Joji Mochida

Tokai University, Hiratuka, Kanagawa, Japan

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Publications (156)381.75 Total impact

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    ABSTRACT: Purpose To examine the impact that neuropathic or nociceptive pain has on the quality of life (QOL) in patients with low back pain (LBP) using the Japanese Orthopedic Association Back Pain Evaluation Questionnaire (JOABPEQ) and the Japanese version of the PainDETECT Questionnaire (PDQ-J). Methods Between June 2012 and December 2013, 650 new patients were treated at our institution for LBP. All patients between the ages of 20 and 79 were asked to complete a set of questionnaires including the PDQ-J, a pain visual analog scale (VAS), the JOABPEQ, and the Short Form 36 (SF-36). Based on the PDQ-J scores, participants were classified into three groups: a neuropathic pain group, a nociceptive pain group, and an intermediate mixed pain group. Among them, patients with clear neuropathic and nociceptive LBP were selected. To investigate the differences between neuropathic and nociceptive LBP, diagnosis of spinal disorder, prevalence, age, gender, duration of symptoms, VAS scores, and self-reported general health (SF-36 and JOABPEQ) were compared between the neuropathic and nociceptive pain groups. Results Of 650 patients with LBP, 331 completed the questionnaires and were enrolled in the study. There were 193 men (58.3 %) and 138 women (41.7 %) with a mean age of 54.5 years (range 20–79 years). From the PDQ-J survey, 49 patients (15 %) were classified as having neuropathic pain, and 190 (58 %) were categorized as having nociceptive pain. Patients in the neuropathic pain group had significantly higher VAS scores and lower SF-36 and JOABPEQ scores compared to the nociceptive pain group. Conclusion We examined the impact of nociceptive or neuropathic LBP on QOL. A comparison of JOABPEQ scores between LBP patients assessed by PDQ-J as having neuropathic pain or nociceptive pain suggests that neuropathic pain affects the social and psychological well-being of LBP patients.
    European Spine Journal 12/2014; · 2.47 Impact Factor
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    ABSTRACT: Mobilization and homing of bone marrow-derived cells (BMCs) play a pivotal role in healing and regeneration of various tissues. However, the cellular response of BMCs in avascular tissue such as the intervertebral disc (IVD) has not been studied in detail. One of the main obstacles to this is a lack of a suitable mouse disc degeneration model. To establish a reproducible disc degeneration mouse model suitable for analyzing the cellular response of the disc microenvironment and to determine whether BMCs are recruited into the IVD. An experimental animal study of disc degeneration investigating the potential of BMCs in endogenous repair of the IVD. We transplanted whole bone marrow cells from mice ubiquitously expressing enhanced green fluorescent protein into lethally irradiated mice. IVD degeneration was induced through uneven loading by creating a loop in the tail of these mice. The vertebral bone-disc-vertebral bone units were harvested, and BMCs were identified by immunohistochemistry. A new disc degeneration model was established in the mouse. Applying this model in the bone marrow chimeric mice increased the number of BMCs in the peripheral bone marrow and vascular canals in the end plate, and some were found in the IVD. The migration of BMCs was related to the severity of IVD degeneration. While providing a new disc degeneration model in mice, the current study provide evidence to suggest that although BMCs are recruited during disc degeneration, only a limited number of BMCs migrate to the IVD, presumably because of its avascular nature. This fact provides important elements for developing new treatments as many growth factors and compounds are being tested, both in investigational levels and clinical trials to nourish resident endogenous cells during the degenerative process. Copyright © 2014 Elsevier Inc. All rights reserved.
