Antonios G Mikos
Rice University
Project start date: 2008-04-10
Project end date: 2013-02-28
Sponsored Links Excellgen http://Excellgen.com
IN SITU HARDENING CELLULAR CONSTRUCTS FOR CRANIOFACIAL BONE REGENERATION
Antonios G Mikos
Rice University, 6100 Main, Houston, Tx 77005-1892
Grant 5R01DE017441-03 from National Institute Of Dental & Craniofacial Research
Keywords: Area; Biocompatible Materials; Biomaterials; Birth Defects; Blood Coagulation Factor IV; Body Temperature; Bone; Bone Formation; Bone Regeneration; Bone Tissue; Bone and Bones; Bones and Bone Tissue; Ca++ element; Calcium; Calcium Binding; Calcium-Binding Domain; Calvaria; Cancers; Cell Differentiation; Cell Differentiation process; Cell Survival; Cell Viability; Cells; Cephalic; Characteristics; Chemicals; Coagulation Factor IV; Common Rat Strains; Congenital Abnormality; Congenital Anatomic Abnormality; Congenital Anatomical Abnormality; Congenital Defects; Congenital Deformity; Congenital Malformation; Cranial; Defect; Drug Formulations; Encapsulated; Factor IV; Formulation; Formulations, Drug; Gel; Goals; Hydrogels; In Situ; In Vitro; Injectable; Injection of therapeutic agent; Injections; Injury; Investigation; Kinetic; Kinetics; Knowledge; Lead; Malignant Neoplasms; Malignant Tumor; Mammals, Rats; Marrow; Mechanics; Methods; Modeling; Molecular Genetic Abnormality; Natural regeneration; Operation; Operative Procedures; Operative Surgical Procedures; Osteogenesis; Patients; Pb element; Polymers; Porosity; Property; Property, LOINC Axis 2; QOL; Quality of life; Rat; Rattus; Regeneration; Site; Staging; Stromal Cells; Surgical; Surgical Interventions; Surgical Procedure; Swelling; Testing; Tissue Engineering; Tissue Grafts; Wound Healing; Wound Repair; ing; biocompatibility; biomaterial compatibility; bone; bone repair; calvarial; clinical applicability; clinical application; craniofacial; craniofacial repair; craniofacies; cross-link; crosslink; density; engineered tissue; experience; functional group; heavy metal Pb; heavy metal lead; in vivo; malignancy; mineralization; neoplasm/cancer; novel; osteoblast differentiation; regenerate; regenerative; surgery; tissue grafting; tissue repair
Project start date: 2008-04-10
Project end date: 2013-02-28
Budget start date: 1-MAR-2010
Budget end date: 28-FEB-2011
PFA/PA: PA-07-070
5R01DE017441-03 (2010): $326362
Grants awarded to Antonios G Mikos
IN SITU POLYMERIZABLE GELS FOR DENTAL TISSUE ENGINEERING
Antonios G Mikos, Professor
Chemical Engineeringrice University
6100 S Main
houston, Tx 770051892
Grant 5R01DE013031-04 from National Institute Of Dental & Craniofacial Research IRG: ZHL1
Abstract: The ultimate goal of this research proposal is to develop novel in situ polymerizable, biodegradable hydrogels for treating dental defects with guided bone regeneration. Based on a novel highly unsaturated linear copolyester developed in our laboratory, poly(propylene fumarate-co- ethylene glycol) (P(PF-co-EG)), these hydrogels will have a covalently bound osteopontin (OPN) peptide for enhancing bone growth while diminishing fibrous infiltration. The peptide will be appropriately spaced from the hydrogel network to provide for sufficient interaction between the peptide and osteoblast receptors. The unsaturated linear P(PF-co-EG)-co-OPN peptide copolyester will be crosslinked in situ via an addition polymerization with N-vinyl pyrrolidi. Additional components of an injectable formulation will include gelatin porogen, a radical polymerization initiator, and an accelerator. A three-step approach will be followed to engineer optimal in situ polymerizable hydrogels using novel video microscopy and image analysis techniques developed in our laboratory. The first step will involve the production of thin films of hydrogels exhibiting minimal or no adhesion of either fibroblasts or osteoblasts. After choosing the polymer compositions that eliminate non-specific cell adhesion, bioactive thin films will then be produced and evaluated. Finally, the hydrogel/peptide formulations that minimize adhesion and migration of fibroblasts while promoting adhesion and migration of osteoblasts will be tested using a three-dimensional cell/polymer model developed in our laboratory to determine the optimal porosity and pore size of hydrogel scaffolds for maximum bone formation in vitro. The efficacy of the optimized peptide composite formulation to form new bone in an orthotopic site to restore osseous continuity will be tested using an acute segmental critical-size long-bone defect model in rats. New bone formation and graft consolidation to host bone will be assessed radiographically as a function of time. Light and fluorescence microscopy will allow quantitative and qualitative analyses of the extent, character and dynamics of new bone formation. The mechanical properties of the grafted bones will be measured to verify restoration of the integrity of the reconstituted region under functional loads. The proposed project will provide clinically valuable information regarding new injectable dental biomaterials for guided bone regeneration. It will lead to a major advance in treating dental defects using biocompatible and biodegradable polymers which are becoming particularly important because of the renewed concern for the safety of non-degradable implants and the potential for disease transmission with allografts
Keywords: biomaterial development /preparation, bone regeneration, bone transplantation, dental material, gel, tissue engineering biomaterial evaluation, cell adhesion, cell migration, copolymer, dental implant, fibroblast, oral facial restoration material, osteoblast, osteopontin, polymer fluorescence microscopy, laboratory rat, light microscopy, medical implant science, radiography
