Hatakeyama J1, Inoue S2, Yakuwa T2, Nomura M2,3, Wakimoto Y2, Li C2, Wakigawa T1, Tsubaki T1, Kinoshita S1, Sakai Y4, Akisue T5, Moriyama H5
1Kobe University School of Medicine Faculty of Health Sciences, Physical Therapy Major, Kobe, Japan, 2Kobe University Graduate School of Health Sciences, Department of Rehabilitation Science, Kobe, Japan, 3Research Fellow of Japan Society for the Promotion of Science, Tokyo, Japan, 4Kobe University Graduate School of Medicine, Division of Rehabilitation Medicine, Kobe, Japan, 5Kobe University, Life and Medical Sciences Area, Health Sciences Discipline, Kobe, Japan
Background: Spinal cord injury (SCI) causes a degeneration of muscles and joint components (e.g., fibrosis, increase in the connective tissue, and adhesions and shorting of synovial membrane). These degenerations are one of the causes of joint contractures, which are a major complication in the patients with SCI and decrease activities of daily living and quality of life. Positioning, stretching, and physical therapy are advocated to prevent and treat these contractures; however, we are often confronted with patients who have contractures. Recently, transcutaneous carbon dioxide (CO2) therapy were developed, verifying the efficacy of this therapeutic application for various disorders such as acceleration of fracture healing and muscle injury repair. However, whether CO2 therapy improves the degeneration of muscles and joint components after SCI remains unclear.
Purpose: We aimed to examine the efficacy of CO2 therapy on degeneration of muscles and joint component as a prevention and treatment regimen for the degeneration of muscles and joint components after SCI.
Methods: Thirty-six 10-week-old male Wistar rats were randomized into 3 groups: control without interventions, untreated after SCI, and treated with CO2 therapy after SCI (CO2). The hindlimb of CO2 group was exposed CO2 gas for 20 min once daily. As a preventive intervention, CO2 group was treated with CO2 from the first postoperative day for 2 or 4 weeks. On the other hand, as a therapeutic intervention, CO2 group was treated from the 15th postoperative day for 2 or 4 weeks. At the end of the interventions, semitendinosus was extracted and stained with picrosirius red for the quantification of fibrosis. Total RNA was isolated from semitendinosus, and mRNA expression of fibrosis markers, collagen types I and TGF-b, were assessed by real-time PCR. In addition, undecalcified frozen section was prepared from the knee joint, and each section was stained with hematoxylin and eosin to measure the posterior synovium length. Fibrosis of joint capsule was evaluated by immunohistochemical staining of collagen type I and TGF-b.
Results: Histologically, there was the trend toward increase muscle fibrosis after 2 and 4 weeks in the SCI groups. The CO2 treatment decreased the mRNA expression of collagen types I in the preventive and therapeutic interventions for 4 weeks. The synovial intima length was decreased after SCI, and CO2 treatment did not improve the shortening of the length. The CO2 treatment decreased TGF-β immunolabeling in the preventive intervention for 2 weeks., although the staining intensity of collagen types Ⅰ showed no differences among the groups at all-time points.
Conclusion(s): The CO2 treatment partially improved the gene expression of muscle fibrosis, and further it tended to decrease the expression of TGF-β in the joint capsules. Therefore, CO2 therapy may be effective treatment for the fibrosis of muscle and joint capsules after SCI.
Implications: CO2 therapy is easy and non-invasive treatment and has potential to be a novel therapeutic approach for the degenerations of muscles and joint components leading to contractures after SCI.
Keywords: Spinal cord injury, contractures, CO2 therapy
Funding acknowledgements: This study was supported by the Japan Society for the Promotion of Science (17K19908).
Purpose: We aimed to examine the efficacy of CO2 therapy on degeneration of muscles and joint component as a prevention and treatment regimen for the degeneration of muscles and joint components after SCI.
Methods: Thirty-six 10-week-old male Wistar rats were randomized into 3 groups: control without interventions, untreated after SCI, and treated with CO2 therapy after SCI (CO2). The hindlimb of CO2 group was exposed CO2 gas for 20 min once daily. As a preventive intervention, CO2 group was treated with CO2 from the first postoperative day for 2 or 4 weeks. On the other hand, as a therapeutic intervention, CO2 group was treated from the 15th postoperative day for 2 or 4 weeks. At the end of the interventions, semitendinosus was extracted and stained with picrosirius red for the quantification of fibrosis. Total RNA was isolated from semitendinosus, and mRNA expression of fibrosis markers, collagen types I and TGF-b, were assessed by real-time PCR. In addition, undecalcified frozen section was prepared from the knee joint, and each section was stained with hematoxylin and eosin to measure the posterior synovium length. Fibrosis of joint capsule was evaluated by immunohistochemical staining of collagen type I and TGF-b.
Results: Histologically, there was the trend toward increase muscle fibrosis after 2 and 4 weeks in the SCI groups. The CO2 treatment decreased the mRNA expression of collagen types I in the preventive and therapeutic interventions for 4 weeks. The synovial intima length was decreased after SCI, and CO2 treatment did not improve the shortening of the length. The CO2 treatment decreased TGF-β immunolabeling in the preventive intervention for 2 weeks., although the staining intensity of collagen types Ⅰ showed no differences among the groups at all-time points.
Conclusion(s): The CO2 treatment partially improved the gene expression of muscle fibrosis, and further it tended to decrease the expression of TGF-β in the joint capsules. Therefore, CO2 therapy may be effective treatment for the fibrosis of muscle and joint capsules after SCI.
Implications: CO2 therapy is easy and non-invasive treatment and has potential to be a novel therapeutic approach for the degenerations of muscles and joint components leading to contractures after SCI.
Keywords: Spinal cord injury, contractures, CO2 therapy
Funding acknowledgements: This study was supported by the Japan Society for the Promotion of Science (17K19908).
Topic: Neurology: spinal cord injury; Musculoskeletal: lower limb; Disability & rehabilitation
Ethics approval required: Yes
Institution: Kobe University
Ethics committee: Institutional Animal Care and Use Committee
Ethics number: P160506
All authors, affiliations and abstracts have been published as submitted.
