ObjectiveTo review the application of silk fibroin scaffold in bone tissue engineering. MethodsThe related literature about the application of silk fibroin scaffold in bone tissue engineering was reviewed, analyzed, and summarized. ResultsSilk fibroin can be manufactured into many types, such as hydrogel, film, nano-fiber, and three-dimensional scaffold, which have superior biocompatibility, slow biodegradability, nontoxic degradation products, and excellent mechanical strength. Meanwhile these silk fibroin biomaterials can be chemically modified and can be used to carry stem cells, growth factors, and compound inorganic matter. ConclusionSilk fibroin scaffolds can be widely used in bone tissue engineering. But it still needs further study to prepare the scaffold in accordance with the requirement of tissue engineering.
OBJECTIVE: To observe the effects of silks on attachment, shape and function of chondrocytes cultured in vitro. METHODS: The silks from silk worm cocoons were digested by trypsin and coated with polylactic acid to from three dimensional scaffolds for rabbit rib chondrocyte culture. The growth and shape of chondrocytes were observed with phase contrast microscopy, scanning electron microscopy. RESULTS: The chondrocytes were adhered to silks slowly after chondrocytes were seeded into silk scaffolds and cells fixed on silks well 1 or 2 days later. Cells began to proliferate after 3 days and multiplicative growth was observed on the 6th day. Microholes of silk scaffolds were filled with chondrocytes 2 weeks later. Scanning electron microscopy showed that there was a lot of extracellular matrix surrounding cells. CONCLUSION: Silks are ideal for attachment, growth and function maintenance of chondrocytes, and silks can be used as scaffolds for chondrocytes in three dimensional culture.
Objective To summarize the latest developments in silk protein fiber as biomaterials and their applications in tissue engineering. Methods Recent original literature on silk protein fiber as biomaterials were reviewed, illustrating the properties of silk protein fiber biomaterials. Results The silk protein fiber has the same functions of supporting the cell adhesion, differentiation and growth as native collagen, and is renewed as novel biomaterials with good biocompatibility, unique mechanical properties and is degradable over a longer time. Conclusion Silk protein-fiber can be used as asuitable matrix for three dimensional cell culture in tissue engineering. It has a great potential applications in other fields.
Objective To investigate the diagnostic efficacy of silkworm larvae plasma (SLP) colorimetry in the accurate diagnosis of periprosthetic joint infection (PJI). Methods Ninety healthy male New Zealand white rabbits were used for knee arthroplasty with Swanson prosthesis. Then they were randomly divided into 3 groups according to different pathogenic bacteria: group A (Staphylococcus aureus group), group B (Staphylococcus epidermidis group) and group C (Escherichia coli group), with 30 rats in each group. The PJI model was prepared by knee injection with 1 mL of pathogenic bacteria of different concentrations. Samples were taken before inoculation and at 7, 14, and 21 days after inoculation, and based on the 2018 PJI Philadelphia International Consensus diagnostic criteria, the success rate of modeling among 3 groups of experimental animals was determined. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic efficiency of SLP colorimetry were calculated. Results At 21 days after inoculation, 26, 18, and 23 rabbits in groups A, B, and C were diagnosed as infection, respectively. The success rates of modeling were 86.7%, 60.0%, and 76.7%, respectively, showing no significant difference among the 3 groups (χ2=5.724, P=0.073). The results of PJI colorimetry showed that 1 false-positive animal (specificity 75.0%) appeared in group A at 7 days, and the specificity of SLP increased to 100.0% over time (on 14 and 21 days); on 14 and 21 days, another animal appeared false-negative results (sensitivity decreased from 100.0% to 96.2%). One false-positive animal appeared in group B at 7 days (specificity 91.7%), the specificity returned to 100.0% over time; 1 and 4 false-negative animals appeared at 14 and 21 days, respectively (sensitivity 94.4% and 83.3%, respectively). In group C, two false-positive animals (specificity 71.4%) were found at 7 days, and then returned to 100.0%. The diagnostic efficiency of groups A and C was very high at 21 days (96.7% and 100.0%), even for the low virulence Staphylococcus epidermidis in group B, the diagnostic efficiency could be maintained at 90.0% (21 days), and the overall diagnostic efficiency was very good (95.6%). Conclusion SLP colorimetry has high sensitivity, specificity, and diagnostic efficiency in the diagnosis of PJI, which is a potential diagnostic method.
Test of tissue tolerance to domastic prosthetic materials (carbon fiber mesh, siliconized velvet, silk cloth and dacron cloth) as a subcutaneos transplant was performed in the adcominal wall of rabbit. These implants and their surroundding tissues were excied for studies at second , fourth, eighth and the twelfth weeks after operation. Ratio of fibroblast count to inflammatory cells count which is a common parameter of tissue tolerance was calculated in these four groups. The result shows that fibroblastic cell reaction elicited by carbon fiber mesh is the greates among the four prosthetic materials, the second one is dasron cloth. The inflammatory cell reaction elicited by silk is the greatest among the four materials, the second is carbon fiber mesh, and the dacron cloth the least. Tissure tolerance of dacron cloth is the best in the four prosthetic materials for implantation while sick is the worst.
