The Experts below are selected from a list of 1098 Experts worldwide ranked by ideXlab platform
Peter E. M. Butler - One of the best experts on this subject based on the ideXlab platform.
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Optimizing the Decellularization Process of an upper limb skeletal muscle; implications for muscle tissue engineering
Artificial organs, 2019Co-Authors: Anish Naik, Michelle Griffin, Matthew Szarko, Peter E. M. ButlerAbstract:Upper limb muscle reconstruction is required following cancer resection, trauma, and congenital deformities. Current surgical reconstruction of the muscle involves local, regional and free flaps. However, muscle reconstruction is not always possible due to the size of the defect and functional donor site morbidity. These challenges could be addressed with the production of scaffolds composed of an extracellular matrix (ECM) derived from decellularized human skeletal muscle. This study aimed to find an optimal technique to decellularize a flexor digitorum superficialis muscle. The first two protocols were based on a detergent only (DOT) and a detergent-enzymatic protocol (DET). The third protocol avoided the use of detergents and proteolytic enzymes (NDNET). The decellularized scaffolds were characterized using qualitative techniques including histological and immunofluorescent staining and quantitative techniques assessing deoxyribonucleic acid (DNA), glycosaminoglycan (GAG), and collagen content. The DOT protocol consisting of 2% SDS for 4 hours was successful at decellularizing human FDS, as shown by DNA content assay and nuclei immunofluorescence staining. The DOT protocol maintained the microstructure of the scaffolds as shown by Masson's trichrome staining and collagen and GAG content. DET and NDNET protocols maintained the ECM, but were unsuccessful in removing all DNA content after two cycles of Decellularization. Decellularization of skeletal muscle is a viable option for muscle reconstruction using a detergent only technique for upper limb defects. Further testing in vivo will assess the effectiveness of decellularized scaffolds for upper limb muscle skeletal tissue engineering.
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optimizing the Decellularization Process of human maxillofacial muscles for facial reconstruction using a detergent only approach
Journal of Tissue Engineering and Regenerative Medicine, 2019Co-Authors: Anish Naik, Michelle Griffin, Matthew Szarko, Peter E. M. ButlerAbstract: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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Optimising the Decellularization of human elastic cartilage with trypsin for future use in ear reconstruction.
Scientific reports, 2018Co-Authors: Shafiq Rahman, Anish Naik, Michelle Griffin, Matthew Szarko, Peter E. M. ButlerAbstract:Decellularized scaffolds can induce chondrogenic differentiation of stem cells. This study compares different methods to optimise the Decellularization of auricular cartilage. The Process consisted of an initial 12 hour dry freeze thaw which froze the cartilage specimens in an empty tube at −20 °C. Samples were allowed to thaw at room temperature followed by submersion in phosphate buffer solution in which they were frozen at −20 °C for a 12 hour period. They were then allowed to thaw at room temperature as before. Protocol A subsequently involved subjecting specimens to both deoxyribonuclease and sodium deoxycholate. Protocol B and C were adaptations of this using 0.25% trypsin (7 cycles) and a 0.5 molar solution of ethylenediaminetetraacetic acid (3 hours for each cycle) respectively as additional steps. Trypsin accelerated the Decellularization Process with a reduction in DNA content from 55.4 ng/μL (native) to 17.3 ng/μL (P-value
Tugba Sezgin Arslan - One of the best experts on this subject based on the ideXlab platform.
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A Novel Protocol to Generate Decellularized Bovine Spinal Cord Extracellular Matrix-Based Scaffolds (3D-dCBS).
Bio-protocol, 2019Co-Authors: Yavuz Emre Arslan, Burcu Efe, Tugba Sezgin ArslanAbstract:Extracellular matrix (ECM)-based tissue engineering scaffolds have an essential role in promoting tissue regeneration. Nerve tissue engineering aims at facilitating the repair of permanent damage to the peripheral and central nervous systems, which are difficult to heal. For this purpose, a variety of biomaterials are being developed consisting of numerous synthetic and/or natural polymers to provide axonal reinnervation and to direct the growth of axons. Here, we present a novel protocol that enables to fabricate a 3-dimensional (3D) decellularized scaffold derived from the bovine spinal cord (BSC) ECM (3D-dCBS) for neural tissue engineering applications. In this protocol, a viscous ECM-derived gel from BSC is prepared, molded, and chemically crosslinked with EDC/NHS (3D-CBS) before Decellularization Process. Decellularization of 3D-CBS is performed with 1% SDS to attain 3D-dCBS. As compared with other available methods, our protocol is a novel Decellularization method that preserves a more significant part of the ECM. We believe that the mentioned protocol has the potential to produce a bioengineered scaffold from spinal cord tissue with desired geometry for regenerative medicine applications related to neural tissue engineering.
