The aim of this study is to prepare poly-L-lactide(PLLA)electrospun nanofibrous scaffolds coated with hippocampal neuron-derived extracellular matrix(N-ECM)and construct a novel neural tissue engineering scaffold.Neon...The aim of this study is to prepare poly-L-lactide(PLLA)electrospun nanofibrous scaffolds coated with hippocampal neuron-derived extracellular matrix(N-ECM)and construct a novel neural tissue engineering scaffold.Neonatal rat hippocampal neurons were seeded on PLLA nanofibers,and then decellularized to derive a cell-free extracellular matrix loaded N-ECM/PLLA modified scaffolds.The morphology and ingredients of N-ECM/PLLA were observed by scanning electron microscopy(SEM)and immunofluorescence staining respectively,and the cytocompatibility of the composite scaffolds was characterized by cell count kit-8(CCK-8)assay.The N-ECM was clearly identified loading on scaffolds when being imaged via SEM and immunofluorescence staining results showed that the N-ECM was made up of fibronectin and laminin.Most importantly,compared with tissue culture polystyrene and pure scaffolds,N-ECM/PLLA scaffolds could effectively facilitate the proliferation of rat adrenal neuroma cells(PC12 cells),indicating their better cell compatibilities.Based on the combination of N-ECM and PLLA biomaterials,the present study has fabricated a unique and versatile neural tissue engineering scaffold,offering a new thought for future neural tissue engineering.展开更多
The effects of various cartilage extracellular matrix on the construction of rabbit growth plate cartilage tissue in vitro were studied. The results show that collagen, proteoglycan and hyaluronic acid can promote the...The effects of various cartilage extracellular matrix on the construction of rabbit growth plate cartilage tissue in vitro were studied. The results show that collagen, proteoglycan and hyaluronic acid can promote the growth of cultured chondrocytes but the effects of various cartilage extracellular matrix(ECM)on chondrocyte differentiation are different. Collagen can promote the hypertrophy of chondrocytes while proteoglycan and hyaluronic acid inhibit the transition of mature chondrocytes into hypertrophied chondrocytes.展开更多
Functional repair of injured tissue in the adult central nervous system (CNS) still remains a big challenge for current biomed- ical research and its upcoming clinical translation. The axonal regeneration of the adu...Functional repair of injured tissue in the adult central nervous system (CNS) still remains a big challenge for current biomed- ical research and its upcoming clinical translation. The axonal regeneration of the adult CNS is generally low, and it is addi- tionally restricted after injury by the presence of inhibitory mol- ecules, generated by the glial scar.展开更多
AIM: To improve osteogenic differentiation and attachment of cells.METHODS: An electronic search was conducted inPub Med from January 2004 to December 2013. Studies which performed smart modifications on conventional ...AIM: To improve osteogenic differentiation and attachment of cells.METHODS: An electronic search was conducted inPub Med from January 2004 to December 2013. Studies which performed smart modifications on conventional bone scaffold materials were included. Scaffolds with controlled release or encapsulation of bioactive molecules were not included. Experiments which did not investigate response of cells toward the scaffold(cell attachment, proliferation or osteoblastic differentiation) were excluded. RESULTS: Among 1458 studies, 38 met the inclusion and exclusion criteria. The main scaffold varied extensively among the included studies. Smart modifications included addition of growth factors(group Ⅰ-11 studies), extracellular matrix-like molecules(group Ⅱ-13 studies) and nanoparticles(nano-HA)(group Ⅲ-17 studies). In all groups, surface coating was the most commonly applied approach for smart modification of scaffolds. In group I, bone morphogenetic proteins were mainly used as growth factor stabilized on polycaprolactone(PCL). In group Ⅱ, collagen 1 in combination with PCL, hydroxyapatite(HA) and tricalcium phosphate were the most frequent scaffolds used. In the third group, nano-HA with PCL and chitosan were used the most. As variable methods were used, a thorough and comprehensible compare between the results and approaches was unattainable.CONCLUSION: Regarding the variability in methodology of these in vitro studies it was demonstrated that smart modification of scaffolds can improve tissue properties.展开更多