    The spine journal: official journal of the North American Spine Society 11/2014; · 2.90 Impact Factor
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    ABSTRACT: The purpose of this study was to determine whether radiographic findings associated with thoracolumbar burst fractures could also indicate the presence of posterior ligamentous complex (PLC) injuries, which were identified through short-tau inversion-recovery (STIR)-weighted MRI. Sixty-four patients were surgically treated for thoracolumbar burst fractures between April 2007 and February 2014 at our institution. Twenty-four patients were excluded from this study because of the lack of STIR-weighted MRIs, and therefore 40 patients were included in this study. The patients were divided into two groups based upon the integrity of the PLC, which was evaluated using STIR-weighted MRI: a P group with a PLC injury and a C group without such injury. The following radiographic parameters were evaluated: loss of vertebral body height (LOVBH), local kyphosis (LK), vertebral body translation, canal compromise (sagittal transverse ratio, STR), interlaminar distance (ISD), supraspinous distance (SSD) and interspinous distance (ISD). Frankel scale score and total severity score (load sharing and thoracolumbar injury classification systems, respectively) were also evaluated. Preoperative STIR-weighted MRI showed that 25 patients had a PLC injury (P group: 15 men and 10 women), and 15 patients did not have a PLC injury (C group: 8 men and 7 women). More patients in the P group had an LK>20°: 14 patients in the P group and 1 patient in the C group (p<0.01). The % SSD differed between the P and C groups (118.8%±53.4% and 88.0%±24.3%, respectively; p<0.05). Multivariate logistic analysis showed that an LK>20° was a risk factor for PLC injury in patients with thoracolumbar burst fractures (odds ratio, 55.5 [95% confidence interval, 1.30-2360.1]; p<0.05). These results demonstrate that while LOVBH, vertebral body translation, and canal compromise do not correlate significantly with the presence of a PLC injury in patients with thoracolumbar fractures, an LK>20° and increased % SSD are associated with a PLC injury. Copyright © 2014 Elsevier Ltd. All rights reserved.
    Injury 10/2014; · 2.46 Impact Factor
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    ABSTRACT: IntroductionAngiogenesis is an important factor in the development of osteoarthritis (OA). We investigated the efficacy of bevacizumab, an antibody against vascular endothelial growth factor and an inhibitor of angiogenesis, in the treatment of OA using a rabbit model of anterior cruciate ligament transection.Methods First, we evaluated the response of gene expression and histology of the normal joint to bevacizumab treatment. Next, in a rabbit model of OA induced by anterior cruciate ligament transection, we used macroscopic and histological evaluations and real-time polymerase chain reaction (PCR) to examine the responses to intravenous (systemic) administration of bevacizumab (OAB IV group). We also investigated the efficacy of intra-articular (local) administration of bevacizumab in OA-induced rabbits (OAB IA group).ResultsHistologically, bevacizumab had no negative effect in normal joints. Bevacizumab did not increase the expression of genes for catabolic factors in the synovium, subchondral bone, or articular cartilage, but it increased the expression of collagen type 2 in the articular cartilage. Macroscopically and histologically, the OAB IV group exhibited a reduction in articular cartilage degeneration and less osteophyte formation and synovitis compared with the control group (no bevacizumab; OA group). Real-time PCR showed significantly lower expression of catabolic factors in the synovium in the OAB IV group compared with the OA group. In articular cartilage, expression levels of aggrecan, collagen type 2, and chondromodulin-1 were higher in the OAB IV group than in the OA group. Histological evaluation and assessment of pain behaviour showed a superior effect in the OAB IA group compared with the OAB IV group 12 weeks after administration of bevacizumab, even though the total dosage given to the OAB IA group was half that received by the OAB IV group.Conclusions Considering the dosage and potential adverse effects of bevacizumab, the local administration of bevacizumab is a more advantageous approach than systemic administration. Our results suggest that intra-articular bevacizumab may offer a new therapeutic approach for patients with post-traumatic OA.
    Arthritis research & therapy. 09/2014; 16(5):427.
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    ABSTRACT: In anatomic anterior cruciate ligament (ACL) reconstruction, several pitfalls in creating the femoral bone tunnels at the correct position are of great concern. Our new method, the tibia rotational (TR) technique, may contribute to resolving these. The purpose of this study is to describe further details about the TR technique in anatomic double-bundle ACL reconstruction. Both anteromedial and posterolateral femoral bone tunnels were drilled through a posterolateral tibial bone tunnel using tibial rotation without deep knee flexion. When it is difficult to reach the mark with the rigid guide pin, the narrow curved TR technique guide and the flexible drill system allow drilling femoral bone tunnels in the correct position. The TR technique offers the technical ease required for widespread acceptance while prioritizing the fundamental goals of an anatomic double-bundle ACL reconstruction.
    Arthroscopy techniques. 08/2014; 3(4).
