

Adipose-derived mesenchymal stromal cells (ASC) transplant to recover the optimal tissue structure/function relationship is a promising strategy to regenerate tissue lesions. Because filling local tissue defects by injection alone is often challenging, designing adequate cell carriers with suitable characteristics is critical for in situ ASC delivery. The aim of this study was to optimize the generation phase of a platelet-lysate-based fibrin hydrogel (PLFH) as a proper carrier for in situ ASC implantation and (1) to investigate in vitro PLFH biomechanical properties, cell viability, proliferation and migration sustainability, and (2) to comprehensively assess the local in vivo PLFH/ASC safety profile (local tolerance, ASC fate, biodistribution and toxicity). We first defined the experimental conditions to enhance physicochemical properties and microscopic features of PLFH as an adequate ASC vehicle. When ASC were mixed with PLFH, in vitro assays exhibited hydrogel supporting cell migration, viability and proliferation. In vivo local subcutaneous and subgingival PLFH/ASC administration in nude mice allowed us to generate biosafety data, including biodegradability, tolerance, ASC fate and engraftment, and the absence of biodistribution and toxicity to non-target tissues. Our data strongly suggest that this novel combined ATMP for in situ administration is safe with an efficient local ASC engraftment, supporting the further development for human clinical cell therapy.

Cervical tracheal rupture with persistence of a pseudotrachea in a dog.

A dog was examined because of acute onset of respiratory distress following a cervical dog bite. Physical examination revealed a deep ventral cervical bite wound associated with localized mild subcutaneous emphysema. Thoracic radiographs showed moderate pneumomediastinum. Medical management consisting of oxygen therapy, antibiotics, and anti-inflammatories was initiated. After 2 days, respiratory distress suddenly worsened. Tracheoscopy showed a discontinuity between the tracheal rings of the cervical trachea; however, the inner tracheal wall appeared intact. Computed tomography scan revealed a ~3-cm complete rupture of all layers of the trachea. Surgical resection and anastomosis of the trachea were performed successfully. Follow-up 15 days after surgery showed complete resolution of respiratory signs, as well as subcutaneous emphysema. A mild ventral angulation of the trachea at the surgical site was noticed on thoracic radiographs. This is the first case report of a pseudotrachea in a dog. Persistence of a pseudotrachea may initially result in only minor clinical signs responsive to medical therapy despite tracheal rupture. In the presence of a pseudotrachea, tracheal rupture may be difficult to identify with tracheoscopy alone. Therefore, CT scan should be proposed in every patient with suspected tracheal trauma. Key clinical message: This case report highlights the importance of including a tracheal rupture in the differential diagnosis of cervical subcutaneous emphysema, even if the amount is small and not associated with significant respiratory signs. The presence of a pseudotrachea may result in less severe clinical signs than expected based on the actual degree of tracheal injury; however, the clinical status may rapidly deteriorate and become life-threatening. This case report also underlines the importance of a CT scan as a complement to tracheoscopy, which may not be sufficient to identify a tracheal rupture in the presence of a pseudotrachea.

