Use of Modern Cryomethods in Combined Surgical Treatment of Patients with Purulent Chronic Wounds

The paper considers the possibilities of using in medical practice various cryomethods and techniques of cryopreservation of biological objects to treat the patients with purulent chronic wounds (PCW). The variety of methods and approaches to apply cold in medical practice: general and local cryotherapy, cryosurgery etc. has been emphasized. As a result of using each of these methods, regenerative processes begin, which promote the healing of PCW. Particular attention is paid to cryotechniques, ensuring the preservation of cord blood at low temperatures. Cord blood cells and serum have been shown to have immune modulatory and trophic-stimulating therapeutic effects, that is extremely important when treating the patients with PCW. The issue of combined use of cord blood serum and innovative vacuum therapy for the patients with PCW has been considered.

Assessment of the Dynamics of Temperature Changes in the Knee Joint Area in Response to Selected Cooling Agents in Thermographic Tests

Although local cryotherapy (LC) is performed with various cooling agents (CAg) such as ice, water, and gasses, in clinical practice, it is mostly performed with cooling gasses. Presently, LC with cooling gasses is very popular but the inference about the thermal (stimulus) effect on the tissues is mainly based on research carried out using ice packs. The proposed objective of the study was to evaluate the dynamics of temperature changes in the knee joint area in response to a 3-min exposure to liquid nitrogen vapors (LNVs), cold air (CA) and ice bag (IB). The study group included 23 healthy volunteers with an average age of 26.67 ± 4.56. The exposed (ROIE) and contralateral (ROINE) areas of the knee joint after exposure to CAg were observed. Immediately after 3 min of LC, the ROIE temperature dropped by 10.11 ± 0.91 °C after LNV, 7.59 ± 0.14 °C after IB and 6.76 ± 1.3 °C after CA. Significant tissue cooling was maintained up to 15 min after LNV (p < 0.01), 10 min after IB (p < 0.05) and 5 min after CA (p < 0.05). LC causes significant temperature changes both in ROIE and ROINE. The greatest cooling potential was demonstrated for LNV and the lowest for CA.

The effect of cryotherapy on pain during local and total application

Background: analgesic effect of total and local cryotherapy in people older than 55 years, in respondents with various diagnoses, most often with degenerative diseases, especially with gonarthrosis, coxarthrosis, spondylosis, osteoarthritis and polyarthrosis. Objective: the aim of the survey was to process information from the field of total and local cryotherapy and to analyze the information obtained using questionnaires. We process the percentage effect of total and local cryotherapy on the intensity of painful symptoms, feeling, tolerance and satisfaction of respondents after completing ten therapies. Materials and methods: the group of respondents consisted of 35 men and 65 women aged 55 to 81 years. A total of 100 respondents qualified for the survey. 50% of respondents participated in total cryotherapy and 50% of respondents participated in local cryotherapy. Results: we processed the results using MS Excel and IBM SPSS. The Modified Laitinen Pain Questionnaire was used. Due to established hypotheses and the nature of the data, we used the Kolmogorov – Smirnov test, the non-parametric Mann – Whitney U-test and the Wilcoxon nonparametric test. The results show that with the help of local and total cryotherapy, the intensity of painful symptoms is lower. Conclusions: the performed analysis of the data by means of a questionnaire showed that local and total cryotherapy alleviates pain and has an analgesic effect.

1H-NMR-Based Analysis for Exploring Knee Synovial Fluid Metabolite Changes after Local Cryotherapy in Knee Arthritis Patients

Rehabilitation using cryotherapy has widely been used in inflammatory diseases to relieve pain and decrease the disease activity. The aim of this study was to explore the metabolite changes in inflammatory knee-joint synovial fluids following local cryotherapy treatment (ice or cold CO2). We used proton nuclear magnetic resonance (1H NMR) spectroscopy to assess the metabolite patterns in synovial fluid (SF) in patients with knee arthritis (n = 46) before (D0) and after (D1, 24 h later) two applications of local cryotherapy. Spectra from aqueous samples and organic extracts were obtained with an 11.75 Tesla spectrometer. The metabolite concentrations within the SF were compared between D1 and D0 using multiple comparisons with the application of a false discovery rate (FDR) adjusted at 10% for each metabolite. A total of 32 metabolites/chemical structures were identified including amino acids, organic acids, fatty acids or sugars. Pyruvate, alanine, citrate, threonine was significantly higher at D1 vs D0 (p < 0.05). Tyrosine concentration significantly decreases after cryotherapy application (p < 0.001). We did not observe any effect of gender and cooling technique on metabolite concentrations between D0 and D1 (p > 0.05). The present study provides new insight into a short-term effect of cold stimulus in synovial fluid from patients with knee arthritis. Our observations suggest that the increased level of metabolites involved in energy metabolism may explain the underlying molecular pathways that mediate the antioxidant and anti-inflammatory capacities of cryotherapy.