Recently,tissue engineering (TE)is one of the fast growing research fields due the accessibility of extra-molecular matrix (ECM)at cellular and molecular level with valuable potential prospective of hydrogels.The enha...Recently,tissue engineering (TE)is one of the fast growing research fields due the accessibility of extra-molecular matrix (ECM)at cellular and molecular level with valuable potential prospective of hydrogels.The enhancement in the production of hydrogel-based cellular scaffolds with the structural composition of ECM has been accelerated with involvement of rapid prototyping techniques.Basically,the recreation of ECM has been derived from naturally existed or synthetic hydrogelbased polymers.The rapid utilization of hydrogels in TE puts forward the scope of bioprinfing for the fabrication of the functional biological tissues,cartilage,skin and artificial organs.The main focus of the researchers is on biofabrication of the biomaterials with maintaining the biocompatibility,biodegradability and increasing growth efficiency.In this review, biological development in the structure and cross-linking connections of natural or synthetic hydrogels are discussed.The methods and design criteria that influence the chemical and mechanical properties and interaction of seeding cells before and after the implantations are also demonstrated.The methodology of bioprinting techniques along with recent development has also been reviewed.In the end,some capabilities and shortcomings are pointed out for further development of hydrogels-based scaffolds and selection of bioprinting technology depending on their application.展开更多
The objective of this study was to evaluate the biocompatibility of vessel extracellular matrix (VECM) from rabbit and to discuss the feasibility of vessel extracellular matrix as a matrix for urethral reconstructio...The objective of this study was to evaluate the biocompatibility of vessel extracellular matrix (VECM) from rabbit and to discuss the feasibility of vessel extracellular matrix as a matrix for urethral reconstruction. Primary cultured bladder smooth muscle cells isolated from New Zealand rabbits were implanted on VECM .The effects of VECM on rabbit bladder smooth muscle cells (RBSMCs) metabolic activity, attachment, proliferation were monitored in vitro with the aid of an inverted light microscope and a scanning electron microscope. The cell viability was monitored by MTT(methythiazolye tetrazolium bromide) after 1, 3, 5 days seeding. The in vivo tissue response to VECM was investigated by implanting them into the subcutaneous of rabbits. VECM exhibited a nontoxic and bioactive effect on RBSMCs. RBSMCs could be attached to and proliferated on VECM and maintained their morphologies. MTT assay showed RBSMCs cultured with the extracts of VECM were not significantly different from those of negative controls. In vivo, VECM demonstrated a favorable tissue compatibility without tissue necrosis, fibrosis and other abnormal response. VECM exhibited nontoxic and bioactive effects on RBSMC. It is a suitable material for urethral reconstruction.展开更多
Extracellular matrix( ECM) plays a prominent role in establishing and maintaining an appropriate microenvironment for tissue regeneration. The aims of this study were to construct a tissue engineered scaffold by recon...Extracellular matrix( ECM) plays a prominent role in establishing and maintaining an appropriate microenvironment for tissue regeneration. The aims of this study were to construct a tissue engineered scaffold by reconstituting osteoblast cell-derived ECM( O-ECM) on the electrospun nanofibrous scaffold,and further to evaluate its subsequent application for promoting the proliferation of bone marrow mesenchymal stem cells( BMSCs). To engineer a biomimetic scaffold, calvarial osteoblasts and electrospun poly-llactic acid( PLLA) nanofibers were prepared and subjected to decellularize for O-ECM deposition. To evaluate and characterize the O-ECM/PLLA scaffold, the morphology was examined and several specific mark proteins of osteoblasts matrix were evaluated.Furthermore,the cell counting kit-8( CCK-8) assay was used to detect the proliferation of the BMSCs cultivated on the O-ECM/PLLA scaffold. The results indicated O-ECM/PLLA scaffold was loaded with Collagen I, Fibronectin, and Laminin, as the composition of the marrow ECM. After decellularization,O-ECM deposition was observed in O-ECM/PLLA scaffold. Moreover,the O-ECM/PLLA scaffold could significantly enhance the proliferation of BMSCs,suggesting better cytocompatibility compared to the other groups tested. Taken together,a biomimetic scaffold based on the joint use of O-ECM and PLLA biomaterials,which represents a promising approach to bone tissue engineering, facilitates the expansion of BMSCs in vitro.展开更多