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    ABSTRACT: After traumatic spinal cord injury (SCI), endoplasmic reticulum (ER) stress exacerbates secondary injury, leading to expansion of demyelination and reduced remyelination due to oligodendrocyte precursor cell (OPC) apoptosis. Although recent studies have revealed that amiloride controls ER stress and leads to improvement in several neurological disorders including SCI, its mechanism is not completely understood. Here, we used a rat SCI model to assess the effects of amiloride on functional recovery, secondary damage expansion, ER stress-induced cell death and OPC survival. Hindlimb function in rats with spinal cord contusion significantly improved after amiloride administration. Amiloride significantly decreased the expression of the pro-apoptotic transcription factor CHOP in the injured spinal cord and significantly increased the expression of the ER chaperone GRP78, which protects cells against ER stress. In addition, amiloride treatment led to a significant decrease in ER stress-induced apoptosis and a significant increase of NG2-positive OPCs in the injured spinal cord. Furthermore, in vitro experiments performed to investigate the direct effect of amiloride on OPCs revealed that amiloride reduced CHOP expression in OPCs cultured under ER stress. These results suggest that amiloride controls ER stress in SCI and inhibits cellular apoptosis, contributing to OPC survival. The present study suggests that amiloride may be an effective treatment to reduce ER stress-induced cell death in the acute phase of SCI.
    European Journal of Neuroscience 06/2014; · 3.75 Impact Factor
  • Yoshiyasu Uchiyama, Joji Mochida
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    ABSTRACT: It is difficult to target low-intensity pulsed ultrasound (LIPUS) to the thick portion of soft tissue at the femoral shaft, proximal humerus, spine, and hip joint. A femoral targeting method using ultrasonography (US) was therefore employed to examine the clinical effect of treatment.
    Journal of orthopaedic trauma. 06/2014; 28(6):S5.
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    ABSTRACT: Oxidative stress, caused by the over production of reactive oxygen species (ROS), has been shown to contribute to cell damage associated with neurotrauma and neurodegenerative diseases. ROS mediates cell damage either through direct oxidation of lipids, proteins and DNA or by acting as signaling molecules to trigger cellular apoptotic pathways. The 78 kDa glucose-regulated protein (GRP78) is an ER chaperone that has been suggested to protect cells against ROS-induced damage. However, the protective mechanism of GRP78 remains unclear. In this study, we used C6 glioma cells transiently overexpressing GRP78 to investigate the protective effect of GRP78 against oxidative stress (hydrogen peroxide)-induced injury. Our results showed that the overexpression of GRP78 significantly protected cells from ROS-induced cell damage when compared to non-GRP78 overexpressing cells, which was most likely due to GRP78-overexpressing cells having higher levels of glutathione (GSH) and quinone oxidoreductase 1 (NQO1), two antioxidants that protect cells against oxidative stress. Although hydrogen peroxide treatment increased lipid peroxidation in non-GRP78 overexpressing cells, this increase was significantly reduced in GRP78-overexpressing cells. Overall, these results indicate that GRP78 plays an important role in protecting glial cells against oxidative stress via regulating the expression of GSH and NQO1.
    PLoS ONE 01/2014; 9(1):e86951. · 3.53 Impact Factor
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    ABSTRACT: Loss of vital functions in the somatic motor and sensory nervous systems can be induced by severe peripheral nerve transection with a long gap following trauma. In such cases, autologous nerve grafts have been used as the gold standard, with the expectation of activation and proliferation of graft-concomitant Schwann cells associated with their paracrine effects. However, there are a limited number of suitable sites available for harvesting of nerve autografts due to the unavoidable sacrifice of other healthy functions. To overcome this problem, the potential of skeletal muscle-derived multipotent stem cells (Sk-MSCs) was examined as a novel alternative cell source for peripheral nerve regeneration. Cultured/expanded Sk-MSCs were injected into severely crushed sciatic nerve corresponding to serious neurotmesis. After 4 weeks, engrafted Sk-MSCs preferentially differentiated into not only Schwann cells, but also perineurial/endoneurial cells, and formed myelin sheath and perineurium/endoneurium, encircling the regenerated axons. Increased vascular formation was also observed, leading to a favorable blood supply and waste product excretion. In addition, engrafted cells expressed key neurotrophic and nerve/vascular growth factor mRNAs; thus, endocrine/paracrine effects for the donor/recipient cells were also expected. Interestingly, skeletal myogenic capacity of expanded Sk-MSCs was clearly diminished in peripheral nerve niche. The same differentiation and tissue reconstitution capacity of Sk-MSCs was sufficiently exerted in the long nerve gap bridging the acellular conduit, which facilitated nerve regeneration/reconnection. These effects represent favorable functional recovery in Sk-MSC-treated mice, as demonstrated by good corduroy walking. We also demonstrated that these differentiation characteristics of the Sk-MSCs were comparable to native peripheral nerve-derived cells, whereas the therapeutic capacities were largely superior in Sk-MSCs. Therefore, Sk-MSCs can be a novel/suitable alternative cell source for healthy nerve autografts.
