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1.
World J Orthop ; 6(10): 754-61, 2015 Nov 18.
Article in English | MEDLINE | ID: mdl-26601056

ABSTRACT

In this editorial, the authors tried to evaluate the present state of cellular therapy in orthopedic field. The topics the authors try to cover include not only the clinical trials but the various research areas as well. Both the target diseases for cellular therapy and the target cells were reviewed. New methods to activate the cells were interesting to review. Most advanced clinical trials were also included because several of them have advanced to phase III clinical trials. In the orthopedic field, there are many diseases with a definite treatment gap at this time. Because cellular therapies can regenerate damaged tissues, there is a possibility for cellular therapies to become disease modifying drugs. It is not clear whether cellular therapies will become the standard of care in any of the orthopedic disorders, however the amount of research being performed and the number of clinical trials that are on-going make the authors believe that cellular therapies will become important treatment modalities within several years.

2.
BMC Musculoskelet Disord ; 16: 141, 2015 Jun 10.
Article in English | MEDLINE | ID: mdl-26059549

ABSTRACT

BACKGROUND: A localized non-surgical delivery of allogeneic human chondrocytes (hChonJ) with irradiated genetically modified chondrocytes (hChonJb#7) expressing transforming growth factor-ß1 (TGF-ß1) showed efficacy in regenerating cartilage tissue in our pre-clinical studies and human Phase I and II clinical trials. These previous observations led us to investigate the molecular mechanisms of the cartilage regeneration. METHODS: Genetically modified TGF-ß1preprotein was evaluated by monitoring cell proliferation inhibition activity. The effect of modified TGF-ß1 on chondrocytes was evaluated based on the type II collagen mRNA levels and the amount of glycosaminoclycan (GAG) formed around chondrocytes, which are indicative markers of redifferentiated chondrocytes. Among the cartilage matrix components produced by hChonJb#7 cells, type II collagen and proteoglycan, in addition to TGF-ß1, were also tested to see if they could induce hChonJ redifferentiation. The ability of chondrocytes to attach to artificially induced defects in rabbit cartilage was tested using fluorescent markers. RESULTS: Throughout these experiments, the TGF-ß1 produced from hChonJb#7 was shown to be equally as active as the recombinant human TGF-ß1. Type II collagen and GAG production were induced in hChonJ cells by TGF-ß1 secreted from the irradiated hChonJb#7 cells when the cells were co-cultured in micro-masses. Both hChonJ and hChonJb#7 cells could attach efficiently to the defect area in the rabbit cartilage. CONCLUSIONS: This study suggests that the mixture (TG-C) of allogeneic human chondrocytes (hChonJ) and irradiated genetically modified human chondrocytes expressing TGF-ß1 (hChonJb#7) attach to the damaged cartilage area to produce type II collagen-GAG matrices by providing a continuous supply of active TGF-ß1.


Subject(s)
Chondrocytes/pathology , Chondrogenesis , Collagen Type II/metabolism , Glycosaminoglycans/metabolism , Transforming Growth Factor beta1/metabolism , Animals , COS Cells , Cartilage Diseases/metabolism , Cartilage Diseases/pathology , Cartilage Diseases/therapy , Cartilage, Articular/metabolism , Cartilage, Articular/pathology , Cartilage, Articular/surgery , Cell Adhesion , Chlorocebus aethiops , Chondrocytes/drug effects , Chondrocytes/transplantation , Chondrogenesis/drug effects , Coculture Techniques , Disease Models, Animal , Femur/metabolism , Femur/pathology , Femur/surgery , Genetic Therapy/methods , Hep G2 Cells , Humans , Infant , Mutation , Rabbits , Regeneration , Transfection , Transforming Growth Factor beta1/genetics , Transforming Growth Factor beta1/pharmacology , Up-Regulation
3.
Cytotherapy ; 14(2): 247-56, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22242865

ABSTRACT

UNLABELLED: BACKGROUND AIMS. TissueGene-C (TG-C) represents a cell-mediated gene therapy for localized delivery of allogeneic chondrocytes expressing transforming growth factor (TGF)-ß1 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-ß1-expressing chondrocytes (hChonJb#7) in order to help fill in the defect and as target cells for the actions of the expressed TGF-ß1. 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.


Subject(s)
Chondrocytes/cytology , Chondrocytes/transplantation , Genetic Therapy/adverse effects , Knee Injuries/therapy , Patient Safety , Transforming Growth Factor beta1/metabolism , Aged , Cartilage/pathology , Female , Humans , Injections, Intra-Articular , Knee Injuries/pathology , Knee Joint/pathology , Male , Middle Aged , Regeneration , Transforming Growth Factor beta1/genetics
4.
Curr Gene Ther ; 11(3): 218-28, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21453283

ABSTRACT

There have been major changes since the incidents of leukemia development in X-SCID patients after the treatments using retroviral gene therapy. Due to the risk of oncogenesis caused by retroviral insertional activation of host genes, most of the efforts focused on the lentiviral therapies. However, a relative clonal dominance was detected in a patient with ß-thalassemia Major, two years after the subject received genetically modified hematopoietic stem cells using lentiviral vectors. This disappointing result of the recent clinical trial using lentiviral vector tells us that the current and most advanced vector systems does not have enough safety. In this review, various safety features that have been tried for the retroviral gene therapy are introduced and the possible new ways of improvements are discussed. Additional feature of chromatin insulators, co-transduction of a suicidal gene under the control of an inducible promoter, conditional expression of the transgene only in appropriate target cells, targeted transduction, cell type-specific expression, targeted local administration, splitting of the viral genome, and site specific insertion of retroviral vector are discussed here.


