Effect of double growth factor release on cartilage tissue engineering (original) (raw)
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Dual growth factor-releasing nanoparticle/hydrogel system for cartilage tissue engineering
Journal of Materials Science: Materials in Medicine, 2010
In order to induce the chondrogenesis of mesenchymal stem cells (MSCs) in tissue engineering, a variety of growth factors have been adapted and encouraging results have been demonstrated. In this study, we developed a delivery system for dual growth factors using a gelation rate controllable alginate solution (containing BMP-7) and polyion complex nanoparticles (containing TGF-b 2) to be applied for the chondrogenesis of MSCs. The dual growth factors (BMP-7/TGF-b 2)-loaded nanoparticle/hydrogel system showed a controlled release of both growth factors: a faster release of BMP-7 and a slower release of TGF-b 2 , ca., approximately 80 and 30% release at the end of an incubation period (21 days), respectively, which may be highly desirable for chondrogenic differentiation of MSCs. On the contrary, the release of each growth factor from the dual growth factors-loaded hydrogel (without the nanoparticles) was much slower than that of the nanoparticle/hydrogel system, approximately 36% (BMP-7) and 16% (TGF-b 2) for 21 days, and this is more than likely attributed to the aggregation between growth factors during the hydrogel fabrication step. The nanoparticle/hydrogel system with separate growth factor loading may provide desirable growth factor delivery kinetics for cartilage regeneration, as well as the chondrogenesis of MSCs.
Sequential release of bioactive IGF-I and TGF-β1 from PLGA microsphere-based scaffolds
Biomaterials, 2008
Growth factors have become an important component for tissue engineering and regenerative medicine. Insulin-like growth factor-I (IGF-I) and transforming growth factor-beta1 (TGF-b 1 ) in particular have great significance in cartilage tissue engineering. Here, we describe sequential release of IGF-I and TGF-b 1 from modular designed poly(L,D-lactic-co-glycolic acid) (PLGA) scaffolds. Growth factors were encapsulated in PLGA microspheres using spontaneous emulsion, and in vitro release kinetics was characterized by ELISA. Incorporating BSA in the IGF-I formulations decreased the initial burst from 80% to 20%, while using uncapped PLGA rather than capped decreased the initial burst of TGF-b 1 from 60% to 0% upon hydration. The bioactivity of released IGF-I and TGF-b 1 was determined using MCF-7 proliferation assay and HT-2 inhibition assay, respectively. Both growth factors were released for up to 70 days in bioactive form. Scaffolds were fabricated by fusing bioactive IGF-I and TGF-b 1 microspheres with dichloromethane vapor. Three scaffolds with tailored release kinetics were fabricated: IGF-I and TGF-b 1 released continuously, TGF-b 1 with IGF-I released sequentially after 10 days, and IGF-I with TGF-b 1 released sequentially after 7 days. Scaffold swelling and degradation were characterized, indicating a peak swelling ratio of 4 after 7 days of incubation and showing 50% mass loss after 28 days, both consistent with scaffold release kinetics. The ability of these scaffolds to release IGF-I and TGF-b 1 sequentially makes them very useful for cartilage tissue engineering applications.
2004
As cartilage tissue has limited repair capacities, tissue engineering has emerged as a promising alternative for cartilage repair. The scaffold is a primary component of the tissue engineering design, yet little information exists regarding the effects of polymer and scaffold properties on tissue growth. In this study, we have developed a novel scaffold, PLG microspheres, for use in cartilage tissue engineering, which has the capacity for alterations in polymer and scaffold. We examined the effects of molecular weight, hydrophobic capping, delivery of Mg(OH) 2 , microsphere size, and controlled release of IGF-I. Our findings demonstrated that polymer parameters distinctively affect tissue and matrix output. Specifically, micro spheres with high molecular weight polymer produced tissue with high GAG content and tissue mass in vivo and in vitro, while micro spheres with capped polymer induced steady tissue and matrix accumulation, but may have precluded cell attachment. Release of buf...