Rupture trachéale cervicale avec persistance d'une pseudotrachée chez un chien. Un chien a été présenté pour une dyspnée aiguë modérée consécutive à des morsures cervicales par un autre chien. L'examen clinique révéla une plaie cervicale ventrale profonde associée à un emphysème sous-cutané localisé léger. Les radiographies thoraciques ont montré un pneumomédiastin modéré. Un traitement médical consistant en une oxygénothérapie, des antibiotiques et des anti-inflammatoires a été initié. Après deux jours, la dyspnée s'aggrava brutalement. Une trachéoscopie révéla une discontinuité entre les anneaux trachéaux malgré la persistance d'une paroi trachéale interne intègre. L'examen par tomodensitométrie montra une rupture trachéale cervicale complète dans toute son épaisseur, sur 3 cm de long. Une chirurgie de résection-anastomose de la trachée a été réalisée avec succès.Il s'agit de la première description de pseudotrachée chez un chien. La persistance d'une pseudotrachée peut initialement ne provoquer que des signes cliniques mineurs, notamment un emphysème sous-cutané léger et une dyspnée répondant au traitement médical, malgré une lésion trachéale en réalité importante. Par conséquent, un examen par tomodensitométrie de la trachée doit être envisagé chez tous les patients pour lesquels un traumatisme trachéal est suspecté.Message clinique clé :Ce cas souligne l'importance d'inclure une rupture trachéale dans le diagnostic différentiel de l'emphysème souscutané cervical, et cela même s'il n'est présent qu'en petite quantité et associé à faibles signes cliniques respiratoires. La persistance d'une pseudotrachée peut entraîner des signes cliniques moins importants qu'une rupture trachéale complète, cependant l'état respiratoire de l'animal peut rapidement s'aggraver et devenir une urgence vitale.Ce cas souligne de plus l'importance de l'examen par tomodensitométrie en complément de la trachéoscopie, qui peut parfois s'avérer insuffisante pour le diagnostic des ruptures trachéales, en particulier en présence d'une pseudotrachée.(Traduit par les auteurs).

Tissue Regeneration: The Dark Side of Opioids.

Opioids are regarded as among the most effective analgesic drugs and their use for the management of pain is considered standard of care. Despite their systematic administration in the peri-operative period, their impact on tissue repair has been studied mainly in the context of scar healing and is only beginning to be documented in the context of true tissue regeneration. Indeed, in mammals, growing evidence shows that opioids direct tissue repair towards scar healing, with a loss of tissue function, instead of the regenerative process that allows for recovery of both the morphology and function of tissue. Here, we review recent studies that highlight how opioids may prevent a regenerative process by silencing nociceptive nerve activity and a powerful anti-inflammatory effect. These data open up new perspectives for inducing tissue regeneration and argue for opioid-restricted strategies for managing pain associated with tissue injury.

Influence of extracorporeal membrane oxygenation on the pharmacokinetics of ceftolozane/tazobactam: an ex vivo and in vivo study.

BACKGROUND: Extracorporeal membrane oxygenation (ECMO) is increasingly used in intensive care units and can modify drug pharmacokinetics and lead to under-exposure associated with treatment failure. Ceftolozane/tazobactam is an antibiotic combination used for complicated infections in critically ill patients. Launched in 2015, sparse data are available on the influence of ECMO on the pharmacokinetics of ceftolozane/tazobactam. The aim of the present study was to determine the influence of ECMO on the pharmacokinetics of ceftolozane-tazobactam. METHODS: An ex vivo model (closed-loop ECMO circuits primed with human whole blood) was used to study adsorption during 8-h inter-dose intervals over a 24-h period (for all three ceftolozane/tazobactam injections) with eight samples per inter-dose interval. Two different dosages of ceftolozane/tazobactam injection were studied and a control (whole blood spiked with ceftolozane/tazobactam in a glass tube) was performed. An in vivo porcine model was developed with a 1-h infusion of ceftolozane-tazobactam and concentration monitoring for 11 h. Pigs undergoing ECMO were compared with a control group. Pharmacokinetic analysis of in vivo data (non-compartmental analysis and non-linear mixed effects modelling) was performed to determine the influence of ECMO. RESULTS: With the ex vivo model, variations in concentration ranged from - 5.73 to 1.26% and from - 12.95 to - 2.89% respectively for ceftolozane (concentrations ranging from 20 to 180 mg/l) and tazobactam (concentrations ranging from 10 to 75 mg/l) after 8 h. In vivo pharmacokinetic exploration showed that ECMO induces a significant decrease of 37% for tazobactam clearance without significant modification in the pharmacokinetics of ceftolozane, probably due to a small cohort size. CONCLUSIONS: Considering that the influence of ECMO on the pharmacokinetics of ceftolozane/tazobactam is not clinically significant, normal ceftolozane and tazobactam dosing in critically ill patients should be effective for patients undergoing ECMO.