BACKGROUND:Tissue engineered acellular nerves are good autologous nerve substitutes. Acellular peripheral nerves prepared using a conventional chemical extraction method cause a great deal of damage to nerve structur...BACKGROUND:Tissue engineered acellular nerves are good autologous nerve substitutes. Acellular peripheral nerves prepared using a conventional chemical extraction method cause a great deal of damage to nerve structures, and the allograff affects the nerve regeneration following transplantation.OBJECTIVE:To prepare peripheral nerve grafts through an acellular tissue engineering method, and observe their histology, ultrastructure, protein components and histocompatibility.DESIGN, TIME AND SETTING:A randomized, controlled, in vivo nerve tissue engineering experiment was performed at the Department of Biochemistry and Molecular Biology, Shenyang Medical College, China, from September 2006 to June 2007.MATERIALS:Triton X-100, Pepstatin A, Aprotinin and Leupeptin were purchased from Sigma, USA; Tris (hydroxymethyl) aminomethane was purchased from Gibco, USA.METHODS:The bilateral sciatic nerves of Wistar rats were harvested, treated with 0.05 mol/L Tris-HCI buffer, followed by proteinase inhibitor and Triton X-100 to prepare acellular peripheral nerves. The nerves were implanted in the quadriceps femoris muscle of healthy Wistar rats.MAIN OUTCOME MEASURES:Tissue structure and ultrastructure of acellular peripheral nerves were observed by optical microscopy and scanning electron microscopy. Growth associated proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Nerve allograft and the surrounding muscles were observed by hematoxylin-eosin staining.RESULTS:Acellular treatment eliminated Schwann cells, epineurium or perineurium cells, myelin sheaths and axons of nerve fibers in normal peripheral nerves, while the spatial structure, comprising basement membrane tubes of Schwann cells and the extracellular matrix of perineurium and nerve fascicles was maintained. Protein bands at the region of 30 kD were no longer visible, had slightly decreased at 43 kD and remained unchanged at 65 kD. Following implantation for 7 days, epineurium cells were absorbed. However, increased fibroblasts, decreased newly-generated capillaries and maturation of granulation tissue were observed.CONCLUSION:The acellular nerve allograft prepared through the use of a hypotonic, acellular method displays good histocompatibility, eliminates immune substances and retains growth associated proteins that induce the growth of the neural axis. In addition, this method provides an ideal scaffold to construct artificial nerves.展开更多
Despite the considerable advancements in fabricating polymeric-based scaffolds for tissue engineering,the clinical transformation of these scaffolds remained a big challenge because of the difficulty of simulating nat...Despite the considerable advancements in fabricating polymeric-based scaffolds for tissue engineering,the clinical transformation of these scaffolds remained a big challenge because of the difficulty of simulating native organs/tissues'microenvironment.As a kind of natural tissue-derived biomaterials,decellularized extracellular matrix(dECM)-based scaffolds have gained attention due to their unique biomimetic properties,providing a specific microenvironment suitable for promoting cell proliferation,migration,attachment and regulating differentiation.The medical applications of dECM-based scaffolds have addressed critical challenges,including poor mechanical strength and insufficient stability.For promoting the reconstruction of damaged tissues or organs,dif-ferent types of dECM-based composite platforms have been designed to mimic tissue microenvironment,including by integrating with natural polymer or/and syntenic polymer or adding bioactive factors.In this review,we summarized the research progress of dECM-based composite scaffolds in regenerative medicine,highlighting the critical challenges and future perspectives related to the medical application of these composite materials。展开更多
Decellularised extracellular matrix(dECM)is a biomaterial derived from natural tissues that has attracted considerable attention from tissue engineering researchers due to its exceptional biocompatibility and malleabi...Decellularised extracellular matrix(dECM)is a biomaterial derived from natural tissues that has attracted considerable attention from tissue engineering researchers due to its exceptional biocompatibility and malleability attributes.These advantageous properties often facilitate natural cell infiltration and tissue reconstruction for regenerative medicine.Due to their excellent fluidity,the injectable hydrogels can be administered in a liquid state and subsequently formed into a gel state in vivo,stabilising the target area and serving in a variety of ways,such as support,repair,and drug release functions.Thus,dECM-based injectable hydrogels have broad prospects for application in complex organ structures and various tissue injury models.This review focuses on exploring research advances in dECM-based injectable hydrogels,primarily focusing on the applications and prospects of dECM hydrogels in tissue engineering.Initially,the recent developments of the dECM-based injectable hydrogels are explained,summarising the different preparation methods with the evaluation of injectable hydrogel properties.Furthermore,some specific examples of the applicability of dECM-based injectable hydrogels are presented.Finally,we summarise the article with interesting prospects and challenges of dECM-based injectable hydrogels,providing insights into the development of these composites in tissue engineering and regenerative medicine.展开更多