Project start date: 1998-09-01
Project end date: 2003-06-30
5R01DE013031-04 (2001): $286440
5R01DE013031-03 (2000): $274413
5R01DE013031-02 (1999): $274413
INJECTABLE BIOMATERIALS FOR BONE TISSUE ENGINEERING
Antonios G Mikos, Professor
Rice University
6100 S Main
houston, Tx 770051892
Grant 5R01AR044381-04 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases IRG: SB
Abstract: The ultimate goal of this research proposal is to develop injectable orthopaedic biomaterials for treating skeletal defects with guided bone ingrowth into biodegradable polymer conduits. The biomaterial is based on a highly unsaturated linear polyester with covalently bound peptide moieties for enhancing cellular attachment to the polymer. The biomaterial also serves as a carrier for bone growth factors to stimulate the bone regeneration cascade. The proposed work involves attachment of the RGDS adhesive peptide sequence to a novel polymer, poly(propylene fumarate) (PPF), developed in our laboratory. The peptide sequence will be appropriately spaced from the polymeric backbone to provide for sufficient interaction between the peptide and stromal osteoblast receptors. The fabrication of new injectable biodegradable composite formulations will be investigated for the development of polymeric conduits for guided bone regeneration. The composite formulation will be based on PPF-co-RGDS. This unsaturated linear polyester will be crosslinked via an addition polymerization with N-vinyl pyrrolidi. Additional components of the composite formulation will include a water soluble salt for initial porosity, a calcium phosphate matrix for formation of an osteoconductive scaffold for new bone growth, a microparticle carrier for the bone growth factor TGF-Beta1, a radical polymerization initiator and an accelerator. The combined effects of the peptide surface concentration and growth factor dose on new bone formation will be determined in vitro using a three-dimensional biodegradable polymer/stromal osteoblast model developed in our laboratory. The efficacy of the optimized peptide/growth factor composite to form new bone in an orthotopic site to restore osseous continuity will be tested using an acute segmental critical-size long- bone defect model in rats. New bone formation and graft consolidation to host bone will be assessed radiographically as a function of time. Light and fluorescence microscopy will allow quantitative and qualitative analyses of the extent, character and dynamics of new bone formation. The mechanical properties of the grafted bones will be measured to verify restoration of the integrity of the reconstituted region under functional loads. The proposed project will provide clinically valuable information regarding new injectable orthopaedic biomaterials for bone repair and replacement. It will lead to a major advance in treating skeletal defects using biocompatible and biodegradable polymers which are becoming particularly important because of the renewed concern for the safety of non-degradable implants and the potential for disease transmission with allografts
Keywords: biomaterial development /preparation, biomaterial evaluation, biotransformation, bone regeneration, polymer, tissue engineering calcium phosphate, cell adhesion molecule, crosslink, osteoblast, polymerization, pyridi, synthetic peptide, transforming growth factor X ray spectrometry, fluorescence microscopy, laboratory rat, light microscopy, nuclear magnetic resonance spectroscopy, radiography
Project start date: 1996-12-15
Project end date: 2001-11-30
5R01AR044381-04 (2000): $192121
5R01AR044381-03 (1999): $185485
1R01AR044381-01 (1997): $172937
Injectable Cellular Composites For Cartilage Engineering
Antonios G Mikos, Professor
Rice University 6100 S Main Houston, Tx 770051892
Grant 5R01AR048756-04 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases IRG: ZRG1
Abstract: The ultimate goal of this research proposal is to develop an injectable cell-polymer composite that will aid in the repair of tissue in osteochondral defects. Based on a novel hydrogel material developed in our laboratory, oligo(poly(ethylene glycol) fumarate), these constructs will be photocrosslinked in situ in the presence of mesenchymal stem cells (MSCs). A three-step approach will be followed to engineer optimal injectable constructs using a combination of cells, biodegradable scaffolds, and spatially and temporally graded release of a growth factor. The first step will be to determine how hydrogel mesh size (distance between crosslinks) affects differentiation of embedded MSCs in vitro and in a rabbit osteochondral defect model. The hydrogel system will then be modified through the synthesis of crosslinking molecules with peptide sequences that are degradable by matrix metalloproteinases found in articular cartilage. The optimal initial mesh size, determined previously, will be used as the basis for study of the effect of different crosslinker concentrations on degradation of the hydrogel due to differentiation of embedded MSCs in vitro and in vivo. Finally, a bilayered construct will be created from the biodegradable hydrogel with the optimal crosslinker concentration in which a gradient of TGF-beta1, released in a controlled manner in response to enzymes produced as neotissue forms, is established to promote cartilage formation in the top half of the construct and bone formation in the lower half. Efficacy of tissue formation from this construct will be tested in vitro and in vivo. The proposed project will provide clinically valuable information regarding new composite constructs for improved repair of osteochondral defects, thus providing a method to generate cartilage repair tissue, which will not degenerate over time, a major limitation with current techniques.
