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![A: the biotin–avidin system, the biotin was conjugated to cell membrane, and the avidin was immobilized to the chitosan substrate. The bond formation between biotin and avidin mediated chondrocyte adhesion to the chitosan surfaces. B: The biotin-conjugated anti-CD44 antibody–avidin binding system, where the biotin–avidin was used as a bridge connecting the chitosan substrate and the antibodies, and the monoclonal anti-CD44 antibodies were used as the bridge connecting the chondrocytes and the biotin. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]](profile/Yuhui-Ruan/publication/236581550/figure/fig5/AS:281661611757577@1444164769212/A-the-biotin-avidin-system-the-biotin-was-conjugated-to-cell-membrane-and-the-avidin.png)
A: the biotin–avidin system, the biotin was conjugated to cell membrane, and the avidin was immobilized to the chitosan substrate. The bond formation between biotin and avidin mediated chondrocyte adhesion to the chitosan surfaces. B: The biotin-conjugated anti-CD44 antibody–avidin binding system, where the biotin–avidin was used as a bridge connecting the chitosan substrate and the antibodies, and the monoclonal anti-CD44 antibodies were used as the bridge connecting the chondrocytes and the biotin. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]
Source publication
The clinical need for improved treatment options for patients with cartilage injuries has motivated tissue-engineering studies aimed at the in vitro generation of cell-based implants with functional properties. The success of tissue-engineered repair of cartilage may depend on the rapid and efficient adhesion of transplanted cells to the scaffold....
Contexts in source publication
Context 1
... Meanwhile, the BA system can combine with solid materials, such as the chitosan cartilage tissue-engineering scaffold used in this study. 15 The mechanism of the BA system for improving chondrocyte adhesion is illustrated in Figure 7(A). The biotin was conjugated to cell membrane, [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.] and the avidin was immobilized to the chitosan substrate. ...
Context 2
... the current study, the combination of the BA system and monoclonal anti-CD44 antibodies was applied for enhancing chondrocyte adhesion and cartilage regeneration in porous scaffolds [ Fig. 7(B)]. The BA system was used as a bridge to connect the chitosan substrate and the antibodies, and the monoclonal anti-CD44 antibodies were used as the bridge to connect the chondrocytes and the biotin. Consequently, the endocytosis process was solved by avoiding direct contact of biotin to the chondrocytes. The monoclonal anti-CD44 ...
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INTRODUCTION In the design of engineered tissues, the processes of biomaterial degeneration and neotissue synthesis combine to affect the mechanical state of the multi-phase composite. Cell-polymer constructs as used in engineered cartilage, may consist of chondrocytes seeded on scaffolds of biodegradable polymers [Wilson et al., 2002]. Synthetic s...
Citations
... [132] Affinity peptides can bind with specific cells, scaffolds, and cytokines associated with cartilage regeneration. Using anti-CD44 antibody, biotin-avidin binding system [133], chondrocyte affinity peptides, [134] E7 peptide, [135], etc., cell adhesion was greatly improved. A TGF-β affinity peptide which can recruit TGF-β to the impaired region [136,137] was ligated to the PA nanofibers, promoted in vitro chondrogenic differentiation of hMSCs and cartilage repair in rabbits. ...
The healing of osteochondral defects (OCD) resulting from injury, osteochondritis, or osteoarthritis in middle- and old-age individuals, bearing lesions in the cartilage and bone, pain, and loss of joint function, presents challenges to the clinical practitioners because of non-regenerative cartilage and the limitations of current therapies. Bioactive peptide-based osteochondral (OC) tissue regeneration is becoming more popular because it does not have the immunogenicity, misfolding, or denaturation problems associated with original proteins. Periodically, reviews are published on the regeneration of bone and cartilage; however, none of them address the simultaneous healing of these tissues in the complicated heterogeneous environment of the OC interface. As regulators of cell adhesion, proliferation, differentiation, angiogenesis, immunomodulation, and antibacterial activity, potential therapeutic strategies for OC defects utilizing bone and cartilage-specific peptides should be examined and investigated. Therefore, the main goal of this review is to study how they contribute to the healing of OC defects, either alone or in conjunction with other peptides and biomaterials.
... The system enabled the efficient improvement of cell adhesion mediated by the avidin-biotin interaction. In addition, the anti-CD44 antibody attachment to CD44 increased the mRNA expression of chondrogenic markers and glycosaminoglycans synthesis and enhanced cell proliferation and viability [81]. These observations are in accordance with our in vitro results. ...
