Using Hemolysis as a Novel Method for Assessment of Cytotoxicity and Blood Compatibility of Decellularized Heart Tissues

Momtahan, Nima; Panahi, Tayyebeh; Poornejad, Nafiseh; Stewart, Michael G.; Vance, Brady R.; Struk, Jeremy A.; Castleton, Arthur A.; Roeder, Beverly L.; Sukavaneshvar, Sivaprasad; Cook, Alonzo D.

doi:10.1097/mat.0000000000000373

Cited by 16 publications

(17 citation statements)

References 33 publications

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“…Because hemocompatibility of any biomaterial is extremely important, we conducted the hemolysis assay to exclude the presence of residual toxic chemicals within the decellularized scaffolds. Momtahan et al have reported the efficacy of this assay to determine the blood compatibility of decellularized cardiac tissue . Our decellularized kidney showed a non‐hemolytic effect and good blood compatibility.…”

Section: Discussionmentioning

confidence: 64%

“…The hemolysis rate which may occur because of chemicals used during decellularization, was measured according Momtahan et al Briefly, fresh blood was collected from healthy live pigs at slaughterhouse and directly transferred to the laboratory. Erythrocytes were separated by centrifuging at 2000 g for 15 min, followed by dilution in 1× PBS to create an erythrocyte suspension with 2 × 10 9 cells mL −1 .…”

Section: Methodsmentioning

confidence: 99%

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Biocompatibility and hemocompatibility of efficiently decellularized whole porcine kidney for tissue engineering

Hussein

Saleh

Ahmed

et al. 2018

J Biomedical Materials Res

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Whole kidney decellularization is a promising approach in regenerative medicine for engineering a functional organ. The reaction of the potential host depends on the biocompatibility of these decellularized constructs. Despite the proven ability of decellularized kidney scaffolds to guide cell attachment and growth, little is known about biocompatibility and hemocompatibility of these scaffolds. Our aim is to prepare decellularized kidneys of a clinically relevant size and evaluate its biocompatibility and hemocompatibility. Porcine kidneys were cannulated via the renal artery, and then perfused with 0.1% sodium dodecyl sulfate solution. Hematoxylin and eosin as well as DAPI staining confirmed cellular clearance from native kidneys in addition to preservation of the microstructure. SEM confirmed the absence of any cellular content within the scaffold, which is maintained in a well-organized 3D architecture. Decellularized kidneys retained the intact renal vasculature upon examination with contrast radiography. The essential structural extracellular matrix molecules were well-preserved. Scaffolds were susceptible to enzymatic degradation upon collagenase treatment. Scaffolds showed a good hemocompatibility when exposed to porcine blood. Decellularization was efficient to remove 97.7% of DNA from native kidneys in addition to the immunogenic and pathogenic antigens. Scaffolds did not induce the human immune response in vitro. Decellularized kidneys were non-cytotoxic to pig kidney cells (PKs). PKs were able to grow and proliferate within the decellularized renal scaffolds with maintaining a higher function than cells grown as monolayers. Thus, we have developed a rapid decellularization technique for generating biocompatible kidney scaffolds that represents a step toward development of a transplantable organ. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2034-2047, 2018.

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Section: Discussionmentioning

confidence: 64%

Section: Methodsmentioning

confidence: 99%

Biocompatibility and hemocompatibility of efficiently decellularized whole porcine kidney for tissue engineering

Hussein

Saleh

Ahmed

et al. 2018

J Biomedical Materials Res

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“…Accumulated data showed that long-term SDS treatment destroys the structure of ECM, and limits the following adhesion, survival and proliferation of seeding cells [28, 29]. Based on published literatures, we employed pretreatment/SDS/TritonX-100 three-step-method with shorter SDS treatment to obtain sECM.…”

Section: Discussionmentioning

confidence: 99%

Skeletal Extracellular Matrix Supports Cardiac Differentiation of Embryonic Stem Cells: a Potential Scaffold for Engineered Cardiac Tissue

