The aim of this study was to fabricate biomatrix/polymer hybrid scaffolds using an electrospinning technique. Then tissue engineered heart valves were engineered by seeding mesenchymal stromal cells （MSCs） onto the ...The aim of this study was to fabricate biomatrix/polymer hybrid scaffolds using an electrospinning technique. Then tissue engineered heart valves were engineered by seeding mesenchymal stromal cells （MSCs） onto the scaffolds. The effects of the hybrid scaffolds on the proliferation of seed cells, formation of extracellular matrix and mechanical properties of tissue engineered heart valves were investigated. MSCs were obtained from rats. Porcine aortic heart valves were decellularized, coated with poly（3-hydroxybutyrate-co-4-hydroxybutyrate） using an electrospinning technique, and reseeded and cultured over a time period of 14 days. In control group, the decellularized valve scaffolds were reseeded and cultured over an equivalent time period. Specimens of each group were examined histologically （hematoxylin-eosin [HE] staining, immunohistostaining, and scanning electron microscopy）, biochemically （DNA and 4-hydroxyproline） and mechanically. The results showed that recellularization was comparable to the specimens of hybrid scaffolds and controls. The specimens of hybrid scaffolds and controls revealed comparable amounts of cell mass and 4-hydroxyproline （P〉0.05）. However, the specimens of hybrid scaffolds showed a significant increase in mechanical strength, compared to the controls （P〈0.05）. This study demonstrated the superiority of the hybrid scaffolds to increase the mechanical strength of tissue engineered heart valves. And compared to the decellularized valve scaffolds, the hybrid scaffolds showed similar effects on the proliferation of MSCs and formation of extracellular matrix. It was believed that the hybrid scaffolds could be used for the construction of tissue engineered heart valves.展开更多
A formalin-treated polyvinyl-alcohol (PVF) sponge is convenient as a scaffold because its configuration is easily modified. However, coating the sponge with an adhesive chemical agent is necessary to attach bone marro...A formalin-treated polyvinyl-alcohol (PVF) sponge is convenient as a scaffold because its configuration is easily modified. However, coating the sponge with an adhesive chemical agent is necessary to attach bone marrow cells (BMCs) to the sponge structure. Moreover, it was considered that a hybrid scaffold composed of a sponge and enveloped cylindrical porous hydroxyapatite (HA) would be convenient. In this study, the effect of leucine (Leu) coating on a PVF sponge was examined for osteogenesis on an HA/PVF hybrid scaffold by rat BMCs (rBMCs). In an in vivo assessment, the sponge immersed in Leu solution (10 mg/ml) was inserted into the hollow center of cylindrical HA. The sponge received 1.5 × 106 rBMCs obtained from male Fischer 344 rats. The hybrid scaffolds were then implanted subcutaneously of syngeneic rats for 6 weeks. In vitro assessment of Leu to hard tissue formation with coating on the well or addition in rBMC culture medium was also performed in a 6-well plate for 2 weeks. In vivo examinations showed the excellent effect of Leu coating on PVF sponge. Leu-coated PVF sponge in the scaffolds showed marked new bone formation in the pores by histological examination. Leu-coated PVF sponge showed a high quantity of osteocalcine (OC). HA might prevent the release of rBMCs from PVF as a barrier. In in vitro examinations, the quantity of OC in rBMC culture with and without the addition of Leu in culture medium showed no significant difference. However, addition of Leu showed significant ALP activity level in culture medium. Leu coating in culture plate wells showed no influence on the quantity of OC. It was concluded from the results that Leu might prevent the emigration of rBMCs to the outside of the scaffold and promote the differentiation of cells to osteoblasts in the scaffold.展开更多