Effect of sedation with butorphanol on variables pertaining to the ophthalmic examination in dogs.

OBJECTIVE: To evaluate whether sedation with intramuscular butorphanol can interfere with different variables of the ocular examination in dogs. ANIMALS: Twenty-two beagles without ophthalmic abnormalities. PROCEDURES: Each dog was examined 20 min prior to and again just before administration of butorphanol to establish baseline data. The globe and nictitating membrane position was evaluated, and the following were recorded: menace response, dazzle reflex, corneal blink reflex, phenol red thread tear test (PRT), Schirmer tear test-1 (STT-1), pupil size (PS) measurement, and rebound tonometry. Then, butorphanol was injected intramuscularly at a dose of 0.2 mg/kg and these procedures were repeated 10, 20, 30, and 45 min postadministration. A sedation score graded 0 to 3 was also established at these time points. Statistical analyses were performed on quantitative data using ANOVA. RESULTS: The sedative effect was not associated with any changes in globe and nictitating membrane position; did not affect the results of the menace response, dazzle reflex, and corneal blink reflex; and had no significant effect on PRT values. However, butorphanol administration was associated with a statistically significant decrease in STT-1 and PS values (P < 0.005), and a statistically significant increase in IOP (P < 0.05). All these variations remained in the range of normal values. CONCLUSIONS AND CLINICAL RELEVANCE: Butorphanol administered intramuscularly at 0.2 mg/kg provided a degree of sedation allowing eye examination, but was found to interfere with STT-1, PS, and IOP values among the diagnostic tests studied. However, these values remained within normal limits.

Anatomical and ultrasonographic study of the femoral nerve within the iliopsoas muscle in beagle dogs and cats.

OBJECTIVES: An ultrasound (US)-guided ventral suprainguinal approach to block the femoral nerve (FN) within the iliopsoas muscle (IPM) has recently been described in dogs. The goal of the present study was to provide the operator with additional information to locate the FN within the IPM in dogs and cats using US. STUDY DESIGN: The study was carried out in three phases: a dissection of the FN (phase 1); an in vivo US-assisted nerve study (phase 2), and an anatomical cross-sectional study (phase 3). ANIMALS: Nine healthy adult beagle dogs and nine healthy adult cats. METHODS: Dissections were performed to investigate the anatomical characteristics of the FN and its related structures in one dog and one cat. Ultrasound scans of the left and right FN were performed in eight dogs and eight cats. The FN diameter and the distance between the FN and the external iliac artery (EIA) in US images and in anatomical cryosections were measured. RESULTS: The median FN diameter did not differ significantly between cats and dogs (1.1 mm versus 1.0 mm) or between the two techniques (US versus anatomical cross-sectional study) (1.1 mm versus 1.1 mm in dogs; 1.0 mm versus 1.1 mm in cats). The US and anatomical measurements of the median distances between the FN and EIA differed significantly between dogs and cats (8.2 mm versus 5.8 mm by US; 5.7 mm versus 4.8 mm in the anatomical study). CONCLUSIONS AND CLINICAL RELEVANCE: The distance between the EIA and FN is reproducible in beagle dogs and cats and can be used in locating the FN within the IPM.

Postoperative opioid-free analgesia in dogs undergoing tibial plateau leveling osteotomy: a feasibility study.

