G. Lehoczky (Basel, CH)
University Hospital of Basel Departments of Surgery and of BiomedicinePresenter Of 1 Presentation
P075 - Preclinical Studies Using Rapidly Isolated Human Nasal Chondrocytes for a Single-stage Arthroscopic Cartilage Regeneration Therapy
Abstract
Purpose
Nasal chondrocytes (NCs) have a higher and more reproducible chondrogenic capacity than articular chondrocytes and the engineered cartilage tissue they generate in vitro was demonstrated to be safe in clinical applications. Here we aimed at determining the feasibility for a single-stage application of NCs for cartilage regeneration under minimally invasive settings.
Methods and Materials
Fresh nasal septal cartilage grafts were digested according to a new, rapid protocol. Cell yield, viability, proliferative and chondrogenic capacity were assessed. The unexpanded (p0) NCs were delivered in a degradable polyethylene glycol (PEG) gel enriched with pooled human platelet lysate (hPL) extract, in low density (300.000 cells/mL gel), and cultured in vitro and in vivo. Histological, immunofluorescence and biochemical analyses were carried out at day 0, after 1 and 4 weeks. For in vivo tests, an ectopic human osteochondral model was used to create full thickness cartilage lesions of 4 mm diameter, filled with low-density, p0 NCs in hPL-PEG gel (without in vitro pre-culturing), and implanted subcutaneously in nude mice for 8 weeks.
Results
"Rapidly isolated" NCs show similar viability, chondrogenic and proliferative capacity compared to standard digestion; however, cell yield is even higher after the new protocol (mean 4.8x106 ± 3.5x106 vs. 3.3x106 ±2.7x106 cells/gram of tissue, p= 0.0135). NCs embedded in hPL-PEG gels efficiently proliferated (total of 6.33 ±0.51 population doublings in 4 weeks) and generated tissue rich in hyalin cartilage-specific glycosaminoglycans (GAG) and type II collagen, with GAG/DNA ratio of mean 12.3 (±7.9). NC-hPL-PEG gels developed into hyaline-like cartilage tissues upon ectopic in vivo implantation and morphologically integrated with surrounding native cartilage and bone tissues in situ.
Conclusion
Our findings collectively indicate that this could be a feasible strategy for cartilage repair and now requires further validation in orthotopic in vivo models.
Presenter Of 1 Presentation
P075 - Preclinical Studies Using Rapidly Isolated Human Nasal Chondrocytes for a Single-stage Arthroscopic Cartilage Regeneration Therapy
Abstract
Purpose
Nasal chondrocytes (NCs) have a higher and more reproducible chondrogenic capacity than articular chondrocytes and the engineered cartilage tissue they generate in vitro was demonstrated to be safe in clinical applications. Here we aimed at determining the feasibility for a single-stage application of NCs for cartilage regeneration under minimally invasive settings.
Methods and Materials
Fresh nasal septal cartilage grafts were digested according to a new, rapid protocol. Cell yield, viability, proliferative and chondrogenic capacity were assessed. The unexpanded (p0) NCs were delivered in a degradable polyethylene glycol (PEG) gel enriched with pooled human platelet lysate (hPL) extract, in low density (300.000 cells/mL gel), and cultured in vitro and in vivo. Histological, immunofluorescence and biochemical analyses were carried out at day 0, after 1 and 4 weeks. For in vivo tests, an ectopic human osteochondral model was used to create full thickness cartilage lesions of 4 mm diameter, filled with low-density, p0 NCs in hPL-PEG gel (without in vitro pre-culturing), and implanted subcutaneously in nude mice for 8 weeks.
Results
"Rapidly isolated" NCs show similar viability, chondrogenic and proliferative capacity compared to standard digestion; however, cell yield is even higher after the new protocol (mean 4.8x106 ± 3.5x106 vs. 3.3x106 ±2.7x106 cells/gram of tissue, p= 0.0135). NCs embedded in hPL-PEG gels efficiently proliferated (total of 6.33 ±0.51 population doublings in 4 weeks) and generated tissue rich in hyalin cartilage-specific glycosaminoglycans (GAG) and type II collagen, with GAG/DNA ratio of mean 12.3 (±7.9). NC-hPL-PEG gels developed into hyaline-like cartilage tissues upon ectopic in vivo implantation and morphologically integrated with surrounding native cartilage and bone tissues in situ.
Conclusion
Our findings collectively indicate that this could be a feasible strategy for cartilage repair and now requires further validation in orthotopic in vivo models.