Muscle-contraction training can enhance the efficacy of cell transplantation treatment for Duchenne Muscular Dystrophy (DMD)
Nana Takenaka-Ninagawa 1, Yuta Itoh 2, Keisuke Kawakami 3, Hidetoshi Sakurai 1.
1Center for iPS Cell Research and Application (CiRA), Kyoto University, 2Faculty of Rehabilitation Science, Nagoya Gakuin University, 3Oita University Faculty of Welfare and Health Sciences.
Muscle stem cell transplantation therapy is one of the hopeful therapies for intractable muscular diseases such as Duchenne muscular dystrophy (DMD). In adult skeletal muscles, satellite cells act as a stem cell with regenerating damaged myofibers. We have been investigating generation of muscle stem cells from human iPS cells. Although, a lot of reports showed effects of cell transplantation therapy toward DMD, the best way of cell transplantation into skeletal muscle and valid evaluating method to assess the efficacy of cell therapy have not been developed yet. Therefore we tried to establish the most stable and efficient method of cell transplantation in skeletal muscle by using human immortalized myogenic progenitor cell (Hu5/KD3), and the best way to assess the functional recovery of cell transplanted muscle of DMD model mice (DMD-null/NSG). Moreover, we are also trying to establish an effective rehabilitation programs to promote the effect of cell transplantation therapy toward DMD. These newly developed techniques could be a fundamental steps for establishment of cell therapy towards DMD patients.
Assessment of therapeutic value of osteoarthritis rehabilitation by molecular biological and biochemical approach
Takashi Ohtsuki 1, Kanae Kumagishi 2,4, Akira Shinaoka 2, Keiichiro Nishida 2, Aiji Ootsuka 2, Kenichiro Sakata 3, Syunsuke Sakata 3, Kenji Kawamura 4, Satoshi Hirohata 1.
1Department of Medical Technology, Okayama University Graduate School of Health Sciences, 2Department of Human Morphology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama university, 3Sakata Orthopedic Hospital, 4Department of Physical Therapy, Kibi International university.
Backgrounds: Osteoarthritis is one of a major joint disease. Popular conservative therapies are physiotherapy and hyaluronan (HA) treatment. But, little was known about the impact of combined therapy. We compared the impact of combined therapy by molecular biological and biochemical analysis
Methods: The patients were divided 4 groups. Group 1 underwent physiotherapy (PT) alone as control. Group 2 underwent both PT and HA treatment. Group 3 underwent PT with Shaking board (SB) treatment. Group 4 underwent PT, SB and HA treatment. Urine collagen type II C-telopeptide (CTXII) was determined by Enzyme linked immunosorvent assay. Knee joint range of Motion (ROM), Visual analogue scale (VAS) and time up & go test (TUG) were measured. Shaking board GB 700 (OG Wellness, Okayama, Japan)
Results: CTXII concentration reduced in primary stage patients (Kellgren Lawrence grade I and II) but did not reduced in the progress stage (KL grade III-IV). CTXII concentration decreased in group 4.
Conclusion: Therapeutic impact was thought to be effective in the patients with the primary stage. The best therapy was PT with SB and HA treatment.
Scaffold-free Bio 3D Conduits for the peripheral nerve regeneration
Junichi Tajino 1, Akira Ito 2, Hirofumi Yurie 2, Ryosuke Ikeguchi 2, Shizuka Akieda 3, Manami Tsuji 3, and Tomoki Aoyama 1.
1Department of Physical Therapy, Human Health Sciences, Graduate School of Medicine, Kyoto University, Japan, 2Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Japan, 3Cyfuse Biomedical K.K., Japan.
Introduction: Functional restoration after the peripheral nerve injuries remains challenging. Although autologous nerve grafting (autograft) would be the gold standard, it has several shortcomings such as limited supply, sacrifice of the donor site function etc. Tube-like materials (nerve conduits) have been developed to bridge the disconnected nerves. However, synthetic materials don’t match autografts in regenerative capacity. Nor they do in infectious safeness. To address these difficulties, we focused on Bio 3D printing technology that enables us to build biocompatible nerve conduits with no foreign materials.
