Study on the feasibility of tissue engineering cartilage constructed by cartilage particle and bionic sol matrix composite

Abstract

Objective To explore the feasibility of constructing tissue engineered cartilage by using the cartilage particle as seed cells and collagenⅡ- hyaluronic acid- chondroitin sulfate(COLⅡ- HA- CS) bionic matrix sol. Methods Cartilage particles were acquired from 5 months old Guizhou minipig knee joint; particles with different diameters were acquired by sieve filtration.The proliferation and adhesion of chondrocytes were evaluated at day 1,3,and 7.The bionic sol matrix at final concentration of 10 g/L was prepared by dissolving COLⅡ- HA- CS(mass ratio:8.0∶1.5∶0.5)in 0.15 mol/L HCl; The quality of prepared gel was examined.The cartilage particles with the diameter of strongest proliferation rate was seeded into COLⅡ/ HA/CSsol mix at 5×107/L the in vitro results were evaluated by Living cell immunofluorescence and histological observation after 1 week. Results Cartilage particles with different sizes all developed visible chondrocytes adherence and proliferation, and particle size less than 120μm showed strongest chondrocytes proliferation ability, the difference is statistically significant(P< 0.05). Neutralized COLⅡ/HA/CS bionic sol matrix showed good flowability at low temperature;and transferred to gel at 37 ℃for 6 min.After 1 week the cultured cartilage particles/gel complex showed transparent chondroid appearance, smooth surface and certain level of flexibility; Immunofluorescence observation showed good chondrocytes activity; Histological observation demonstrated chondrocytes were evenly distributed with good viability. Conclusion It reached the best proliferation of chondrocytes when particle size was less than 120μm; Cartilage particles and COLⅡ/ HA/CSbionic matrix sol are feasible to culture and construct bionic cartilage in vitro. Key words: Cartilage particles; Bionic matrix sol; Tissue engineering cartilage