Method to obtain PRP and L-PRP for the use in routine medical practice of orthopedic traumatologist

The paper presents the results of experimental study of two methods for blood processing to obtain platelet-rich plasma (PRP). Due to the widespread clinical use of platelet-rich plasma in orthopedics and traumatology, a differential approach to the use of PRP and leukocyte- and platelet-rich plasma (L-PRP) has become relevant. Most commercial kits for PRP preparation significantly increase the cost of treatment. In addition, the main attention is focused on the concentration of platelets in the final product and the marketing features of a particular kit (separating gel, tube shape, anticoagulant, activator, etc.). The role of leukocytes in the features of the regenerative effect of platelet-rich plasma stays out of focus. The blood of volunteers was processed in two different ways and the cellular composition of the obtained products was analyzed. Sample No. 1 was used as a control for the baseline blood cell composition. Sample No. 2 was centrifuged in a sterile plastic tube, platelet rich plasma was collected manually. Sample No. 3 was centrifuged in a sterile modified syringe, platelet rich plasma was collected using a closed technique by means of connectors together with a rich layer. The results of statistical analysis showed that there were obtained totally different biological products with different concentrations of platelets and leukocytes. The first sample (control) had platelet concentration of (228.69 ± 39.15) × 109/l and leukocyte concentration of (5.18 ± 1.32) × 109/l. In the second sample (tube + manual sampling), platelet concentration was (429.38 ± 79.92) × 109/l and leukocyte concentration — (0.85 ± 0.34) × 109/l. The third sample (syringe plunger + closed vacuum collection) had platelet concentration of (541.15 ± 85.49) × 109/l and leukocytes of (6.56 ± 1.92) × 109/l. The data are given without regard to –12.5% deviation on dilution with citrate dextrose. As the result of the work, orthopedic traumatologists can use the simplest non-commercial methods for obtaining PRP and L-PRP in their practice depending on nosology and clinical tasks.

Changed inflammatory markers after application of 4DryField PH for adhesion prevention in gynecological surgery

Introduction The development of peritoneal adhesions and the effects of different antiadhesion agents on such mechanisms are not fully understood. Temporary rises of the C-reactive protein (CRP) level have been reported after antiadhesion agent application. We present the changes of inflammation markers observed after use of a starch-based polysaccharide certified for adhesion prevention and hemostasis 4DF (4DryField® PH). Method Retrospective comparative analysis of inflammation markers in 40 patients undergoing laparoscopic adhesiolysis with or without adhesion prophylaxis was conducted. Statistical comparisons were performed by means of paired or unpaired t tests (for normally distributed continuous data), Wilcoxon matched pairs signed-rank tests or Mann–Whitney tests (for not-normally distributed continuous data), Mantel–Cox tests (for continuous data describing time intervals), and Fisher’s exact tests (for discrete data). Results The maximum post-operative CRP level was significantly elevated in the 4DF group (87 vs. 29%; p < 0.001), whereas leukocyte concentration and body temperature did not differ between groups. No signs of infection were detected in any of the patients and CRP levels spontaneously dropped to normal values within few days. No side effects or complications were observed in both groups. In second-look surgeries performed for other diagnoses 1–56 weeks after the first interventions, no remnants of 4DF or any peritoneal inflammatory reactions were observed. Conclusion The starch-based polysaccharide 4DF can be considered safe and does not induce inflammatory reactions of clinical significance. Further studies regarding 4DF degradation are recommended and, apart from macrophage migration, could also examine corresponding markers such as IL-6 and PCT.