Keywords: articular cartilage, biotechnology, injection /infusion, tissue engineering, tissue support frame, biodegradable product, cell differentiation, crosslink, enzyme mechanism, mesenchyme, metalloprotein, osteochondritis, polyethylene glycol, polymer, stem cell, transforming growth factor, enzyme linked immunosorbent assay, histology, immunocytochemistry, laboratory rabbit, morphometry, tissue /cell culture
Project start date: 2003-04-01
Project end date: 2008-03-31
5R01AR048756-04 (2006): $311886
5R01AR048756-03 (2005): $347384
5R01AR048756-02 (2004): $347671
1R01AR048756-01A1 (2003): $347971
Sponsored Links Excellgen http://Excellgen.com
2R01AR048756-06A2 (2010): $334183
BONE REGENERATION BY OSTEOBLAST TRANSPLANTATION
Antonios G Mikos, Professor
Rice University 6100 S Main Houston, Tx 770051892
Grant 5R01AR042639-10 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases IRG: ZRG1
Abstract: The ultimate goal of this research project is to develop osteoinductive osteoblast/polymer constructs for bone regeneration and repair. We will investigate the effects of flow and fluid shear, as well as scaffold pore architecture and surface composition on the proliferation and osteogenic differentiation of marrow stromal cells seeded in three-dimensional biodegradable polymer scaffolds and cultured in vitro in a flow perfusion bioreactor developed in our laboratory. We will determine the effects of these parameters on the secretion and deposition of osteoinductive and angiogenic growth factors, such as bone morphogenetic protein-2 and vascular endothelial growth factor, the secretion of bone matrix proteins, such as osteocalcin, and the deposition of mineralized extracellular matrix. We will also investigate the effects of fluid shear and in vitro culture period on mineralized tissue formation after implantation of the cell/polymer constructs in the mesentery of rats and will elucidate the contributions of the mineralized extracellular matrix generated in vitro and the differentiated cells existing in the cell/polymer constructs on the construct osteoinductivity in vivo. Lastly, we will evaluate the ability of optimized cell/polymer constructs created in vitro under flow perfusion culture and loaded with deposited osteoinductive extracellular matrix and secreted angiogenic factors to form new bone in an orthotopic site and restore osseous continuity using a critical size cranial defect model in rats. New bone formation will be assessed radiographically and histomorphometrically as a function of time. Light and fluorescence microscopy will allow quantitative and qualitative analyses of the extent, character, and dynamics of new bone formation. The mechanical properties of the grafted bones will be measured to verify restoration of the integrity of the reconstituted region. This project will provide clinically valuable information regarding new tissue-inducing orthopaedic biomaterials which are becoming particularly important because of the renewed concern for the safety of non-degradable implants and the potential for disease transmission with allografts.