Aim: To characterize the feasibility of the surgical replacement of the penile tunica albuginea (TA) and to evaluate the value of a porcine bladder acellular matrix (BAM) graft. Methods: Acellular matrices were ...Aim: To characterize the feasibility of the surgical replacement of the penile tunica albuginea (TA) and to evaluate the value of a porcine bladder acellular matrix (BAM) graft. Methods: Acellular matrices were constructed from pigs' bladders by cell lysis, and then examined by scanning electron microscopy (SEM). Expression levels of the mRNA of the vascular endothelial growth factor (VEGF) receptor, fibroblast growth factor (FGF)-1 receptor, neuregulin, and brain-derived neurotrophic factor (BDNF) in the acellular matrix and submucosa of the pigs' bladders were determined through the reverse transcription-polymerase chain reaction (PCR). A 5 mm× 5 mm square was excised from the penile TA of nine rabbits. The defective TA was then covered in porcine BAM. Equal numbers of animals were sacrificed and histochemically examined at 2, 4 and 6 months after implantation. Results: SEM of the BAM showed collagen fibers with many pores. VEGF receptor, FGF-1 receptor and neuregulin mRNA were expressed in the porcine BAM; BDNF mRNA was not detected. Two months after implantation, the graft sites exhibited excellent healing without contracture, and the fusion between the graft and the neighboring normal TA appeared to be well established. There were no significant histological differences between the implanted tunica and the normal control tunica at 6 months after implantation. Conclusion: The porcine BAM graft resulted in a structure which was sufficiently like that of the normal TA. This implantation might be considered applicable to the reconstruction of the TA in conditions such as trauma or Peyronie's disease.展开更多
Therapeutic approaches for musculoskeletal tissue regeneration commonly employ growth factors(GFs)to influence neighboring cells and promote migration,proliferation,or differentiation.Despite promising results in prec...Therapeutic approaches for musculoskeletal tissue regeneration commonly employ growth factors(GFs)to influence neighboring cells and promote migration,proliferation,or differentiation.Despite promising results in preclinical models,the use of inductive biomacromolecules has achieved limited success in translation to the clinic.The field has yet to sufficiently overcome substantial hurdles such as poor spatiotemporal control and supraphysiological dosages,which commonly result in detrimental side effects.Physiological presentation and retention of biomacromolecules is regulated by the extracellular matrix(ECM),which acts as a reservoir for GFs via electrostatic interactions.Advances in the manipulation of extracellular proteins,decellularized tissues,and synthetic ECM-mimetic applications across a range of biomaterials have increased the ability to direct the presentation of GFs.Successful application of biomaterial technologies utilizing ECM mimetics increases tissue regeneration without the reliance on supraphysiological doses of inductive biomacromolecules.This review describes recent strategies to manage GF presentation using ECM-mimetic substrates for the regeneration of bone,cartilage,and muscle.展开更多
The present review aims to illustrate the strategies that are being implemented to regenerate or bioengineer livers for clinical purposes.There are two general pathways to liver bioengineering and regeneration.The fir...The present review aims to illustrate the strategies that are being implemented to regenerate or bioengineer livers for clinical purposes.There are two general pathways to liver bioengineering and regeneration.The first consists of creating a supporting scaffold,either synthetically or by decellularization of human or animal organs,and seeding cells on the scaffold,where they will mature either in bioreactors or in vivo.This strategy seems to offer the quickest route to clinical translation,as demonstrated by the development of liver organoids from rodent livers which were repopulated with organ specific cells of animal and/or human origin.Liver bioengineering has potential for transplantation and for toxicity testing during preclinical drug development.The second possibility is