Whole rat stomach decellularisation using a detergent-enzymatic protocol

Zambaiti, Elisa; Scottoni, Federico; Rizzi, Eleonora; Russo, Simone; Deguchi, Kentaro; Eaton, Simon; Pellegata, Alessandro Filippo; Coppi, Paolo De

doi:10.1007/s00383-018-4372-8

Cited by 14 publications

(8 citation statements)

References 28 publications

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“…Large holes and fragmented areas were often observed demonstrating that tissue morphology was not maintained. The use of detergent-enzymatic decellularized protocols has shown to be successful for the decellularization of muscle (Piccoli, Trevisan, Maghin, Franzin, & Pozzobon, 2018;Urciuolo et al, 2018;Zambaiti et al, 2018). However, there is a higher risk of ECM degradation and compromised mechanical and structural loss.…”

Section: Discussionmentioning

confidence: 99%

Optimizing the decellularization process of human maxillofacial muscles for facial reconstruction using a detergent‐only approach

Naik

Szarko

Butler

2019

J Tissue Eng Regen Med

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Trauma, congenital diseases, and cancer resection cause muscle deformities of the human facial muscle. Muscle defects are either treated with local or distal flaps if direct closure is not possible. However, such surgical interventions are limited by donor morbidity and limited tissue availability. Decellularized scaffolds provide alternative strategies for replacing and restoring missing facial muscle by creating scaffolds that mimic the native tissue. This study aimed to develop a protocol to decellularize human zygomaticus major muscle (ZMM) and masseter muscle (MM). Three protocols were assessed including a detergent‐only treatment (DOT), detergent‐enzymatic treatment (DET) protocol, and a third nondetergent nonenzymatic treatment protocol. Scaffolds were then characterized via histological, immunofluorescent, and quantitative techniques to assess which protocol provided optimal decellularization and maintenance of the extracellular matrix (ECM). The results demonstrated three cycles of DOT protocol consisting of 2% sodium dodecyl sulfate for 4 hr was optimal for decellularization for both ZMM and MM. After three cycles, DNA content was significantly reduced compared with native ZMM and MM (p < .05) with preservation of collagen and glycosaminoglycan content and ECM on histological analysis. DET and nondetergent nonenzymatic treatment protocols were unsuccessful in decellularizing the ZMM and MM with residual DNA content after four cycles and caused ECM disruption on histological analysis. All protocols did not impair the mechanical properties and supported human fibroblast growth. In conclusion, the DOT protocol is effective in producing human decellularized muscle scaffolds that maintain the ECM. Further investigation of detergent only decellurization techniques should be explored as a first step to create effective scaffolds for muscle tissue engineering.

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Section: Discussionmentioning

confidence: 99%

Optimizing the decellularization process of human maxillofacial muscles for facial reconstruction using a detergent‐only approach

Naik

Szarko

Butler

2019

J Tissue Eng Regen Med

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“…Whereas the isolation of exosomes from body fluids has been well described and faithfully repeated, [35][36][37] MBV must first be dissociated from the parent ECM before isolation for subsequent investigation or use. The harvesting of ECM from source tissues typically involves the use of detergents, enzymes, and/or mechanical forces to disrupt and remove cells and cell remnants [38][39][40][41] with subsequent dissolution of the remaining ECM to release and isolate the MBV. The MBV must survive these biophysical manipulations with preservation of functional surface moieties, lipid membrane integrity, and intravesicular cargo.…”

Section: Introductionmentioning

confidence: 99%

Matrix-Bound Nanovesicles: The Effects of Isolation Method upon Yield, Purity, and Function