3D printing/bioprinting are promising techniques to fabricate scaffolds with well controlled and patient-specific structures and architectures for bone tissue engineering.In this study,we developed a composite bioink ...3D printing/bioprinting are promising techniques to fabricate scaffolds with well controlled and patient-specific structures and architectures for bone tissue engineering.In this study,we developed a composite bioink consisting of silk fibroin(SF),gelatin(GEL),hyaluronic acid(HA),and tricalcium phosphate(TCP)and 3D bioprinted the silk fibroin-based hybrid scaffolds.The 3D bioprinted scaffolds with dual crosslinking were further treated with human platelet-rich plasma(PRP)to generate PRP coated scaffolds.Live/Dead and MTT assays demonstrated that PRP treatment could obviously promote the cell growth and proliferation of human adipose derived mesenchymal stem cells(HADMSC).In addition,the treatment of PRP did not significantly affect alkaline phosphatase(ALP)activity and expression,but significantly upregulated the gene expression levels of late osteogenic markers.This study demonstrated that the 3D printing of silk fibroin-based hybrid scaffolds,in combination with PRP post-treatment,might be a more efficient strategy to promote osteogenic differentiation of adult stem cells and has significant potential to be used for bone tissue engineering.展开更多
Bio-mechanically active scaffolds for tissue engineering combining hydrophilic polymeric matrix and nano-diamond fillers properties are presented and discussed in this paper. The resulting scaffolding materials reveal...Bio-mechanically active scaffolds for tissue engineering combining hydrophilic polymeric matrix and nano-diamond fillers properties are presented and discussed in this paper. The resulting scaffolding materials revealed re-markable mechanical and biological properties to be exploited in advanced biomedical applications. The novel hybrid material is based on 2 and 5 vol-ume % of detonation nano-diamond particles in a hydrophilic poly-(hydroxyl- ethyl-methacrylate) matrix. According to its mechanical and biological properties, the nanocomposite shows a hybrid nature. The base analytical procedures for the preparation of the hybrid nanocomposites and some preliminary mechanical characteristics are presented. The proposed hybrid system has been considered for potential biomimetic, osteoconductive and osteoinductive scaffolds application in bio-mechanically active bone scaffolds for osteoblast, and stem cell differentiation and growth. These more rigid hybrid nano-composites are predicted to possess improved mechanical strength overcoming the mechanical weaknesses of traditional hydrogels clinically utilized for bone regeneration.展开更多
The inertness of synthetic polymer materials and the insufficient mechanical strength of reprocessed decellu-larized extracellular matrix(dECM)limited their promotive efforts on tissue regeneration.Here,we prepared a ...The inertness of synthetic polymer materials and the insufficient mechanical strength of reprocessed decellu-larized extracellular matrix(dECM)limited their promotive efforts on tissue regeneration.Here,we prepared a hybrid scaffold composed of PCL microfibers and human placental extracellular matrix(pECM)nanofibers by co-electrospinning,which was grafted with heparin and further absorbed with IL-4.The hybrid scaffold with improved hemocompatibility firstly switched macrophages to anti-inflammatory phenotype(increased by 18.1%)and then promoted migration,NO production,tube formation of endothelial cells(ECs),and migration and maturation of vascular smooth muscle cells(VSMCs),and ECM deposition in vitro and in vivo.ECs coverage rate increased by 8.6%and the thickness of the smooth muscle layer was 1.8 times more than PCL grafts at 12 wks.Our study realized the complementary advantages of synthetic polymer materials and dECM materials,and opened intriguing perspectives for the design and construction of small-diameter vascular grafts(SDVGs)and immune-regulated materials for other tissue regeneration.展开更多