Keywords: biomaterial development /preparation, bone regeneration, bone transplantation, cell transplantation, osteoblast, polymer, tissue engineering, biomaterial compatibility, biomaterial evaluation, bone marrow, bone morphogenetic protein, extracellular matrix, fluidity, mechanical stress, osteocalcin, perfusion, tissue support frame, vascular endothelial growth factor, bioengineering /biomedical engineering, histology, immunocytochemistry, laboratory rat, microscopy, radioimmunoassay, tissue /cell culture
Project start date: 1996-04-15
Project end date: 2007-06-30
5R01AR042639-10 (2006): $204712
5R01AR042639-09 (2005): $209638
5R01AR042639-08 (2004): $247982
2R01AR042639-06A1 (2002): $247987
FLOW PERFUSION BIOREACTOR FABRICATION OF BIOACTIVE POLYMER/ECM HYBRID CONSTRUCTS
Antonios G Mikos
Rice University, 6100 Main, Houston, Tx 77005-1892
Grant 5R01AR057083-02 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases
Abstract: The overall goal of the proposed research is to apply flow perfusion bioreactor culture of mesenchymal stem cells (MSCs) toward the fabrication of bioactive, biodegradable polymer/extracellular matrix (ECM) hybrid constructs for tissue engineering. The present proposal focuses upon the development and application of this innovative approach to fabricate bi-layered constructs for the repair of osteochondral defects. It is hypothesized that flow perfusion bioreactor culture of MSCs upon electrospun poly(5-caprolactone) (PCL) nanofiber scaffolds in medium augmented with osteogenic or chondrogenic supplements will produce bioactive polymer/ECM hybrid constructs with an ECM component containing osteogenic or chondrogenic factors, respectively, and that the character of the ECM is influenced by the culture conditions and the properties of the scaffold. It is further hypothesized that, following decellularization and implantation, these acellular osteogenic and chondrogenic polymer/ECM hybrid constructs will direct the differentiation of host progenitor cells toward the generation of bone or cartilage tissue, respectively. It is hypothesized that scaffolds composed of nanofibers will result in the deposition of ECM containing more osteogenic or chondrogenic factors than ECM deposited on microfiber scaffolds, as nanofiber scaffolds more closely approximate the scale of native ECM molecules and, due to the smaller pore size, produce increased shear stress at a given flow rate. The effects of the applied shear stress, the architecture of the scaffold (microfibers vs. nanofibers), and the culture conditions on the generated osteogenic and chondrogenic hybrid constructs will be investigated by monitoring the presence of molecules characteristic of the respective tissue types (e.g., collagen type I for bone and collagen type II for cartilage). Further, the culture duration for ECM generation will be modulated to examine the effect of the maturity of the ECM component of the decellularized hybrid constructs upon the osteoblastic and chondrocytic differentiation of subsequently seeded MSCs in vitro (as measured by differentiation markers such as alkaline phosphatase activity, calcium and glycosaminoglycan content, and the presence of collage types I and II) and upon tissue formation in vivo in an osteochondral defect in a rabbit model (as measured by histology and histomorphometry). Finally, acellular bi-layered polymer/ECM hybrid constructs will be fabricated with an osteogenic layer and a chondrogenic layer and then implanted in a rabbit osteochondral defect model to assess the potential of the constructs to influence the spatial differentiation of progenitor cells of the host to form bone and cartilage in the respective layers. This novel approach to fabricate acellular bioactive degradable tissue engineering constructs containing ECM rich in growth factors produced by cells under engineered conditions in vitro presents tremendous potential for application in the guided regeneration of a wide range of tissues. A significant clinical need exists for novel implant materials capable of promoting the repair and regeneration of injured or compromised tissues, such as damaged articular cartilage. Indeed, as cartilage has a limited natural capacity to repair itself, damage to articular cartilage and underlying bone often leads to considerable clinical problems that afflict million of people worldwide, including pain, limited mobility and osteoarthritis. The research project presented in this proposal seeks to apply advanced cell culture technologies to fabricate biologically active implant materials that can promote cells within the recipient to regenerate or repair specific damaged tissues, in this case articular cartilage and underlying bone