Objectives: This study was designed to prospectively evaluate the feasibility of an opioid-free anesthesia protocol and describe the quality of recovery and management of postoperative analgesia in dogs after a tibial plateau leveling osteotomy (TPLO). Methods: In total, 20 dogs presented for TPLO were included. After premedication with intravenous (IV) medetomidine (0.005-0.007 mg/kg) and midazolam (0.2 mg/kg), the dogs were anesthetized using ketamine (2 mg/kg) and propofol and maintained with isoflurane and ketamine CRI (0.6 mg/kg/h). Sciatic and femoral nerve blocks were performed with bupivacaine 0.5% (0.087 +/- 0.01 and 0.09 +/- 0.02 mL/kg, respectively). Meloxicam (0.2 mg/kg IV) was administered intraoperatively, after osteotomy. Fentanyl (0.002 mg/kg IV) was administered intraoperatively, as rescue analgesia in the case of sustained increase in cardiorespiratory variables. Two pain scores (French 4A-VET and Glasgow short form) were performed at conscious sternal recumbency and 2, 4, 6, 8, 12, and 20 h after extubation and compared to baseline using a Friedman test followed by a Nemenyi post-hoc test. The time taken for the first food intake and urination was reported. Results: Intraoperative opioid-free anesthesia was feasible in 11 dogs, whereas 9 dogs received fentanyl once during arthrotomy. No opioid postoperative rescue analgesia was required. Food intake occurred within 6 h, and all dogs were discharged after 24 h without any complication. Conclusion: Total opioid-free postoperative analgesia was achieved in all dogs, with adequate recoveries. Although opioid-free anesthesia was feasible in 55% of the population, a single dose of fentanyl was necessary in 45% of the dogs during arthrotomy.

Morphological comparative study of canine and feline diaphragms: Effect of bodyweight, body condition score, gender, age and breed.

The canine diaphragm is a muscular and tendinous dome-like plate and is largely involved in digestive and respiratory functions. Very few studies compared morphology of the diaphragm between dogs and cats and no studies have investigated the effects of individual factors on this morphology. So the aim of this study was to (1) study the effects of individual factors on the morphology of the diaphragm and (2) to compare its morphology between cats and dogs. Surface measurements of 86 diaphragms were performed. The tendinous centre (TC), the lumbar part of the peripheral muscular (LP) and the sternal and costal parts of the peripheral muscular (SCPM) were measured. Measurement ratios (surface of anatomical part of the diaphragm/total surface of the diaphragm) were used for statistical analysis (TC%S, LP%S, SCPM%S). The SCPM%S are significantly lower, and the LP%S are significantly higher when bodyweight increases in dogs and cats. The TC%S are significantly lower when the body condition score of dogs increases. The SCPM%S are significantly higher, and therefore, the TC%S and LP%S lower in brachycephalic animals as opposed to mesocephalic animals. The TC%S are significantly higher in dogs than in cats and the SCPM%S are significantly higher in cats than in dogs. This study proposed a cartography of the diaphragm morphology in cats and dogs taking into account individual animal factors. Significant differences in the diaphragm morphology between cats and dogs and between mesocephalic and brachycephalic animals were found. Further studies are necessary to confirm these results and to investigate the consequences of these variations.

Effectiveness and Safety of a Prolonged Hemodynamic Support by the IVAC2L System in Healthy and Cardiogenic Shock Pigs.

BACKGROUND: Mechanical circulatory supports are used in case of cardiogenic shock (CS) refractory to conventional therapy. Several devices can be employed, but are limited by their availability, benefit risk-ratio, and/or cost. AIMS: To investigate the feasibility, safety, and effectiveness of a long-term support by a new available device (IVAC2L) in pigs. METHODS: Experiments were carried out in male pigs, divided into healthy (n = 6) or ischemic CS (n = 4) groups for a median support time of 34 and 12 h, respectively. IVAC2L was implanted under fluoroscopic and TTE guidance under general anesthesia. CS was induced by surgical ligation of the left anterior descending artery. An ipsilateral lower limb reperfusion was created with the Solopath® system. Reperfusion was started after 1 h of support in healthy pigs and upon IVAC2L insertion in CS pigs. Hemodynamic and biological parameters were monitored before and during the whole period of support in each group. RESULTS: Occurrence of an ipsilateral lower limb ischemia was systematic in healthy and CS pigs in a few minutes after IVAC2L implantation, and could be reversed by the arterial reperfusion, as demonstrated by distal transcutaneous pressure in oxygen (TcPO2) and lactate normalization. IVAC2L support decreased pulmonary capillary wedge pressure (PCWP) (15.3 ± 0.3 vs. 7.5 ± 0.9 mmHg, p < 0.001), increased systolic blood pressure (SBP) (70 ± 4.5 vs. 101.3 ± 3.1 mmHg, p < 0.01), and cardiac output (CO) (4.0 ± 0.3 vs. 5.2 ± 0.6 l/min, p < 0.05) in CS pigs; at CS onset and after 12 h of support, without effects on heart rate or pulmonary artery pressure (PAP). Non-sustained ventricular arrhythmias were frequent at implantation (50%). A non-significant hemolysis was observed under support in CS pigs. Bleedings were frequent at the insertion and/or operating sites (30%). CONCLUSION: Long-term support by IVAC2L is feasible and associated with a significant hemodynamic improvement in a porcine model. These preclinical data open the door for a study of IVAC2L in human ischemic CS, keeping in mind the need for systematic reperfusion of the lower limb and the associated risk of bleeding.

