Modeling autosomal recessive cutis laxa type 1C in mice reveals distinct functions for Ltbp-4 isoforms

Recent studies have revealed an important role for LTBP-4 in elastogenesis. Its mutational inactivation in humans causes autosomal recessive cutis laxa type 1C (ARCL1C), which is a severe disorder caused by defects of the elastic fiber network. Although the human gene involved in ARCL1C has been discovered based on similar elastic fiber abnormalities exhibited by mice lacking the short Ltbp-4 isoform (Ltbp4S(-/-)), the murine phenotype does not replicate ARCL1C. We therefore inactivated both Ltbp-4 isoforms in the mouse germline to model ARCL1C. Comparative analysis of Ltbp4S(-/-) and Ltbp4-null (Ltbp4(-/-)) mice identified Ltbp-4L as an important factor for elastogenesis and postnatal survival, and showed that it has distinct tissue expression patterns and specific molecular functions. We identified fibulin-4 as a previously unknown interaction partner of both Ltbp-4 isoforms and demonstrated that at least Ltbp-4L expression is essential for incorporation of fibulin-4 into the extracellular matrix (ECM). Overall, our results contribute to the current understanding of elastogenesis and provide an animal model of ARCL1C.Peer reviewe

Modeling autosomal recessive cutis laxa type 1C in mice reveals distinct functions for Ltbp-4 isoforms

Recent studies have revealed an important role for LTBP-4 in elastogenesis. Its mutational inactivation in humans causes autosomal recessive cutis laxa type 1C (ARCL1C), which is a severe disorder caused by defects of the elastic fiber network. Although the human gene involved in ARCL1C has been discovered based on similar elastic fiber abnormalities exhibited by mice lacking the short Ltbp-4 isoform (Ltbp4S(-/-)), the murine phenotype does not replicate ARCL1C. We therefore inactivated both Ltbp-4 isoforms in the mouse germline to model ARCL1C. Comparative analysis of Ltbp4S(-/-) and Ltbp4-null (Ltbp4(-/-)) mice identified Ltbp-4L as an important factor for elastogenesis and postnatal survival, and showed that it has distinct tissue expression patterns and specific molecular functions. We identified fibulin-4 as a previously unknown interaction partner of both Ltbp-4 isoforms and demonstrated that at least Ltbp-4L expression is essential for incorporation of fibulin-4 into the extracellular matrix (ECM). Overall, our results contribute to the current understanding of elastogenesis and provide an animal model of ARCL1C.Peer reviewe

Zero-Heat-Flux Thermometry for Non-Invasive Measurement of Core Body Temperature in Pigs

Hypothermia is a severe, unpleasant side effect during general anesthesia. Thus, temperature surveillance is a prerequisite in general anesthesia settings during experimental surgeries. The gold standard to measure the core body temperature (T-core) is placement of a Swan-Ganz catheter in the pulmonary artery, which is a highly invasive procedure. Therefore, T-core is commonly examined in the urine bladder and rectum. However, these procedures are known for their inaccuracy and delayed record of temperatures. Zero-heat-flux (ZHF) thermometry is an alternative, non-invasive method quantifying T-core in human patients by applying a thermosensoric patch to the lateral forehead. Since the porcine cranial anatomy is different to the human's, the optimal location of the patch remains unclear to date. The aim was to compare three different patch locations of ZHF thermometry in a porcine hypothermia model. Hypothermia (33.0 degrees C T-core) was conducted in 11 anesthetized female pigs (26-30kg). T-core was measured continuously by an invasive Swan-Ganz catheter in the pulmonary artery (T-pulm). A ZHF thermometry device was mounted on three different defined locations. The smallest average difference between T-pulm and T-ZHF during stable temperatures was 0.21 +/- 0.16 degrees C at location A, where the patch was placed directly behind the eye. Also during rapidly changing temperatures location A showed the smallest bias with 0.48 +/- 0.29 degrees C. Location A provided the most reliable data for T-core. Therefore, the ZHF thermometry patch should be placed directly behind the left temporal corner of the eye to provide a non-invasive method for accurate measurement of T-core in pigs