    PLoS ONE 01/2014; 9(3):e91257. · 3.53 Impact Factor
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    ABSTRACT: Release of chemotactic factors in response to tissue damage has been described for different musculoskeletal tissues, including the intervertebral disc (IVD). This study investigated the chemoattractants that are released by induced degenerative IVDs and may be involved in recruiting mesenchymal stem cells (MSCs). Bovine caudal discs were cultured within a bioreactor and loaded under conditions that mimicked physiological or degenerative settings. Between days 4-6, medium was replaced by PBS, which was subsequently used for proteomic, ELISA and immunoprecipitation analyses of secreted chemokines and cytokines. A Boyden chamber assay was used to observe human MSC migration towards native and chemokine depleted media. Gene expression levels of chemokine receptors in human MSCs were analysed, and CCL5 was localised in bovine and human IVD by immunohistochemistry. Proteomic analysis revealed the presence of CCL5 and CXCL6 within conditioned media. Higher concentrations of CCL5 were found in the degenerative media, and a relationship was found between interleukin-1β and CCL5 concentration. Chemokine immunoprecipitation showed that MSCs had a significantly reduced chemotactic migration towards CCL5-immunoprecipitated and CCL5/CXCL6 co-immunoprecipitated media, whilst CXCL6 depletion did not change MSC chemotaxis. MSCs showed a significant increase in mRNA expression of the CCL5 receptors, CCR1 and CCR4, upon culture in degenerative media. Furthermore, CCL5 was identified in bovine and human disc tissue by immunohistochemistry. Hence, CCL5 may be a key chemoattractant that is produced and released by the intervertebral disc cells. Therefore, these factors could be used to enhance stem/progenitor cell mobilisation in regenerative therapies for early stages of disc degeneration.
    European cells & materials 01/2014; 27:124-36. · 4.56 Impact Factor
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    ABSTRACT: In our previous studies, we have demonstrated effective regeneration of cartilage through the creation and application of layered cell sheets that combine both chondrocytes and synovial cells. In this study, we were able to demonstrate that cells derived from cell sheets can survive for long periods after transplantation into rat knee joints having osteochondral defects. We established a method for generating cell sheets from firefly luciferase-expressing chondrocytes obtained from transgenic Lewis rats, and carried out allogenic transplantation of these cell sheets into wild-type Lewis rats. We then administered luciferin and monitored the survival of the transplanted cells by using bioluminescence imaging (BLI). Our data showed that the transplanted cells survived and could be detected for more than 21 months, which was longer than expected. Furthermore, the BLI data showed that the transplanted cells remained in the knee joint and did not migrate to other parts of the body, thus confirming the safety of the cell sheets. In this study, we monitored the duration of survival of cell sheets composed of only chondrocytes, only synovial cells, or both chondrocytes and synovial cells, and found that all three types of cell sheets survived for an extended period of time.
    Biomaterials 12/2013; · 8.31 Impact Factor
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    ABSTRACT: Cartilage damage is typically treated by chondrocyte transplantation, mosaicplasty, or microfracture. Recent advances in tissue engineering have prompted research on techniques to repair articular cartilage damage using a variety of transplanted cells. We studied the repair and regeneration of cartilage damage using layered chondrocyte sheets prepared in a temperature-responsive culture dish. We previously reported achieving robust tissue repair when covering only the surface layer of partial-thickness defects with layered chondrocyte sheets in domestic rabbits. We also reported good Safranin O staining and integration with surrounding tissue in a minipig model of full-thickness cartilaginous defects in the knee joint. We have continued our studies using human chondrocytes obtained from patients under IRB approval, and have confirmed the safety and efficacy of chondrocyte sheets, and have submitted a report to the Ministry of Health, Labour, and Welfare in Japan. In 2011, the Ministry gave us approval to perform a clinical study of joint repair using cell sheets. We have just started implanting cell sheets in patients at Tokai University Hospital. Anat Rec, 2013. © 2013 Wiley Periodicals, Inc.
