In vitro evaluation of bovine pericardium after a soft decellularization approach for use in tissue engineering

Abstract

Abbreviations: ECM, extracellular matrix; FBS, fetal bovine serum; FITC, fluorescein isothiocyanate; H&E, hematoxylin and eosin; hASC, human adipose-derived stem cell; PBS, phosphate-buffered saline; PFA, paraformaldehyde; PUCPR, Pontifical Catholic University of Paraná; SDS, sodium dodecyl sulfate; SEM, scanning electron microscopy. AbstractPericardial membrane derived from bovine heart tissues is a promising source of material for use in tissue-engineering applications. However, tissue processing is required for its use in humans due to the presence of animal antigens. Therefore, the purpose of this study was to evaluate the structural integrity and biocompatibility of the bovine pericardium (BP) after a soft decellularization process with a 0.1% sodium dodecyl sulfate (SDS) solution, with the aim to remove xenoantigens and preserve extracellular matrix (ECM) bioactivity. The decellularization process promoted a mean reduction of 77% of the amount of DNA in the samples in which cell nuclei staining was undetectable. The ECM content was maintained as mostly preserved after decellularization as well as its biomechanical properties. In addition, the decellularization protocol has proven to be efficient in removing the xenoantigen alphagal, which is responsible for immune rejection. The decellularized BP was noncytotoxic in vitro and allowed human adipose-derived stem cell (hASC) adhesion. Finally, after 7 days in culture, the tissue scaffold became repopulated by hASCs, and after 30 days, the ECM protein pro-collagen I was seen in the scaffold. Together, these characteristics indicated that soft BP decellularization with 0.1% SDS solution allows the acquirement of a bioactive scaffold suitable for cell repopulation and potentially useful for regenerative medicine. K E Y W O R D Sbovine pericardium, decellularization, extracellular matrix, tissue engineering

Structural assessments in decellularized extracellular matrix of porcine semilunar heart valves: Evaluation of cell niches

“…Quantification of the nuclear material residue was performed as previously validated by our group . In brief, the DNA was extracted from 10.0 mg of freeze‐dried tissue using proteinase K in extraction buffer (1% SDS, Tris HCl 50 mmol/L, NaCl 100 mmol/L, EDTA 0.1 mol/L—Sigma), followed by nucleic acid precipitation by the addition of acetate buffer (potassium acetate, 3.0 mol/L) and isopropanol p.a.…”

“…The evaluations of ECMS cytocompatibility were carried out by seeding and culturing human adipose stromal cells (ADSC; Human Adipose‐Derived Stem Cells—Lonza Clonetics ® ) in the scaffolds for 7 days. The protocol for cell seeding and culturing was previously described by our group . Briefly, decellularized ECMS samples (n = 3) were cut into pieces of approximately 3 mm 2 and kept overnight at 4°C in DMEM supplemented with 100 U/mL penicillin, 1 mg/mL streptomycin.…”

“…Compound, Sakura‐4583) to be sliced at 15 μm for analysis with nuclei DAPI staining. Additionally, the seeded scaffolds were treated for SEM analysis as described previously . The samples were analyzed under a JEOL JSM 6010PLUS‐LA (Microscopy Facility RPT07C PDTIS/Carlos Chagas Institute, FIOCRUZ‐PR) at 20 kV acceleration voltage.…”

Processed Tissue–Derived Extracellular Matrices: Tailored Platforms Empowering Diverse Therapeutic Applications

“…While native dECM‐based constructs have been widely explored as scaffolds for cardiac and skeletal muscle regeneration, the employment of different biomanufacturing methods to this material have expanded its applicability in this field, generally allowing the control over scaffold properties, dimensions, delivery route, and its simple supplementation with favorable factors and cells. Processed dECM‐based engineered cardiac tissues have been investigated most extensively for the treatment of ischemic injury.…”

“…The use of tissue‐derived decellularized extracellular matrices (dECM) has been dated back to the beginning of the 20th century, and was since followed and improved by an increasing number of groups revealing the endless potential of this unique biomaterial, isolated from the heart, liver, pancreas, brain, bone, small intestinal submucosa, and urinary bladder, among other tissues . Numerous dECM‐based products have been successfully translated into the clinic for a myriad of medical applications, as has been excellently reviewed elsewhere …”