2022
Researchers have scrutinized cartilage tissue regeneration to handle the deficiency of cartilage restoration capacity. This investigation proposed to compose an innovative nanocomposite biomaterial that enhances growth factor delivery to the injured cartilage site. Here, we describe the design and development of the biocompatible PLGA-collagen / PLGA-PEG-PLGA nanocomposite scaffold containing TGF-β1. PLGA-PEG-PLGA nanoparticles were employed as a delivery system embedding TGF-β1 as an articular cartilage repair therapeutic agent. This study evaluates various physicochemical aspects of fabricated scaffolds by 1HNMR, FT-IR, SEM, BET, and DLS methods. The physicochemical features of the developed scaffolds, including porosity, density, degradation, swelling ratio, mechanical properties, morphologies, BET, ELIZA, and cytotoxicity were assessed. SEM images displayed suitable cell adhesion and distribution of hDPSCs throughout the scaffolds. The cell viability was investigated with the MT...
Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2019
Degeneration of articular cartilage due to damages, diseases, or age‐related factors can significantly decrease the mobility of the patients. Various tissue engineering approaches which take advantage of stem cells and growth factors in a three‐dimensional constructs have been used for reconstructing articular tissue. Proliferative impact of basic fibroblast growth factor (bFGF) and chondrogenic differentiation effect of transforming growth factor‐beta 1 (TGF‐β1) over mesenchymal stem cells have previously been verified. In this study, silk fibroin (SF) and of poly(ethylene glycol) dimethacrylate (PEGDMA) were used to provide a versatile platform for preparing hydrogels with tunable mechanical, swelling and degradation properties through physical and chemical crosslinking as a microenvironment for chondrogenic differentiation in the presence of bFGF and TGF‐β1 releasing nanoparticles (NPs) for the first time. Scaffolds with compressive moduli ranging from 95.70 ± 17.82 to 338.05 ± 3...
Pharmaceutics, 2021
Mesenchymal stem cells (MSCs) have been extensively used in the tissue regeneration therapy. Ex vivo therapy with well-differentiated osteogenic cells is known as an efficient treatment for musculoskeletal diseases, including rheumatoid diseases. However, along with its high cost, the current therapy has limitations in terms of restoring bone regeneration procedures. An efficient process for the cell differentiation to obtain a large number of functionalized osteogenic cells is necessary. Therefore, it is strongly recommended to develop strategies to produce sufficient numbers of well-differentiated osteogenic cells from the MSCs. In general, differentiation media with growth factors have been used to facilitate cell differentiation. In the present study, the poly (lactic-co-glycolic acid) (PLGA) nanoparticles incorporating the growth factors were included in the media, resulting in releasing growth factors (dexamethasone and β-glycerophosphate) in the media in the controlled manner...
Knee Surgery, Sports Traumatology, Arthroscopy, 2012
Purpose Due to the attractive properties of poly(L-lactic acid) (PLLA) for tissue engineering, the aim was to determine the growth and differentiation capacity of mesenchymal stromal cells (MSCs) in PLLA scaffolds and their potential use in the treatment of cartilage diseases. Methods MSCs were cultured in PLLA films and thin porous membranes to study adherence and proliferation. Permeability and porosity were determined for the different scaffolds employed. The optimal conditions for cell seeding were first determined, as well as cell density and distribution inside the PLLA. Scaffolds were then maintained in expansion or chondrogenic differentiation media for 21 days. Apoptosis, proliferation and chondrogenic differentiation was assessed after 21 days in culture by immunohistochemistry. Mechanical characteristics of scaffolds were determined before and after cell seeding. Results MSCs uniformly adhered to PLLA films as well as to porous membranes. Proliferation was detected only in monolayers of pure PLLA, but was no longer detected after 10 days. Mechanical characterization of PLLA scaffolds showed differences in the apparent compression elastic modulus for the two sizes used. After determining high efficiencies of seeding, the production of extracellular matrix (ECM) was determined and contained aggrecan and collagens type I and X. ECM produced by the cells induced a twofold increase in the apparent elastic modulus of the composite. Conclusions Biocompatible PLLA scaffolds have been developed that can be efficiently loaded with MSCs. The scaffold supports chondrogenic differentiation and ECM deposition that improves the mechanics of the scaffold. Although this improvement does not met the expectations of a hyaline-like cartilage ECM, in part due to the lack of a mechanical stimulation, their potential use in the treatment of cartilage pathologies encourages to improve the mechanical component.