    The Anatomical Record Advances in Integrative Anatomy and Evolutionary Biology 12/2013; · 1.34 Impact Factor
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    ABSTRACT: Study design:Focus group studyObjective:To investigate cell-specific endoplasmic reticulum (ER) stress reactions in contusive spinal cord by evaluating the expression of the glucose-regulated protein 78 (GRP78) and C/EBP homologous transcription factor protein (CHOP) using immunohistochemical staining.Setting:Data were analysed at Tokai University School of Medicine in Japan.Methods:The authors generated rat spinal cord injury (SCI) models using an IH-Impactor (100 kdyne(LI), 200 kdyne (HI)). Rats were killed at 1, 3, 5, 7 and 14 days post operation (dpo). Spinal cord sections were prepared and the expression ratio of GRP78 and CHOP was evaluated in oligodendrocyte precursor cells (OPCs) (NG2+), oligodendrocytes (OLs) (APC+), neurons (NeuN+) and astrocytes (GFAP+) using double immunohistochemical staining. We examined an area 8 mm distal from SCI-epicenter.Results:Compared with the sham group, both injured groups had higher GRP78 expression ratio in contused spinal cord at 1 dpo. GRP78 expression ratio was highest in GFAP+ cells of both groups, and lowest in NG2+ cells. Although GRP78 was expressed strongly immediately after SCI in the both groups, increased CHOP expression was observed only in the HI group. The CHOP expression in NG2+ cells was significantly higher than that observed in GFAP+ cells at 5 dpo.Conclusion:Although the ER stress response contributes to cell survival in the low-stress SCI conditions, the ER stress response induces an apoptotic cascade in high-stress SCI conditions. The ER response varies according to cell type, with the highest observed in astrocytes, and the lowest observed in oligodendrocyte precursor cells.Spinal Cord advance online publication, 22 October 2013; doi:10.1038/sc.2013.118.
    Spinal Cord 10/2013; · 1.90 Impact Factor
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    ABSTRACT: Development of tissue-engineered materials to treat anterior cruciate ligament (ACL) injury has been limited by the lack of phenotypic markers. We investigated the feasibility of inducing ACL regeneration using cell sheet technology based on the expression of tenomodulin (TNMD) as an early phenotypic marker of ligaments. ACL remnants, the synovium surrounding cruciate ligaments (SCL), the synovium surrounding the infrapatellar fat pads (SIF), and subcutaneous fat tissue (SCF) were obtained from patients undergoing ACL reconstruction or total knee arthroplasty. ACL cell sheets and SCL-derived cell sheets were fabricated successfully A three-dimensional bioengineered ACL was generated by combining triple-layered ACL cell sheets with a bioabsorbable mesh composite. Immunohistochemical examination showed that TNMD was expressed in human ACL fibers, triple-layered ACL cell sheets, ACL remnants, SCL, and SIF, but not in SCF. Real-time PCR showed that TNMD mRNA was expressed at substantially higher levels in the ACL, SCL, and SIF than in the SCF. These results suggest that TNMD is a specific marker of the human ACL and that ACL sheets have a phenotype similar to that of the ACL. The greater expression of TNMD in the SCL- and SIF- suggests that the synovium is a potential cell source for ACL regeneration. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013.
