Chul-Won Ha

Mesenchymal stem/progenitor cells developed in cultures from UC blood

Background Whether umbilical cord blood (UCB) serves as a source of mesenchymal stem/progenitor cells (MSPC) is controversial. MSPC are the best candidates for cellular therapy of orthopedic skeletal tissues. In order to explore the possibility of UCB as a useful source of MSPC, we identified, expanded in culture, and characterized MSPC from UCB harvests on a large scale. Methods Mononuclear cells isolated from UCB harvests (n=411) were cultured in media supplemented with 10% FBS. MSPC-like cells cultured from each UCB harvest were expanded ex vivo by successive subcultivation. UCB harvests with a more than 1000-fold expanding capacity (n=9) were examined for surface Ag phenotypes and in vitro differentiation potentials into osteogenic, chondrogenic and adipogenic lineages. Results Ninety-five out of a total of 411 UCB units (23.1%) generated MSPC-like cells during cultivation. Nine UCB units (2.2%) yielded MSPC with more than 1000-fold expansion capacity. These cells positively expressed MSPC-related Ag, but did not express myeloid, histocompatibility or endothelial Ag. These cells also possessed multiple capacities for osteogenic, chondrogenic and adipogenic differentiation. Discussion Although the incidence of UCB harvests producing MSPC in culture was low, some of them showed a more than 1000-fold expanding capacity, which is enough in cell numbers to be an allogeneic source for cellular therapy. Our results may encourage the use of UCB as an attractive target for allogeneic cellular therapeutic options in tissue engineering.

Percutaneous release of trigger digits

We describe a safe and easy percutaneous technique for release of trigger finger using a specially designed knife. The A1 pulley is sectioned by a blade which has a hooked end. We released, percutaneously, 185 trigger fingers, including 62 which were locked using this technique. Satisfactory results were achieved in 173 (93.5%). There were no significant complications. We recommend this as a safe and effective outpatient procedure for those patients who have not responded successfully to conservative treatment, have longstanding symptoms or severe triggering.

Initial phase I safety of retrovirally transduced human chondrocytes expressing transforming growth factor-beta-1 in degenerative arthritis patients.

Background aims. TissueGene-C (TG-C) represents a cell-mediated gene therapy for localized delivery of allogeneic chondrocytes expressing transforming growth factor (TGF)-β1 directly to the damaged knee joint. Untransduced human chondrocytes (hChonJ cells) have also been incorporated into the TG-C product at a 3:1 ratio with TGF-β1-expressing chondrocytes (hChonJb#7) in order to help fill in the defect and as target cells for the actions of the expressed TGF-β1. Methods. A phase I dose-escalating clinical trial was performed to evaluate the safety and biologic activity of TG-C in patients with advanced osteoarthritis of the knee joint (full thickness cartilage defect) that was refractory to existing non-operative therapies. Following a single intra-articular injection into the joint space of the damaged knee, patients were monitored for safety, and an evaluation was performed to assess the pharmacokinetics and biologic activity of TG-C. Results. There were no treatment-related serious adverse events. Swelling, effusion and minor localized reactions such as warming sensation or itching were observed in a dose-dependent manner at the injection site. Knee evaluation scores seemed to indicate a dose-dependent trend toward efficacy; however, patient numbers were not sufficient to determine statistical significance. Conclusions. Overall, there were no significant safety issues related to the administration of TG-C, with only some minor injection site reactions observed. Additionally, knee scoring analyzes indicated a possibility that TG-C may contribute to improvement of arthritic symptoms. More study is warranted to evaluate further the safety and determine the potential efficacy of TG-C.

Cartilage Repair Using Composites of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells and Hyaluronic Acid Hydrogel in a Minipig Model

The cartilage regeneration potential of human umbilical cord blood‐derived mesenchymal stem cells (hUCB‐MSCs) with a hyaluronic acid (HA) hydrogel composite has shown remarkable results in rat and rabbit models. The purpose of the present study was to confirm the consistent regenerative potential in a pig model using three different cell lines. A full‐thickness chondral injury was intentionally created in the trochlear groove of each knee in 6 minipigs. Three weeks later, an osteochondral defect, 5 mm wide by 10 mm deep, was created, followed by an 8‐mm‐wide and 5‐mm‐deep reaming. A mixture (1.5 ml) of hUCB‐MSCs (0.5 × 107 cells per milliliter) and 4% HA hydrogel composite was then transplanted into the defect on the right knee. Each cell line was used in two minipigs. The osteochondral defect created in the same manner on the left knee was untreated to act as the control. At 12 weeks postoperatively, the pigs were sacrificed, and the degree of subsequent cartilage regeneration was evaluated by gross and histological analysis. The transplanted knee resulted in superior and more complete hyaline cartilage regeneration compared with the control knee. The cellular characteristics (e.g., cellular proliferation and chondrogenic differentiation capacity) of the hUCB‐MSCs influenced the degree of cartilage regeneration potential. This evidence of consistent cartilage regeneration using composites of hUCB‐MSCs and HA hydrogel in a large animal model could be a stepping stone to a human clinical trial in the future.