Relevance: A significant clinical need exists for novel implant materials capable of promoting the repair and regeneration of injured or compromised tissues, such as damaged articular cartilage. Indeed, as cartilage has a limited natural capacity to repair itself, damage to articular cartilage and underlying bone often leads to considerable clinical problems that afflict million of people worldwide, including pain, limited mobility and osteoarthritis. The research project presented in this proposal seeks to apply advanced cell culture technologies to fabricate biologically active implant materials that can promote cells within the recipient to regenerate or repair specific damaged tissues, in this case articular cartilage and underlying bone
Project start date: 2009-04-01
Project end date: 2014-03-31
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
PFA/PA: PA-07-070
5R01AR057083-02 (2010): $330095
PROMOTION OF ALVEOLAR SOCKET HEALING WITH BIOPOLYMERS
Antonios G Mikos
Rice University, 6100 Main, Houston, Tx 77005-1892
Grant 5R01DE015164-05 from National Institute Of Dental & Craniofacial Research
Abstract: The objective of this proposal is to promote the formation of durable bone tissue in critical size alveolar socket defects by the implantation of a biologically active, biodegradable polymer scaffold. Preliminary work in our laboratory has shown that osteogenesis within the healing alveolar bone of a rabbit tooth extraction socket model, a subcritical size defect, correlates with an increase in specific growth factor localizations. We plan to promote bone formation in a critical size alveolar socket defect, which does not heal naturally, by recapitulating those events observed in the healing, subcritical extraction socket defect. The key steps we plan to recapitulate are vascular growth, progenitor cell migration, and bone formation, as these events are most significant in the healing of bone tissue. This will be achieved by implanting a biodegradable poly(propylene fumarate-co-ethylene glycol) [P(PF-co-EG)] based hydrogel scaffold, developed in our laboratory, that also serves as a carrier for the controlled delivery of specific growth factors. First, a biomimetic P(PF-co-EG) hydrogel scaffold will be developed to release fibroblastic growth factor-2, an early factor in wound healing and angiogenesis, during the initial stages of healing to promote vascular growth and maintenance within the defect site. Second, the scaffold will be designed to encourage progenitor cell migration into the defect site by the incorporation of an adhesive osteopontin-derived peptide selective for osteoblastic cells, while enhancing the formation of a wound healing tissue matrix by preventing soft tissue invasion. Third, the scaffold will release transforming growth factor-beta1, a potent differentiating factor, to encourage bone formation by osteoblastic differentiation of progenitor cells within the defect site. The amount of bone formation and quality of the newly formed bone will be assessed by light microscopy, histomorphometry of immunohistochemically stained sections, and micro-computed tomography analysis. This project will provide clinically valuable information regarding new tissue-inducing dental biomaterials for restoring oral function and esthetics to individuals who are afflicted with critical size alveolar socket defects
Keywords: 2ar peptide; Adhesions; Adhesives; Alveolar; Alveolar ridge; Alveolus Dentalis; Animal Model; Animal Models and Related Studies; Animal growth regulators, transforming growth factors; Biocompatible Materials; Biological Preservation; Biology; Biomaterials; Biomimetics; Biopolymers; Blood Vessels; Body Tissues; Bone; Bone Formation; Bone Regeneration; Bone Tissue; Bone and Bones; Bone plates; Bones and Bone Tissue; CAT Scan, X-Ray; CAT scan; CT X Ray; CT scan; Cell Communication and Signaling; Cell Differentiation; Cell Differentiation process; Cell Locomotion; Cell Migration; Cell Movement; Cell Signaling; Cells; Cellular Migration; Clinical; Complex; Computed Tomography; Computerized Axial Tomography (Computerized Tomography); Computerized Tomography, X-Ray; Defect; Dental; Dental Alveolus; Dose; EMI scan; Environment; Esthetics; Eta-1 protein; Eta-1-Op protein; Event; FGF-2; FGF2; Fibroblast Growth Factor 2; Fibroblast Growth Factor, Basic; Fracture; GFAC; Gelatin; Generalized Growth; Goals; Growth; Growth Agents; Growth Factor; Growth Factors, Proteins; Growth Substances; HBGF-2; Healed; Heparin-Binding Growth Factor 2; Heparin-Binding Growth Factor Class II; Hydrogels; Implant; Individual; Intracellular Communication and Signaling; Jaw; Kinetic; Kinetics; Laboratories; Length; Maintenance; Maintenances; Mammals, Rabbits; Methods; Mimetics, Biological; Modeling; Molecular; Mother Cells; Motility; Motility, Cellular; Numbers; Oral; Oryctolagus cuniculus; Osteogenesis; Parodontosis; Pathology; Peptide Signal Sequences; Peptides; Periodontal Diseases; Preservation, Biologic; Preservation, Biological; Prevention; Procidentia; Progenitor Cells; Programs (PT); Programs [Publication Type]; Prolapse; Property; Property, LOINC Axis 2; Prostate Epithelial Cell Growth Factor; Prostatropin; Ptosis; Rabbit, Domestic; Rabbits; Signal Peptide; Signal Sequences; Signal Sequences, Peptide; Signal Transduction; Signal Transduction Systems; Signaling; Site; Staging; Staining method; Stainings; Stains; Stem