Subject(s)
Genetic Therapy/methods , Retroviridae/genetics , Chromatin/metabolism , Genes, Transgenic, Suicide/genetics , Genetic Therapy/trends , Genetic Vectors/genetics , Humans , Mutagenesis, Insertional , beta-Thalassemia/genetics , beta-Thalassemia/therapy
5.
Cytotherapy ; 12(3): 384-93, 2010 May.
Article in English | MEDLINE | ID: mdl-20370350

ABSTRACT

BACKGROUND AIMS: The aim was to evaluate cartilage regeneration in animal models involving induced knee joint damage. Through cell-mediated gene therapy methods, a cell mixture comprising a 3:1 ratio of genetically unmodified human chondrocytes and transforming growth factor beta-1 (TGF-beta1)-secreting human chondrocytes (TG-C), generated via retroviral transduction, resulted in successful cartilage proliferation in damaged regions. METHODS: Non-clinical toxicology assessments for efficacy, biodistribution and local/systemic toxicity of single intra-articular administration of the cell mixture in mice, rabbits and goats was conducted. RESULTS: Administration of the mixture was tolerated well in all of the species. There was evidence of cartilage proliferation in rabbits and goats. As an additional precautionary step, the efficacy of TGF-beta1 secretion in irradiated human chondrocytes was also demonstrated. CONCLUSIONS: Four studies in rabbits and goats demonstrated the safety and efficacy of TG-C following direct intra-articular administration in animal models involving induced knee joint damage. Based on these pre-clinical studies authorization has been received from the USA Food and Drug Administration (FDA) to proceed with an initial phase I clinical study of TG-C for degenerative arthritis.


Subject(s)
Cell Transplantation , Chondrocytes/physiology , Retroviridae , Transduction, Genetic , Transforming Growth Factor beta1/metabolism , Animals , Cartilage/cytology , Cartilage/pathology , Cartilage/physiology , Cells, Cultured , Chondrocytes/cytology , Gene Transfer Techniques , Goats , Humans , Male , Mice , Mice, SCID , Rabbits , Regeneration/physiology , Retroviridae/genetics , Retroviridae/metabolism , Transforming Growth Factor beta1/genetics
6.
Tissue Eng Part A ; 15(10): 2853-63, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19254139

ABSTRACT

Bone morphogenetic protein 2 (BMP2) was selected as a transgene to regenerate osteoporotic bone defects after several BMPs were tested using a bone formation study in nude mice. Human chondrocytes were transduced with a BMP2-containing retroviral vector, and single clones were selected. The cells were characterized over numerous passages for growth and BMP2 expression. The single clones were irradiated and tested for viability. BMP2 expression lasted for 3 weeks before dying off completely after approximately 1 month. Irradiated and non-irradiated transduced chondrocytes successfully healed fractures in osteoporotic rats induced by ovariectomy. The osteoinducing effect of irradiated cells was better than that of their non-irradiated counterparts or a chondrocytes-only control. This study showed that delivering BMP2 from the transduced and irradiated chondrocytes could be an effective and safe method of repairing osteoporotic bone fractures.


Subject(s)
Bone Morphogenetic Protein 2/metabolism , Chondrocytes/metabolism , Chondrocytes/physiology , Fractures, Bone/therapy , Wound Healing/physiology , Animals , Cell Line , Cell- and Tissue-Based Therapy/methods , Chondrocytes/cytology , Enzyme-Linked Immunosorbent Assay , Female , Gamma Rays , Humans , Mice , Mice, Nude , NIH 3T3 Cells , Osteogenesis/radiation effects , Rats , Rats, Sprague-Dawley , Wound Healing/radiation effects
7.
Tissue Eng ; 11(9-10): 1516-26, 2005.
Article in English | MEDLINE | ID: mdl-16259606