Hydrogels are suitable for osteochondral defect regeneration as they mimic the viscoelastic environment of cartilage. However, their biomechanical properties are not sufficient to withstand high mechanical forces. Therefore, we have prepared electrospun poly-ε-caprolactone-chitosan (PCL-chit) and poly(ethylene oxide)-chitosan (PEO-chit) nanofibers, and FTIR analysis confirmed successful blending of chitosan with other polymers. The biocompatibility of PCL-chit and PEO-chit scaffolds was tested; fibrochondrocytes and chondrocytes seeded on PCL-chit showed superior metabolic activity. The PCL-chit nanofibers were cryogenically grinded into microparticles (mean size of about 500 µm) and further modified by polyethylene glycol–biotin in order to bind the anti-CD44 antibody, a glycoprotein interacting with hyaluronic acid (PCL-chit-PEGb-antiCD44). The PCL-chit or PCL-chit-PEGb-antiCD44 microparticles were mixed with a composite gel (collagen/fibrin/platelet rich plasma) to improve its biomechanical properties. The storage modulus was higher in the composite gel with microparticles compared to fibrin. The Eloss of the composite gel and fibrin was higher than that of the composite gel with microparticles. The composite gel either with or without microparticles was further tested in vivo in a model of osteochondral defects in rabbits. PCL-chit-PEGb-antiCD44 significantly enhanced osteogenic regeneration, mainly by desmogenous ossification, but decreased chondrogenic differentiation in the defects. PCL-chit-PEGb showed a more homogeneous distribution of hyaline cartilage and enhanced hyaline cartilage differentiation.
... This method was first performed using a chondrocytespecific antibody. Lin et al. later used anti-CD44 antibody and the biotin-avidin binding system to improve cell adhesion and cartilage repair [31]. However, the high cost and difficulty of antibody loading in this system make it impracticable. ...
Cartilage repair after degeneration or trauma continues to be a challenge both in the clinic and for scientific research due to the limited regenerative capacity of this tissue. Cartilage tissue engineering, involving a combination of cells, scaffolds, and growth factors, is increasingly used in cartilage regeneration. Due to their ease of synthesis, robustness, tunable size, availability of functional groups, and activity, peptides have emerged as the molecules with the most potential in drug development. A number of peptides have been engineered to regenerate cartilage by acting as scaffolds, functional molecules, or both. In this paper, we will summarize the application of peptides in cartilage tissue engineering and discuss additional possibilities for peptides in this field.
... The incorporation of GMs had little influence on the scaffold morphology for its relative smaller size. Scaffolds must be 100-200 lm in diameter and approximately 90% porous so that sufficient volume and surface can be provided for cell growth for skin tissue engineering [42]. The results suggested that the three-dimensional GS/GM/Coll-CNCs scaffold have potential for skin regeneration. ...
In this study, a kind of antibacterial collagen (Coll)/cellulose nanocrystals (CNCs) scaffolds was prepared and characterized, which can be applied in skin trauma as a promising dressing. In order to achieve the goal of antibacterial function for a period of long time, the Gentamycin sulfate (GS) was absorbed in the prepared gelatin microsphere (GMs) then doped in the porous Coll–CNCs freeze-drying scaffold. The CNCs had a width of 20–40 nm and length of 150–350 nm from electron microscopy (TEM), retained the basic chemical structure of cellulose observed from Fourier transform infrared, and increased the machinery properties of Coll scaffold from thermal property analysis. From the SEM, the mean pore size of the compound scaffolds was about 120 μm with or without GMs. The in vitro release profile of GS showed a sustained release of this GS/GMs/Coll–CNCs composite scaffold. The biological compatibility experiments showed the composite scaffold low cellular cytotoxicity of NIH-3T3 cells. The significant inhibition zones indicated the antibacterial efficacy of the composite scaffold against Escherichia coli and Staphylococcus aureus.
... our results showed that the ability of chondrocytes to adhere to scaffolds prepared by the CD44 monoclonal antibody biotinavidin (CBa) binding technique led to improved cartilage tissue engineering. 27 However, in vivo repair of cartilage defects with the tissue-engineered cartilage constructed with the biotin-conjugated anti-CD44 antibody-avidin binding system, especially for defects in the weightbearing area, remains to be explored. Thus, the current study was designed to repair full-thickness articular cartilage defects in the weight-bearing area in a porcine model, and to investigate whether the CBa binding technique could provide better tissue-engineered cartilage for clinical applications. ...
... 24 In our previous study, the combination of the Ba system and monoclonal anti-CD44 antibodies was applied in order to enhance the adhesion of chondrocytes and the regeneration of cartilage in porous scaffolds, and we found that the CBa technique can enhance the adhesion between chondrocytes and 3D chitosan scaffold in vitro. 27 In the current study, we found that the rate of detachment was greatly reduced by the CBa technique at four hours after implantation. Real-time PCR results at one week after implantation showed an increase in the levels of expression of Col II, aggrecan and SoX9 in the CBa group. ...
... This is consistent with our previous in vitro studies. 27 The pigs in this study recovered well post-implantation without local complications. The surgical process is simple and safe. ...
Objectives:
The lack of effective treatment for cartilage defects has prompted investigations using tissue engineering techniques for their regeneration and repair. The success of tissue-engineered repair of cartilage may depend on the rapid and efficient adhesion of transplanted cells to a scaffold. Our aim in this study was to repair full-thickness defects in articular cartilage in the weight-bearing area of a porcine model, and to investigate whether the CD44 monoclonal antibody biotin-avidin (CBA) binding technique could provide satisfactory tissue-engineered cartilage.