Urothelial toxicity of esketamine in the treatment of depression

Rationale Ketamine is the first widely used substance with rapid-onset antidepressant action. However, there are uncertainties regarding its potential urothelial toxicity, particularly after repeated application. In the context of rising recreational ketamine use, severe side effects affecting the human urinary tract have been reported. It is assumed that ketamine interacts with bladder urothelial cells and induces apoptosis. Objectives This study aimed to assess whether single or repeated doses of esketamine used in an antidepressant indication are associated with urinary toxicity. Methods We included male and female inpatients with a current episode of depression and a diagnosis of recurrent depressive disorder, bipolar disorder or schizoaffective disorder according to ICD-10 criteria (n = 25). The esketamine treatment schedule involved a maximum of 3× weekly dosing at 0.25–0.5 mg/kg i.v. or s.c. The primary outcome was the change in urine toxicity markers (leukocytes, erythrocytes, protein and free haemoglobin). Description of demographic, clinical and laboratory data was conducted using means, standard deviations, frequencies and percentages. Changes in urinary toxicity markers over time were evaluated using linear mixed models with gender as a covariate. Results The participants received an average of 11.4 (SD 8) esketamine treatments, and an average number of 11.2 (SD 8) urine samples were analysed over the course of treatment. Neither urinary leukocyte concentration (F(20; 3.0) = 3.1; p = 0.2) nor erythrocyte concentration (F(20;2.2) = 4.1; p = 0.2) showed a significant trend towards increase during the course of esketamine treatment. Similarly, free haemoglobin and protein concentrations, which were analysed descriptively, did not display a rise during treatment. There was a significant improvement in depression ratings after esketamine treatment (p < 0.001). Conclusions This study is, to the best of our knowledge, the first to focus on urothelial toxicity of esketamine used in antidepressant indication and dose. The results indicate that the use of single or repeated doses of esketamine is unlikely to cause urothelial toxicity. The results are in need of confirmation as sample size was small.

Hematology and serum biochemistry of free-range brown-throated sloths in two urban areas in Brazil

Background and Aim: The brown-throated sloth (Bradypus variegatus) is widely distributed in three biomes: The Amazon, Atlantic Forest, and Caatinga. Some subpopulations are isolated in urban areas in Brazil, usually in squares and small woods. Due to the scarcity of reference values, an investigation was carried out on the hematology and blood biochemistry of brown-throated sloths from urban areas. Materials and Methods: Blood was collected by venipuncture in the femoral vein from 19 brown-throated sloths for hematological and biochemical analyses, living in two municipalities; Teófilo Otoni (TO) (Minas Gerais State) and Rio Tinto (RT) (Paraíba state), in the Atlantic Forest biome, Brazil. The samples were analyzed in specialized veterinary laboratories using automatic cell counters and slide fixation staining methods. Results: The two subpopulations of brown-throated sloths had no significant differences in most hematological values, with the exception of a higher leukocyte concentration (p<0.01) in the RT sub-population. The difference in leukocyte concentration suggests an idiosyncratic effect, as the animals were healthy and the stress of the capture was acute, not chronic. The concentrations of urea, creatinine, and alkaline phosphatase (ALP) were similar (p<0.05) in brown-throated sloths from TO and RT. Likewise, the liver enzyme concentrations (ALP, alanine transaminase [ALT], and aspartate transaminase [AST]) did not differ between the two subpopulations (p<0.05). Conclusion: Compared to another study, brown-throated sloths from TO and RT have higher plasma concentrations of ALT and ALP, suggestive of a hepatic overload. Hematological and blood biochemical findings of TO and RT can be used as clinical reference values for brown-throated sloths living in an urban environment.