Influence of different carrier materials on biphasic calcium phosphate induced bone regeneration

Objectives Biphasic calcium phosphate (BCP) is a bioceramic material successfully used in alloplastic bone augmentation. Despite many advantages, a disadvantage of BCP seems to be a difficult application and position instability. The aim of this study was to determine how different carrier materials influence BCP-induced quantitative and qualitative bone regeneration. Materials and methods A total of 70 critical size defects were set in the frontal bone of 14 domestic pigs (5 each) and filled randomly with either BCP alone (BCP), BCP in combination with nano-hydroxyapatite (BCP + NHA), BCP embedded in native porcine type I/III collagen blocks (BCP + C), autologous bone (AB), or were left empty (ED). Specimens were harvested after 4 and 8 weeks and were evaluated histologically as well as histomorphometrically. Results Significantly lowest rate of new bone formation was found in ED (p = < 0.001) and BCP + NHA groups (p = 0.05). After 8 weeks, the highest percentage of new bone formation was observed in the BCP + C group. Fibrous matrix was detected highest in BCP alone. The lowest residual bone substitute material was found in BCP + C after 8 weeks. Conclusions BCP-induced bone regeneration is indeed affected by different carrier types. Surface morphology and bioactive characteristics influence osseointegration and new bone formation in vivo. The combination of type I/III collagen seems most suitable for qualitative and quantitative bone regeneration

Critical-size Defect Augmentation Using Sintered and Non-Sintered Bovine Bone Matrix - An Experimental Controlled Study in Minipigs

Purpose: Xenogeneic bone substitute materials are often used for augmentation of larger bone defects. Purification methods for these materials vary, mainly in terms of temperature. The aim of this study was to determine in vivo how sintering affects quantitative and qualitative bone regeneration of 2 bovine augmentation materials. Methods: A total of 56 critical size defects were set at the frontal bone of 14 domestic pigs (4 each) and filled randomly with either bovine, sintered hydroxyapatite (BO), bovine, non-sintered hydroxyapatite (BOS), local autologous bone (AB) or left empty. All defects were additionally covered with a collagen membrane. Specimens were harvested after 4 and 8 weeks and were evaluated histologically and histomorphometrically. Results: Histologically new bone could be seen in every group. Significantly highest new bone formation was found in AB. No significant difference could be detected between BO and BOS. Conclusions: According to the results of this study, sintered bone substitute material remains histologically distinguishable but does not affect quantitative and qualitative bone regeneration. (C) 2021 The American Association of Oral and Maxillofacial Surgeons

A new device to prevent fascial retraction in the open abdomen - proof of concept in vivo

BackgroundAn open abdomen is often necessary for survival of patients after peritonitis, compartment syndrome, or in damage control surgery. However, abdominal wall retraction relieves delays and complicates abdominal wall closure. The principle of the newly fascia preserving device (FPD) is the application of anteriorly directed traction on both fascial edges over an external support through a longitudinal beam to relieve increased abdominal pressure and prevent fascial retraction.MethodsTwelve pigs were randomly divided into two groups. Both groups underwent midline laparotomy under general anesthesia. Group one was treated with the new device, group two served as controls. The tension for closing the abdominal fascia was measured immediately after laparotomy as well as at 24 and 48h. Vital parameters and ventilation pressure were recorded. Post mortem, all fascial tissues were histologically examined.ResultsAll pigs demonstrated increases in abdominal circumference. In both groups, forces for closing the abdomen increased over the observation period. Concerning the central closing force after 24h we saw a significant lower force in the FPD group (14.43N) vs. control group (21.6 +/- 5.7N, p<0.001). By testing the main effects using an ANOVA analysis we found a significant group related effect concerning closing force and abdominal circumference of the FDP-group vs. control group (p<0.001; p<0.001). The placement of the device on chest and pelvis did not influence vital parameters and ventilation pressure. Histologic exam detected no tissue damage.Conclusions This trial shows the feasibility to prevent fascial retraction during the open abdomen by using the new device. Thus, it is expected that an earlier closure of the abdominal wall will be possible, and a higher rate of primary closure will be attained

