Evaluation of midazolam-ketamine with dexmedetomidine and fentanyl for injectable anaesthesia in dogs

dexmedetomidine and fentanyl for injectable anaesthesia in dogs. Vet. arhiv 83, 509

Prosudba učinka midazolam-ketamina s deksmedetomidinom i fentanilom za injekcijsku anesteziju u pasa.

A prospective randomized blinded study was conducted on 12 clinically healthy adult dogs of both sexes (mean weight of 18.34 ± 0.78 kg) divided into three groups (n = 4). The animals received 0.4 mg/kg midazolam and 10 μg/kg dexmedetomidine (group A), 0.4 mg/kg midazolam and 20 μg/kg dexmedetomidine (group B) and 0.4 mg/kg midazolam + 20 μg/kg dexmedetomidine + 4 μg/kg fentanyl (group C) intramuscularly, using separate syringes. Ten minutes later Ketamine was administered intravenously in all the groups. A significantly (P<0.05) shorter weak time (onset of sedation) and down time (onset of recumbency) were recorded in animals in group C as compared to the animals of groups A and B. Muscle relaxation was excellent in group C. The pedal reflex was abolished up to 30 min in groups A and B and up to 60 min in group C. Intubation was only possible in groups B and C. The anaesthetic induction dose of ketamine was minimal in group C. Standing recovery time was shortest in the animals of group C. Respiratory rate (RR) decreased significantly (P<0.05) throughout the observation period, but rectal temperature (RT) decreased significantly (P<0.05) towards the end of the observation period in all the groups. Heart rate decreased significantly (P<0.05) in the animals of group B. Mean arterial pressure (MAP) was maintained within the physiological range in all the groups. It was concluded that dexmedetomidine (10 μg/kg)-midazolam-ketamine can produce anaesthesia for about 20 min in dogs. Increasing the dose of dexmedetomidine did not enhance the duration of anaesthesia, but the further addition of fentanyl not only reduced the induction dose of ketamine but also increased the duration of anaesthesia up to 50 min. Dexmedetomidine-midazolam-fentanyl-ketamine can be used for prolonged duration of injectable anaesthesia in dogs.Poduzeto je prospektivno istraživanje na 12 slučajno odabranih klinički zdravih pasa i kuja (prosječne tjelesne mase 18,34 ± 0,78 kg) podijeljenih u tri skupine (n = 4). Životinjama skupine A bio je intramuskularno primijenjen midazolam u dozi od 0,4 mg/kg i deksmedetomidin u dozi od 10 μg/kg. Životinjama skupine B bio je i/m primijenjen midazolam u dozi od 0,4 mg/kg i deksmedetomidin 20 μg/kg, a životinje skupine C primile su i/m 0,4 mg/kg midazolama, 20 μg/kg deksmedetomidina i 4 μg/kg fentanila. Deset minuta nakon toga svim je životinjama intravenski bio ubrizgan ketamin. Značajno (P<0,05) kraće vrijeme smirivanja (nastup sedacije) i vrijeme lijeganja ustanovljeno je u životinja skupine C u usporedbi sa skupinama A i B. Opuštanje mišićja bilo je izvrsno u skupini C. Nožni refleks nestao je nakon 30 minuta u skupinama A i B, a nakon 60 minuta u skupini C. Intubacija je bila moguća samo u životinja skupine B i C. Doza ketamina potrebna za početak anestezije bila je najmanja u životinja skupine C. Vrijeme potrebno za ponovno ustajanje bilo je najkraće u životinja skupine C. Frekvencija disanja značajno se smanjila (P<0,05) u čitavom razdoblju promatranja, dok se rektalna temperatura u svih životinja značajno smanjila (P<0,05) na kraju razdoblja promatranja. Frekvencija bila znatno se smanjila (P<0,05) u životinja skupine B. Srednji arterijski tlak bio je u fiziološkim granicama u svih životinja. Može se zaključiti da kombinacija deksmedetomidin (10 μg/kg)-midazolam-ketamin može u pasa dovesti do anestezije za oko 20 minuta. Povećanje doze deksmedetomidina nije povećalo trajanje anestezije. Ipak, daljnja primjena fentanila ne samo da je smanjila početnu dozu ketamina već je povećala trajanje anestezije na 50 minuta. Deksmedetomidin-midazolam-fentanil-ketamin mogu se rabiti za produženo trajanje injekcijske anestezije u pasa