to induce liver regeneration of dead or resected tissue by manipulating cell pathways.In fact,it is well known that the liver has peculiar regenerative potential which allows hepatocyte hyperplasia after amputation of liver volume.Infusion of autologous bone marrow cells,which aids in liver regeneration,into patients was shown to be safe and to improve their clinical condition,but the specific cells responsible for liver regeneration have not yet been determined and the underlying mechanisms remain largely unknown.A complete understanding of the cell pathways and dynamics and of the functioning of liver stem cell niche is necessary for the clinical translation of regenerative medicine strategies.As well,it will be crucial to elucidate the mechanisms through which cells interact with the extracellular matrix,and how this latter supports and drives cell fate.展开更多
基金Fundamental Research Funds for the Central Universities,China(No.16D110520)
文摘The aim of this study is to prepare poly-L-lactide(PLLA)electrospun nanofibrous scaffolds coated with hippocampal neuron-derived extracellular matrix(N-ECM)and construct a novel neural tissue engineering scaffold.Neonatal rat hippocampal neurons were seeded on PLLA nanofibers,and then decellularized to derive a cell-free extracellular matrix loaded N-ECM/PLLA modified scaffolds.The morphology and ingredients of N-ECM/PLLA were observed by scanning electron microscopy(SEM)and immunofluorescence staining respectively,and the cytocompatibility of the composite scaffolds was characterized by cell count kit-8(CCK-8)assay.The N-ECM was clearly identified loading on scaffolds when being imaged via SEM and immunofluorescence staining results showed that the N-ECM was made up of fibronectin and laminin.Most importantly,compared with tissue culture polystyrene and pure scaffolds,N-ECM/PLLA scaffolds could effectively facilitate the proliferation of rat adrenal neuroma cells(PC12 cells),indicating their better cell compatibilities.Based on the combination of N-ECM and PLLA biomaterials,the present study has fabricated a unique and versatile neural tissue engineering scaffold,offering a new thought for future neural tissue engineering.
基金This work was supported by a grant from Foundation of Education Department of China (NO.02190) and the Science and Technology Foundation of Guang-zhou City (NO.2002 Z1 -E0032)
文摘The effects of various cartilage extracellular matrix on the construction of rabbit growth plate cartilage tissue in vitro were studied. The results show that collagen, proteoglycan and hyaluronic acid can promote the growth of cultured chondrocytes but the effects of various cartilage extracellular matrix(ECM)on chondrocyte differentiation are different. Collagen can promote the hypertrophy of chondrocytes while proteoglycan and hyaluronic acid inhibit the transition of mature chondrocytes into hypertrophied chondrocytes.
基金supported by MEYS of the Czech Republic,No.LO1309
文摘Functional repair of injured tissue in the adult central nervous system (CNS) still remains a big challenge for current biomed- ical research and its upcoming clinical translation. The axonal regeneration of the adult CNS is generally low, and it is addi- tionally restricted after injury by the presence of inhibitory mol- ecules, generated by the glial scar.
文摘AIM: To improve osteogenic differentiation and attachment of cells.METHODS: An electronic search was conducted inPub Med from January 2004 to December 2013. Studies which performed smart modifications on conventional bone scaffold materials were included. Scaffolds with controlled release or encapsulation of bioactive molecules were not included. Experiments which did not investigate response of cells toward the scaffold(cell attachment, proliferation or osteoblastic differentiation) were excluded. RESULTS: Among 1458 studies, 38 met the inclusion and exclusion criteria. The main scaffold varied extensively among the included studies. Smart modifications included addition of growth factors(group Ⅰ-11 studies), extracellular matrix-like molecules(group Ⅱ-13 studies) and nanoparticles(nano-HA)(group Ⅲ-17 studies). In all groups, surface coating was the most commonly applied approach for smart modification of scaffolds. In group I, bone morphogenetic proteins were mainly used as growth factor stabilized on polycaprolactone(PCL). In group Ⅱ, collagen 1 in combination with PCL, hydroxyapatite(HA) and tricalcium phosphate were the most frequent scaffolds used. In the third group, nano-HA with PCL and chitosan were used the most. As variable methods were used, a thorough and comprehensible compare between the results and approaches was unattainable.CONCLUSION: Regarding the variability in methodology of these in vitro studies it was demonstrated that smart modification of scaffolds can improve tissue properties.
