The main objective of this study was to evaluate the intensity and character of the inflammatory reaction caused by an innovative polyester-polyurethane vascular prosthesis implanted into the abdominal aorta of 9 Beagle dogs aged 1-3 years. At 6 and 12 months post implantation the prostheses were removed and tissues samples were examined using 2 methods: histology and immunohistochemistry (IHC). Histology slides stained with hematoxylin and eosin (H&E) were evaluated for the intensity of inflammation by observing the density of inflammatory cells and graded 1 to 4 (1- light inflammation, 4 – severe inflammation). The pro-inflammatory mediator tumor necrosis factor-alpha (TNFα) and two anti-inflammatory mediators, interleukin 1 receptor antagonist (IL1ra), and interleukin 10 (IL10), were also assessed in the tissue samples by IHC methods. Mean (n=5) inflammation grade in H&E slides at 6 months post-implantation (6Mpost) was 2 and mean (n=4) inflammation grade at 12 months (12Mpost) was almost 3. IHC staining showed that TNFα and IL1ra in tissue samples obtained from 6Mpost dogs were expressed at the same intensity indicating equal pro- and anti-inflammatory cytokine levels. However, in the 12Mpost tissues TNFα was expressed more intensely than IL1ra and IL10. Moreover, in 2 dogs at 12Mpost, there were signs of infection assessed on the basis of neutrophil infiltration in the prostheses. In conclusion, the assessment of pro-inflammatory mediators such as TNFα and anti-inflammatory mediators, such as IL1ra and IL10, can help to interpret the intensity of the inflammatory process directed at synthetic prostheses.
Combined retrograde tracing and double-labelling immunofluorescence were used to investi- gate the distribution and chemical coding of neurons in aorticoerenal (ARG) and testicular (TG) ganglia supplying the urinary bladder apex (UBA) in the juvenile male pig (n=4, 12 kg. body weight). Retrograde fluorescent tracer Fast Blue (FB) was injected into the wall of the bladder apex under pentobarbital anesthesia. After three weeks all the pigs were deeply anesthetized and transcardially perfused with 4% buffered paraformaldehyde. TG and ARG were collected and processed for double-labelling immunofluorescence. The presence of tyrosine hydroxylase (TH) or dopamine beta-hydroxylase (DBH), neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL), nitric oxide synthase (NOS) and vesicular acetylcholine transporter (VAChT) were inves- tigated. The cryostat sections were examined with a Zeiss LSM 710 confocal microscope equipped with adequate filter blocks. The TG and ARG were found to contain many FB-positive neurons projecting to the UBA (UBA-PN). The UBA-PN were distributed in both TG and ARG. The majority were found in the left ganglia, mostly in TG. Immunohistochemistry disclosed that the vast majority of UBA-PN were noradrenergic (TH- and/or DBH-positive). Many noradrenergic neurons also contained immunoreactivity to NPY, SOM or GAL. Most of the UBA-PN were supplied with varicose VAChT-, or NOS- IR (immunoreactive) nerve fibres. This study has revealed a relatively large population of differently coded ARG and TG neu- rons projecting to the porcine urinary bladder. As judged from their neurochemical organization these nerve cells constitute an important element of the complex neuro-endocrine system involved in the regulation of the porcine urogenital organ function.
The present study investigated the expression of androgen receptor (AR) in neurons of the anterior pelvic ganglion (APG) and celiac-superior mesenteric ganglion (CSMG; ganglion not involved in the innervation of reproductive organs) in the male pig with quantitative real-time PCR (qPCR) and immunohistochemistry. qPCR investigations revealed that the level of AR gene expression in the APG tissue was approximately 2.5 times higher in the adult (180-day-old) than in the juvenile (7-day-old) boars. Furthermore, in both the adult and juvenile animals it was sig- nificantly higher in the APG than in CSMG tissue (42 and 85 times higher, respectively). Immu- nofluorescence results fully confirmed those obtained with qPCR. In the adult boars, nearly all adrenergic (DβH-positive) and the majority of non-adrenergic neurons in APG stained for AR. In the juvenile animals, about half of the adrenergic and non-adrenergic neurons were AR-posi- tive. In both the adult and juvenile animals, only solitary CSMG neurons stained for AR. The present results suggest that in the male pig, pelvic neurons should be considered as an element of highly testosterone-dependent autonomic circuits involved in the regulation of urogenital func- tion, and that their sensitization to androgens is a dynamic process, increasing during the prepu- bertal period.