Methods: F344–rnu/rnu rats with immune deficiency (9-10 wks old) were used in two groups (Conduit, Control). The right side sciatic nerve of each rat was bisected in the middle-thigh level and 5mm gap was created. For Conduit group, Bio 3D conduit solely made of human fibroblast was interposed between the nerve ends. For Control group, the cut nerve was left with no grafting. Eight weeks after the surgery, nerve regeneration and functional recoveries were evaluated.
Results and Discussion: Histological and morphometric evaluation showed enhanced nerve regeneration in Conduit group as well as greater recovery in electrophysiological and kinematical evaluation. Taken together, Bio 3D conduit might promote nerve regeneration after injury.
Effect of low-intensity pulsed ultrasound after mesenchymal stromal cells injection to treat osteochondral defect
S. Yamaguchi 1,2, T. Aoyama 1, A. Ito 2,3, M. Nagai 4, J. Tajino 1, H. Iijima 1,2, X. Zhang 1, W. Kiyan 1, H. Kuroki 1.
1Department of Physical Therapy, Graduated of School Medicine, Kyoto University, Japan, 2Japan Society for the Promotion of Science, Japan, 3Department of Orthopaedic surgery, Gradate of School Medicine, Kyoto University, Japan, 4 Congenital Anomaly Research Center, Gradate of School Medicine, Kyoto University, Japan.
We investigated the effect of low-intensity pulsed ultrasound (LIPUS) treatment combined with mesenchymal stromal cell (MSC) injection for cartilage repair and subchondral bone reconstitution to treat rat knee osteochondral defect. An osteochondral defect was created on both femur grooves of Wistar rats (n=28). Four weeks later, bone marrow MSCs were injected into the right knee joint, and phosphate buffer saline were injected into the left knee as control. The rats were divided into 2 intervention groups: without or with LIPUS irradiation. Two days after injection, the rats with LIPUS were subjected to LIPUS treatment, 20 min/day, 5 days/week, to both knee joints. After 4 and 8 weeks intervention, the rats were euthanized, femora were removed for evaluation. Subchondral bone reconstitution was evaluated as bone volume (BV)/tissue volume (TV) by micro-computerized tomography analysis. Cartilage repair was evaluated histologically based on the Wakitani cartilage repair score. MSC injection improved the cartilage repair score and LIPUS irradiation improved BV/TV. Combination treatment promoted both cartilage repair and BV/TV improvement. Thus, MSC injection combined with LIPUS irradiation is more effective than either treatment alone in promoting concurrent cartilage repair and subchondral reconstitution.
Functional Assessment in Patients with Knee Osteoarthritis Treated by Different Preparation Protocols of Platelet Rich Plasma: Preliminary Data
Carolina Mie K. Onodera 1,2, Marco C. Uchida 1, Ricardo A. C. Sampaio 1, Priscila Y. S. Sampaio 1, Aline Urban Paffaro 2, José Fabio Fabio Santos Duarte Lana 2, Joyce M. A. Bizzacchi 2.
1Department of Adapted Physical Activity; School of Physical Education, University of Campinas, Campinas-SP, Brazil, 2Hematology-Hemotherapy Center, University of Campinas, Campinas-SP, Brazil.
Background: Platelet-rich plasma (PRP) is an autologous therapy able to induce healing in soft tissue in Osteoarthritis (OA). The main objective of this study was to assess physical functioning comparing Platelet Poor Plasma (PPP), Platelet Rich Plasma Poor Leucocytes (P-PRP) and Platelet Rich Plasma Rich in Leucocytes (L-PRP) in knee OA treatment during a 3-month follow-up.
Methods: A total of 24 patients with knee OA (II, III, IV-Kellgren and Lawrence grading scale) were enrolled in this study. Patients were randomized in PPP, P-PRP, L-PRP groups. Outcome measures included Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC-function), 30 seconds chair test, usual walking speed, stair climb test. A paired t test was used to compare the differences between the pre and post interventions results in different groups.