文摘Recently,tissue engineering (TE)is one of the fast growing research fields due the accessibility of extra-molecular matrix (ECM)at cellular and molecular level with valuable potential prospective of hydrogels.The enhancement in the production of hydrogel-based cellular scaffolds with the structural composition of ECM has been accelerated with involvement of rapid prototyping techniques.Basically,the recreation of ECM has been derived from naturally existed or synthetic hydrogelbased polymers.The rapid utilization of hydrogels in TE puts forward the scope of bioprinfing for the fabrication of the functional biological tissues,cartilage,skin and artificial organs.The main focus of the researchers is on biofabrication of the biomaterials with maintaining the biocompatibility,biodegradability and increasing growth efficiency.In this review, biological development in the structure and cross-linking connections of natural or synthetic hydrogels are discussed.The methods and design criteria that influence the chemical and mechanical properties and interaction of seeding cells before and after the implantations are also demonstrated.The methodology of bioprinting techniques along with recent development has also been reviewed.In the end,some capabilities and shortcomings are pointed out for further development of hydrogels-based scaffolds and selection of bioprinting technology depending on their application.
基金the Key Program of Education Department of Hubei Province(No.301130697)
文摘The objective of this study was to evaluate the biocompatibility of vessel extracellular matrix (VECM) from rabbit and to discuss the feasibility of vessel extracellular matrix as a matrix for urethral reconstruction. Primary cultured bladder smooth muscle cells isolated from New Zealand rabbits were implanted on VECM .The effects of VECM on rabbit bladder smooth muscle cells (RBSMCs) metabolic activity, attachment, proliferation were monitored in vitro with the aid of an inverted light microscope and a scanning electron microscope. The cell viability was monitored by MTT(methythiazolye tetrazolium bromide) after 1, 3, 5 days seeding. The in vivo tissue response to VECM was investigated by implanting them into the subcutaneous of rabbits. VECM exhibited a nontoxic and bioactive effect on RBSMCs. RBSMCs could be attached to and proliferated on VECM and maintained their morphologies. MTT assay showed RBSMCs cultured with the extracts of VECM were not significantly different from those of negative controls. In vivo, VECM demonstrated a favorable tissue compatibility without tissue necrosis, fibrosis and other abnormal response. VECM exhibited nontoxic and bioactive effects on RBSMC. It is a suitable material for urethral reconstruction.
基金Shanghai Municipal Natural Science Foundation,China(No.15ZR1400500)the Fundamental Research Funds for the Central Universities,China(Nos.16D110520,EG2017011)
文摘Extracellular matrix( ECM) plays a prominent role in establishing and maintaining an appropriate microenvironment for tissue regeneration. The aims of this study were to construct a tissue engineered scaffold by reconstituting osteoblast cell-derived ECM( O-ECM) on the electrospun nanofibrous scaffold,and further to evaluate its subsequent application for promoting the proliferation of bone marrow mesenchymal stem cells( BMSCs). To engineer a biomimetic scaffold, calvarial osteoblasts and electrospun poly-llactic acid( PLLA) nanofibers were prepared and subjected to decellularize for O-ECM deposition. To evaluate and characterize the O-ECM/PLLA scaffold, the morphology was examined and several specific mark proteins of osteoblasts matrix were evaluated.Furthermore,the cell counting kit-8( CCK-8) assay was used to detect the proliferation of the BMSCs cultivated on the O-ECM/PLLA scaffold. The results indicated O-ECM/PLLA scaffold was loaded with Collagen I, Fibronectin, and Laminin, as the composition of the marrow ECM. After decellularization,O-ECM deposition was observed in O-ECM/PLLA scaffold. Moreover,the O-ECM/PLLA scaffold could significantly enhance the proliferation of BMSCs,suggesting better cytocompatibility compared to the other groups tested. Taken together,a biomimetic scaffold based on the joint use of O-ECM and PLLA biomaterials,which represents a promising approach to bone tissue engineering, facilitates the expansion of BMSCs in vitro.