Tissue Engineering, 2006
Despite the widespread role of transforming growth factor-β3 (TGFβ3) in wound healing and tissue regeneration, its long-term controlled release has not been demonstrated. Here, we report microencapsulation of TGFβ3 in poly-d-l-lactic-co-glycolic acid (PLGA) microspheres and determine its bioactivity. The release profiles of PLGA-encapsulated TGFβ3 with 50:50 and 75:25 PLA:PGA ratios differed throughout the experimental period. To compare sterilization modalities of microspheres, bFGF was encapsulated in 50:50 PLGA microspheres and subjected to ethylene oxide (EO) gas, radiofrequency glow discharge (RFGD), or ultraviolet (UV) light. The release of bFGF was significantly attenuated by UV light, but not significantly altered by either EO or RFGD. To verify its bioactivity, TGFβ3 (1.35 ng/mL) was control-released to the culture of human mesenchymal stem cells (hMSC) under induced osteogenic differentiation. Alkaline phosphatase staining intensity was markedly reduced 1 week after exposing hMSC-derived osteogenic cells to TGFβ3. This was confirmed by lower alkaline phosphatase activity (2.25 ± 0.57 mU/mL/ng DNA) than controls (TGFβ3-free) at 5.8 ± 0.9 mU/mL/ng DNA (p < 0.05). Control-released TGFβ3 bioactivity was further confirmed by lack of significant differences in alkaline phosphatase upon direct addition of 1.35 ng/mL TGFβ3 to cell culture (p > 0.05). These findings provide baseline data for potential uses of microencapsulated TGFβ3 in wound healing and tissue-engineering applications.
Collagen scaffolds for nonviral IGF-1 gene delivery in articular cartilage tissue engineering
Gene therapy, 2007
This study investigated the use of a type II collagenglycosaminoglycan (CG) scaffold as a nonviral gene delivery vehicle for facilitating gene transfer to seeded adult articular chondrocytes to produce an elevated, prolonged and local expression of insulin-like growth factor (IGF)-1 for enhancing cartilage regeneration. Gene-supplemented CG (GSCG) scaffolds were synthesized by two methods: (1) soaking a pre-cross-linked CG scaffold in a plasmid solution followed by a freeze-drying process, and (2) chemically cross-linking the plasmid DNA to the scaffold. Two different plasmid solutions were also compared: (1) naked plasmid IGF-1 alone, and (2) plasmid IGF-1 with a lipid transfection reagent. Plasmid release studies revealed that cross-linking the plasmid to the CG scaffold prevented passive bolus release of plasmid and resulted in vector release controlled by scaffold degradation. In chondrocyte-seeded GSCG scaffolds, prolonged and elevated IGF-1 expression was enhanced by using the cross-linking method of plasmid incorporation along with the addition of the transfection reagent. The sustained level of IGF-1 overexpression resulted in significantly higher amounts of tissue formation, chondrocyte-like cells, GAG accumulation, and type II collagen production, compared to control scaffolds. These findings demonstrate that CG scaffolds can serve as nonviral gene delivery vehicles of microgram amounts of IGF-1 plasmid (o10 mg per scaffold) to provide a locally sustained therapeutic level of overexpressed IGF-1, resulting in enhanced cartilage formation.
Differential Effects of Growth Factors on Tissue-Engineered Cartilage
Tissue Engineering, 2002
The effects of four regulatory factors on tissue-engineered cartilage were examined with specific focus on the ability to increase construct growth rate and concentrations of glycosaminoglycans (GAG) and collagen, the major extracellular matrix (ECM) components. Bovine calf articular chondrocytes were seeded onto biodegradable polyglycolic acid (PGA) scaffolds and cultured in medium with or without supplemental insulin-like growth factor (IGF-I), interleukin-4 (IL-4), transforming growth factor-b1 (TGF-b1) or platelet-derived growth factor (PDGF). IGF-I, IL-4, and TGF-b1 increased construct wet weights by 1.5-2.9-fold over 4 weeks of culture and increased amounts of cartilaginous ECM components. IGF-I (10-300 ng/mL) maintained wet weight fractions of GAG in constructs seeded at high cell density and increased by up to fivefold GAG fractions in constructs seeded at lower cell density. TGF-b1 (30 ng/mL) increased wet weight fractions of total collagen by up to 1.4-fold while maintaining a high fraction of type II collagen (79 6 11% of the total collagen). IL-4 (1-100 ng/mL) minimized the thickness of the GAG-depleted region at the construct surfaces. PDGF (1-100 ng/mL) decreased construct growth rate and ECM fractions. Different regulatory factors thus elicit significantly different chondrogenic responses and can be used to selectively control the growth rate and improve the composition of engineered cartilage.