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A novel method for constructing an acellular 3D biomatrix from bovine spinal cord for neural tissue engineering applications
Biotechnology Progress, 2019Co-Authors: Yavuz Emre Arslan, Tugba Sezgin ArslanAbstract:In this study, we aimed at generating 3-dimensional (3D) decellularized bovine spinal cord extracellular matrix-based scaffolds (3D-dCBS) for neural tissue engineering applications. Within this scope, bovine spinal cord tissue pieces were homogenized in 0.1 M NaOH and this viscous mixture was molded to attain 3D bioscaffolds. After resultant bioscaffolds were chemically crosslinked, the Decellularization Process was conducted with detergent, buffer, and enzyme solutions. Nuclear remnants in the native tissue and 3D-dCBS were determined with DNA content analysis and agarose gel electrophoresis. Afterward, 3D-dCBS were biochemically characterized in depth via glycosaminoglycan (GAG) content, hydroxyproline (HYP) assay, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Cellular survival of human adipose-derived mesenchymal stem cells (hAMSCs) on the 3D-dCBS for 3rd, 7th, and 10th days was assessed via MTT assay. Scaffold and cell/scaffold constructs were also evaluated with scanning electron microscopy and histochemical studies. DNA contents for native and 3D-dCBS were respectively found to be 520.76 ± 18.11 and 28.80 ± 0.20 ng/mg dry weight (n = 3), indicating a successful Decellularization Process. GAG content, HYP assay, and SDS-PAGE results proved that the extracellular matrix was substantially preserved during the Decellularization Process. In conclusion, it is believed that the novel Decellularization method may allow fabricating 3D bioscaffolds with desired geometry from soft nervous system tissues.
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Trans-differentiation of human adipose-derived mesenchymal stem cells into cardiomyocyte-like cells on decellularized bovine myocardial extracellular matrix-based films.
Journal of materials science. Materials in medicine, 2018Co-Authors: Yavuz Emre Arslan, Tugba Sezgin Arslan, Yusuf Furkan Galata, Burak DerkusAbstract:In this study, we aimed at fabricating decellularized bovine myocardial extracellular matrix-based films (dMEbF) for cardiac tissue engineering (CTE). The Decellularization Process was carried out utilizing four consecutive stages including hypotonic treatment, detergent treatment, enzymatic digestion and decontamination, respectively. In order to fabricate the dMEbF, dBM were digested with pepsin and gelation Process was conducted. dMEbF were then crosslinked with N-hydroxysuccinimide/1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide (NHS/EDC) to increase their durability. Nuclear contents of native BM and decellularized BM (dBM) tissues were determined with DNA content analysis and agarose-gel electrophoresis. Cell viability on dMEbF for 3rd, 7th, and 14th days was assessed by MTT assay. Cell attachment on dMEbF was also studied by scanning electron microscopy. Trans-differentiation capacity of human adipose-derived mesenchymal stem cells (hAMSCs) into cardiomyocyte-like cells on dMEbF were also evaluated by histochemical and immunohistochemical analyses. DNA contents for native and dBM were, respectively, found as 886.11 ± 164.85 and 47.66 ± 0.09 ng/mg dry weight, indicating a successful Decellularization Process. The results of glycosaminoglycan and hydroxyproline assay, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), performed in order to characterize the extracellular matrix (ECM) composition of native and dBM tissue, showed that the BM matrix was not damaged during the proposed method. Lastly, regarding the histological study, dMEbF not only mimics native ECM, but also induces the stem cells into cardiomyocyte-like cells phenotype which brings it the potential of use in CTE.
Yavuz Emre Arslan - One of the best experts on this subject based on the ideXlab platform.
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A Novel Protocol to Generate Decellularized Bovine Spinal Cord Extracellular Matrix-Based Scaffolds (3D-dCBS).