In order to provide a biomimetic natural extracellular matrix microenvironment with excellent mechanical capacity for tissue regeneration,a novel porous hybrid glycidyl methacrylate-modified silk fibroin/poly(L-lactic...In order to provide a biomimetic natural extracellular matrix microenvironment with excellent mechanical capacity for tissue regeneration,a novel porous hybrid glycidyl methacrylate-modified silk fibroin/poly(L-lactic acid-co-ε-caprolactone)–polyethylene glycol diacrylate(SFMA/P(LLA-CL)–PEGDA)hybrid three-dimensional(3D)nanofibrous scaffolds was successfully fabricated through the combination of 3D nanofibrous platforms and divinyl PEGDA based photocrosslinking,and then further improved water resistance by ethanol vapor post-treatment.Scanning electron microscopy and micro-computed tomography results demonstrated significant PEGDA hydrogel-like matrices bonded nanofibers,which formed a 3D structure similar to that of“steel bar(nanofibers)‒cement(PEGDA)”,with proper pore size,high porosity,and high pore connectivity density.Meanwhile,the hybrid 3D nanofibrous scaffolds showed outstanding swelling properties as well as improved compressive and tensile properties.Furthermore,these hybrid 3D nanofibrous scaffolds could provide a biocompatible microenvironment,capable of inducing the material‒cell hybrid and regulating human umbilical vein endothelial cells proliferation.They thus present significant potential in tissue regeneration.展开更多
Articular cartilage injury is a common disease in the field of orthopedics.Because cartilage has poor self-repairing ability,medical intervention is needed.Using melt electro-writing(MEW)technology,tissue engineering ...Articular cartilage injury is a common disease in the field of orthopedics.Because cartilage has poor self-repairing ability,medical intervention is needed.Using melt electro-writing(MEW)technology,tissue engineering scaffolds with high porosity and high precision can be prepared.However,ordinary materials,especially natural polymer materials,are difficult to print.In this study,gelatin was mixed with poly(lactic-co-glycolic acid)to prepare high-concentration and high-viscosity printer ink,which had good printability and formability.A composite scaffold with full-layer TGF-β1 loading mixed with hydroxyapatite was prepared,and the scaffold was implanted at the cartilage injury site;microfracture surgery was conducted to induce the mesenchyme in the bone marrow.Quality stem cells thereby promoted the repair of damaged cartilage.In summary,this study developed a novel printing method,explored the molding conditions based on MEW printing ink,and constructed a bioactive cartilage repair scaffold.The scaffold can use autologous bone marrow mesenchymal stem cells and induce their differentiation to promote cartilage repair.展开更多
基金supported by grants from National Natural Sciences Foundation of China (No.30571839,30600608 and 30872540)National High Technology Research and Development Program ("863" Program) of China (No.2009AA-03Z420)
文摘The aim of this study was to fabricate biomatrix/polymer hybrid scaffolds using an electrospinning technique. Then tissue engineered heart valves were engineered by seeding mesenchymal stromal cells （MSCs） onto the scaffolds. The effects of the hybrid scaffolds on the proliferation of seed cells, formation of extracellular matrix and mechanical properties of tissue engineered heart valves were investigated. MSCs were obtained from rats. Porcine aortic heart valves were decellularized, coated with poly（3-hydroxybutyrate-co-4-hydroxybutyrate） using an electrospinning technique, and reseeded and cultured over a time period of 14 days. In control group, the decellularized valve scaffolds were reseeded and cultured over an equivalent time period. Specimens of each group were examined histologically （hematoxylin-eosin [HE] staining, immunohistostaining, and scanning electron microscopy）, biochemically （DNA and 4-hydroxyproline） and mechanically. The results showed that recellularization was comparable to the specimens of hybrid scaffolds and controls. The specimens of hybrid scaffolds and controls revealed comparable amounts of cell mass and 4-hydroxyproline （P〉0.05）. However, the specimens of hybrid scaffolds showed a significant increase in mechanical strength, compared to the controls （P〈0.05）. This study demonstrated the superiority of the hybrid scaffolds to increase the mechanical strength of tissue engineered heart valves. And compared to the decellularized valve scaffolds, the hybrid scaffolds showed similar effects on the proliferation of MSCs and formation of extracellular matrix. It was believed that the hybrid scaffolds could be used for the construction of tissue engineered heart valves.