cells; TGF-Beta 1; TGF-Beta1; TGFB1; TGFB1 protein, human; Testing; Tissue Growth; Tissues; Tomodensitometry; Tomography, Xray Computed; Tooth; Tooth Extraction; Tooth Socket; Tooth structure; Transforming Growth Factor Beta 1; Transforming Growth Factors; Trauma; Tumor Growth Factors; Vascularization; Week; Work; Wound Healing; Wound Repair; X-Ray Computed Tomography; alveolar bone; alveolar bone of maxilla; alveolar process of maxilla; alveolar socket; alveolar supporting bone; angiogenesis; bFGF; base; biodegradable polymer; biological signal transduction; bioresorbable polymer; bone; bone fracture; bone repair; bone sialoprotein 1; bone sialoprotein I; catscan; cell motility; computed axial tomography; computerized axial tomography; computerized tomography; degradable polymer; design; designing; early T-lympocyte activation-1 protein; healing; human TGFB1 protein; implantation; light microscopy; migration; model organism; neovascularization; ontogeny; osteopontin; p(PF-co-EG); periodontal disorder; periodontium disease; periodontium disorder; phosphoprotein I, 2aR; poly(propylene fumarate-co-ethylene glycol); preservation; prevent; preventing; programs; protein signal sequence; scaffold; scaffolding; secreted phosphoprotein 1; sialoprotein 1; size; socket wall; soft tissue; teeth; tissue repair; transforming growth factor beta1; vascular
Project start date: 2004-04-01
Project end date: 2010-03-31
Budget start date: 1-APR-2008
Budget end date: 31-MAR-2010
5R01DE015164-05 (2008): $0
5R01DE015164-04 (2007): $275340
5R01DE015164-03 (2006): $283564
Sponsored Links Excellgen http://Excellgen.com
5R01DE015164-02 (2005): $290388
1R01DE015164-01A1 (2004): $290388
DEVELOP INJECTABLE And DEGRADABLE BIOMATERIALS BASED ON POLYPROPYLENE FUMARATE
Antonios G Mikos, Professor
University Of Washington Office Of Sponsored Programs Seattle, Wa 98105
Grant 5P41RR001296-180042 from National Center For Research Resources
Abstract: The aim of this project is to develop injectable and degradable biomaterials for orthopaedic applications. ESCA will be used to determine the surface composition of these poly(propylene fumarate)-based biomaterials. The possible unsaturated carbonyl group existing at the surface would allow direct modification of the surface with peptide chains. Parallel cell attachment studies will be done with the surface analysis.
Keywords: bioengineering /biomedical engineering, biological product, biomaterial, biomedical resource, health care, prosthesis, technology /technique, medical rehabilitation related tag
REGULATED OSTEOCHONDROGENESIS OF HUMAN MESENCHYMAL STEM CELLS USING GENE DELIVERY
Antonios G Mikos
Rice University, 6100 Main, Houston, Tx 77005-1892
Grant 5R21AR056076-02 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases
Abstract: Regeneration of articular cartilage damaged either by disease or injury is a complex problem that remains a significant clinical challenge despite extensive orthopaedic research. Although there are numerous analgesics, therapeutic strategies and surgical procedures developed to address this health concern, most of the therapies are short-lived and non-remedial. The global objective of this project is to repair osteochondral defects with a multilayered construct presenting spatially controlled chondrogenic and osteogenic properties and generated from human mesenchymal stem cells (hMSCs). To this end, the following specific aims are proposed (1) to develop 3D controlled release systems that deliver plasmid encoded transcription factors at rates and concentrations appropriate for osteogenic and chondrogenic differentiation of MSCs and (2) to evaluate the controlled release systems created in Specific Aim 1 for their potential to generate bi-layered chondrogenic and osteogenic constructs. Osteochondral differentiation of MSCs in 3D scaffolds will be achieved through gene delivery of transcription factors Sox-5, Sox-6, Sox-9 (Sox trio) for chondrogenesis, and Runx-2 for osteogenesis. Each of these plasmids will be complexed with a novel polymeric gene delivery vector developed in our laboratory (a conjugate of branched polyethylenimine and hyaluronic acid) to increase transfection efficiency. Initially, the effect of plasmid concentration and exposure duration on osteochondral differentiation will be examined to identify the appropriate target release profiles. Electrospun, co-axial fiber mesh scaffolds will then be fabricated with the vector-plasmid complexes embedded in the core of the polymer fibers and well-established fabrication variables will be utilized to achieve the desired release profiles. Evaluation of the individual layers for the envisioned final bi-layered construct will be based upon the osteogenic and chondrogenic differentiation of the seeded MSCs and the generation of extracellular matrix similar to the native tissue. The studies will involve hMSCs to evaluate the initial feasibility of the proposed approach for ultimate clinical application in humans, while parallel studies will be conducted using rabbit MSCs to evaluate the appropriateness