Porous chitosan-gelatin scaffold containing plasmid DNA encoding transforming growth factor-beta1 for chondrocytes proliferation.

Cartilage defects as a result of disease or injury have a very limited ability to heal spontaneously. Recently, tissue engineering and local therapeutic gene delivery systems have been paid much attention in the cartilage natural healing process. Gene-activated matrix (GAM) blends these two strategies, serving as local bioreactor with therapeutic agents expression and also providing a structural template to fill the lesion defects for cell adhesion, proliferation and synthesis of extracellular matrix (ECM). In the current study, we used chitosan-gelatin complex as biomaterials to fabricate three-dimensional scaffolds and plasmid DNA were entrapped in the scaffolds encoding transforming growth factor-beta1 (TGF-beta1), which has been proposed as a promoter of cartilage regeneration for its effect on the synthesis of matrix molecules and cell proliferation. The plasmid DNA incorporated in the scaffolds showed a burst release in the first week and a sustained release for the other 2 weeks. The gene transfectd into chondrocytes expresses TGF-beta1 protein stably in 3 weeks. The histological and immunohistochemical results confirmed that the primary chondrocytes cultured into the chitosan-gelatin scaffold maintained round and owned characters of high secretion of specific ECM. From this study, it can be concluded that this gene-activated chitosan-gelatins matrix has a potential in the application of cartilage defects regeneration.

[1]  J. Hughes,et al.  In Situ Gel Formulations for Gene Delivery: Release and Myotoxicity Studies , 2000, Pharmaceutical development and technology.

[2]  D. Rosen,et al.  Differentiation of rat mesenchymal cells by cartilage-inducing factor. Enhanced phenotypic expression by dihydrocytochalasin B. , 1986, Experimental cell research.

[3]  Y Ikada,et al.  Neovascularization effect of biodegradable gelatin microspheres incorporating basic fibroblast growth factor. , 1999, Journal of biomaterials science. Polymer edition.

[4]  L. Chandler,et al.  Matrix‐enabled gene transfer for cutaneous wound repair , 2000, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[5]  T. Ochiya,et al.  Biomaterials for gene delivery: atelocollagen-mediated controlled release of molecular medicines. , 2001, Current gene therapy.

[6]  A. Davis,et al.  FRESH OSTEOCHONDRAL ALLOGRAFTS FOR POST-TRAUMATIC OSTEOCHONDRAL DEFECTS OF THE KNEE , 1997 .

[7]  L Sedel,et al.  Effects of chitosan on rat knee cartilages. , 1999, Biomaterials.

[8]  J. Buckwalter Articular cartilage injuries. , 2002, Clinical orthopaedics and related research.

[9]  T. Ochiya,et al.  New delivery system for plasmid DNA in vivo using atelocollagen as a carrier material: the Minipellet , 1999, Nature Medicine.

[10]  W. B. van den Berg,et al.  Growth factors and cartilage repair. , 2001, Clinical orthopaedics and related research.

[11]  Tim Hardingham,et al.  Tissue engineering: chondrocytes and cartilage , 2002, Arthritis research.

[12]  V. Goldberg,et al.  Hyaluronan‐based polymers in the treatment of osteochondral defects , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[13]  S. De,et al.  Osteochondral defects in the knee. A treatment using lateral patella autografts. , 2000 .

[14]  R. LaPrade Autologous chondrocyte implantation was superior to mosaicplasty for repair of articular cartilage defects in the knee at one year. , 2003, The Journal of bone and joint surgery. American volume.

[15]  S. Goldstein,et al.  Combination of Local and Systemic Parathyroid Hormone Enhances Bone Regeneration , 2003, Clinical orthopaedics and related research.

[16]  T J Ebner,et al.  Sustained release of nerve growth factor from biodegradable polymer microspheres. , 1992, Neurosurgery.

[17]  K. Paigen,et al.  A simple, rapid, and sensitive DNA assay procedure. , 1980, Analytical biochemistry.

[18]  David J. Mooney,et al.  DNA delivery from polymer matrices for tissue engineering , 1999, Nature Biotechnology.

[19]  M. Longaker,et al.  Transforming Growth Factor Beta Superfamily Members: Role in Cartilage Modeling , 2000, Plastic and Reconstructive Surgery.

[20]  H J Mankin,et al.  Articular cartilage: tissue design and chondrocyte-matrix interactions. , 1998, Instructional course lectures.

[21]  P. Robbins,et al.  Vector systems for gene transfer to joints. , 2000, Clinical orthopaedics and related research.

[22]  Y. Tabata,et al.  Controlled release of plasmid DNA from cationized gelatin hydrogels based on hydrogel degradation. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[23]  L. Hangody,et al.  A prospective, randomised comparison of autologous chondrocyte implantation versus mosaicplasty for osteochondral defects in the knee. , 2004, The Journal of bone and joint surgery. British volume.

[24]  A. Domard,et al.  Relation between the physicochemical characteristics of collagen and its interactions with chitosan: I. , 1993, Biomaterials.

[25]  O. Johnell,et al.  Effect of transforming growth factor‐β and platelet‐derived growth factor‐BB on articular cartilage in rats , 1996, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[26]  D. Hungerford,et al.  Chitosan supports the expression of extracellular matrix proteins in human osteoblasts and chondrocytes. , 2000, Journal of biomedical materials research.

[27]  R Langer,et al.  Controlled and modulated release of basic fibroblast growth factor. , 1991, Biomaterials.

[28]  D. E. Ashhurst,et al.  The expression of collagen mRNAs in normally developing neonatal rabbit long bones and after treatment of neonatal and adult rabbit tibiae with transforming growth factor-β2 , 1995, The Histochemical Journal.

[29]  M. Ochi,et al.  Current concepts in tissue engineering technique for repair of cartilage defect. , 2001, Artificial organs.

[30]  K. Yip,et al.  Controlled release of plasmid DNA , 1997 .

[31]  T. Niidome,et al.  Gene Therapy Progress and Prospects: Nonviral vectors , 2002, Gene Therapy.

[32]  L. Barrett,et al.  Sustained Effects of Gene-Activated Matrices after CNS Injury , 2001, Molecular and Cellular Neuroscience.

[33]  W. Mark Saltzman,et al.  Controlled DNA Delivery Systems , 1999, Pharmaceutical Research.

[34]  W. B. van den Berg,et al.  Stimulation of articular cartilage repair in established arthritis by local administration of transforming growth factor-beta into murine knee joints. , 1998, Laboratory investigation; a journal of technical methods and pathology.

[35]  Jeffrey Bonadio,et al.  Localized, direct plasmid gene delivery in vivo: prolonged therapy results in reproducible tissue regeneration , 1999, Nature Medicine.

[36]  J. Massagué,et al.  Transforming growth factor-beta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. , 1986, The Journal of biological chemistry.

[37]  J. Vacanti,et al.  Tissue engineering : Frontiers in biotechnology , 1993 .

[38]  Gregory F Payne,et al.  Enzyme-catalyzed gel formation of gelatin and chitosan: potential for in situ applications. , 2003, Biomaterials.