Angiotensin II type 1 receptor localizes at the blood-bile barrier in humans and pigs

Animal models and clinical studies suggest an influence of angiotensin II (AngII) on the pathogenesis of liver diseases via the renin-angiotensin system. AngII application increases portal blood pressure, reduces bile flow, and increases permeability of liver tight junctions. Establishing the subcellular localization of angiotensin II receptor type 1 (AT1R), the main AngII receptor, helps to understand the effects of AngII on the liver. We localized AT1R in situ in human and porcine liver and porcine gallbladder by immunohistochemistry. In order to do so, we characterized commercial anti-AT1R antibodies regarding their capability to recognize heterologous human AT1R in immunocytochemistry and on western blots, and to detect AT1R using overlap studies and AT1R-specific blocking peptides. In hepatocytes and canals of Hering, AT1R displayed a tram-track-like distribution, while in cholangiocytes AT1R appeared in a honeycomb-like pattern; i.e., in liver epithelia, AT1R showed an equivalent distribution to that in the apical junctional network, which seals bile canaliculi and bile ducts along the blood-bile barrier. In intrahepatic blood vessels, AT1R was most prominent in the tunica media. We confirmed AT1R localization in situ to the plasma membrane domain, particularly between tight and adherens junctions in both human and porcine hepatocytes, cholangiocytes, and gallbladder epithelial cells using different anti-AT1R antibodies. Localization of AT1R at the junctional complex could explain previously reported AngII effects and predestines AT1R as a transmitter of tight junction permeability

Esophageal Heat Exchanger Versus Water-Circulating Cooling Blanket for Targeted Temperature Management

To date, the optimal cooling device for targeted temperature management (TTM) remains unclear. Water-circulating cooling blankets are broadly available and quickly applied but reveal inaccuracy during maintenance and rewarming period. Recently, esophageal heat exchangers (EHEs) have been shown to be easily inserted, revealed effective cooling rates (0.26-1.12 degrees C/h), acceptable deviations from target core temperature (<0.5 degrees C), and rewarming rates between 0.2 and 0.4 degrees C/h. The aim of this study was to compare cooling rates, accuracy during maintenance, and rewarming period as well as side effects of EHEs with water-circulating cooling blankets in a porcine TTM model. Mean core temperature of domestic pigs (n = 16) weighing 83.2 +/- 3.6 kg was decreased to a target core temperature of 33 degrees C by either using EHEs or water-circulating cooling blankets. After 8 hours of maintenance, rewarming was started at a goal rate of 0.25 degrees C/h. Mean cooling rates were 1.3 +/- 0.1 degrees C/h (EHE) and 3.2 +/- 0.5 degrees C/h (blanket, p < 0.0002). Mean difference to target core temperature during maintenance ranged between +/- 1 degrees C. Mean rewarming rates were 0.21 +/- 0.01 degrees C/h (EHE) and 0.22 +/- 0.02 degrees C/h (blanket, n.s.). There were no differences with regard to side effects such as brady- or tachycardia, hypo- or hyperkalemia, hypo- or hyperglycemia, hypotension, shivering, or esophageal tissue damage. Target temperature can be achieved faster by water-circulating cooling blankets. EHEs and water-circulating cooling blankets were demonstrated to be reliable and safe cooling devices in a prolonged porcine TTM model with more variability in EHE group

Time delay of T_ZHF compared to T_pulm during the cooling phase.

<p>Time delay of T_ZHF compared to T_pulm during the cooling phase.</p