Bio-protocol, 2019Co-Authors: Yavuz Emre Arslan, Burcu Efe, Tugba Sezgin ArslanAbstract:Extracellular matrix (ECM)-based tissue engineering scaffolds have an essential role in promoting tissue regeneration. Nerve tissue engineering aims at facilitating the repair of permanent damage to the peripheral and central nervous systems, which are difficult to heal. For this purpose, a variety of biomaterials are being developed consisting of numerous synthetic and/or natural polymers to provide axonal reinnervation and to direct the growth of axons. Here, we present a novel protocol that enables to fabricate a 3-dimensional (3D) decellularized scaffold derived from the bovine spinal cord (BSC) ECM (3D-dCBS) for neural tissue engineering applications. In this protocol, a viscous ECM-derived gel from BSC is prepared, molded, and chemically crosslinked with EDC/NHS (3D-CBS) before Decellularization Process. Decellularization of 3D-CBS is performed with 1% SDS to attain 3D-dCBS. As compared with other available methods, our protocol is a novel Decellularization method that preserves a more significant part of the ECM. We believe that the mentioned protocol has the potential to produce a bioengineered scaffold from spinal cord tissue with desired geometry for regenerative medicine applications related to neural tissue engineering.
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A novel method for constructing an acellular 3D biomatrix from bovine spinal cord for neural tissue engineering applications
Biotechnology Progress, 2019Co-Authors: Yavuz Emre Arslan, Tugba Sezgin ArslanAbstract:In this study, we aimed at generating 3-dimensional (3D) decellularized bovine spinal cord extracellular matrix-based scaffolds (3D-dCBS) for neural tissue engineering applications. Within this scope, bovine spinal cord tissue pieces were homogenized in 0.1 M NaOH and this viscous mixture was molded to attain 3D bioscaffolds. After resultant bioscaffolds were chemically crosslinked, the Decellularization Process was conducted with detergent, buffer, and enzyme solutions. Nuclear remnants in the native tissue and 3D-dCBS were determined with DNA content analysis and agarose gel electrophoresis. Afterward, 3D-dCBS were biochemically characterized in depth via glycosaminoglycan (GAG) content, hydroxyproline (HYP) assay, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Cellular survival of human adipose-derived mesenchymal stem cells (hAMSCs) on the 3D-dCBS for 3rd, 7th, and 10th days was assessed via MTT assay. Scaffold and cell/scaffold constructs were also evaluated with scanning electron microscopy and histochemical studies. DNA contents for native and 3D-dCBS were respectively found to be 520.76 ± 18.11 and 28.80 ± 0.20 ng/mg dry weight (n = 3), indicating a successful Decellularization Process. GAG content, HYP assay, and SDS-PAGE results proved that the extracellular matrix was substantially preserved during the Decellularization Process. In conclusion, it is believed that the novel Decellularization method may allow fabricating 3D bioscaffolds with desired geometry from soft nervous system tissues.
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Trans-differentiation of human adipose-derived mesenchymal stem cells into cardiomyocyte-like cells on decellularized bovine myocardial extracellular matrix-based films.
Journal of materials science. Materials in medicine, 2018Co-Authors: Yavuz Emre Arslan, Tugba Sezgin Arslan, Yusuf Furkan Galata, Burak DerkusAbstract:In this study, we aimed at fabricating decellularized bovine myocardial extracellular matrix-based films (dMEbF) for cardiac tissue engineering (CTE). The Decellularization Process was carried out utilizing four consecutive stages including hypotonic treatment, detergent treatment, enzymatic digestion and decontamination, respectively. In order to fabricate the dMEbF, dBM were digested with pepsin and gelation Process was conducted. dMEbF were then crosslinked with N-hydroxysuccinimide/1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide (NHS/EDC) to increase their durability. Nuclear contents of native BM and decellularized BM (dBM) tissues were determined with DNA content analysis and agarose-gel electrophoresis. Cell viability on dMEbF for 3rd, 7th, and 14th days was assessed by MTT assay. Cell attachment on dMEbF was also studied by scanning electron microscopy. Trans-differentiation capacity of human adipose-derived mesenchymal stem cells (hAMSCs) into cardiomyocyte-like cells on dMEbF were also evaluated by histochemical and immunohistochemical analyses. DNA contents for native and dBM were, respectively, found as 886.11 ± 164.85 and 47.66 ± 0.09 ng/mg dry weight, indicating a successful Decellularization Process. The results of glycosaminoglycan and hydroxyproline assay, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), performed in order to characterize the extracellular matrix (ECM) composition of native and dBM tissue, showed that the BM matrix was not damaged during the proposed method. Lastly, regarding the histological study, dMEbF not only mimics native ECM, but also induces the stem cells into cardiomyocyte-like cells phenotype which brings it the potential of use in CTE.
Amelia Seifalian - One of the best experts on this subject based on the ideXlab platform.