Quijano

Naranjo

El-Mossier

et al. 2020

Tissue Engineering Part C: Methods

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Identification of matrix-bound nanovesicles (MBV) as ubiquitous components of the extracellular matrix (ECM) raises questions regarding their biologic functions and their potential theranostic application. Unlike liquid-phase extracellular vesicles (e.g., exosomes), MBV are tightly bound to the ECM, which makes their isolation and harvesting more challenging. The indiscriminate use of different methods to harvest MBV can alter or disrupt their structural and/or functional integrity. The objective of the present study was to compare the effect of various MBV harvesting methods upon yield, purity, and biologic activity. Combinations of four methods to solubilize the ECM (collagenase [COL], liberase [LIB], or proteinase K [PK] and nonenzymatic elution with potassium chloride) and four isolation methods (ultracentrifugation, ultrafiltration [UF], density barrier, and size exclusion chromatography [SEC]) were used to isolate MBV from urinary bladder-derived ECM. All combinations of solubilization and isolation methods allowed for the harvesting of MBV, however, distinct differences were noted. The highest yield, purity, cellular uptake, and biologic activity were seen with MBV isolated by a combination of liberase or collagenase followed by SEC. The combination of proteinase K and UF was shown to have detrimental effects on bioactivity. The results show the importance of selecting appropriate MBV harvesting methods for the characterization and evaluation of MBV and for analysis of their potential theranostic application.

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“…This process begins with a harvested organ which is recently derived from the brain-dead patient and is placed into a specialized vessel to generate the pale white matrix, i.e., extracellular matrix (ECM) with a complex mixture of structural and functional proteins to preserve structural entities, such as collagen, elastin, glycosaminoglycan (GAG), and fibronectin which function as the scaffold. The matrix can be obtained by employing the decellularization agents which include chemical, enzymatic, and physical methods [30][31][32][33][34][35][36][37][38][39][40][41][42][43].…”

Section: Decellularization: Methods Of Whole-organ Decellularizationmentioning

confidence: 99%

3D Printing: Advancement in Biogenerative Engineering to Combat Shortage of Organs and Bioapplicable Materials

Parihar

Pandita

Kumar

et al. 2021

Regen. Eng. Transl. Med.

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Organ or cell transplantation is medically evaluated for end-stage failure saving or extending the lives of thousands of patients who are suffering from organ failure disorders. The unavailability of adequate organs for transplantation to meet the existing demand is a major challenge in the medical field. This led to day-day-increase in the number of patients on transplant waiting lists as well as in the number of patients dying while on the queue. Recently, technological advancements in the field of biogenerative engineering have the potential to regenerate tissues and, in some cases, create new tissues and organs. In this context, major advances and innovations are being made in the fields of tissue engineering and regenerative medicine which have a huge impact on the scientific community is three-dimensional bioprinting (3D bioprinting) of tissues and organs. Besides this, the decellularization of organs and using this as a scaffold for generating new organs through the recellularization process shows promising results. This review discussed about current approaches for tissue and organ engineering including methods of scaffold designing, recent advances in 3D bioprinting, organs regenerated successfully using 3D printing, and extended application of 3D bioprinting technique in the field of medicine. Besides this, information about commercially available 3D printers has also been included in this article. Lay SummaryToday's need for organs for the transplantation process in order to save a patient's life or to enhance the survival rate of diseased one is the prime concern among the scientific community. Recent, advances in the field of biogenerative engineering have the potential to regenerate tissues and create organs compatible with the patient's body. In this context, major advances and innovations are being made in the fields of tissue engineering and regenerative medicine which have a huge impact on the scientific community is three-dimensional bioprinting (3D bioprinting) of tissues and organs. Besides this, the decellularization of organs and using this as a scaffold for generating new organs through the recellularization process shows promising results. This review dealt with the current approaches for tissue and organ engineering including methods of scaffold designing, recent advances in 3D bioprinting, organs regenerated successfully using 3D printing, and extended application of 3D bioprinting technique in the field of medicine. Furthermore, information about commercially available 3D printers has also been included in this article.

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Whole rat stomach decellularisation using a detergent-enzymatic protocol

Cited by 14 publications

References 28 publications

Optimizing the decellularization process of human maxillofacial muscles for facial reconstruction using a detergent‐only approach

Optimizing the decellularization process of human maxillofacial muscles for facial reconstruction using a detergent‐only approach

Matrix-Bound Nanovesicles: The Effects of Isolation Method upon Yield, Purity, and Function

3D Printing: Advancement in Biogenerative Engineering to Combat Shortage of Organs and Bioapplicable Materials

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