Comparative Evaluation of Masson's Trichrome and Picrosirius Red Staining for Digital Collagen Quantification Using ImageJ in Rabbit Wound Healing Research

The therapeutic potential of Pluronic F127 (PF127) hydrogel loaded with adipose-derived stromal vascular fraction (AdSVF), mesenchymal stem cells (AdMSC), and conditioned media (AdMSC-CM) for repairing full-thickness skin wounds was evaluated using a rabbit model. The rabbits were randomly divided into eight groups with six animals each and treatment was given as per the predetermined protocol (3 doses at one-week interval): Group A (Control), Group B (AdSVF), Group C (AdMSC), Group D (AdMSC-CM), Group E (PF127), Group F (AdSVF + PF127), Group G (AdMSC + PF127), and Group H (AdMSC-CM + PF127). Skin tissue samples were collected from the healing wounds on day 28 for staining and collagen quantification. Collagen density (Area %) was quantified using tissue sections stained with Masson's Trichrome (MT) and Picrosirius Red (PSR) stain using the Colour Deconvolution plugin of ImageJ and RGB stack method, respectively. These techniques function based on separating different colour channels in the stained tissue sections to isolate the collagen fibers and then quantifying them through thresholding and image analysis. Across the treatment groups, both staining methods generally showed a trend of increased collagen density compared to the control group. For most groups, PSR staining consistently indicated slightly lower collagen densities than MT staining. However, the overall trends were similar in both staining. The comparison between PSR and MT staining methods revealed that both techniques effectively assess collagen density in healing wounds. However, there were subtle differences in the absolute values obtained, with PSR staining tending to yield slightly lower collagen density measurements than MT. These differences can be attributed to the distinct mechanisms of these staining methods. Therefore, both staining methods can digitally quantify collagen density in wound healing research

Prosudba učinkovitosti celomske tekućine kišne gujavice na cijeljenje rana u punoj debljini kože kunića.

The present study was conducted on 16 New Zealand White rabbits of 10-12 months of age, to evaluate the healing potential of earthworm coelomic fluid in full thickness skin wounds. Under xylazine-ketamine anaesthesia, four rectangular full thickness excisional skin wounds, measuring 2×2 cm2 were created on the dorsum of each animal and designated as groups I, II, III and IV. Wounds were treated by topical coelomic fluid (I), 0.5 % povidone-iodine and coelomic fluid (II), 0.5 % povidone-iodine (III) and normal saline (IV). Healing was evaluated on the basis of gross and histomorphological parameters. The mean wound area was significantly lesser (P<0.05) in the wounds of groups I and II as compared to groups III and IV, up to 21 days. Out of 16 wounds, nine wounds in group I (56.25 %) and 11 wounds in group II (68.75 %) healed completely by day 21, but none in groups III and IV. Histomorphological studies showed more mature and densely placed collagen and better epithelialization in groups I and II as compared to groups III and IV. It was concluded that coelomic fluid of the earthworm Eisenia foetida can accelerate healing of full-thickness skin wounds in rabbits.Istraživanje je provedeno na 16 novozelandskih bijelih kunića u dobi od 10 do 12 mjeseci s ciljem da se prosudi mogući učinak celomske tekućine kišne gujavice na cijeljenje rana u punoj debljini kože u kunića. Kunići su bili podijeljeni u četiri skupine označene I, II, III i IV. Četiri pravokutne ekscizijske kožne rane u punoj debljini veličine 2x2 cm načinjene su na leđima svake životinje pod anestezijom ksilazin-ketaminom. Rane su bile obrađene celomskom tekućinom (skupina I), 0,5 %-tnim povidon-jodom i celomskom tekućinom (skupina II), 0,5 %-tnim povidon-jodom (skupina III) i fiziološkom otopinom (skupina IV). Cijeljenje rana bilo je prosuđivano na osnovi patoanatomskih i patohistoloških nalaza. Prosječna površina rana 21 dan nakon ekscizije bila je značajno manja (P<0,05) u skupinama I i II u usporedbi s površinom skupina III i IV. Devet od 16 rana u skupini I (56,25 %) i 11 u skupini II (68,75 %) u potpunosti su zacijelile 21 dan nakon ekscizije, dok nijedna nije zacijelila u skupini III i IV. Patohistološki nalaz pokazao je više zrelog i gušće raspoređenog kolagena te bolju epitelizaciju u skupinama I i II u usporedbi sa skupinama III i IV. Može se zaključiti da celomska tekućina kišne gujavice Eisenia foetida može ubrzati cijeljenje rana u punoj debljini kože u kunića