of the envisioned application of the rabbit animal model in the translational pre-clinical development of this novel approach to osteochondral tissue regeneration. The novelty of this proposal is that it utilizes (a) MSCs to circumvent the traditional problems associated with the limited availability and maintenance of differentiated cell types, (b) transcription factors encoded in plasmids to provide a broader phenotypic induction of MSCs and (c) polymeric scaffolds to provide a three dimensional support lattice for cell proliferation/differentiaton and the controlled release of gene delivery vectors. Although this proposal addresses a specific clinical need, the impact of these studies is not limited to cartilage or even orthopaedic applications. The principles proposed herein provide a means to circumvent the problems associated with the isolation and maintenance of differentiated cells. Transcription factors will potentially facilitate the use of hMSCs in a number of tissue engineering applications. Furthermore, the novel scaffold fabrication technique permits regulated delivery of multiple gene delivery vectors while providing a 3D support for cell growth. These techniques have wide-ranging applications in tissue engineering and can potentially be used to create multilayer scaffolds that mimic the zonal architecture of cartilage and other complex tissues. This project proposes an innovative technology and approach for creating osteochondral constructs for the repair of damaged articular cartilage and subchondral bone. A combination of developmental biological signals will be utilized in a novel controlled release system to deliver differential signals to induce human mesenchymal stem cells into appropriate osteochondral phenotypes. This system will increase the scope of techniques available to tissue engineers for generating anisotropic tissues with multiple cell types
Keywords: Address; Affect; Age; Analgesic Agents; Analgesic Drugs; Analgesic Preparation; Analgesics; Animal Model; Animal Models and Related Studies; Anodynes; Antinociceptive Agents; Antinociceptive Drugs; Architecture; Articulation; Biological; Body Tissues; Bone; Bone Formation; Bone and Bones; Bones and Bone Tissue; Caliber; Cartilage; Cartilage, Articular; Cartilagenous Tissue; Cell Communication and Signaling; Cell Growth in Number; Cell Multiplication; Cell Proliferation; Cell Signaling; Cell Survival; Cell Viability; Cell-Extracellular Matrix; Cells; Cellular Expansion; Cellular Growth; Cellular Proliferation; Chondrocytes; Chondrogenesis; Classification; Clinical; Complex; Defect; Development; Developmental Gene; Diameter; Disease; Disorder; ECM; Engineering / Architecture; Evaluation; Extracellular Matrix; Fiber; Fibroblasts; GFAC; Gene Delivery; Gene Expression; Gene, Developmental; Generations; Genes, Reporter; Growth Agents; Growth Factor; Growth Factors, Proteins; Growth Substances; Health; Human; Human, General; Hyaluronic Acid; Individual; Injury; Intracellular Communication and Signaling; Joints; Kinetic; Kinetics; Laboratories; Length; Life; Limb Development; Maintenance; Maintenances; Mammals, Rabbits; Man (Taxonomy); Man, Modern; Measures; Mediator; Mediator of Activation; Mediator of activation protein; Mesenchymal Progenitor Cell; Mesenchymal Stem Cells; Methods; Methods and Techniques; Methods, Other; Natural regeneration; Operation; Operative Procedures; Operative Surgical Procedures; Organ; Orthopedic; Orthopedic Surgical Profession; Orthopedics; Oryctolagus cuniculus; Osteogenesis; Pathway interactions; Pattern; Phenotype; Plasmid Cloning Vector; Plasmid Vector; Plasmids; Polymers; Programs (PT); Programs [Publication Type]; Property; Property, LOINC Axis 2; Rabbit, Domestic; Rabbits; Regeneration; Reporter Genes; Research; SOX9 protein; Signal Transduction; Signal Transduction Systems; Signaling; Structure; Structure of articular cartilage; Supporting Cell; Surgical; Surgical Interventions; Surgical Procedure; System; System, LOINC Axis 4; Systematics; Techniques; Therapeutic; Time; Tissue Engineering; Tissues; Transfection; analgesia; articular cartilage; base; biological signal transduction; bone; cell growth; cell type; chemical conjugate; clinical applicability; clinical application; controlled release; design; designing; disease/disorder; elderly patient; engineered tissue; experiment; experimental research; experimental study; innovative technologies; interest; model organism; multipotent cell; new approaches; novel; novel approaches; novel strategies; novel strategy; older patient; osteochondral; osteochondral repair; osteochondral tissue; osteogenic; pathway; plasmid DNA; pre-clinical; preclinical; programs; public health relevance; regenerate; regenerate new tissue; regenerating damaged tissue; regenerative; repair; repaired; research study; scaffold; scaffolding; sox-9; surgery; tissue regeneration; transcription factor; vector