    Journal of Biomedical Materials Research Part A 09/2013; · 2.83 Impact Factor
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    ABSTRACT: Type II odontoid fracture is the most frequent individual fracture in elderly people. An older person usually sustains a Type II odontoid fracture in a fall from standing or a seated height. A relationship between osteoarthritis in the upper cervical spine and Type II odontoid fracture has been reported. However, to our knowledge, few reports have investigated statistically whether disproportionate degeneration between joints influences the susceptibility to fracture. The purpose of this study was to assess predisposition to Type II odontoid fracture in the elderly. Retrospective review of elderly patients sustained Type II odontoid fracture and other axis fractures. Thirty-eight patients aged 65 years and older with axis fractures. Evaluation of computed tomography findings by focusing on osteoporosis and the disproportion in degeneration between each of the upper cervical joints (atlantooccipital, atlantoodontoid, and lateral atlantoaxial joints). Seventeen patients had a Type II odontoid fracture, and 21 patients had other axis fractures. Using the computed tomography findings, we classified osteoporosis at the dens-body junction and the severity of degenerative changes in the atlantoodontoid, atlantooccipital, and lateral atlantoaxial joints as none, mild, moderate, or severe. The proportion of patients with moderate or severe osteoporosis and degenerative changes in each joint and that of patients with disproportionate degenerative changes between joints (difference in grade of ≥2 levels between joints) were compared statistically. The authors report no conflict of interest concerning the materials or methods used in this study or the findings specified in this article. Patients with osteoporosis and with disproportionate degenerative changes between the atlantoodontoid and lateral atlantoaxial joints were significantly more likely to have a Type II odontoid fracture than other axis fractures. These two factors were also assessed in multivariate logistic analysis. The disproportionate degenerative change between the atlantoodontoid and lateral atlantoaxial joint remained significant, even after adjusting for osteoporosis. Older patients with the dens fixed to the atlas because of degeneration of the atlantoodontoid joint and a smooth lateral atlantoaxial joint seem to sustain Type II odontoid fractures because, during a simple fall, the rotation of the head produces torque force on the osteoporotic dens-body junction, which acts as the rotatory center. The presence of the disproportionate osteoarthritic degeneration between the atlantoodontoid and lateral atlantoaxial joints predisposes older people to a Type II odontoid fracture.
    The spine journal: official journal of the North American Spine Society 09/2013; · 2.90 Impact Factor
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    ABSTRACT: It has been shown that coculture of bone marrow-derived stromal cells (BMSCs) with intervertebral disc (IVD) nucleus pulposus (NP) cells significantly activates the biological characteristics of NP cells in animal models and in humans. We therefore predicted that activated NP cells would be a useful graft source for cellular transplantation therapy in the treatment of degenerative IVDs. However, the activation protocol is based on fresh isolation and activation of NP cells, which limits the timing of clinical application. Cell transplantation therapy could be offered to more patients than is now possible if activated NP cells could be transplanted as and when required by the condition of the patient. No study has investigated the effect of cryopreservation on NP cells after enzymatic isolation. We investigated the effects of cryopreservation of canine and human NP cells in both cell and tissue form before coculture with autologous BMSCs. Cell viability, proliferation, glycosaminoglycan production, aggrecan transcriptional activity, colony generation, and gene expression profile of the cells after cryopreservation and subsequent coculture were analyzed. The influence of cryopreservation on cell chromosomal abnormalities and tumorigenesis was also studied. The results showed that there were no clear differences between the noncryopreserved and cryopreserved cells in terms of cell viability, proliferation capacity, and capacity to synthesize extracellular matrix. Furthermore, the cells showed no apparent chromosomal abnormalities or tumorigenic ability and exhibited similar patterns of gene expression. These findings suggest that by using cryopreservation, it may be possible to transplant activated NP cells upon request for patients' needs.
    BioResearch open access. 08/2013; 2(4):273-82.
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    ABSTRACT: There is considerable interest in using cell sheets for the treatment of various lesions as part of regenerative medicine therapy. Cell sheets can be prepared in temperature-responsive culture dishes and applied to injured tissue. For example, cartilage-derived cell sheets are currently under preclinical testing for use in treatment of knee cartilage injuries. The additional use of cryopreservation technology could increase the range and practicality of cell sheet therapies. To date, however, cryopreservation of cell sheets has proved impractical. Here we have developed a novel and effective method for cryopreserving fragile chondrocyte sheets. We modified the vitrification method previously developed for cryopreservation of mammalian embryos to vitrify a cell sheet through use of a minimum volume of vitrification solution containing 20% dimethyl sulfoxide, 20% ethylene glycol, 0.5 M sucrose, and 10% carboxylated poly-L-lysine. The principal feature of our method is the coating of the cell sheet with a viscous vitrification solution containing permeable and non-permeable cryoprotectants prior to vitrification in liquid nitrogen vapor. This method prevented fracturing of the fragile cell sheet even after vitrification and rewarming. Both the macro- and microstructures of the vitrified cell sheets were maintained without damage or loss of major components. Cell survival in the vitrified sheets was comparable to that in non-vitrified samples. We have shown here that it is feasible to vitrify chondrocyte cell sheets and that these sheets retain their normal characteristics upon thawing. The availability of a practical cryopreservation method should make a significant contribution to the effectiveness and range of applications of cell sheet therapy.
