OBJECTIVE: To sum up the clinical results of bio-derived bone transplantation in orthopedics with tissue engineering technique. METHODS: From January 2000 to May 2002, 52 cases with various types of bone defect were treated with tissue engineered bone, which was constructed in vitro by allogeneous osteoblasts from periosteum (1 x 10(6)/ml) with bio-derived bone scaffold following 3 to 7 days co-culture. Among them, there were 7 cases of bone cyst, 22 cases of non-union or malunion of old fracture, 15 cases of fresh comminuted fracture of bone defect, 4 cases of spinal fracture and posterior route spinal fusion, 3 cases of bone implant of alveolar bone, 1 case of fusion of tarsotarsal joint. The total weight of tissue engineered bone was 349 g in all the cases, averaged 6.7 g in each case. RESULTS: All the cases were followed up after operation, averaged in 18.5 months. The wound in all the case healed by first intention, but 1 case with second intention. Bone union was completed within 3 to 4.5 months in 50 cases, but 2 cases of delayed union. Six cases were performed analysis of CD3, CD4, CD8, ICAM-1 and VCAM-1 before and after operation, and no obvious abnormities were observed. CONCLUSION: Bio-derived tissue engineered bone has good osteogenesis. No obvious rejection and other complications are observed in the clinical application.
To summarize the medium-term cl inical result of bio-derived bone transplantation in orthopedics with tissue engineering technique. Methods From December 2000 to June 2001, 10 cases of various types of bone defect were treated with tissue engineered bone, which was constructed in vitro by allogenous osteoblasts from periosteum (1 × 106/ mL) with bio-derived bone scaffold following 3 to 7 days co-culture. Six men and 4 women were involved in this study, aged from 14 to 70 years with a median of 42 years. Among them, there were 2 cases of bone cyst, 1 case of non-union of old fracture, 6 cases of fresh comminuted fracture with bone defect, and 1 case of chronic suppurative ostemyel itis. The total weight of tissue engineered bone was 3-15 g in all the cases, averaged 7.3 g in each case. Results The wound in all the case healed by first intention. For 7 year follow up, bone union was completed within 3.0 to 4.5 months in 9 cases, but loosening occurred and the graft was taken out 1 year after operation in 1 case. The X-ray films showed that 9 cases achieved union except one who received resection of the head of humerus. No obvious abnormities were observed, and the function of affected l imbs met daily l ife and work. Conclusion Bio-derived tissue engineered bone has good osteogenesis. No obvious rejection and other compl ications are observed in the cl inical appl ication.
Objective To manufacture a poly (lactic-co-glycolic acid) (PLGA) scaffold by low temperature deposition three-dimensional (3D) printing technology, prepare a PLGA/decellularized articular cartilage extracellular matrix (DACECM) cartilage tissue engineered scaffold by combining DACECM, and further investigate its physicochemical properties. Methods PLGA scaffolds were prepared by low temperature deposition 3D printing technology, and DACECM suspensions was prepared by modified physical and chemical decellularization methods. DACECM oriented scaffolds were prepared by using freeze-drying and physicochemical cross-linking techniques. PLGA/DACECM oriented scaffolds were prepared by combining DACECM slurry with PLGA scaffolds. The macroscopic and microscopic structures of the three kinds of scaffolds were observed by general observation and scanning electron microscope. The chemical composition of DACECM oriented scaffold was analyzed by histological and immunohistochemical stainings. The compression modulus of the three kinds of scaffolds were measured by biomechanical test. Three kinds of scaffolds were embedded subcutaneously in Sprague Dawley rats, and HE staining was used to observe immune response. The chondrocytes of New Zealand white rabbits were isolated and cultured, and the three kinds of cell-scaffold complexes were prepared. The growth adhesion of the cells on the scaffolds was observed by scanning electron microscope. Three kinds of scaffold extracts were cultured with L-929 cells, the cells were cultured in DMEM culture medium as control group, and cell counting kit 8 (CCK-8) was used to detect cell proliferation. Results General observation and scanning electron microscope showed that the PLGA scaffold had a smooth surface and large pores; the surface of the DACECM oriented scaffold was rough, which was a 3D structure with loose pores and interconnected; and the PLGA/DACECM oriented scaffold had a rough surface, and the large hole and the small hole were connected to each other to construct a vertical 3D structure. Histological and immunohistochemical qualitative analysis demonstrated that DACECM was completely decellularized, retaining the glycosaminoglycans and collagen typeⅡ. Biomechanical examination showed that the compression modulus of DACECM oriented scaffold was significantly lower than those of the other two scaffolds (P<0.05). There was no significant difference between PLGA scaffold and PLGA/DACECM oriented scaffold (P>0.05). Subcutaneously embedded HE staining of the three scaffolds showed that the immunological rejections of DACECM and PLGA/DACECM oriented scaffolds were significantly weaker than that of the PLGA scaffold. Scanning electron microscope observation of the cell-scaffold complex showed that chondrocytes did not obviously adhere to PLGA scaffold, and a large number of chondrocytes adhered and grew on PLGA/DACECM oriented scaffold and DACECM oriented scaffold. CCK-8 assay showed that with the extension of culture time, the number of cells cultured in the three kinds of scaffold extracts and the control group increased. There was no significant difference in the absorbance (A) value between the groups at each time point (P>0.05). Conclusion The PLGA/DACECM oriented scaffolds have no cytotoxicity, have excellent physicochemical properties, and may become a promising scaffold material of tissue engineered cartilage.