文摘A formalin-treated polyvinyl-alcohol (PVF) sponge is convenient as a scaffold because its configuration is easily modified. However, coating the sponge with an adhesive chemical agent is necessary to attach bone marrow cells (BMCs) to the sponge structure. Moreover, it was considered that a hybrid scaffold composed of a sponge and enveloped cylindrical porous hydroxyapatite (HA) would be convenient. In this study, the effect of leucine (Leu) coating on a PVF sponge was examined for osteogenesis on an HA/PVF hybrid scaffold by rat BMCs (rBMCs). In an in vivo assessment, the sponge immersed in Leu solution (10 mg/ml) was inserted into the hollow center of cylindrical HA. The sponge received 1.5 × 106 rBMCs obtained from male Fischer 344 rats. The hybrid scaffolds were then implanted subcutaneously of syngeneic rats for 6 weeks. In vitro assessment of Leu to hard tissue formation with coating on the well or addition in rBMC culture medium was also performed in a 6-well plate for 2 weeks. In vivo examinations showed the excellent effect of Leu coating on PVF sponge. Leu-coated PVF sponge in the scaffolds showed marked new bone formation in the pores by histological examination. Leu-coated PVF sponge showed a high quantity of osteocalcine (OC). HA might prevent the release of rBMCs from PVF as a barrier. In in vitro examinations, the quantity of OC in rBMC culture with and without the addition of Leu in culture medium showed no significant difference. However, addition of Leu showed significant ALP activity level in culture medium. Leu coating in culture plate wells showed no influence on the quantity of OC. It was concluded from the results that Leu might prevent the emigration of rBMCs to the outside of the scaffold and promote the differentiation of cells to osteoblasts in the scaffold.
基金supported by National Institutes of Health(R01 AR073225)to Dr.Bin Duan(R21AI140026)to Drs Patrick Reid and Bin Duan+2 种基金Chinese Universities Scientific Fund(CUSF-DH-D-2016008)China Scholarship Council,Doctoral Program of Xi'an Polytechnic University(BS201902)to Dr.Liang Weisupported by state funds from the Nebraska Research Initiative(NRI)and the University of Nebraska Foundation,and institutionally by the Office of the Vice Chancellor for Research.
文摘3D printing/bioprinting are promising techniques to fabricate scaffolds with well controlled and patient-specific structures and architectures for bone tissue engineering.In this study,we developed a composite bioink consisting of silk fibroin(SF),gelatin(GEL),hyaluronic acid(HA),and tricalcium phosphate(TCP)and 3D bioprinted the silk fibroin-based hybrid scaffolds.The 3D bioprinted scaffolds with dual crosslinking were further treated with human platelet-rich plasma(PRP)to generate PRP coated scaffolds.Live/Dead and MTT assays demonstrated that PRP treatment could obviously promote the cell growth and proliferation of human adipose derived mesenchymal stem cells(HADMSC).In addition,the treatment of PRP did not significantly affect alkaline phosphatase(ALP)activity and expression,but significantly upregulated the gene expression levels of late osteogenic markers.This study demonstrated that the 3D printing of silk fibroin-based hybrid scaffolds,in combination with PRP post-treatment,might be a more efficient strategy to promote osteogenic differentiation of adult stem cells and has significant potential to be used for bone tissue engineering.
文摘Bio-mechanically active scaffolds for tissue engineering combining hydrophilic polymeric matrix and nano-diamond fillers properties are presented and discussed in this paper. The resulting scaffolding materials revealed re-markable mechanical and biological properties to be exploited in advanced biomedical applications. The novel hybrid material is based on 2 and 5 vol-ume % of detonation nano-diamond particles in a hydrophilic poly-(hydroxyl- ethyl-methacrylate) matrix. According to its mechanical and biological properties, the nanocomposite shows a hybrid nature. The base analytical procedures for the preparation of the hybrid nanocomposites and some preliminary mechanical characteristics are presented. The proposed hybrid system has been considered for potential biomimetic, osteoconductive and osteoinductive scaffolds application in bio-mechanically active bone scaffolds for osteoblast, and stem cell differentiation and growth. These more rigid hybrid nano-composites are predicted to possess improved mechanical strength overcoming the mechanical weaknesses of traditional hydrogels clinically utilized for bone regeneration.
