As the components of proteoglycans, glycosaminoglycans (GAGs) are linear polysaccharides consisting of hexose and uronic acid units linked by β-1,3-glycosidic bond. GAGs mainly distribute in extracellular matrix and on cell surfaces. They guide many biological processes, such as proliferation of cells, transmission of signals and mediation of inflammation. Because of their large molecular weights, GAGs have limited biological functions in vitro. However, the appearance of chondroitinase ABC (ChSase ABC), which can lyse polysaccharides, solves the difficulties. Based on our work, we summarized the classification and the crystal structure of ChSase ABC, as well as other recent research progress on ChSase ABCs. The separation and purification methods of ChSase ABC and construction of engineering bacteria are illustrated. The stability and immobilization are also analyzed by taking account of the characterization of ChSase ABC. Finally, problems and future prospect of the ChSase ABC study are summarized.
Bacteria ; Chondroitin ABC Lyase ; chemistry ; Extracellular Matrix ; chemistry ; Glycosaminoglycans ; chemistry ; Proteoglycans ; chemistry

Contributing to organ formation and tissue regeneration, extracellular matrix (ECM) constituents provide tissue with three-dimensional (3D) structural integrity and cellular-function regulation. Containing the crucial traits of the cellular microenvironment, ECM substitutes mediate cell-matrix interactions to prompt stem-cell proliferation and differentiation for 3D organoid construction in vitro or tissue regeneration in vivo. However, these ECMs are often applied generically and have yet to be extensively developed for specific cell types in 3D cultures. Cultured cells also produce rich ECM, particularly stromal cells. Cellular ECM improves 3D culture development in vitro and tissue remodeling during wound healing after implantation into the host as well. Gaining better insight into ECM derived from either tissue or cells that regulate 3D tissue reconstruction or organ regeneration helps us to select, produce, and implant the most suitable ECM and thus promote 3D organoid culture and tissue remodeling for in vivo regeneration. Overall, the decellularization methodologies and tissue/cell-derived ECM as scaffolds or cellular-growth supplements used in cell propagation and differentiation for 3D tissue culture in vitro are discussed. Moreover, current preclinical applications by which ECM components modulate the wound-healing process are reviewed.
Cell Differentiation ; Cell Proliferation ; Decellularized Extracellular Matrix ; Extracellular Matrix/metabolism* ; Humans ; Mesenchymal Stem Cells ; Tissue Engineering/methods* ; Tissue Scaffolds/chemistry*

OBJECTIVE: The prepare decellularized extracellular matrix (ECM) scaffold materials derived from human cervical carcinoma tissues for 3D culture of cervical carcinoma cells. METHODS: Fresh human cervical carcinoma tissues were treated with sodium lauryl ether sulfate (SLES) solution to prepare decellularized ECM scaffolds. The scaffolds were examined for ECM microstructure and residual contents of key ECM components (collagen, glycosaminoglycan, and elastin) and genetic materials by pathological staining and biochemical content analysis. In vitro 3D culture models were established by injecting cultured cervical cancer cells into the prepared ECM scaffolds. The cells in the recellularized scaffolds were compared with those in a conventional 2D culture system for cell behaviors including migration, proliferation and epithelial-mesenchymal transition (EMT) wsing HE staining, immunohistochemical staining and molecular biological technology analysis. Resistance to 5-fluorouracil (5-Fu) of the cells in the two culture systems was tested by analyzing the cell apoptosis rates via flow cytometry. RESULTS: SLES treatment effectively removed cells and genetic materials from human cervical carcinoma tissues but well preserved the microenvironment structure and biological activity of ECM. Compared with the 2D culture system, the 3D culture models significantly promoted proliferation, migration, EMT and 5-Fu resistance of human cervical cancer cells. CONCLUSION The decellularized ECM scaffolds prepared using human cervical carcinoma tissues provide the basis for construction of in vitro 3D culture models for human cervical cancer cells.
Female ; Humans ; Decellularized Extracellular Matrix ; Extracellular Matrix ; Uterine Cervical Neoplasms ; Tissue Scaffolds/chemistry* ; Carcinoma ; Fluorouracil/pharmacology* ; Tissue Engineering ; Tumor Microenvironment

Pericellular matrix (PCM) is a narrow tissue region surrounding chondrocytes, which "chondron" with its enclosed cells. A number of studies suggested that PCM is rich in proteoglycans, collagen and fibronectin, and plays an important role in regulating microenvironment of chondrocytes. Direct measures of PCM properties through micropipette aspiration technique showed that PCM was different from mechanical property of chondrocytes and nature extracellular matrix. However, the function of PCM is not clear, and need further study.
Animals ; Biomechanical Phenomena ; Chondrocytes ; chemistry ; cytology ; metabolism ; Extracellular Matrix ; chemistry ; metabolism ; Humans

Cartilage has poor self-recovery because of its characteristics of no blood vessels and high extracellular matrix. In clinical treatment, physical therapy or drug therapy is usually used for mild cartilage defects, and surgical treatment is needed for severe ones. In recent years, cartilage tissue engineering technology provides a new way for the treatment of cartilage defects. Compared with the traditional surgical treatment, cartilage tissue engineering technology has the advantages of small wound and good recovery. The application of microcarrier technology in the design of tissue engineering scaffolds further expands the function of scaffolds and promotes cartilage regeneration. This review summarized the main preparation methods and development of microcarrier technology in recent years. Subsequently, the properties and specific application scenarios of microcarriers with different materials and functions were introduced according to the materials and functions of microcarriers used in cartilage repair. Based on our research on osteochondral integrated layered scaffolds, we proposed an idea of optimizing the performance of layered scaffolds through microcarriers, which is expected to prepare bionic scaffolds that are more suitable for the structural characteristics of natural cartilage.
Cartilage ; Extracellular Matrix/chemistry* ; Technology ; Tissue Engineering/methods* ; Tissue Scaffolds/chemistry*