Hong

Yin

Sun

et al. 2018

Cell Physiol Biochem

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Background/Aims: Decellularized cardiac extracellular matrix (cECM) has been widely considered as an attractive scaffold for engineered cardiac tissue (ECT), however, its application is limited by immunogenicity and shortage of organ donation. Skeletal ECM (sECM) is readily available and shows similarities with cECM. Here we hypothesized that sECM might be an alternative scaffold for ECT strategies. Methods: Murine ventricular tissue and anterior tibial muscles were sectioned into 300 mm-thick, and then cECM and sECM were acquired by pretreatment/SDS/TritonX-100 three-step-method. Acellularity and morphological properties of ECM was assessed. SECM was recellularized with murine embryonic stem cells (mESCs) or mESC-derived cardiomyocytes (mESC-CMs), and was further studied by biocompatibility assessment, immunofluorescent staining, quantitative real-time PCR and electrophysiological experiment. Results: The relative residual contents of DNA, protein and RNA of sECM were comparable with cECM. The morphological properties and microstructure of sECM were similar to cECM. SECM supported mESCs to adhere, survive, proliferate and differentiate into functional cardiac microtissue with both electrical stimulated response and normal adrenergic response. Purified mESC-CMs also could adhere, survive, proliferate and form a sECM-based ECT with synchronized contraction within 6 days of recellularization. Conclusion: ECMs from murine skeletal muscle support survival and cardiac differentiation of mESCs, and are suitable to produce functional ECT patch. This study highlights the potential of patient specific of sECM to replace cECM for bioengineering ECT.

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“…There are different decellularizing agents such as sodium dodecyl sulfate (SDS), surfactants, triton X-100, enzymatic and mechanical agents (12). The previous study in porcine heart tissue has shown that SDS may potentially damage ECM proteins (13). In addition, the residue of detergent in a scaffold may induce apoptosis, cellular dysfunction, and thrombosis (13).…”

Section: Introductionmentioning

confidence: 99%

“…The previous study in porcine heart tissue has shown that SDS may potentially damage ECM proteins (13). In addition, the residue of detergent in a scaffold may induce apoptosis, cellular dysfunction, and thrombosis (13). Another detergent for decellularization is sodium lauryl ether sulfate (SLES) that have been used to decellularize rat heart (14).…”

Section: Introductionmentioning

confidence: 99%

Decellularization of Kidney Tissue: Comparison of Sodium Lauryl Ether Sulfate and Sodium Dodecyl Sulfate for Allotransplantation in Rat

Keshvari¹,

Afshar²,

Daneshi³

et al. 2020

Preprint

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Chronic kidney diseases (CKD) and end stage renal disease (ESRD) are growing threats worldwide. Tissue engineering is a new hope to surpass the current limitations such as the shortage of donor. To do so, the first step would be fabrication of an intact decellularized kidney scaffold. In the current study, an automatic decellularization device was developed to perfuse and decellularize male rats' kidneys using both sodium lauryl ether sulfate (SLES) and sodium dodecyl sulfate (SDS) and to compare their efficacy in kidney decellularization and post-transplantation angiogenesis. After anesthesia, kidneys were perfused with either 1% SDS solution for 4 h or 1% SLES solution for 6 h. The decellularized scaffolds were stained with hematoxylin and eosin (H&E), periodic acid Schiff (PAS), Masson’s trichrome, and alcian blue to determine cell removal and glycogen, collagen and glycosaminoglycans (GAGs) contents, respectively. Moreover, scanning electron microscopy (SEM) was performed to evaluate the cell removal and preservation of microarchitecture of both SDS and SLES scaffolds. Additionally, DNA quantification assay was applied for all groups in order to measure residual DNA in the scaffolds and normal kidney. In order to demonstrate biocompatibility and bioactivity of the decellularized scaffolds, allotransplantation was performed in back muscle and angiogenesis was evaluated. Complete cell removal in both SLES and SDS groups was observed in SEM and DNA quantification assays. Moreover, the extracellular matrix (ECM) architecture of rat kidney in the SLES group was significantly preservation better than the SDS group was shown. The formation of blood capillaries and vessels were observed in the kidney allotransplantations in both SLES and SDS decellularized kidneys. In conclusion, we demonstrated that both SLES and SDS could be promising tools in kidney tissue engineering. The better preservation of ECM than SDS, introduces SLES as the solvent of choice for kidney decellularization. ¬¬

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Using Hemolysis as a Novel Method for Assessment of Cytotoxicity and Blood Compatibility of Decellularized Heart Tissues

Cited by 16 publications

References 33 publications

Biocompatibility and hemocompatibility of efficiently decellularized whole porcine kidney for tissue engineering

Biocompatibility and hemocompatibility of efficiently decellularized whole porcine kidney for tissue engineering

Skeletal Extracellular Matrix Supports Cardiac Differentiation of Embryonic Stem Cells: a Potential Scaffold for Engineered Cardiac Tissue

Decellularization of Kidney Tissue: Comparison of Sodium Lauryl Ether Sulfate and Sodium Dodecyl Sulfate for Allotransplantation in Rat

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