Appetite-stimulating effect of gabapentin vs mirtazapine in healthy cats post-ovariectomy.

OBJECTIVES: The aim of the study was to evaluate the appetite-stimulating effect of gabapentin by comparing it with mirtazapine in healthy cats in the first 8 h after ovariectomy surgery. METHODS: This double-masked, placebo-controlled, prospective clinical trial included 60 healthy cats presented to the hospital for ovariectomy: 20 received gabapentin, 21 received mirtazapine and 19 received a placebo immediately before and 6 h after surgery. Food was offered at 2, 4, 6 and 8 h post-ovariectomy. After each meal, food intake was measured. Data were analysed using repeated-measure ANOVA and a linear mixed-model analysis. Post-hoc Tukey's honest significant difference test was performed for multiple comparisons. RESULTS: Food intake increased in both treatment groups vs placebo. No statistically significant difference was found between cats treated with gabapentin or mirtazapine. CONCLUSIONS AND RELEVANCE: Cats receiving gabapentin ate more than cats in the placebo group. Thirty percent of cats in the gabapentin group covered their resting energy requirements, while none of the cats in the placebo group did. Gabapentin and mirtazapine produced similar effects on food intake.

In vivo pharmacodynamic interactions between two drugs used in orthostatic hypotension--midodrine and dihydroergotamine.

A combination of midodrine and dihydroergotamine (DHE) is frequently used clinically in patients suffering from severe orthostatic hypotension (OH). Whereas midodrine acts as a selective, peripheral alpha1-receptor agonist, DHE displays complex pharmacology and can behave as an alpha-adrenergic receptor agonist or antagonist. Surprisingly, the consequences of such a combination on blood pressure have never been investigated. The present study was performed in order to evaluate the pressor effects induced by the administration of both midodrine and DHE in old conscious dogs (n = 6) in experimental condition reproducing autonomic failure-related baroreflex dysfunction (atropine 0.1 mg/kg). For this purpose, we first studied the relative potency and intrinsic activity of each agonist and noradrenaline (NA) for the alpha1-adrenergic receptor. The orders of potency obtained in our study were 0.35, 11 and 400 microg/kg for NA, DHE and midodrine, and intrinsic activity: NA > midodrine > DHE. These results strongly suggest that DHE really acts in vivo as an alpha1-adrenoceptor partial agonist. Afterwards, the pressor effects of coadministration of midodrine (0.4 mg/kg) and DHE (15 microg/kg) were investigated: in one setting, midodrine was first administered, followed by DHE; in another, DHE was first administered, followed by midodrine. Our results show that in conscious dogs, the combination of midodrine and DHE leads to near-complete abolition of the pressor effect induced by the first administered drug. This in vivo proof of such antagonistic effects on blood pressure could explain clinical observations of worsening of OH in humans administered midodrine plus DHE. Although in vivo results obtained in conscious healthy dogs need to be experimentally and clinically confirmed in humans suffering from OH, these results strongly suggest that a midodrine-DHE combined treatment should be avoided in clinical practice.

Heated intravenous fluids alone fail to prevent hypothermia in cats under general anaesthesia.