Methods:
Cartilage defects were created in the load-bearing region of the lateral femoral condyle of mini-type pigs. The defects were repaired with traditional tissue-engineered cartilage, tissue-engineered cartilage constructed with the biotin-avidin (BA) technique, tissue-engineered cartilage constructed with the CBA technique and with autologous cartilage. The biomechanical properties, Western blot assay, histological findings and immunohistochemical staining were explored.
Results:
The CBA group showed similar results to the autologous group in biomechanical properties, Moran's criteria, histological tests and Wakitani histological scoring.
Conclusions:
These results suggest that tissue-engineered cartilage constructed using the CBA technique could be used effectively to repair cartilage defects in the weight-bearing area of joints.Cite this article: H. Lin, J. Zhou, L. Cao, H. R. Wang, J. Dong, Z. R. Chen. Tissue-engineered cartilage constructed by a biotin-conjugated anti-CD44 avidin binding technique for the repairing of cartilage defects in the weight-bearing area of knee joints in pigs. Bone Joint Res 2017;6:-295. DOI: 10.1302/2046-3758.65.BJR-2016-0277.
The healing of osteochondral defects (OCDs) that result from injury, osteochondritis, or osteoarthritis and bear lesions in the cartilage and bone, pain, and loss of joint function in middle- and old-age individuals presents challenges to clinical practitioners because of non-regenerative cartilage and the limitations of current therapies. Bioactive peptide-based osteochondral (OC) tissue regeneration is becoming more popular because it does not have the immunogenicity, misfolding, or denaturation problems associated with original proteins. Periodically, reviews are published on the regeneration of bone and cartilage separately; however, none of them addressed the simultaneous healing of these tissues in the complicated heterogeneous environment of the osteochondral (OC) interface. As regulators of cell adhesion, proliferation, differentiation, angiogenesis, immunomodulation, and antibacterial activity, potential therapeutic strategies for OCDs utilizing bone and cartilage-specific peptides should be examined and investigated. The main goal of this review was to study how they contribute to the healing of OCDs, either alone or in conjunction with other peptides and biomaterials.
Cartilages are unique in the family of connective tissues in that they contain a high concentration of the glycosaminoglycans, chondroitinsulfate and keratan sulfate attached to the core protein of the proteoglycan, aggrecan. Multiple aggrecan molecules are organized in the extracellular matrix via a domain-specific molecular interaction with hyaluronan and a link protein, and these high molecular weight aggregates are immobilized within the collagen and glycoproteinnetwork. The high negative charge density of glycosaminoglycans provides hydrophilicity, high osmotic swelling pressure and conformational flexibility, which together function to absorb fluctuations in biomechanical stresses on cartilage during movement of an articular joint. We have summarized information on the history and current knowledge obtained by biochemical and genetic approaches, on cell-mediated regulation of aggrecan metabolism and its role in skeletal development, growth as well as during the development of joint disease. In addition, we describe the pathways for hyaluronan metabolism, with particular focus on the role as a “metabolicrheostat” during chondrocyte responses in cartilage remodeling in growth and disease.
Future advances in effective therapeutic targeting of cartilage loss during osteoarthritic diseases of the joint as an organ as well as in cartilage tissue engineering would benefit from ‘big data’ approaches and bioinformatics, to uncover novel feed-forward and feed-back mechanisms for regulating transcription and translation of genes and their integration into cell-specific pathways.
Gelatin (GA), hyaluronic acid (HA) and cellulose nanocrystals (CNC) are promising materials for skin wound care. In this study the GA-HA-CNC hydrogels were prepared by cross-linking and freeze-drying. The composition and mechanism of GA-HA-CNC hydrogels were confirmed by FTIR. The morphology and pore size were obtained by SEM. We accessed the physical property from rheological results and swelling ratio. NIH-3T3 cells were inoculated into the hydrogels and cultured for different days, then we analyzed the cytotoxicity of the prepared hydrogels by CCK-8 methods and live/dead pictorial diagram using staining kits. FTIR revealed the combination between GA, HA and CNC was attributed to the amide bond and hydrogen bonding. SEM results showed that the drying GA-HA-CNC hydrogels were spongy, with the pore diameter about 80-120μm. CNC significantly enhanced the property of the hydrogels and play a vital role according to the rheology and swelling results. The cells culture results showed that NIH-3T3 cells can attached to, grow, and proliferate well on the GA-HA-CNC hydrogels. In conclusion, the natural GA-HA-CNC hydrogel has great potential for the skin wound repair.
Avidin is a glycoprotein with antinutritional property, which should be limited in daily food. We developed an affinity biosensor system based on resonance Rayleigh scattering (RRS) and using affinity biotin labeling Au nanoparticles (AuNPs). This method was selective and sensitive for quick avidin detection due to the avidin–biotin affinitive interaction. Under optimal conditions, RRS intensity of biotin–AuNPs increase linearly with an increasing concentration of avidin from 5 to 160 ng/mL. The lower limit of detection was 0.59 ng/mL. This rapid and selective avidin detection method was used in synthetic samples and egg products with recoveries of between 102.97 and 107.92%, thereby demonstrating the feasible and practical application of this assay.