    BMC Biotechnology 07/2013; 13(1):58. · 2.17 Impact Factor
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    ABSTRACT: Conventional culture methods using temperature-responsive culture dishes require 4-5 weeks to prepare layered chondrocyte sheets that can be used in articular cartilage repair and regeneration. This study investigated whether the use of synovial tissue obtained from the same joint as the chondrocyte nutritive supply source could more quickly facilitate the preparation of chondrocyte sheets. After culturing derived synoviocytes and chondrocytes together (i.e. combined culture or co-culture) on temperature-responsive inserts, chondrocyte growth was assessed and a molecular analysis of the chondrocyte sheets was performed. Transplantable tissue could be obtained more quickly using this method (average 10.5 days). Real-time polymerase chain reaction and immunostaining of the three-layer chondrocyte sheets confirmed the significant expression of genes critical to cartilage maintenance, including type II collagen (COL2), aggrecan-1 and tissue metallopeptidase inhibitor 1. However, the expression of COL1, matrix metalloproteinase 3 (MMP3), MMP13 and A-disintegrin and metalloproteinase with thrombospondin motifs 5 was suppressed. The adhesive factor fibronectin-1 (FN1) was observed in all sheet layers, whereas in sheets generated using conventional preparation methods positive FN1 immunostaining was observed only on the surface of the sheets. The results indicate that synoviocyte co-cultures provide an optimal environment for the preparation of chondrocyte sheets for tissue transplantation and are particularly beneficial for shortening the required culture period. Copyright © 2013 John Wiley & Sons, Ltd.
    Journal of Tissue Engineering and Regenerative Medicine 07/2013; · 4.43 Impact Factor
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    ABSTRACT: Aim: To cover the large tissue deficits associated with significant loss of function following surgery, a 3D gel-patch-like nerve-vascular reconstitution system was developed using the skeletal muscle-derived multipotent stem cell (Sk-MSC) sheet pellet. Materials & methods: The Sk-MSC sheet pellet was prepared from GFP transgenic mice by the collagenase extraction and 7 days expansion cell culture, and transplanted into a severe muscle damage model with large disruptions to muscle fibers, blood vessels and peripheral nerves. Results: At 4 weeks after transplantation, engrafted cells contributed to nerve-vascular regeneration associated with cellular differentiation into Schwann cells, perineurial/endoneurial cells, vascular endothelial cells and pericytes. However, skeletal myogenic differentiation was scarcely observed. Paracrine effects regarding donor cells/tissues could also be expected, because of the active expression of neurogenic and vasculogenic factor mRNAs in the sheet pellet. Conclusion: These results indicate that the vigorous skeletal myogenic potential of Sk-MSCs was clearly reduced in the sheet pellet preparation and this method may be a useful adjuvant for nerve-vascular regeneration in various tissue engineering applications.
    Regenerative Medicine 07/2013; 8(4):437-51. · 3.87 Impact Factor

Publication Stats

2k Citations
381.75 Total Impact Points

Institutions

  • 1993–2014
    • Tokai University
      • Department of Orthopaedic Surgery
      Hiratuka, Kanagawa, Japan
  • 2011
    • Konkuk University
      • Department of Laboratory Medicine
      Sŏul, Seoul, South Korea
  • 2008–2011
    • Thomas Jefferson University
      • Department of Orthopaedic Surgery
      Philadelphia, PA, United States
  • 2010
    • Jichi Medical University
      • Department of Anesthesiology and Critical Care Medicine
      Totigi, Tochigi, Japan
  • 2007–2009
    • Numazu City Hospital
      Sizuoka, Shizuoka, Japan
  • 2005–2006
    • National Defense Medical College
      Tokorozawa, Saitama, Japan