SBIR-STTR Award

Resurfacing of articular cartilage with cold stored osteochondral allografts is employed clinically for repair of trauma and osteoarthritis-induced articular cartilage surface damage. Chondrocyte viability of transplanted articular cartilage is accepted as one of the determinants of outcome following osteochondral allograft transplantation. We have previously developed an ice-free vitrification method of cryopreservation that maintains excellent chondrocyte viability in animal model and human articular cartilage. The chondrocytes survive vitrification due to the absence of ice formation during cooling and warming of 1-3mL samples. However, it has not been possible to rewarm larger samples due to ice nucleation during rewarming that results in loss of chondrocyte viability. The innovation in this proposal relates to a new rewarming method that does not have the limitations of boundary convection warming that should be effective for samples up to 50mL in volume. This rewarming method utilizes radio frequency induced heating of magnetic iron nanoparticles. We will optimize nanowarming of full thickness osteocartilage storage for maintenance of both chondrocyte viability and extracellular matrix integrity. This objective will be developed in two specific aims to optimize nanowarming variables and scaleup from 5mL to 30-50mL volumes. Nanowarmed chondrocyte viability, chemistry, and biomaterial properties will be compared with untreated fresh control tissue. The nanowarming conditions that provides the best preservation of chondrocytes with minimal if any cartilage biomaterial changes will be selected for further investigation in vivo and translation to human cartilage in a subsequent Phase II SBIR application.

Public Health Relevance Statement:
Both literature review and an independent survey indicate that there is a significant need for a cartilage preservation solution for clinical and research applications that maintains both chondrocyte viability and cartilage biomaterial properties. The impact of this research will be optimized preservation of both articular cartilage chondrocytes and biomaterial properties making transplants more effective in vivo. Commercialization of this cartilage storage technology will result in increased utilization of banked allogeneic cartilage for reconstruction of articular cartilage defects in younger patients. The cryopreservation formulation and magnetic iron particles will also be commercialized for cartilage storage for research and future tissue engineered cartilage products.

Project Terms:
Allogenic; Allografting; Animal Model; Animals; articular cartilage; attenuation; Beavers; Biocompatible Materials; Biological Assay; Biological Preservation; biomaterial compatibility; bone; Caliber; Cardiovascular system; Cartilage; cartilage allograft; cartilage repair; Cell Culture Techniques; Cell Survival; Cells; Chemistry; Chondrocytes; Clinical; Clinical Research; commercialization; Complex; Convection; Coupling; Cryopreservation; Data; Defect; Degenerative polyarthritis; Effectiveness; Ensure; Evaluation; Extracellular Matrix; Family suidae; Food Processing; Formulation; Freedom; Freezing; Frequencies; Future; Harvest; Heating; Human; IACUC; Ice; implantation; In Vitro; in vivo; innovation; Investigation; Iron; Life; Magnetism; Maintenance; Metabolic; Methods; nanoparticle; osteochondral tissue; Outcome; particle; Patients; Permeability; Pharmacopoeias; Phase; Plants; Plug-in; Property; radio frequency; reconstruction; repaired; Research; response; Review Literature; Rewarming; Sample Size; Sampling; Small Business Innovation Research Grant; Surface; Surveys; Technology; Testing; Thick; Time; Tissue Engineering; Tissue Sample; Tissue Viability; Tissues; translation to humans; Transplantation; Trauma; Trypan Blue; United States

Resurfacing of articular cartilage with cold stored osteochondral allografts is employed clinically for repair of trauma and osteoarthritis-induced articular cartilage surface damage. Chondrocyte viability of transplanted articular cartilage is accepted as one of the determinants of outcome following osteochondral allograft transplantation. We have previously developed an ice-free vitrification method of cryopreservation that maintains excellent chondrocyte viability in animal model and human articular cartilage. The chondrocytes survive vitrification due to the absence of ice formation during cooling and warming of 1-3mL samples. However, it had not been possible to rewarm larger samples due to ice nucleation during rewarming that results in loss of chondrocyte viability prior to our Phase I studies. The innovation in this proposal relates to a new rewarming method that does not have the limitations of boundary convection warming that should be effective for samples up to 50mL in volume. This rewarming method utilizes radio frequency induced heating of magnetic iron nanoparticles. In Phase I we demonstrated the effectiveness of ice-free vitrification combined with nanowarming for large full thickness osteochondral tissues in 50 mL volumes in 80 seconds with maintenance of both chondrocyte viability and extracellular matrix integrity. In Phase II we propose development of a porcine model, further optimization of ice-free vitrification and nanowarming and in vivo evaluation in a porcine model in three specific aims. Nanowarmed chondrocyte viability, chemistry, and biomaterial properties will be compared with untreated fresh control tissues both before and after transplant. The nanowarming conditions that provides the best preservation of chondrocytes with minimal if any cartilage biomaterial changes will be selected for further investigation and translation to human cartilage in a subsequent Phase IIb SBIR application.

Public Health Relevance Statement:
Both literature review and an independent survey indicate that there is a significant need for a cartilage preservation solution for clinical and research applications that maintains both chondrocyte viability and cartilage biomaterial properties. The impact of this research will be optimized preservation of both articular cartilage chondrocytes and biomaterial properties making transplants more effective in vivo. Commercialization of this cartilage storage technology will result in increased utilization of banked allogeneic cartilage for reconstruction of articular cartilage defects in patients. The cryopreservation formulation and magnetic iron particles will also be commercialized for cartilage storage for research and future tissue engineered cartilage products.

NIH Spending Category:
Arthritis; Bioengineering; Biotechnology; Nanotechnology; Organ Transplantation; Osteoarthritis; Transplantation

Project Terms:
Allogenic; Allografting; Animal Model; articular cartilage; attenuation; Beavers; Biocompatible Materials; biomaterial compatibility; Cardiovascular system; Cartilage; cartilage cell; Cell Survival; Cells; Chemistry; Chondrocytes; Clinical; Clinical Research; commercialization; Contralateral; Convection; Coupling; Cryopreservation; Custom; Data; Defect; Degenerative polyarthritis; Development; Effectiveness; Ensure; Evaluation; Extracellular Matrix; Family suidae; Farming environment; Food Processing; Formulation; Freedom; Freezing; Frequencies; Future; Heating; Histology; Human; IACUC; Ice; Implant; implantation; improved; In Vitro; in vivo; in vivo evaluation; innovation; Investigation; Iron; Lead; Magnetism; Maintenance; Metabolic; Methods; Miniature Swine; Modeling; nanoparticle; nanowarming; osteochondral tissue; Outcome; particle; Patients; Permeability; Phase; phase 1 study; Plants; preclinical study; preservation; Property; radio frequency; reconstruction; Recovery; repaired; Research; response; Review Literature; Rewarming; Sample Size; Sampling; Small Business Innovation Research Grant; success; Surface; Surveys; Technology; Testing; Thick; Time; Tissue Engineering; Tissue Viability; Tissues; translation to humans; transplant model; Transplantation; Trauma