Results: Our results demonstrated a statistically significant increase in functional tests (p 0.05) in groups treated with P-PRP and L-PRP when compared to PPP treatment; WOMAC scores showed no significant differences in any group after 3-month follow-up.
Conclusion: Our preliminary results suggest that P-PRP and L-PRP are promising to treat knee OA; in addition, functional tests could be more objective to detect differences in the treatment using PRP.
Electrical stimulation drives chondrogenesis of mesenchymal stem cells in the absence of exogenous growth factors
Hyuck Joon Kwon 1, Gyu Seock Lee 1, Honggu Chun 2.
1Department of Physical Therapy and Rehabilitation, College of Health Science, Eulji University, Gyeonggi, Korea, 2Department of Bio-convergence Engineering, Korea University, Seoul, Korea.
Mesenchyme stem cells (MSCs) have high therapeutic potential for cartilage regeneration. We previously uncovered the role of ATP oscillations for prechondrogenic condensation in MSC chondrogenesis. In this study, we have tried to induce ATP oscillations which are crucial for prechondrogenic condensation in chondrogenesis by using electrical stimulation (ES) as a physical inducer. In this study, we found that ES generated ATP and calcium oscillations and induced MSCs to be condensed into the aggregation and subsequently differentiate into chondrocytes without addition of exogenous growth factors, displaying high expression levels of chondrogenic markers such as type II collagen, aggrecan and Sox9. The actions of ES for ATP oscillations and chondrogenesis depended on extracellular ATP signaling via P2X4 receptor. In addition, ES induced the significant increase in expression of TGF-β1 and BMP2, and inhibition of either TGF-β or BMP signaling prevented the ES-induced upregulated expression of chondrogenic markers, which indicates that ES stimulates chondrogenesis via TGF-β and BMP signalings. However, inhibition of TGF-β signaling blocked ES-induced condensation, while inhibition of BMP signaling did not, which reveals that TGF-β signaling, but not BMP signaling, mediates ES-induced condensation. In conclusion, these findings advance our understanding of the effects of ES on chondrogenesis and their mechanisms of action, and contribute to developing electrotherapeutic strategies for cartilage repair using MSCs.
Exercise therapy for knee osteoarthritis
Hirotaka Iijima 1,2, Tomoki Aoyama 1, Akira Ito 2,3, Junichi Tajino 1, Shoki Yamaguchi 1,2, Momoko Nagai 4, Hiroshi Kuroki 1.
1Department of Physical Therapy, Human Health Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan, 2Japan Society for the Promotion of Science, Tokyo, Japan, 3Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan, 4Congenital Anomaly Research Center, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Background: We have been researching the effects of physical activity including exercise intervention on articular cartilage-subchondral bone unit as well as functional performance, with the ultimate goal of developing effective and financially viable treatments in patients with knee OA. Recently, we conducted two researches from clinical and basic aspect using biological and epidemiological approaches as follows.
Methods and Results: [STUDY1]: Of 207 patients with radiographic Kellgren/Lawrence grade ≥1, 137 (66.2%) patients with knee OA were inactive ( 5000 steps/day: sedentary lifestyle) due to knee specific impairment and other factors. Walking additional 1000 steps/day was associated with improved self-reported and performance-based physical function, particularly in patients with lower physical function. [STUDY2] We found that physiological level exercise intervention increases expression of bone morphogenetic proteins (BMPs) and prevents the progression of cartilage-subchondral bone lesions in a post-traumatic rat knee model (n = 60). Intraarticular injection of Gremlin-1, an inhibitor of BMPs, was likely to cancel the effects of physiological level exercise on cartilage-subchondral bone lesions, suggesting that exercise intervention may act as a disease-modifying therapy via increase of BMPs expression.
Conclusion: These findings indicate that increased physical activity might prevent osteoarthritic changes in knee joint as well as enhance functional improvement in patients with knee OA.