Objectives The objective was to evaluate the clinical efficiacy of a constant rate infusion of heated fluid as the sole means of preventing intraoperative hypothermia in cats. Methods This randomised, prospective, clinical study was conducted at a university teaching veterinary hospital. Female cats (American Society of Anesthesiologists [ASA] grade I) undergoing elective surgery by laparotomy under general anaesthesia (acepromazine 0.05 mg/kg SC; morphine 0.2 mg/kg IV; propofol IV titrated, isoflurane 2% in 100% oxygen) were randomised in two groups. Both groups were infused with fluid (NaCl 0.9%, 5 ml/kg/h) either at room temperature (control group) or prewarmed at 43°C (warmed group) using an Astoflo Plus eco (Stihler Electronic) fluid heating device. No other heating device was used. Temperature, heart rate, respiratory rate and SpO2 were evaluated after induction (T0) and every 15 mins for 1 h (T15, T30, T45, T60). Mean arterial blood pressure was recorded every 30 mins (T0, T30 and T60). Results Thirty-four female cats (ASA grade I) were enrolled in the study. There was no difference in age, weight, propofol dose or room temperature (22.4 ± 1.1°C vs 22.0 ± 1.5°C; P = 0.363) between control and warmed groups, respectively. In both groups, oesophageal temperature significantly decreased during anaesthesia ( P <0.0001). The temperature decrease after 1 h was -3.6 ± 0.7°C in the warmed group and was not significantly different from the control group (-3.4 ± 0.7°C; P = 0.307). The slopes of the temperature decrease did not significantly differ between the two groups (-0.058 ± 0.013°C/min vs -0.060 ± 0.010°C/min for the control and warmed groups, respectively; P = 0.624). Conclusions and relevance This study provides clinical evidence that a constant rate infusion of heated fluid alone fails to prevent intraoperative hypothermia in cats. The low infusion rate (5 ml/kg/h) could partly explain the ineffectiveness of this active warming device in minimising or delaying the onset of intraoperative hypothermia.

Intravenous lipid emulsion therapy in 20 cats accidentally overdosed with ivermectin.

OBJECTIVE: To describe the outcome of 20 cats treated with intravenous lipid emulsion (IVLE) after an accidental parenteral ivermectin overdose. CASE SERIES SUMMARY: Twenty adult cats presented after receiving a 4 mg/kg accidental subcutaneous overdose of ivermectin. After admission, two IVLE treatments were initiated in asymptomatic cats: a single bolus (1.5 mL/kg; n = 16) versus a bolus followed by a 30-minute constant rate infusion (0.25 mL/kg/min; n = 4). Six out of the 16 cats that received only the single bolus developed clinical signs of ivermectin intoxication. Based on the severity of the clinical signs and their duration (approximately 48 hours), these 6 cats were retrospectively considered either moderately (n = 3) or severely (n = 3) intoxicated by ivermectin. Cats with a low body condition score (BCS) had more severe signs of ivermectin toxicity. Additional IVLE was administered until clinical resolution was complete. Median (min to max) cumulative dose of IVLE per cat was 4.5 (3.0-4.5) mL/kg for 36 (12-36) hours and 19.5 (7.5-37.5) mL/kg for 96 (72-168) hours for moderately and severely intoxicated cats, respectively. NEW OR UNIQUE INFORMATION PROVIDED: Our series describes the treatment of accidental ivermectin parenteral overdose in 20 cats with early initiation of IVLE therapy. An early bolus followed by a 30-minute constant rate infusion of IVLE appeared to mitigate the signs of ivermectin toxicosis in cats compared to a single treatment bolus. Our observations also suggest that cats with a low BCS given only a bolus of IVLE treatment were more likely to develop signs of ivermectin intoxication and require a greater amount of IVLE for the resolution of clinical signs. Based on our observations, BCS appears to influence the severity of ivermectin toxicity with a low BCS being associated with more severe signs of ivermectin toxicity.