【Abstract】 Objective To summarize the latest developments in silk fibroin as biomaterials and its appl icationsin tissue engineering. Methods The recent original l iterature on silk fibroin as biomaterials were extensively reviewed,illustrating the properties and appl ications of silk fibroin biomaterials in tissue engineering. Results Silk fibroinas biomaterials had good biocompatibil ity and degradabil ity. It supported the cell adhesion differentiation and growth. It was used for artificial l igament, vessel, bone, nerve and so on. After modification, silk fibroin could be extensively used in tissue engineering. Conclusion Silk fibroin is a good biomaterial, which has a great potential appl ications in tissue engineering.
Objective To observe the effect of dynamic mechanical loading on the proliferation, differentiation, and specific gene expression of MC3T3-E1 cells that on three-dimensional (3D) biomimetic composite scaffolds prepared by low temperature 3D printing technology combined with freeze-drying. Methods The silk fibroin, collagen type Ⅰ, and nano-hydroxyapatite (HA) were mixed at a mass ratio of 3∶9∶2 and were used to prepare the 3D biomimetic composite scaffolds via low temperature 3D printing technology combined with freeze-drying. General morphology of 3D biomimetic composite scaffold was observed. Micro-CT was used to observe the pore size and porosity of the scaffolds, and the water swelling rate, stress, strain, and elastic modulus were measured. Then, the MC3T3-E1 cells were seeded on the 3D biomimetic composite scaffolds and the cell-scaffold composites were randomly divided into 2 groups. The experimental group was subjected to dynamic mechanical loading (3 500 με, 1 Hz, 15 minutes per day); the control group was not subjected to loading treatment. After 7 days and 14 days, the cell-scaffold composites of 2 groups were harvested to observe the growth of cells on the scaffolds by HE staining and scanning electron microscope. And the gene and protein expressions of collagen type Ⅰ, BMP-2, and osteocalcin (OCN) were measured by real-time fluorescent quantitative PCR and Western blot. Results The 3D biomimetic composite scaffold was a white cubic grid. Micro-CT detection showed the pore network structure in the scaffold material with good pore connectivity. The diameters of large pore and micro-aperture were (506.37±18.63) μm and (62.14±17.35) μm, respectively. The porosity was 97.70%±1.37%, and the water absorption swelling rate was 1 341.97%±64.41%. Mechanical tests showed that the compression displacement of the scaffold was (0.376±0.004) mm, the compressive stress was (0.016±0.002) MPa, and the elastic modulus was (162.418±18.754) kPa when the scaffold was compressed to 10%. At 7 days and 14 days, HE staining and scanning electron microscope observation showed that the cells grew inside the scaffold, mainly distributed around the scaffold pore wall. The cells in experimental group were more than control group, and the cells morphology changed from shuttle to flat. There was no significant difference in the cell counting between 2 groups at 14 days after 200-fold microscopy (t=–2.024, P=0.080), but significant differences were found between 2 groups at different time points under different magnifications (P<0.05). Real-time fluorescent quantitative PCR showed that the mRNA relative expressions of collagen type Ⅰ and OCN in experimental group were significantly higher than those in control group at 7 and 14 days (P<0.05). However, the mRNA relative expression of BMP-2 showing no significant difference between 2 groups (P>0.05). The protein relative expressions of collagen type Ⅰ, BMP-2, and OCN in experimental group were significantly higher than those in control group at 7 and 14 days (P<0.05). Conclusion After dynamic mechanical loading, the expressions of BMP-2, collagen type Ⅰ, and OCN in MC3T3-E1 cells inoculated into 3D biomimetic composite scaffolds are significantly up-regulated, indicating that appropriate mechanical loads favor osteoblast differentiation of MC3T3-E1 cells.
ObjectiveThe properties and characteristics of different types of silk fibroin (SF) drug-loaded sustained-release carriers and their effects on the drug release behavior were reviewed, and the existing problems and development prospects of SF drug-loaded sustained-release carriers in tissue engineering drug delivery system were discussed.MethodsThe literatures about drug-loaded SF sustained-release carriers in recent years were extensively consulted, and the types of sustained-release carriers, characteristics of drug release, range of applications, advantages and disadvantages, and solutions were summarized and analyzed.ResultsAt present, the SF drug-loaded sustained-release carriers are mainly divided into SF microparticles, SF scaffolds, SF membranes, SF hydrogels, SF nanofibers, SF sponges, and so on. These types of SF drug-loaded sustained-release carriers have their own advantages and problems, of which the most prominent problem is the burst release of drugs at the initial stage. While, the initial burst release of drugs can be effectively solved by improving the preparation process and adjusting the material ratio. Different types of drug-loaded sustained-release carriers can be prepared by combining different materials to achieve different application scopes and drug release behaviors under different conditions.ConclusionSF is a good drug-loaded carrier for tissue engineering, the burst release of drugs at the initial stage can be solved by improving the preparation process and changing the material structure; through the combination of the advantages of various types of SF drug-loaded sustained-release carriers, it is expected to prepare SF drug-loaded sustained-release carriers that meet different clinical needs.