THE IMPACT OF VETOM 20.76 BASED ON PREDATORY FUNGUS ARTHROBOTRYS OLIGOSPORA ON THE LEUCOCYTES IN THE GEESE BLOOD

The paper demonstrates the results on the effect of new specimen Vetom 20.76 on concentration of leukocytes in the blood of geese on the basis of the predatory fungus Artusbotus oligospora. In order to achieve the goal of the experiment, one control group and six experimental groups were arranged on the principle of paired analogues. Each group contained 10 geese aged 1 month. The geese from the experimental groups received Vetom 20.76 in different doses in the morning with water once a day: the geese of the 1st experimental group - dose of 0.5 ppm/kg of live weight during 15 days; 2nd experimental group - 1 ppm/kg of live weight during 15 days; 3rd experimental group - 2 ppm/kg of live weight during 15 days, 4th experimental group - 0.5 ppm/kg live weight during 30 days, 5th experimental group - 1 ppm/kg live weight during 30 days and 6th - 2 ppm/kg live weight during 30 days. The geese of control group didn’t receive the specimen. The concentration of leukocytes in the blood of experimental geese increases in the period of specimen application as well as in the period of its aftereffect. If Vetom 20.76 is prescribed for 15 days, the effect of leukopoiesis stimulation finishes on the 30th day. If the specimen is applied during 30 days, the leukocytes in the blood continue to increase up to the 60th day. This long-term application of Vetom 20.76 dosed 0.5ppm/kg increases leucocytes within the physiological norm. Application of higher doses (1 and 2 ppm/kg) the leukocyte concentration conforms to the physiological norm

Does the Difference in Leukocyte Concentration of PRP Affect the Short-Term Follow-Up Results in Cases Diagnosed with Early Stage Knee Osteoarthritis?

This prospective study was conducted for the clinical evaluation of pain severity and knee functionality following PRP injections with different leukocyte (WBC) concentrations applied to cases diagnosed with knee osteoarthritis. A total of 109 patients were included in the study. According to the leukocyte content the PRP injections were prepared as low-leukocyte content PRP (PPRP) and high concentration leukocyte content PRP (L-PRP). Patients were divided into 2 groups. Group 1 (n=44) received low-leukocyte content PRP and Group II (n =65) received high-leukocyte content PRP. The patients were evaluated clinically with Visual Analog Scale (VAS) and Knee Society Score (KSS). The changes in the PLT levels of the L-PRP group after the procedure compared to the levels prior to the procedure were found to be statistically significantly greater than the changes in the PPRP group. The mean VAS score of all the cases before treatment was 9.05±0.91 and this score decreased to 3.71±1.46 within 12 months. The increases in the mean Knee Society Score (KSS) values were determined as 16.92±1.97 within 6 months and 16.89±2.97 within 12 months in the P-PRP group and 19.71±1.24 within 6 months and 19.86±0.42 within 12 months in the L-PRP group. The most important aspect of this study is that, in contrast to many other studies, the results continued after the 6th month and were reported to be good in the 12th month. It was also recorded that L-PRP was clinically superior to P-PRP in the treatment of early stage knee osteoarthritis.

Development of an algorithm for optimizing the synthesis of leukocyte polypeptides

The article presents experimental data on the influence of various factors on the synthesis of leukocyte polypeptides. It was shown that the productive ability of leukocytes depends on the method of sampling the donor raw materials and the used blood containers. The obtained experimental data indicate a favorable effect of cytapheresis during harvesting of raw materials on polypeptides produced by leukocytes. Assessment of the level of specific activity of polypeptides obtained from raw materials by the method of cytapheresis indicates an increase of 2.4 times. It was also found that the use of the cytapheresis method made it possible to obtain a leukoerythromass with a leukocyte concentration 4 times higher than using the classical method of taking raw materials. The results of the influence of the culture regime of leukocyte cells on the level of specific activity of the synthesized polypeptides are presented. It was shown that the suspension mode of cultivation, in which the level of specific activity of polypeptides increases 1.3 times, is optimal for the synthesis of leukocyte polypeptides. The conditions for the synthesis of polypeptides by leukocytes were optimized. It was established by calculation that the leukocyte concentration of 8 billion/l and the mixing rate of the culture medium of 32 rpm were optimal at the stage of biosynthesis. The effect of L-dextrose and maltose on the residual moisture content of polypeptides in cryoprotectant concentrations from 2.5 to 10% was studied. In a series of experiments, it was found that maltose at a concentration of 2.5% is the optimal cryoprotectant for lyophilization of leukocyte polypeptides. According to the results of the experiments, an algorithm for optimizing the synthesis of leukocyte polypeptides was developed, which consists in using the cytapheresis method in the preparation of raw materials and “Terumo” hemakontainers, observing the optimal parameters during the cultivation of leukocytes, and lyophilization of the finished product using a cryoprotectant of maltose at a concentration of 2.5%.