基金National Natural Science Foundation of China(NSFC)projects(81972063,81701840,32201122)Innovative Research Group Project(81921004)+1 种基金Tianjin Key Medical Discipline(Specialty)Construction Project(TJYXZDXK-043A)China Postdoctoral Science Fundation(2022M711705).
文摘The inertness of synthetic polymer materials and the insufficient mechanical strength of reprocessed decellu-larized extracellular matrix(dECM)limited their promotive efforts on tissue regeneration.Here,we prepared a hybrid scaffold composed of PCL microfibers and human placental extracellular matrix(pECM)nanofibers by co-electrospinning,which was grafted with heparin and further absorbed with IL-4.The hybrid scaffold with improved hemocompatibility firstly switched macrophages to anti-inflammatory phenotype(increased by 18.1%)and then promoted migration,NO production,tube formation of endothelial cells(ECs),and migration and maturation of vascular smooth muscle cells(VSMCs),and ECM deposition in vitro and in vivo.ECs coverage rate increased by 8.6%and the thickness of the smooth muscle layer was 1.8 times more than PCL grafts at 12 wks.Our study realized the complementary advantages of synthetic polymer materials and dECM materials,and opened intriguing perspectives for the design and construction of small-diameter vascular grafts(SDVGs)and immune-regulated materials for other tissue regeneration.
基金This work was sponsored by the Jiaxing Public Welfare Research Project(Grant Nos.2021AY10062 and 2021AY10063)the Zhejiang Provincial Natural Science Foundation of China(Grant Nos.LY20C100003 and LY22C100001)the National Undergraduate Training Program for Innovation and Entrepreneurship(Grant No.202010354009).
文摘In order to provide a biomimetic natural extracellular matrix microenvironment with excellent mechanical capacity for tissue regeneration,a novel porous hybrid glycidyl methacrylate-modified silk fibroin/poly(L-lactic acid-co-ε-caprolactone)–polyethylene glycol diacrylate(SFMA/P(LLA-CL)–PEGDA)hybrid three-dimensional(3D)nanofibrous scaffolds was successfully fabricated through the combination of 3D nanofibrous platforms and divinyl PEGDA based photocrosslinking,and then further improved water resistance by ethanol vapor post-treatment.Scanning electron microscopy and micro-computed tomography results demonstrated significant PEGDA hydrogel-like matrices bonded nanofibers,which formed a 3D structure similar to that of“steel bar(nanofibers)‒cement(PEGDA)”,with proper pore size,high porosity,and high pore connectivity density.Meanwhile,the hybrid 3D nanofibrous scaffolds showed outstanding swelling properties as well as improved compressive and tensile properties.Furthermore,these hybrid 3D nanofibrous scaffolds could provide a biocompatible microenvironment,capable of inducing the material‒cell hybrid and regulating human umbilical vein endothelial cells proliferation.They thus present significant potential in tissue regeneration.
基金This work was supported by Shanghai Ninth People’s Hospital(grant number XK2019013)National Natural Science Foundation of China(No.81802131,82002293)China Postdoctoral Science Foundation(No.2019T120347).
文摘Articular cartilage injury is a common disease in the field of orthopedics.Because cartilage has poor self-repairing ability,medical intervention is needed.Using melt electro-writing(MEW)technology,tissue engineering scaffolds with high porosity and high precision can be prepared.However,ordinary materials,especially natural polymer materials,are difficult to print.In this study,gelatin was mixed with poly(lactic-co-glycolic acid)to prepare high-concentration and high-viscosity printer ink,which had good printability and formability.A composite scaffold with full-layer TGF-β1 loading mixed with hydroxyapatite was prepared,and the scaffold was implanted at the cartilage injury site;microfracture surgery was conducted to induce the mesenchyme in the bone marrow.Quality stem cells thereby promoted the repair of damaged cartilage.In summary,this study developed a novel printing method,explored the molding conditions based on MEW printing ink,and constructed a bioactive cartilage repair scaffold.The scaffold can use autologous bone marrow mesenchymal stem cells and induce their differentiation to promote cartilage repair.