A technique for intraoperative construction of antibiotic spacers

A technique for intraoperatively creating an antibiotic spacer for two-stage treatment of infected total knee replacements is described. An intraoperative mold is made from the removed components and used to create antibiotic spacers with surface contours similar to those of the original total knee replacement. The spacers restore leg length and knee stability. This allows limited function during the interval before reimplantation of the new total knee replacement. It is a cost-effective and convenient technique for creating a suitably shaped and sized cement spacer for two-stage revision total knee replacement after infection. The clinical results of 12 consecutive patients using this technique with minimum of 2 years followup seem to be at least equal or better than results reported in previous studies. Level of Evidence: Prognostic study, Level II (retrospective study). See the Guidelines for Authors for a complete description of levels of evidence.

Cartilage Regeneration in Osteoarthritic Patients by a Composite of Allogeneic Umbilical Cord Blood-Derived Mesenchymal Stem Cells and Hyaluronate Hydrogel: Results From a Clinical Trial for Safety and Proof-of-Concept With 7 Years of Extended Follow-Up.

Few methods are available to regenerate articular cartilage defects in patients with osteoarthritis. We aimed to assess the safety and efficacy of articular cartilage regeneration by a novel medicinal product composed of allogeneic human umbilical cord blood‐derived mesenchymal stem cells (hUCB‐MSCs). Patients with Kellgren‐Lawrence grade 3 osteoarthritis and International Cartilage Repair Society (ICRS) grade 4 cartilage defects were enrolled in this clinical trial. The stem cell‐based medicinal product (a composite of culture‐expanded allogeneic hUCB‐MSCs and hyaluronic acid hydrogel [Cartistem]) was applied to the lesion site. Safety was assessed by the World Health Organization common toxicity criteria. The primary efficacy outcome was ICRS cartilage repair assessed by arthroscopy at 12 weeks. The secondary efficacy outcome was visual analog scale (VAS) score for pain on walking. During a 7‐year extended follow‐up, we evaluated safety, VAS score, International Knee Documentation Committee (IKDC) subjective score, magnetic resonance imaging (MRI) findings, and histological evaluations. Seven participants were enrolled. Maturing repair tissue was observed at the 12‐week arthroscopic evaluation. The VAS and IKDC scores were improved at 24 weeks. The improved clinical outcomes were stable over 7 years of follow‐up. The histological findings at 1 year showed hyaline‐like cartilage. MRI at 3 years showed persistence of the regenerated cartilage. Only five mild to moderate treatment‐emergent adverse events were observed. There were no cases of osteogenesis or tumorigenesis over 7 years. The application of this novel stem cell‐based medicinal product appears to be safe and effective for the regeneration of durable articular cartilage in osteoarthritic knees. Stem Cells Translational Medicine 2017;6:613–621

Comparison of robot-assisted and conventional total knee arthroplasty: A controlled cadaver study using multiparameter quantitative three-dimensional CT assessment of alignment

Introduction: A functional total knee replacement has to be well aligned, which implies that it should lie along the mechanical axis and in the correct axial and rotational planes. Incorrect alignment will lead to abnormal wear, early mechanical loosening, and patellofemoral problems. There has been increased interest of late in total knee arthroplasty with robotic assistance. This study was conducted to determine whether robot-assisted total knee arthroplasty is superior to the conventional surgical method with regard to the precision of implant positioning. Materials and Methods: Twenty knee replacements, comprising ten robot-assisted procedures and ten conventional operations, were performed on ten cadavers. Two experienced surgeons performed the surgeries. Both procedures on each cadaver were performed by the same surgeon. The choice of which procedure was to be performed first was randomized. Following implantation of the prosthesis, the mechanical axis deviation, femoral coronal angle, tibial coronal angle, femoral sagittal angle, tibial sagittal angle, and femoral rotational alignment were measured via 3D CT scanning. These variables were then compared with the preoperatively planned values. Results: In the knees that underwent robot-assisted surgery, the mechanical axis deviation ranged from −1.94° to 2.13° (mean: −0.21°), the femoral coronal angle from 88.08° to 90.99° (mean: 89.81°), the tibial coronal angle from 89.01° to 92.36° (mean: 90.42°), the tibial sagittal angle from 81.72° to 86.24° (mean: 83.20°), and the femoral rotational alignment from 0.02° to 1.15° (mean: 0.52°) in relation to the transepicondylar axis. In the knees that underwent conventional surgery, the mechanical axis deviation ranged from −3.19° to 3.84° (mean: −0.48°), the femoral coronal angle from 88.36° to 92.29° (mean: 90.50°), the tibial coronal angle from 88.15° to 91.51° (mean: 89.83°), the tibial sagittal angle from 80.06° to 87.34° (mean: 84.50°), and the femoral rotational alignment from 0.32° to 4.13° (mean: 2.76°) in relation to the transepicondylar axis. In the conventional knee replacement group, there were two instances of outliers outside the range of 3° varus/valgus for the mechanical axis deviation. The robot-assisted knee replacements showed significantly superior femoral rotational alignment results compared with conventional surgery (p = 0.006). There was no statistically significant difference between robot-assisted and conventional total knee arthroplasty with regard to the other variables. All the measurements showed high intra-observer and inter-observer reliability. Conclusion: Robot-assisted total knee arthroplasty showed excellent precision in the sagittal and coronal planes of the 3D CT scan. In particular, the robot-assisted technique showed better accuracy in femoral rotational alignment compared to the conventional surgery, despite the fact that the surgeons who performed the operations were more experienced and familiar with the conventional method than with robot-assisted surgery. It can thus be concluded that robot-assisted total knee arthroplasty is superior to conventional total knee arthroplasty.