In vitro Antimicrobial Properties of Pluronic F-127 Injectable Thermoresponsive Hydrogel

Pluronic F-127 (PF-127) hydrogel is a versatile biomaterial with promising applications in drug delivery, tissue engineering, and regenerative medicine. PF-127 has antiadhesive activity that prevents bacterial adhesion by creating a hydrated layer on the bacterial surface. This property makes PF-127 suitable for preventing implant-associated infections. In this study, we aimed to evaluate the antibacterial properties of PF-127 using field isolates of Staphylococcus aureus (Gram-positive bacteria) and Escherichia coli (Gram-negative bacteria) and compare them with different antibiotic standards. The antimicrobial potential was assessed using disk diffusion assays with four standard concentrations (20%, 25%, 30%, and 40%). The test microorganisms were inoculated on agar plates, and sterile filter paper disks infused with PF-127 hydrogels were placed alongside standard antibiotic disks. After incubation, the inhibition zones were measured to determine antimicrobial activity. Our results showed that PF-127 lacked intrinsic antimicrobial activity against S. aureus and E. coli at the tested concentrations. In conclusion, PF-127 hydrogel is a promising neutral carrier hydrogel system for loading antibiotics and antimicrobial compounds. Its unique properties, such as biocompatibility and thermo-responsive behaviour, combined with its antiadhesive activity, make it an ideal candidate for various biomedical applications

Standardization and characterization of adipose-derived stromal vascular fraction from New Zealand white rabbits for bone tissue engineering

Background and Aim: Adipose tissue-derived stromal vascular fraction (SVF) contains a heterogeneous cell population comprising multipotent adipose-derived stem cells. Regenerative therapy using adipose-derived SVF has broad applications in bone tissue engineering due to the superior osteogenic potential of SVF. This study was designed to standardize and characterize adipose-derived SVF obtained from New Zealand white rabbits for bone tissue engineering and other potential applications. Materials and Methods: Ten skeletally mature and clinically healthy adult New Zealand white rabbits were used in this study. The SVF was prepared using surgically resected interscapular adipose tissue following enzymatic digestion with 0.1% collagenase type I solution. The SVF pellet obtained after the final centrifugation step was suspended in a 0.5 mL control solution to obtain ready-to-use adipose-derived SVF. The freshly prepared SVF was characterized based on the total SVF cell count and cell yield per gram of adipose tissue. The SVF cells were enumerated using a hemocytometer. Results: Interscapular adipose tissue depots are ideal for preparing autologous adipose-derived SVF due to the ease of access. The interscapular adipose-derived SVF prepared by enzymatic digestion had an average cell yield of 3.15±0.09×106 cells/g adipose tissue. Freshly prepared SVF had a total cell count of 3.15±0.09×104 cells/μL. Conclusion: The enzymatic digestion of adipose tissue using 0.1% collagenase resulted in better cell yield per gram than methods previously reported in rabbits. The use of adipose-derived SVF can preclude the requirement for an additional culture period. In addition, it may also reduce the risk of extensive cell contamination, which makes it a safe and cost-effective strategy for bone tissue engineering applications