ObjectiveTo describe the research progress of silk-based biomaterials in peripheral nerve repair and provide useful ideals to accelerate the regeneration of large-size peripheral nerve injury. Methods The relative documents about silk-based biomaterials used in peripheral nerve regeneration were reviewed and the different strategies that could accelerate peripheral nerve regeneration through building bioactive microenvironment with silk fibroin were discussed. Results Many silk fibroin tissue engineered nerve conduits have been developed to provide multiple biomimetic microstructures, and different microstructures have different mechanisms of promoting nerve repair. Biomimetic porous structures favor the nutrient exchange at wound sites and inhibit the invasion of scar tissue. The aligned structures can induce the directional growth of nerve tissue, while the multiple channels promote the axon elongation. When the fillers are introduced to the conduits, better growth, migration, and differentiation of nerve cells can be achieved. Besides biomimetic structures, different nerve growth factors and bioactive drugs can be loaded on silk carriers and released slowly at nerve wounds, providing suitable biochemical cues. Both the biomimetic structures and the loaded bioactive ingredients optimize the niches of peripheral nerves, resulting in quicker and better nerve repair. With silk biomaterials as a platform, fusing multiple ways to achieve the multidimensional regulation of nerve microenvironments is becoming a critical strategy in repairing large-size peripheral nerve injury. Conclusion Silk-based biomaterials are useful platforms to achieve the design of biomimetic hierarchical microstructures and the co-loading of various bioactive ingredients. Silk fibroin nerve conduits provide suitable microenvironment to accelerate functional recovery of peripheral nerves. Different optimizing strategies are available for silk fibroin biomaterials to favor the nerve regeneration, which would satisfy the needs of various nerve tissue repair. Bioactive silk conduits have promising future in large-size peripheral nerve regeneration.
ObjectiveTo prepare the small intestinal submucosa (SIS)-silk composite scaffold for anterior cruciate ligament (ACL) reconstruction, and to evaluate its properties of biomechanics, biocompatibility, and the influence on synovial fluid leaking into tibia tunnel so as to provide a better choice in the clinical application of ACL reconstruction. MethodsThe silk was used to remove sericin and then weaved as silk scaffold, which was surrounded cylindrically by SIS to prepare a composite scaffold. The property of biomechanics was evaluated by biomechanical testing system. The cell biocompatibility of scaffolds was evaluated by live/dead staining and the cell counting kit 8 (CCK- 8). Thirty 6-week-old Sprague Dawley rats were randomly assigned to 2 groups (n=15). The silk scaffold (S group) and composite scaffold (SS group) were subcutaneously implanted. At 2, 4, and 8 weeks after implanted, the specimen were harvested for HE staining to observe the biocompatibility. Another 20 28-week-old New Zealand white rabbits were randomly assigned to the S group and SS group (n=20), and the silk scaffold and composite scaffold were used for ACL reconstruction respectively in 2 groups. Furthermore, a bone window was made on the tibia tunnel. At last, the electric resistance of tendon graft in the bone window was measured and recorded at different time points after 5 mL of 10% NaCl or 5 mL of ink solution was irrigated into the joint cavity recspectively. ResultsThe gross observation showed that the composite scaffold consisted of the helical silk bundle inside which was surrounded by SIS. The maximal load of silk scaffold and composite scaffold was respectively (138.62±11.41) N and (137.05±16.95) N, showing no significant difference (P>0.05); the stiffness was respectively (24.65±2.62) N/mm and (24.21±2.39) N/mm, showing no significant difference (P>0.05). The live/dead staining showed that the cells had good activity on both scaffolds. However, the cells on the composite scaffold had better extensibility. In addition, the cell proliferation curve indicated that no significant difference in the absorbance (A) values was founded between groups at various time points (P>0.05). HE staining showed less inflammatory cells and much more angiogenesis in SS group than in S group at 2, 4, and 8 weeks after subcutaneously implanted (P<0.05), indicating good biocompatibility. Additionally, the starting time points of electric resistance decrease and the ink leakage were both significantly later in SS group than in S group (P<0.05). The duration of ink leakage was significantly longer in SS group than in S group (P<0.05). ConclusionThe SIS-silk composite scaffold has excellent biomechanical properties and biocompatibility and early vacularization after in vivo implantation. Moreover, it can reducing the leakage of synovial fluid into tibia tunnel at the early stage of ACL reconstruction. So it is promising to be an ideal ACL scaffold.