Methylation-induced silencing of ASC and the effect of expressed ASC on p53-mediated chemosensitivity in colorectal cancer.

Tumor suppressor p53 is known to play a crucial role in chemosensitivity in colorectal cancer. We previously demonstrated that an apoptosis-associated speck-like protein, ASC, is a p53-target gene which regulates p53-Bax mitochondrial apoptotic pathway. ASC is also known to be a target of methylation-induced gene silencing. An inactivation of ASC might thus cause resistance to chemotherapy, and if this is the case, then the expression of ASC would restore the chemosensitivity. The aim of this study was to clarify this hypothesis. ASC was methylated in 25% of all resected specimens in patients with colorectal cancer; however, ASC methylation did not always correspond to a lack of ASC protein. When expressed in colon cancer cells, in which ASC is absent due to methylation, ASC was found to enhance the chemosensitivity in a p53-dependent manner. In p53-null cells, ASC increased the p53-mediated cell death induced by p53-expressing adenovirus infection. Our data suggest that the methylation-induced silencing of ASC might cause resistance to p53-mediated chemosensitivity in colorectal cancer. The gene introduction of ASC may thus restore such chemosensitivity, and this modality may therefore be a useful new treatment strategy for colorectal cancer.

A placebo-controlled randomised trial to assess the effect of TGF-ß1-expressing chondrocytes in patients with arthritis of the knee

The aim of this study was to assess the effect of injecting genetically engineered chondrocytes expressing transforming growth factor beta 1 (TGF-β1) into the knees of patients with osteoarthritis. We assessed the resultant function, pain and quality of life. A total of 54 patients (20 men, 34 women) who had a mean age of 58 years (50 to 66) were blinded and randomised (1:1) to receive a single injection of the active treatment or a placebo. We assessed post-treatment function, pain severity, physical function, quality of life and the incidence of treatment-associated adverse events. Patients were followed at four, 12 and 24 weeks after injection. At final follow-up the treatment group had a significantly greater improvement in the mean International Knee Documentation Committee score than the placebo group (16 points; -18 to 49, vs 8 points; -4 to 37, respectively; p = 0.03). The treatment group also had a significantly improved mean visual analogue score at final follow-up (-25; -85 to 34, vs -11 points; -51 to 25, respectively; p = 0.032). Both cohorts showed an improvement in Western Ontario and McMaster Osteoarthritis Index and Knee Injury and Osteoarthritis Outcome Scores, but these differences were not statistically significant. One patient had an anaphylactic reaction to the preservation medium, but recovered within 24 hours. All other adverse events were localised and resolved without further action. This technique may result in improved clinical outcomes, with the aim of slowing the degenerative process, leading to improvements in pain and function. However, imaging and direct observational studies are needed to verify cartilage regeneration. Nevertheless, this study provided a sufficient basis to proceed to further clinical testing.

Comparison between high and low molecular weight hyaluronates in knee osteoarthritis patients: open-label, randomized, multicentre clinical trial.

Efficacy and safety of high and low molecular weight hyaluronates in knee osteoarthritis patients were compared in a randomized, open-label trial. Patients in the high molecular weight hyaluronate group were treated once weekly for 3 weeks and in the low molecular weight group once weekly for 5 weeks. We evaluated weight-bearing pain, degree of flexion, swelling and knee tenderness; frequency and amount of rescue medication; patient and investigator global assessment of pain, and safety over 12 weeks after final injection of study medication. Significant improvements in pain and WOMAC-Likert scores were observed in both groups, but not between groups. Knee joint pain improvement was noted in both groups by patients and investigators during follow-up. Close correlation was observed between patient-and investigator-reported data. There was no significant difference in side-effects between the groups. In conclusion, the efficacy and safety of high and low molecular weight hyaluronate are similar.