Regeneration of periodontal tissue is the most promising method for restoring periodontal structures.To find a suitable bioactive three- dimensional scaffold promoting cell proliferation and differentiation is critica...Regeneration of periodontal tissue is the most promising method for restoring periodontal structures.To find a suitable bioactive three- dimensional scaffold promoting cell proliferation and differentiation is critical in periodontal tissue engineering.The objective of this study was to evaluate the biocompatibility of a novel porcine acellular dermal matrix as periodontal tissue scaffolds both in vitro and in vivo.The scaffolds in this study were purified porcine acellular dermal matrix(PADM) and hydroxyapatite-treated PADM(HA-PADM). The biodegradation patterns of the scaffolds were evaluated in vitro.The biocompatibility of the scaffolds in vivo was assessed by implanting them into the sacrospinal muscle of 20 New Zealand white rabbits.The hPDL cells were cultured with PADM or HA-PADM scaffolds for 3,7,14,21 and 28 days.Cell viability assay,scanning electron microscopy(SEM),hematoxylin and eosin(H&E) staining, immunohistochemistry and confocal microscopy were used to evaluate the biocompatibility of the scaffolds.In vitro,both PADM and HA-PADM scaffolds displayed appropriate biodegradation pattern,and also,demonstrated favorable tissue compatibility without tissue necrosis,fibrosis and other abnormal response.The absorbance readings of the WST-1 assay were increased with the time course, suggesting the cell proliferation in the scaffolds.The hPDL cells attaching,spreading and morphology on the surface of the scaffold were visualized by SEM,H&E staining,immnuohjstochemistry and confocal microscopy,demonstrated that hPDL cells were able to grow into the HA-PADM scaffolds and the amount of cells were growing up in the course of time.This study proved that HA-PADM scaffold had good biocompatibility in animals in vivo and appropriate biodegrading characteristics in vitro.The hPDL cells were able to proliferate and migrate into the scaffold.These observations may suggest that HA-PADM scaffold is a potential cell carrier for periodontal tissue regeneration.展开更多
The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells fol-lowing induction with neural di...The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells fol-lowing induction with neural differentiation medium. We performed long-term, continuous observation of cell morphology, growth, differentiation, and neuronal development using several microscopy techniques in conjunction with immunohistochemistry. We examined speciifc neu-ronal proteins and Nissl bodies involved in the differentiation process in order to determine the neuronal differentiation of bone marrow mesenchymal stem cells. The results show that bone marrow mesenchymal stem cells that differentiate on fetal bovine acellular dermal matrix display neuronal morphology with unipolar and bi/multipolar neurite elongations that express neuro-nal-speciifc proteins, includingβIII tubulin. The bone marrow mesenchymal stem cells grown on fetal bovine acellular dermal matrix and induced for long periods of time with neural differen-tiation medium differentiated into a multilayered neural network-like structure with long nerve ifbers that was composed of several parallel microifbers and neuronal cells, forming a complete neural circuit with dendrite-dendrite to axon-dendrite to dendrite-axon synapses. In addition, growth cones with filopodia were observed using scanning electron microscopy. Paraffin sec-tioning showed differentiated bone marrow mesenchymal stem cells with the typical features of neuronal phenotype, such as a large, round nucleus and a cytoplasm full of Nissl bodies. The data suggest that the biological scaffold fetal bovine acellular dermal matrix is capable of supporting human bone marrow mesenchymal stem cell differentiation into functional neurons and the subsequent formation of tissue engineered nerve.展开更多
Composite membranes and sponge scaffolds consisting chitosan (CS) and acellular derm matrix (ADM) in six ratios were prepared by solvent evaporation technique and freeze-drying method, respectively. The composite mate...Composite membranes and sponge scaffolds consisting chitosan (CS) and acellular derm matrix (ADM) in six ratios were prepared by solvent evaporation technique and freeze-drying method, respectively. The composite materials were characterized by water contact angle measurement, scanning electron microscopy (SEM), water absorption and HaCat cells compatibility. The SEM result showed that CS/ADM three-dimensional (3D) micro-porous structures were successfully produced. The water absorption value of all scaffolds was over 18 times of its initial weight, which is high enough for skin regeneration scaffold, but there were no significant differences of water absorption ratio between deionized water and PBS solution for same scaffold (P > 0.05). HaCat cells were distributed uniformly on the surfaces of membrane 4-6, and an almost confluent monolayer was formed on membrane 6 on the fifth day, whereas cells maintained round and spherical in shape on the surface of membrane 1. The results showed that the cell compatibility of pure CS membrane needed to be improved, and addition of ADM realized this purpose. The results of compatibility of HaCat cells on scaffolds showed that the cell proliferated well on the scaffolds 3 and 4. In our study, the cell’s attachment and growth on the composite membranes was mainly determined by the content of the membrane, whereas the cell’s attachment and growth in the scaffolds was determined by both the content and structure of the scaffolds.展开更多
