Comparative characterization of decellularized renal scaffolds for tissue engineering
| Autor: | Krithika Hariharan, Andreas Kurtz, Petra Reinke, Harald Stachelscheid, Michael Westphal, Bella Rossbach, Iris Fischer, Imran Ullah, Nicole Bethke |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Octoxynol Swine medicine.medical_treatment Cell Detergents Induced Pluripotent Stem Cells Biomedical Engineering Bioengineering Kidney Biomaterials Glycosaminoglycan Extracellular matrix 03 medical and health sciences Tissue engineering Precursor cell medicine Animals Humans Glycosaminoglycans Decellularization Tissue Engineering Chemistry Epithelial Cells Cell biology Extracellular Matrix 030104 developmental biology medicine.anatomical_structure Cytokine Collagen |
| Zdroj: | Biomedical materials (Bristol, England). 12(4) |
| ISSN: | 1748-605X |
| Popis: | Native extracellular matrix (ECM) provides scaffolds for tissue engineering with natural architecture and biochemical composition. Maintaining the native ECM in decellularized tissues provides cues for cells, which promote their tissue specific arrangement and function. Several approaches have been used to decellularize ECM from the kidney in order to reestablish renal tissue but their comparability is hampered because methods for decellularization and assessment of ECM vary widely. Therefore, we applied a standardized immersion protocol to decellularize porcine kidney tissue with three detergents Triton X-100, SDS and sodium deoxycholate (SDC) at variable temperatures. For comparative analysis decellularization efficacies, structural preservation, composition and cell attachment and viability were analyzed. Structural ECM-conservation is strongly dependent on decellularization temperature, while preservation of glycosaminoglycans (GAG), collagens and cytokines was affected by the detergents used. GAG and collagens were best maintained by 1% SDS at 4 °C, whereas cytokines were best maintained in 1% SDC at 4 °C. Viability and attachment of human induced pluripotent stem cell derived renal precursor cells were best in SDC-ECM and thus not associated with the degree of GAG and collagen maintenance but the cytokine preservation. Based on structural and functional characteristics, we developed a scoring system that allows intra- and inter-study comparison of decellularization strategies. Application of the scoring system to our experimental data showed that decellularization with 1% SDS at 4 °C provided the highest structural and composition scores, while 1% SDC at 4 °C had lower structural and composition but a significantly better cell performance score. Inclusion of multiple published studies in the scoring matrix for comparison identified the highest structural and composition scores when decellularization was performed with SDS at low concentration, for a short period of time and at low temperature. Furthermore, the scoring system indicated that cell attachment and viability cannot be concluded from any other parameter and should therefore always be included in evaluation of decellularization strategies. |
| Databáze: | OpenAIRE |
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