ABSTRACT
Based on the data of morphological analysis, we performed histological evaluation of rat tissue reaction to subcutaneous implantation of decellularized matrices of intrathoracic organs and tissues. Cell composition of the inflammatory infiltrate was analyzed, and the dynamics of macrophage and T and B lymphocyte content was assessed on days 7 and 14 of the experiment. It was found that the reaction to implantation depended not only on the quality of decellularization and efficiency of removal of antigen molecules, but also on the original histological structure and quality of preimplantation processing of the transplant.
Subject(s)
Diaphragm/ultrastructure , Extracellular Matrix/ultrastructure , Lung/ultrastructure , Tissue Engineering/methods , Tissue Scaffolds , Animals , Antigens, CD/immunology , Antigens, CD/metabolism , B-Lymphocytes/cytology , B-Lymphocytes/immunology , Biomarkers/metabolism , Deoxycholic Acid/chemistry , Deoxycholic Acid/pharmacology , Deoxyribonucleases/chemistry , Deoxyribonucleases/pharmacology , Diaphragm/cytology , Diaphragm/drug effects , Diaphragm/transplantation , Extracellular Matrix/chemistry , Heart/drug effects , Lectins, C-Type/immunology , Lectins, C-Type/metabolism , Lung/cytology , Lung/drug effects , Macrophages/cytology , Macrophages/immunology , Male , Mannose Receptor , Mannose-Binding Lectins/immunology , Mannose-Binding Lectins/metabolism , Rats , Rats, Wistar , Receptors, Cell Surface/immunology , Receptors, Cell Surface/metabolism , Skin , T-Lymphocytes/cytology , T-Lymphocytes/immunology , Transplantation, HomologousABSTRACT
Chronic renal failure (CRF) is one of the most challenging problems of contemporary medicine. Patients with chronic renal failure usually need renal replacement therapy as either hemodialysis, peritoneal dialysis or a kidney transplant. The latter is the most promising option for end-stage kidney disease. However, the shortage of donor organs, the complexity of their delivery, the difficulty in finding an immunologically compatible donor and the need for lifelong immunosuppression triggered advances in modern tissue engineering. In this field, the primary priority is focused on developing bioengineered scaffolds with subsequent recellularization with autologous cells. Using such constructs would allow for solving both ethical and immunological problems of transplantation. The aim of this pilot study was to develop a new method of renal decellularization using small laboratory animals. MATERIALS AND METHODS: The study investigated the morphological structure of the obtained decellularized matrix and quantitatively tested DNA residues in the resulting scaffold. We proposed a new biophysical method for assessing the matrix quality using the EPR spectroscopy and conducted experiments on the matrix recellularization with mesenchymal multipotent stem cells to estimate cytotoxicity, cell viability and metabolic activity. RESULTS: The obtained decellularized renal matrix retained the native tissue architecture after a complete removal of the cell material, had no cytotoxic properties and supported cell adhesion and proliferation. CONCLUSION: All the above suggests that the proposed decellularization protocol is a promising method to produce tissue-engineered kidney constructs with possible clinical application in the foreseeable future.
Subject(s)
Kidney Failure, Chronic/therapy , Kidney/anatomy & histology , Kidneys, Artificial , Tissue Engineering , Tissue Scaffolds , Animals , Cell Separation , Kidney/cytology , RatsABSTRACT
We modified the protocol of obtaining of biological scaffolds of rat lungs based on dynamic recording of specific resistivity of working detergent solution (conductometry) during perfusion decellularization. Termination of sodium deoxycholate exposure after attaining ionic equilibrium plateau did not impair the quality of decellularization and preserved structural matrix components, which was confirmed by morphological analysis and quantitative assay of residual DNA.