ABSTRACT

The purpose of this study was to investigate the efficacy of cartilage regeneration when using a mixture of transforming growth factor-beta1 (TGF-beta1)-producing human chondrocytes (hChon-TGF-beta1) and primary human chondrocytes (hChon) ("mixed cells"), compared with either hChon-TGF-beta1 or hChon cells alone. Specifically, mixed cells or hChon cells were first injected intradermally into the backs of immune-deficient nude mice to test the feasibility of cartilage formation in vivo. Both the mixed cells and the hChon-TGF-beta1 cells alone induced cartilage formation in nude mice, whereas hChon cells alone did not. To further test the efficacy of the cells in generating cartilage, an artificially induced partial thickness defect of the femoral condyle of a rabbit knee joint was loaded with hChon-TGF-beta1 cells with or without mixing additional untransduced hChon cells, and hyaline cartilage regeneration was observed at 4 or 6 weeks. The efficiency of complete filling of the defect and the quality of tissue generated after implanting were evaluated on the basis of a histological grading system modified from O'Driscoll et al. (J. Bone Joint Surg. 70A, 595, 1988). Significantly, mixed cells (14.2 +/- 0.9) produced significantly better results than hChon-TGF-beta1 (9.0 +/- 1.7) or hChon (8.0 +/- 1.8) cells alone. Histological and immunohistochemical staining of the newly repaired tissues produced after treatment with either mixed cells or hChon-TGF-beta1 cells alone showed hyaline cartilage- like characteristics. These results suggest that the implantation of mixed cells may be a clinically efficient method of regenerating hyaline articular cartilage.


Subject(s)
Chondrocytes/cytology , Chondrocytes/physiology , Chondrogenesis , Hyaline Cartilage/physiology , Transforming Growth Factor beta/biosynthesis , Animals , Cells, Cultured , Chondrocytes/metabolism , Chondrocytes/transplantation , Collagen Type II/metabolism , DNA/genetics , Enzyme-Linked Immunosorbent Assay , Femur/injuries , Genetic Vectors , Histological Techniques , Humans , Hyaline Cartilage/injuries , Hyaline Cartilage/metabolism , Immunohistochemistry , Injections, Subcutaneous , Mice , Mice, Nude , Plasmids , Rabbits , Recombinant Proteins/metabolism , Retroviridae/genetics , Reverse Transcriptase Polymerase Chain Reaction , Time Factors , Transforming Growth Factor beta/analysis , Transforming Growth Factor beta/genetics , Transforming Growth Factor beta1 , Transplantation, Heterologous
8.
Tissue Eng ; 11(1-2): 310-8, 2005.
Article in English | MEDLINE | ID: mdl-15738684

ABSTRACT

One of the most important factors in the production of cartilage is transforming growth factor beta1 (TGF-beta1). To obtain sustained release of TGF-beta1, a cell-mediated gene therapy technique was introduced. We infected chondrocytes with a retroviral vector carrying the TGF-beta1 gene. The single clone derivative showed sustained TGF-beta1 secretion. It also showed constitutive type II collagen expression. Whereas the TGF-beta1 protein itself is unable to induce formation of cartilage in vivo, human chondrocytes engineered to express a retroviral vector encoding TGF-beta1 showed cartilage formation in vivo when cells were injected into nude mice intradermally. These data suggest that cell-mediated gene therapy using TGF-beta1 as a transgene would be a promising treatment for osteoarthritis.


Subject(s)
Cell Differentiation , Chondrocytes/metabolism , Genetic Therapy/methods , Transforming Growth Factor beta/metabolism , Animals , Cartilage, Articular/drug effects , Cartilage, Articular/metabolism , Cell Transplantation , Cells, Cultured , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrocytes/transplantation , Collagen Type II/drug effects , Collagen Type II/metabolism , Feasibility Studies , Genetic Vectors , Humans , Mice , Mice, Nude , Retroviridae/genetics , Transforming Growth Factor beta/genetics , Transforming Growth Factor beta/pharmacology , Transforming Growth Factor beta1 , Transgenes , Transplantation, Heterologous
9.
J Virol Methods ; 121(2): 127-36, 2004 Nov.
Article in English | MEDLINE | ID: mdl-15381349

ABSTRACT

Retroviral vectors have been widely used in gene therapy due to their simple genomic structure and high transduction efficiency. We report a construction of Moloney murine sarcoma virus (MoMSV) and Moloney murine leukemia virus (MoMLV) hybrid-based retroviral vectors with significantly improved efficiency of transgene expression after stable incorporation into the host genome. In these vectors, the residual gag gene coding sequence located in the extended region of packaging signal was removed. These vectors, therefore, contain no coding sequence for the gag, pol, or env gene that can be used for homologous recombination with sequences introduced in the packaging system for a recombinant competent retrovirus (RCR) generation. A strong splice acceptor site obtained from the exon/intron junction of either the chimpanzee EF1-alpha gene or the human CMV major immediate early gene was placed downstream of the MoMSV packaging signal (Psi), significantly improving the efficiency of transgene expression. The 5' LTR U3 sequence was replaced with an extended human CMV major immediate early gene enhancer/promoter for a strong expression of full-length messages from the viral backbone, helping to maintain high levels of viral titer. These newly developed retroviral vectors should facilitate RCR-free gene transfer with significantly improved efficacy in clinical gene therapy trials.


Subject(s)
Genetic Vectors , Moloney murine leukemia virus/genetics , Moloney murine sarcoma virus/genetics , Transgenes , Animals , Cytomegalovirus/genetics , Enhancer Elements, Genetic , Gene Expression , Genetic Engineering , Genetic Therapy , Immediate-Early Proteins/genetics , Mice , NIH 3T3 Cells , Peptide Elongation Factor 1/genetics , Promoter Regions, Genetic , Virus Replication
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