ObjectiveTo review the literature on the research status of vascularization of tissue engineered peripheral nerve so as to provide the theoretical basis for the vascularization of tissue engineered peripheral nerve.MethodsThe literature related to the vascularization of peripheral nerve tissue engineering in recent years was reviewed and summarized according to the five aspects of promoting vascularization: local microenvironment and blood supply characteristics of peripheral nerve regeneration, scaffold material modification, seed cells, autologous vascular bundle implantation, and pro-vascular factors.ResultsTissue engineered peripheral nerve has brought a new hope for the repair of peripheral nerve injury, but the repair effect of large nerve defects is not good, which is mainly related to the degree of vascularization of the nerve grafts. So it is particularly important to promote the early vascularization of tissue engineered peripheral nerve. Previous studies have mainly focused on the four aspects of scaffold material modification, seed cells, autologous vascular bundle implantation, and angiogenesis related factors. Recent studies show that the combination of the above two or more factors in the tissue engineered peripheral nerves can better promote the vascularization of tissue engineered peripheral nerves.ConclusionPromoting early vascularization of tissue engineered peripheral nerves can provide timely nutritional support for seed cells on the scaffold, promote axon growth and nerve regeneration, and facilitate the repair of large peripheral nerve defects in clinical practice.
ObjectiveTo manufacture a polycaprolactone (PCL)/type Ⅰ collagen (COL Ⅰ) tissue engineered meniscus scaffold (hereinafter referred to as PCL/COL Ⅰ meniscus scaffold) by three-dimensional (3D) printing with low temperature deposition technique and to study its physicochemical properties.MethodsFirst, the 15% PCL/4% COLⅠ composite solution and 15% PCL simple solution were prepared. Then, 15% PCL/4% COL Ⅰmeniscus scaffold and 15% PCL meniscal scaffold were prepared by using 3D printing with low temperature deposition techniques. The morphology and microstructure of the scaffolds were observed by gross observation and scanning electron microscope. The compression modulus and tensile modulus of the scaffolds were measured by biomechanical test. The components of the scaffolds were analyzed by Fourier transform infrared spectroscopy (FTIR). The contact angle of the scaffold surface was measured. The meniscus cells of rabbits were cultured with the two scaffold extracts and scaffolds, respectively. After cultured, the cell proliferations were detected by cell counting kit 8 (CCK-8), and the normal cultured cells were used as controls. Cell adhesion and growth of scaffold-cell complex were observed by scanning electron microscope.ResultsAccording to the gross and scanning electron microscope observations, two scaffolds had orientated 3D microstructures and pores, but the surface of the PCL/COLⅠ meniscus scaffold was rougher than the PCL meniscus scaffold. Biomechanical analysis showed that the tensile modulus and compression modulus of the PCL/COL Ⅰ meniscus scaffold were not significantly different from those of the PCL meniscus scaffold (P>0.05). FTIR analysis results showed that COL Ⅰ and PCL were successful mixed in PCL/ COL Ⅰ meniscus scaffolds. The contact angle of PCL/COLⅠ meniscus scaffold [(83.19±7.49)°] was significantly lower than that of PCL meniscus scaffold [(111.13±5.70)°] (t=6.638, P=0.000). The results of the CCK-8 assay indicated that with time, the number of cells cultured in two scaffold extracts showed an increasing trend, and there was no significant difference when compared with the control group (P>0.05). Scanning electron microscope observation showed that the cells attached on the PCL/ COL Ⅰ meniscus scaffold more than that on the PCL scaffold.ConclusionPCL/COLⅠmeniscus scaffolds are prepared by 3D printing with low temperature deposition technique, which has excellent physicochemical properties without cytotoxicity. PCL/COLⅠmeniscus scaffold is expected to be used as the material for meniscus tissue engineering.