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development of a cost effective and simple protocol for Decellularization and preservation of human amniotic membrane as a soft tissue replacement and delivery system for bone marrow stromal cells
Advanced Healthcare Materials, 2015Co-Authors: Mazaher Gholipourmalekabadi, Masoud Mozafari, Mohammad Salehi, Amelia Seifalian, Mojgan Bandehpour, Hossein Ghanbarian, Aleksandra M Urbanska, Marzieh Sameni, Ali SamadikuchaksaraeiAbstract:The aim of this study is to develop a simple andcost-effective method for Decellularization and preservation of human amniotic membrane (HAM) as a soft tissue replacement and a delivery system for stem cells. The HAM is decellularized (D) using new chemical and mechanical techniques. The Decellularization scaffold is evaluated histologically and fully characterized. The cell adhesion and proliferation on the scaffold are also investigated and the biocompatibility of D tissues is evaluated in vivo. The histological studies reveal that the cells are successfully removed from the D tissue. The DNA extraction shows more than 95% cell removal (p = 0.001). The in vitro results indicate that the decellularisation Process does not deteriorate the mechanical properties of the tissue, whereas it increases the in vitro biodegradation value (p 0.05). Immunohistochemistry staining indicates that all the tested components remain unchanged within the D tissues. The count of inflammatory cells show that the Decellularization Process slightly increases the biocompatibility of the tissue after 7 days post-surgery. The results indicate that scaffold proves to be reproducible, rapid, and cost-effective, with a potential role for clinical application.
Anish Naik - One of the best experts on this subject based on the ideXlab platform.
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Optimizing the Decellularization Process of an upper limb skeletal muscle; implications for muscle tissue engineering
Artificial organs, 2019Co-Authors: Anish Naik, Michelle Griffin, Matthew Szarko, Peter E. M. ButlerAbstract:Upper limb muscle reconstruction is required following cancer resection, trauma, and congenital deformities. Current surgical reconstruction of the muscle involves local, regional and free flaps. However, muscle reconstruction is not always possible due to the size of the defect and functional donor site morbidity. These challenges could be addressed with the production of scaffolds composed of an extracellular matrix (ECM) derived from decellularized human skeletal muscle. This study aimed to find an optimal technique to decellularize a flexor digitorum superficialis muscle. The first two protocols were based on a detergent only (DOT) and a detergent-enzymatic protocol (DET). The third protocol avoided the use of detergents and proteolytic enzymes (NDNET). The decellularized scaffolds were characterized using qualitative techniques including histological and immunofluorescent staining and quantitative techniques assessing deoxyribonucleic acid (DNA), glycosaminoglycan (GAG), and collagen content. The DOT protocol consisting of 2% SDS for 4 hours was successful at decellularizing human FDS, as shown by DNA content assay and nuclei immunofluorescence staining. The DOT protocol maintained the microstructure of the scaffolds as shown by Masson's trichrome staining and collagen and GAG content. DET and NDNET protocols maintained the ECM, but were unsuccessful in removing all DNA content after two cycles of Decellularization. Decellularization of skeletal muscle is a viable option for muscle reconstruction using a detergent only technique for upper limb defects. Further testing in vivo will assess the effectiveness of decellularized scaffolds for upper limb muscle skeletal tissue engineering.
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optimizing the Decellularization Process of human maxillofacial muscles for facial reconstruction using a detergent only approach
Journal of Tissue Engineering and Regenerative Medicine, 2019Co-Authors: Anish Naik, Michelle Griffin, Matthew Szarko, Peter E. M. ButlerAbstract: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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Optimising the Decellularization of human elastic cartilage with trypsin for future use in ear reconstruction.
Scientific reports, 2018Co-Authors: Shafiq Rahman, Anish Naik, Michelle Griffin, Matthew Szarko, Peter E. M. ButlerAbstract:Decellularized scaffolds can induce chondrogenic differentiation of stem cells. This study compares different methods to optimise the Decellularization of auricular cartilage. The Process consisted of an initial 12 hour dry freeze thaw which froze the cartilage specimens in an empty tube at −20 °C. Samples were allowed to thaw at room temperature followed by submersion in phosphate buffer solution in which they were frozen at −20 °C for a 12 hour period. They were then allowed to thaw at room temperature as before. Protocol A subsequently involved subjecting specimens to both deoxyribonuclease and sodium deoxycholate. Protocol B and C were adaptations of this using 0.25% trypsin (7 cycles) and a 0.5 molar solution of ethylenediaminetetraacetic acid (3 hours for each cycle) respectively as additional steps. Trypsin accelerated the Decellularization Process with a reduction in DNA content from 55.4 ng/μL (native) to 17.3 ng/μL (P-value