Influence of Intermittent Cold Stimulations on CREB and Its Targeting Genes in Muscle: Investigations into Molecular Mechanisms of Local Cryotherapy

Local cryotherapy is widely used as a treatment for sports-related skeletal muscle injuries. The molecular mechanisms are unknown. To clarify these mechanisms, we applied one to three 15-min cold stimulations at 4 °C to various cell lines (in vitro), the tibialis anterior (TA) muscle (ex vivo), and mouse limbs (in vivo). In the in vitro assay, cyclic AMP (cAMP) response element binding protein 1 (CREB1) was markedly phosphorylated (p-CREB1), and the CREB-binding protein (CBP) was recruited to p-CREB-1 in response to two or three cold stimulations. In a reporter assay with the cAMP-responsive element, the signals significantly increased after two to three cold stimulations at 4 °C. In the ex vivo study, CREB-targeting genes were significantly upregulated following two or three cold stimulations. The in vivo experiment disclosed that cold stimulation of a mouse limb for 9 days significantly increased mitochondrial DNA copy number and upregulated genes involved in mitochondrial biogenesis. The results suggest that local cryotherapy increases CREB transcription and upregulates CREB-targeting genes, in a manner dependent on cold stimulation frequency and duration. This information will inform further investigations into local cryotherapy as a treatment for sports-related skeletal muscle trauma.

SAT0021 STRUCTURAL EFFECTS OF LOCAL CRYOTHERAPY IN RHEUMATOID ARTHRITIS: A STUDY IN ADJUVANT-INDUCED ARTHRITIS

Background:The control of joint destruction caused by rheumatoid arthritis (RA) is a key issue in the treatment of this disease. Recent evidence showed that radiographic progression of joint damage occur despite a sharp decrease in disease activity and the use of aggressive Disease Modifying Anti-Rheumatic Drug (DMARD) therapies [1]. Whether alternative treatments such as cryotherapy may have beneficial effects on joint destruction at the early stages of the disease remains to be demonstrated, but such strategy would be of interest as it would not interfere with DMARDs treatment.Objectives:The aim of this study was to evaluate the effect of a 14-days-treatment of local cryotherapy on radiological outcomes in rat adjuvant induced arthritis.Methods:Adjuvant-induced arthritis (AIA) was induced in 6-weeks old male Lewis rats by injection ofMycobacterium butyricumin Freund’s incomplete adjuvant at the basis of the tail. A control group received saline. At the onset of arthritis, AIA rats were treated or not by application of cryotherapy on paws using either a cold spray or ice, twice a day for 14 days. Arthritis score and paws skin temperature was daily monitored. At the end of treatment, radiological exam of hind paws was performed and a score taking into account (swelling, osteoporosis, cartilage destruction, bone erosion, bone destruction and new bone formation) was assigned, according to Ackermanet al[2]. Circulating levels of cytokines (IL-6, IL-1β, TNF-α) was measured by Magpix Luminex kit.Results:Compared to untreated AIA, local cryotherapy significantly reduced the progression of arthritis score, whatever the modality (p<0.05), and to the same extent (reduction of arthritis score at day 24 post-immunization: -38% with cold spray, p<0.01, -37% with ice, p<0.01). Radiological score was significantly reduced by both treatments with no difference between the two treatments (-33% with cold spray, p<0.01, -44% with ice, p<0.01). All the items of the radiological score were equally reduced by ice and cold spray except swelling that was significantly reduced only by ice. Interestingly, the use of the cold spray induced a greater decrease in the skin temperature than the ice treatment (18.32 ± 0.07 °C vs 20.46 ± 0.08 °C, p<0.001). Conversely, cryotherapy did not significantly change the level of cytokines. No correlation was found between radiological score and arthritis score or cytokine levels.Conclusion:These data demonstrated that local cryotherapy had positive effects on structural damage in adjuvant-induced arthritis. The mechanisms involved remain now to be determined. These results suggest that local cryotherapy would be an interesting complement to conventional DMARDs in early RA.References:[1] Ten Klooster, P. M.et al.Radiographic progression can still occur in individual patients with low or moderate disease activity in the current treat-to-target paradigm: real-world data from the Dutch Rheumatoid Arthritis Monitoring (DREAM) registry.Arthritis Res. Ther.21, 237 (2019).[2] Ackerman, N. R.et al.Effects of naproxen on connective tissue changes in the adjuvant arthritic rat.Arthritis Rheum.22, 1365–1374 (1979).Disclosure of Interests:Célian Peyronnel: None declared, Valentin Petitcolin: None declared, Perle Totoson: None declared, Hélène Martin: None declared, Frank Verhoeven: None declared, Céline Demougeot Grant/research support from: With an institutional support from Pfizer.

Influence of Intermittent Cold Stimulations on CREB and Its Targeting Genes in Muscle: Investigations into Molecular Mechanisms of Local Cryotherapy

Local cryotherapy is widely used as a treatment for sports-related skeletal muscle injury. However, its molecular mechanisms are unknown. To clarify these mechanisms, in this study, we applied one to three 15-min cold stimulations at 4 &deg;C to various cell lines (in vitro), the tibialis anterior (TA) muscle (ex vivo), and mouse limbs (in vivo). In the in vitro assay, cAMP response element-binding protein 1 (CREB1) was markedly phosphorylated (as pCREB1) and CREB-binding protein (CBP) was recruited to pCREB-1 in response to two or three cold stimulations. In a reporter assay with the cAMP-responsive element, the signals significantly increased after two to three cold stimulations at 4 &deg;C. In the ex vivo study, CREB-targeting genes were significantly upregulated following two or three cold stimulations. The in vivo experiment disclosed that cold stimulation of a mouse limb for 9 days significantly increased mitochondrial DNA copy number and upregulated genes such as Pgc-1&alpha; involved in mitochondrial biogenesis. The foregoing results suggest that local cryotherapy increases CREB transcription and upregulates CREB-targeting genes in a manner dependent on cold stimulation frequency and duration. This information may serve as an impetus for further investigations into local cryotherapy as a treatment for sports-related skeletal muscle trauma.