文摘BACKGROUND:Tissue engineered acellular nerves are good autologous nerve substitutes. Acellular peripheral nerves prepared using a conventional chemical extraction method cause a great deal of damage to nerve structures, and the allograff affects the nerve regeneration following transplantation.OBJECTIVE:To prepare peripheral nerve grafts through an acellular tissue engineering method, and observe their histology, ultrastructure, protein components and histocompatibility.DESIGN, TIME AND SETTING:A randomized, controlled, in vivo nerve tissue engineering experiment was performed at the Department of Biochemistry and Molecular Biology, Shenyang Medical College, China, from September 2006 to June 2007.MATERIALS:Triton X-100, Pepstatin A, Aprotinin and Leupeptin were purchased from Sigma, USA; Tris (hydroxymethyl) aminomethane was purchased from Gibco, USA.METHODS:The bilateral sciatic nerves of Wistar rats were harvested, treated with 0.05 mol/L Tris-HCI buffer, followed by proteinase inhibitor and Triton X-100 to prepare acellular peripheral nerves. The nerves were implanted in the quadriceps femoris muscle of healthy Wistar rats.MAIN OUTCOME MEASURES:Tissue structure and ultrastructure of acellular peripheral nerves were observed by optical microscopy and scanning electron microscopy. Growth associated proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Nerve allograft and the surrounding muscles were observed by hematoxylin-eosin staining.RESULTS:Acellular treatment eliminated Schwann cells, epineurium or perineurium cells, myelin sheaths and axons of nerve fibers in normal peripheral nerves, while the spatial structure, comprising basement membrane tubes of Schwann cells and the extracellular matrix of perineurium and nerve fascicles was maintained. Protein bands at the region of 30 kD were no longer visible, had slightly decreased at 43 kD and remained unchanged at 65 kD. Following implantation for 7 days, epineurium cells were absorbed. However, increased fibroblasts, decreased newly-generated capillaries and maturation of granulation tissue were observed.CONCLUSION:The acellular nerve allograft prepared through the use of a hypotonic, acellular method displays good histocompatibility, eliminates immune substances and retains growth associated proteins that induce the growth of the neural axis. In addition, this method provides an ideal scaffold to construct artificial nerves.
基金the National Natural Science Foundation of China(NSFC 32271410,32071323 and 81971734)the Science and Technology Projects in Fujian Province(2022FX1,2023Y4008)+1 种基金Scientific Research Funds of Huaqiao University(21BS113)the Open Research Fund of Academy of Advanced Carbon Conversion Technology,Huaqiao University(AACCT0004).
文摘Despite the considerable advancements in fabricating polymeric-based scaffolds for tissue engineering,the clinical transformation of these scaffolds remained a big challenge because of the difficulty of simulating native organs/tissues'microenvironment.As a kind of natural tissue-derived biomaterials,decellularized extracellular matrix(dECM)-based scaffolds have gained attention due to their unique biomimetic properties,providing a specific microenvironment suitable for promoting cell proliferation,migration,attachment and regulating differentiation.The medical applications of dECM-based scaffolds have addressed critical challenges,including poor mechanical strength and insufficient stability.For promoting the reconstruction of damaged tissues or organs,dif-ferent types of dECM-based composite platforms have been designed to mimic tissue microenvironment,including by integrating with natural polymer or/and syntenic polymer or adding bioactive factors.In this review,we summarized the research progress of dECM-based composite scaffolds in regenerative medicine,highlighting the critical challenges and future perspectives related to the medical application of these composite materials。
基金supported by the National Natural Science Foundation of China(Nos.32271410,32071323,and 81971734)the Science and Technology Projects in Fujian Province(Nos.2022FX1,2023Y4008).
文摘Decellularised extracellular matrix(dECM)is a biomaterial derived from natural tissues that has attracted considerable attention from tissue engineering researchers due to its exceptional biocompatibility and malleability attributes.These advantageous properties often facilitate natural cell infiltration and tissue reconstruction for regenerative medicine.Due to their excellent fluidity,the injectable hydrogels can be administered in a liquid state and subsequently formed into a gel state in vivo,stabilising the target area and serving in a variety of ways,such as support,repair,and drug release functions.Thus,dECM-based injectable hydrogels have broad prospects for application in complex organ structures and various tissue injury models.This review focuses on exploring research advances in dECM-based injectable hydrogels,primarily focusing on the applications and prospects of dECM hydrogels in tissue engineering.Initially,the recent developments of the dECM-based injectable hydrogels are explained,summarising the different preparation methods with the evaluation of injectable hydrogel properties.Furthermore,some specific examples of the applicability of dECM-based injectable hydrogels are presented.Finally,we summarise the article with interesting prospects and challenges of dECM-based injectable hydrogels,providing insights into the development of these composites in tissue engineering and regenerative medicine.