Objective To summarize and analyze the research progress of scaffold materials used in tissue engineered meniscus. Methods The classification and bionics design of scaffold materials were summarized by consulting domestic and foreign literature related to the research of tissue engineered meniscus in recent years. Results Tissue engineered meniscus scaffolds can be roughly classified into synthetic polymers, hydrogels, extracellular matrix components, and tissue derived materials. These different materials have different characteristics, so the use of a single material has its unique disadvantages, and the use of a variety of materials composite scaffolds can learn from each other, which is a hot research area at present. In addition to material selection, material processing methods are also the focus of research. At the same time, according to the morphological structure and mechanical characteristics of the meniscus, the bionic design of tissue engineered meniscus scaffolds has great potential. Conclusion At present, there are many kinds of scaffold materials for tissue engineered meniscus. However, there is no material that can completely simulate the natural meniscus, and further research of scaffold materials is still needed.
ObjectiveTo review the methods of improving the mechanical properties of hydrogels and the research progress in bone tissue engineering. MethodsThe recent domestic and foreign literature on hydrogels in bone tissue engineering was reviewed, and the methods of improving the mechanical properties of hydrogels and the effect of bone repair in vivo and in vitro were summarized. ResultsHydrogels are widely used in bone tissue engineering, but their mechanical properties are poor. Improving the mechanical properties of hydrogels can enhance bone repair. The methods of improving the mechanical properties of hydrogels include the construction of dual network structures, inorganic nanoparticle composites, introduction of conductive materials, and fiber network reinforcement. These methods can improve the mechanical properties of hydrogels to various degrees while also demonstrating a significant bone repair impact. ConclusionThe mechanical properties of hydrogels can be effectively improved by modifying the system, components, and fiber structure, and bone repair can be effectively promoted.
Objective To review the current status and problems in the developing scaffolds for the myocardial tissue engineering appl ication. Methods The l iterature concerning the myocardial tissue engineering scaffold in recent years was reviewed extensively and summarized. Results As one of three elements for tissue engineering, a proper scafold is veryimportant for the prol iferation and differentiation of the seeding cells. The naturally derived and synthetic extracellular matrix (ECM) materials aim to closely resemble the in vivo microenvironment by acting as an active component of the developing tissue construct in myocardial tissue engineering. With the advent and continuous refinement of cell removal techniques, a new class of native ECM has emerged with some striking advantages. Conclusion Through using the principle of composite scaffold, computers and other high-technology nano-polymer technology, surface modification of traditional biological materials in myocardial tissue engineering are expected to provide ideal myocardial scaffolds.
Objective To investigate the biocompatibility of p(3HB-co-3HH) and marrow mesenchymal stell cells (MSCs).Methods MSCs were inoculated to p(3HB-co-3HH), and then cultured for 2-4 weeks in vitro and embedded for 2 weeks in vivo. The growth, proliferation, morphology and phenotype properties of MSCs were observed by use of phase contrast microscope, electron microscope, HE staining and staining of type Ⅰ collagen. Results p(3HB-co-3HH) hadgood compatibility. The inoculated MSCs could be well-distributed, attached well and obtain the phenotype of MSCs in p(3HB-co-3HH). After osteogenic inducer were added, MSCs differentiated to osteoblasts and secreted matrix. Type Ⅰ collagen was stained positively by immunohistochemical techenique. Conclusion The above results demonstrate that there is satisfactory biocompatibility betweenp(3HB-co-3HH) and MSCs.
Objective To review the recent advances in the application of graphene oxide (GO) for bone tissue engineering. Methods The latest literature at home and abroad on the GO used in the bone regeneration and repair was reviewed, including general properties of GO, degradation performance, biocompatibility, and application in bone tissue engineering. Results GO has an abundance of oxygen-containing functionalities, high surface area, and good biocompatibility. In addition, it can promote stem cell adhesion, proliferation, and differentiation. Moreover, GO has many advantages in the construction of new composite scaffolds and improvement of the performance of traditional scaffolds. Conclusion GO has been a hot topic in the field of bone tissue engineering due to its excellent physical and chemical properties. And many problems still need to be solved.