基金supported by Chinese post-doctoral fund(20090451410)International cooperation program of science of Shandong Province (201lHZ035)
文摘Regeneration of periodontal tissue is the most promising method for restoring periodontal structures.To find a suitable bioactive three- dimensional scaffold promoting cell proliferation and differentiation is critical in periodontal tissue engineering.The objective of this study was to evaluate the biocompatibility of a novel porcine acellular dermal matrix as periodontal tissue scaffolds both in vitro and in vivo.The scaffolds in this study were purified porcine acellular dermal matrix(PADM) and hydroxyapatite-treated PADM(HA-PADM). The biodegradation patterns of the scaffolds were evaluated in vitro.The biocompatibility of the scaffolds in vivo was assessed by implanting them into the sacrospinal muscle of 20 New Zealand white rabbits.The hPDL cells were cultured with PADM or HA-PADM scaffolds for 3,7,14,21 and 28 days.Cell viability assay,scanning electron microscopy(SEM),hematoxylin and eosin(H&E) staining, immunohistochemistry and confocal microscopy were used to evaluate the biocompatibility of the scaffolds.In vitro,both PADM and HA-PADM scaffolds displayed appropriate biodegradation pattern,and also,demonstrated favorable tissue compatibility without tissue necrosis,fibrosis and other abnormal response.The absorbance readings of the WST-1 assay were increased with the time course, suggesting the cell proliferation in the scaffolds.The hPDL cells attaching,spreading and morphology on the surface of the scaffold were visualized by SEM,H&E staining,immnuohjstochemistry and confocal microscopy,demonstrated that hPDL cells were able to grow into the HA-PADM scaffolds and the amount of cells were growing up in the course of time.This study proved that HA-PADM scaffold had good biocompatibility in animals in vivo and appropriate biodegrading characteristics in vitro.The hPDL cells were able to proliferate and migrate into the scaffold.These observations may suggest that HA-PADM scaffold is a potential cell carrier for periodontal tissue regeneration.
基金supported by a grant from Construction Project of Gansu Provincial Animal Cell Engineering Center,No.0808NTGA013Program for Innovative Research Team in University of Ministry of Education of China,No.IRT13091
文摘The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells fol-lowing induction with neural differentiation medium. We performed long-term, continuous observation of cell morphology, growth, differentiation, and neuronal development using several microscopy techniques in conjunction with immunohistochemistry. We examined speciifc neu-ronal proteins and Nissl bodies involved in the differentiation process in order to determine the neuronal differentiation of bone marrow mesenchymal stem cells. The results show that bone marrow mesenchymal stem cells that differentiate on fetal bovine acellular dermal matrix display neuronal morphology with unipolar and bi/multipolar neurite elongations that express neuro-nal-speciifc proteins, includingβIII tubulin. The bone marrow mesenchymal stem cells grown on fetal bovine acellular dermal matrix and induced for long periods of time with neural differen-tiation medium differentiated into a multilayered neural network-like structure with long nerve ifbers that was composed of several parallel microifbers and neuronal cells, forming a complete neural circuit with dendrite-dendrite to axon-dendrite to dendrite-axon synapses. In addition, growth cones with filopodia were observed using scanning electron microscopy. Paraffin sec-tioning showed differentiated bone marrow mesenchymal stem cells with the typical features of neuronal phenotype, such as a large, round nucleus and a cytoplasm full of Nissl bodies. The data suggest that the biological scaffold fetal bovine acellular dermal matrix is capable of supporting human bone marrow mesenchymal stem cell differentiation into functional neurons and the subsequent formation of tissue engineered nerve.
文摘Composite membranes and sponge scaffolds consisting chitosan (CS) and acellular derm matrix (ADM) in six ratios were prepared by solvent evaporation technique and freeze-drying method, respectively. The composite materials were characterized by water contact angle measurement, scanning electron microscopy (SEM), water absorption and HaCat cells compatibility. The SEM result showed that CS/ADM three-dimensional (3D) micro-porous structures were successfully produced. The water absorption value of all scaffolds was over 18 times of its initial weight, which is high enough for skin regeneration scaffold, but there were no significant differences of water absorption ratio between deionized water and PBS solution for same scaffold (P > 0.05). HaCat cells were distributed uniformly on the surfaces of membrane 4-6, and an almost confluent monolayer was formed on membrane 6 on the fifth day, whereas cells maintained round and spherical in shape on the surface of membrane 1. The results showed that the cell compatibility of pure CS membrane needed to be improved, and addition of ADM realized this purpose. The results of compatibility of HaCat cells on scaffolds showed that the cell proliferated well on the scaffolds 3 and 4. In our study, the cell’s attachment and growth on the composite membranes was mainly determined by the content of the membrane, whereas the cell’s attachment and growth in the scaffolds was determined by both the content and structure of the scaffolds.