文摘Aim: To characterize the feasibility of the surgical replacement of the penile tunica albuginea (TA) and to evaluate the value of a porcine bladder acellular matrix (BAM) graft. Methods: Acellular matrices were constructed from pigs' bladders by cell lysis, and then examined by scanning electron microscopy (SEM). Expression levels of the mRNA of the vascular endothelial growth factor (VEGF) receptor, fibroblast growth factor (FGF)-1 receptor, neuregulin, and brain-derived neurotrophic factor (BDNF) in the acellular matrix and submucosa of the pigs' bladders were determined through the reverse transcription-polymerase chain reaction (PCR). A 5 mm× 5 mm square was excised from the penile TA of nine rabbits. The defective TA was then covered in porcine BAM. Equal numbers of animals were sacrificed and histochemically examined at 2, 4 and 6 months after implantation. Results: SEM of the BAM showed collagen fibers with many pores. VEGF receptor, FGF-1 receptor and neuregulin mRNA were expressed in the porcine BAM; BDNF mRNA was not detected. Two months after implantation, the graft sites exhibited excellent healing without contracture, and the fusion between the graft and the neighboring normal TA appeared to be well established. There were no significant histological differences between the implanted tunica and the normal control tunica at 6 months after implantation. Conclusion: The porcine BAM graft resulted in a structure which was sufficiently like that of the normal TA. This implantation might be considered applicable to the reconstruction of the TA in conditions such as trauma or Peyronie's disease.
基金This work was supported by the National Institutes of Health under award number R01 DE025475 and R01 DE025899 to JKL.
文摘Therapeutic approaches for musculoskeletal tissue regeneration commonly employ growth factors(GFs)to influence neighboring cells and promote migration,proliferation,or differentiation.Despite promising results in preclinical models,the use of inductive biomacromolecules has achieved limited success in translation to the clinic.The field has yet to sufficiently overcome substantial hurdles such as poor spatiotemporal control and supraphysiological dosages,which commonly result in detrimental side effects.Physiological presentation and retention of biomacromolecules is regulated by the extracellular matrix(ECM),which acts as a reservoir for GFs via electrostatic interactions.Advances in the manipulation of extracellular proteins,decellularized tissues,and synthetic ECM-mimetic applications across a range of biomaterials have increased the ability to direct the presentation of GFs.Successful application of biomaterial technologies utilizing ECM mimetics increases tissue regeneration without the reliance on supraphysiological doses of inductive biomacromolecules.This review describes recent strategies to manage GF presentation using ECM-mimetic substrates for the regeneration of bone,cartilage,and muscle.
文摘The present review aims to illustrate the strategies that are being implemented to regenerate or bioengineer livers for clinical purposes.There are two general pathways to liver bioengineering and regeneration.The first consists of creating a supporting scaffold,either synthetically or by decellularization of human or animal organs,and seeding cells on the scaffold,where they will mature either in bioreactors or in vivo.This strategy seems to offer the quickest route to clinical translation,as demonstrated by the development of liver organoids from rodent livers which were repopulated with organ specific cells of animal and/or human origin.Liver bioengineering has potential for transplantation and for toxicity testing during preclinical drug development.The second possibility is to induce liver regeneration of dead or resected tissue by manipulating cell pathways.In fact,it is well known that the liver has peculiar regenerative potential which allows hepatocyte hyperplasia after amputation of liver volume.Infusion of autologous bone marrow cells,which aids in liver regeneration,into patients was shown to be safe and to improve their clinical condition,but the specific cells responsible for liver regeneration have not yet been determined and the underlying mechanisms remain largely unknown.A complete understanding of the cell pathways and dynamics and of the functioning of liver stem cell niche is necessary for the clinical translation of regenerative medicine strategies.As well,it will be crucial to elucidate the mechanisms through which cells interact with the extracellular matrix,and how this latter supports and drives cell fate.