It is currently reported that extracellular matrix, biological scaffolds, conditions of stress, nutrients and metabolic waste play very important roles in tissue-engineered osteochondral composite. In this paper, we have made a review of their effects on such composite.
Cartilage ; chemistry ; physiology ; Chondrocytes ; Connective Tissue ; growth & development ; Extracellular Matrix ; chemistry ; Humans ; Stress, Mechanical ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

Hydrogel is a creative polymeric biomaterial which can resemble extracellular matrix (ECM) in vitro. Hydrogel is also a material with intrinsic bioinert, but it can offer mechanical support and developmental guide for cell growth and new tissue organization by designing physicochemical and biological properties of hydrogels precisely. This review mainly introduces design of hydrogels, properties and applications in tissue engineering and regenerative medicine, drug delivery, stem cell culture and cell therapy.
Biocompatible Materials ; Cell Culture Techniques ; Extracellular Matrix ; Humans ; Hydrogels ; chemistry ; Regenerative Medicine ; Stem Cells ; Tissue Engineering

In the field of plastic and reconstructive surgery, the loss of organs or tissues caused by diseases or injuries has resulted in challenges, such as donor shortage and immunosuppression. In recent years, with the development of regenerative medicine, the decellularization-recellularization strategy seems to be a promising and attractive method to resolve these difficulties. The decellularized extracellular matrix contains no cells and genetic materials, while retaining the complex ultrastructure, and it can be used as a scaffold for cell seeding and subsequent transplantation, thereby promoting the regeneration of diseased or damaged tissues and organs. This review provided an overview of decellularization-recellularization technique, and mainly concentrated on the application of decellularization-recellularization technique in the field of plastic and reconstructive surgery, including the remodeling of skin, nose, ears, face, and limbs. Finally, we proposed the challenges in and the direction of future development of decellularization-recellularization technique in plastic surgery.
Tissue Engineering/methods* ; Tissue Scaffolds/chemistry* ; Surgery, Plastic ; Regenerative Medicine/methods* ; Extracellular Matrix

The present study was designed to investigate the effects of various extracellular matrix (ECM) proteins on the biological characteristics of late endothelial progenitor cells (EPCs). Density gradient centrifugation-isolated rat bone marrow mononuclear cells were cultured in complete M199 medium, which contained 15% fetal calf serum, 10 μg/L vascular endothelial growth factor (VEGF) and 5 μg/L basic fibroblast growth factor (bFGF). EPCs were plated on substrates containing fibronectin (Fn), laminin (Ln) or rat tail tendon collagen (Col), and the corresponding cells were defined as Fn, Ln and Col groups. The 3rd generation EPCs, namely late EPCs, were harvested. The proliferation, adhesion, migration and the ability of forming tubes were assayed using CCK-8, adhesion test, wound healing assay and Matrigel, respectively. The mRNA expressions of endothelial cell differentiation markers, vWF and CD31, were analyzed by real time RT-PCR. The apoptosis was assayed by flow cytometry (FCM). The results showed that cell proliferation ability of Fn and Col groups were higher than that of Ln group; Fn group showed increased adhesion compared to Col and Ln groups (P < 0.01); The migration ability of Fn and Col groups were higher than that of Ln group. Moreover, Fn group showed increased tube formation abilities compared to Col and Ln groups (P < 0.05). Although 24-hour free-serum-induced apoptosis in Ln group was the highest, there was no difference of auto-apoptosis among the three groups. Furthermore, the mRNA expressions of vWF and CD31 exhibited no difference among the three groups. These results suggest the ECM affects the biological functions of late EPCs, which would have a high probability of providing new directions that lead to the development of artificial heart and blood vessels.
Animals ; Cell Proliferation ; Cells, Cultured ; Collagen ; chemistry ; Endothelial Progenitor Cells ; cytology ; Extracellular Matrix ; physiology ; Extracellular Matrix Proteins ; chemistry ; Fibroblast Growth Factor 2 ; chemistry ; Fibronectins ; chemistry ; Rats ; Vascular Endothelial Growth Factor A ; chemistry

A process of preparing bovine cortical bone in order to form materials suitable for biomedical xenograft implants was described. Fresh bone samples cut from the middiaphyseal region of bovine femora were obtained from a local slaughterhouse. The bovine bone collagen matrix (BBCM) of various shapes fabricated from bovine bone by defatting and deproteination procedure may be implanted surgically for various purposes. The bone cubes were first defatted in a mixture of defatting agent; subsequently, the samples were extracted to release noncollagenous proteins, followed by digestion using a proteolytic enzyme to remove the telopeptide portions of collagen and residual noncollagenous proteins. Finally,the samples were dried in vacuum, packed and sterilized by gamma irradiation. The bone specimens were characterized by a suite of analytical techniques involving FTIR spectroscopy, X-ray diffraction spectroscopy, differential scanning calorimetry (DSC), uniaxial tension mechanical tests and scanning electron microscopy (SEM). The result showed that BBCM occurred as a white structure with suitable porosity. It contains reasonable proprotion of mineral and organic components in the original osseous architecture of the bovine bone, which is beneficial to keeping the mechanic property and weaker immunogenicity; therefore, it can serve as a potential bone implantable material and extracellular matrix material in bone tissue engineering.
Animals ; Biomimetic Materials ; chemistry ; therapeutic use ; Bone Substitutes ; chemical synthesis ; chemistry ; Bone and Bones ; chemistry ; Cattle ; Collagen ; chemistry ; Extracellular Matrix ; chemistry ; Tissue Engineering ; methods