| BackgroundCartilage defects caused by congenital malformation,tumor,trauma,and other causes are common clinical diseases.Due to the dense structure of cartilage tissue,no vascular tissue,and low density of chondrocytes,the healing ability of cartilage tissue is extremely limited.Traditional treatment methods include autologous cartilage transplantation,autologous chondrocyte transplantation,allogeneic cartilage transplantation,and micro fracture,which have some shortcomings and their clinical application is limited.In recent years,with the rapid development of tissue engineering,it is expected to become an effective method for the treatment of cartilage defects.As a dual biomimetic material based on bioactive substances and spatial structure,Acellular Cartilage Matrix(ACM)can simulate the extracellular environment of natural cartilage and provide enough space for cell attachment,proliferation,and nutrient exchange.Also,ACM has good histocompatibility,low cytotoxicity,and certain mechanical properties.This makes cartilage acellular matrix an ideal material for cartilage tissue engineering and has a unique advantage in repairing cartilage defects.However,due to the dense structure of cartilage tissue and the low diffusion efficiency of acellular reagents in tissue,it is necessary to increase the action time and concentration of acellular reagents to improve the degree of acellular.But the mechanical properties of acellular cartilage will decrease significantly.And previous studies have shown that changing the concentration of ACM can change the porosity and pore size of the scaffolds.Therefore,further research is needed to optimize the concentration and preparation scheme of ACM scaffolds,and to explore the effects of ACM concentration on the mechanical strength,pore size,and porosity of ACM scaffolds,to provide the experimental basis and reference for the construction of tissue engineering cartilage.ObjectiveTo compare the physical properties of three-dimensional porous scaffolds with different concentrations of ACM;To select a suitable plan for the construction of tissue engineering cartilage was selected.Subject and Method1.Subject:2 bovine knee joints without any osteoarthropathy.2.Method:1)The cartilage particles were obtained by sampling,freezing,and crushing bovine knee cartilage.2)ACM from bovine articular cartilage was obtained by trypsin digestion,nuclease digestion,washing,and leaning of cartilage particles.3)Three-dimensional porous scaffolds were constructed with different concentrations of ACM:group A(100%),group B(60%),group C(30%).4)Gross observation,histological evaluation,DNA quantitative detection,microstructure observation,and measurement,and physical properties of the obtained scaffold materials.5)Statistical analysis of the measurement results.Results1)Histology showed that there was no obvious cell morphology and no residual cell fragments in ACM scaffolds in each group.HE staining and Sirius red staining showed red staining,indicating that collagen was retained to a certain extent.2)DNA quantitative detection showed that the concentration of residual DNA was 1.103±0.174ng/mg,and the acellular cells were completely acellular.3)The results of scanning electron microscope showed that the pore diameter of stents in group A,B,and C decreased significantly with the increase of ACM concentration,and there were significant differences among groups.4)Physical parameters:the stent density,yield limit,and Young's modulus of groups A,B,and C were positively correlated with ACM concentration,while water absorption and porosity were negatively correlated with ACM concentration.ConclusionThe three-dimensional porous scaffolds constructed with ACM at 100%and 60%concentration can be used as an alternative for the construction of tissue engineering cartilage with high water absorption,porosity,pore diameter,and mechanical properties close to normal cartilage tissue. |
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