Relevance: This project proposes an innovative technology and approach for creating osteochondral constructs for the repair of damaged articular cartilage and subchondral bone. A combination of developmental biological signals will be utilized in a novel controlled release system to deliver differential signals to induce human mesenchymal stem cells into appropriate osteochondral phenotypes. This system will increase the scope of techniques available to tissue engineers for generating anisotropic tissues with multiple cell types
Project start date: 2008-09-20
Project end date: 2010-08-31
Budget start date: 1-SEP-2009
Budget end date: 31-AUG-2010
PFA/PA: PA-06-181
5R21AR056076-02 (2009): $196911
1R21AR056076-01A1 (2008): $163361
BONE REGENERATION BY OSTEOBLAST TRANSPLANTATION
Antonios G Mikos, Professor
Rice University 6100 S Main Houston, Tx 770051892
Grant 5R29AR042639-05 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases IRG: ORTH
Abstract: Adapted from investigator s ) The overall goal of this research proposal is to regenerate bone by osteoblast transplantation using synthetic biodegradable polymers. The polymer will provide temporary scaffolding to attached osteoblasts which will subsequently secrete their own extracellular matrix to form a completely natural bone tissue. Two major research thrusts are required to develop technology for cell transplantation. The first deals with appropriate cell culture techniques, and the second addresses the scaffold material and structure. The two are closely related, and examination of one issue will influence the other. The objective of this project is to integrate the two areas and develop polymer-osteoblast constructs to create new bone tissue. The three-dimensional (3-D) culture of osteoblasts attached on poly(alpha-hydroxy ester) foam scaffolds will be attempted to investigate polymer composition and foam morphology effects on the polymer degradation and osteoblast function. Alkaline phosphatase activity, collagen synthesis, and formation of mineral deposits by cultured osteoblasts in 3-D will be used to optimize the polymer scaffold. New processing methods have been developed in laboratory to fabricate 3-D, open-cell poly(alpha-hydroxy ester) foams with reproducible pore sizes and porosities. Preliminary studies also showed that poly(alpha- hydroxy esters) provided a suitable substrate for osteoblast culture and migration. The investigative team will seek to optimize the polymer scaffold and cell seeding density in order to improve phenotypic development of cultured osteoblasts. They will then investigate the efficacy of syngeneic polymer-primary osteoblast and polymer-fresh bone marrow constructs to regenerate bone using the rat segmental long-bone defect model. New bone formation and graft consolidation to host bone will be assessed radiographically as a function of time. Light and fluorescence microscopy is intended to allow quantitative and qualitative analyses of the extent, character and dynamics of new bone formation. The mechanical properties of the grafted bones will be determined to verify restoration of the integrity of the reconstituted region under functional loads. It is suggested by that the proposed project will provide clinically valuable information regarding new cell-based strategies for bone repair and replacement. It is further speculated that it will lead to a major advance in treating skeletal defects using biocompatible and biodegradable polymers. These are becoming particularly important because of the renewed concern for the safety of non-degradable implants and the potential for disease transmission, especially AIDS, with allografts. This work is intended to furnish the technology for the development of other polymer-cell constructs for use in joint surgery, and plastic and reconstructive surgery. Finally, it will attempt to determine the potential of gene therapy with transplantation of genetically-altered bone cells for treating skeletal diseases.
Keywords: biomaterial development /preparation, bone regeneration, bone transplantation, cell transplantation, osteoblast, polymer, tissue engineering, biomaterial compatibility, biomaterial evaluation, mechanical stress, tissue support frame, laboratory rat
Project start date: 1996-04-15
Project end date: 2001-03-31
5R29AR042639-05 (2000): $113534
5R29AR042639-04 (1999): $107993
5R29AR042639-03 (1998): $78656
Sponsored Links Excellgen http://Excellgen.com
5R29AR042639-02 (1997): $104613
Antonios G Mikos
Rice University
Project start date: 2009-04-01
Project end date: 2014-03-31
Injectable Cellular Composites For Cartilage Engineering
Antonios G Mikos, Professor
Chemical Engineeringrice University
6100 S Main
houston, Tx 770051892
Grant 5R01AR048756-05 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases IRG: ZRG1
Project start date: 2003-04-01
Project end date: 2008-09-30
5R01AR048756-05 (2007): $302570
BONE REGENERATION BY OSTEOBLAST TRANSPLANTATION
Antonios G Mikos, Professor
Rice University
6100 S Main
houston, Tx 770051892
Grant 1R29AR042639-01A3 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases IRG: ORTH
Project start date: 1996-04-15
Project end date: 2001-03-31
1R29AR042639-01A3 (1996): $99631