Combinatory approach for developing silk fibroin scaffolds for cartilage regeneration

Ribeiro, Viviana P.; Morais, Alain da Silva; Maia, F. Raquel; Canadas, Raphaël Faustino; Costa, João B.; Oliveira, Ana; Oliveira, Joaquim M.; Reis, Rui L.

doi:10.1016/j.actbio.2018.03.047

Cited by 101 publications

(83 citation statements)

References 62 publications

(106 reference statements)

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“…The blank lower chamber without any scaffolds was served as the control. Next, 200 µL of serum-free DMEM containing 2.0×10 4 HVSMCs was added on the upper chamber. After culturing in the cell incubator for another 24 hours, the cells on the upper surface of the Transwell membrane were scraped by a cotton tip applicator.…”

Section: Hvsmcs' Migration Assaymentioning

confidence: 99%

“…1 Ideal scaffold was commonly designed to be highly porous for cell infiltration, nutrients and oxygen transport, and metabolic waste removal, thereby facilitating the regeneration of functional neotissues. [2][3][4] For instance, the vascular graft was often designed to be porous for enabling the infiltration of vascular smooth muscle cells (SMCs) and regeneration of functional tunica media, 5 thereby endowing the neo-vessel with the contractile function. Moreover, nanofibrous structure resembling native ECM is another important feature that can provide a biomimetic microenvironment for enhanced cell attachment, proliferation, and differentiation.…”

Section: Introductionmentioning

confidence: 99%

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Macroporous nanofibrous vascular scaffold with improved biodegradability and smooth muscle cells infiltration prepared by dual phase separation technique

Wang¹,

Nie²,

Liu³

et al. 2018

IJN

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IntroductionThe fast degradation of vascular graft and the infiltration of smooth muscle cells (SMCs) into the vascular graft are considered to be critical for the regeneration of functional neo-vessels. In our previous study, a novel dual phase separation technique was developed to one-pot prepare macroporous nanofibrous poly(L-lactic acid) (PLLA)/poly(ε-caprolactone) (PCL) vascular scaffold by phase separating the immiscible polymer blend. However, the slow degradation of PLLA/PCL limited cell infiltration. Herein, we hypothesized that poly(lactic-co-glycolic acid) (PLGA) would be miscible with PLLA but immiscible with PCL. Then, PLGA can be introduced into the PLLA/PCL blend to fabricate macroporous nanofibrous scaffold with improved biodegradability by using dual phase separation technique.Materials and methodsThe miscibility of PLGA with PLLA and PCL was evaluated. Then, the PLLA/PLGA/PCL scaffold was prepared by dual phase separation technique. The prepared scaffolds were characterized in terms of the morphology, in vitro degradation, mechanical properties, and cells’ infiltration and viability for human vascular SMCs (HVSMCs). Finally, platelet-derived growth factor-BB (PDGF-BB) was immobilized on the scaffold and its effect on the bioactivity of HVSMCs was studied.ResultsPLGA is miscible with PLLA but immiscible with PCL as hypothesized. The addition of PLGA enlarged the pore size and improved the biodegradability of composite scaffold. Notably, PLLA/PLGA/PCL scaffold with the blend ratio of 30:40:30 possessed improved pore interconnectivity for cells’ infiltration and enough mechanical properties. Moreover, HVSMCs could grow and infiltrate into this scaffold, and surface modification with PDGF-BB on the nanofibrous scaffold enhanced HVSMCs migration and proliferation.ConclusionThis study provides a strategy to expand dual phase separation technique into utilizing ternary even multinary polymer blend to fabricate macroporous nanofibrous scaffold with improved physicochemical properties. The prepared PLLA/PLGA/PCL scaffold would be promising for the regeneration of functional tunica media in vascular tissue engineering.

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Section: Hvsmcs' Migration Assaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Macroporous nanofibrous vascular scaffold with improved biodegradability and smooth muscle cells infiltration prepared by dual phase separation technique

Wang¹,

Nie²,

Liu³

et al. 2018

IJN

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“…P3 BMSCs (4 × 10 5 cells/ Electric field-driven building blocks for introducing multiple gradients to hydrogels RESEARCH ARTICLE well) were seeded on the hydrogel surface in 48-well plates and cultured for 24 h with low glucose DMEM (Gibco, Thermo Fisher, MO, USA) containing 10% FBS and 100 units/mL penicillin-streptomycin (Gibco, Thermo Fisher, MO, USA). Chondrogenic-osteogenic co-culture medium supplemented with high glucose DMEM (Gibco, Thermo Fisher, MO, USA), 10% FBS, 10 ng/mL recombinant human TGF-β3 (PeproTech, Newark, NJ, USA), 100 U/mL penicillin/streptomycin (Gibco, Thermo Fisher, MO, USA), 100 nmol/L dexamethasone (Sigma-Aldrich, St. Louis, MO, USA), 91.5 μg/mL ascorbic acid 2-phosphate (Sigma-Aldrich, St. Louis, MO, USA), 10 mmol/L β-sodium glycerophosphate (Sigma-Aldrich, St. Louis, MO, USA), and 40 lg/mL L-proline (Sigma-Aldrich, St. Louis, MO, USA) was used to culture the cells for 28 days (Ribeiro et al, 2018). The co-culture medium was changed every 2 days.…”

Section: Chondrogenic-osteogenic Differentiation Of Bmscs On Gsnf Hydmentioning

confidence: 99%

“…The co-culture medium was changed every 2 days. Quantitative real time polymerase chain reaction (PCR) was used to measure different osteogenic gene expression markers including Runt-related transcription factor 2 (Runx2), osteocalcin (OCN) and osteocalcin (OPN) (Ding et al, 2017.09;Ribeiro et al, 2018), and different chondrogenic gene expression markers such as Sry-type high mobility group box transcription factor 9 (SOX9), collagen II (COL II), and aggrecan (Acan) (Bhardwaj and Kundu, 2012). At the desired time points, total RNA from cells cultured on hydrogels was isolated using total RNA extraction kit (Tiangen Biotech, Beijing, China).…”

Section: Chondrogenic-osteogenic Differentiation Of Bmscs On Gsnf Hydmentioning

confidence: 99%

Electric field-driven building blocks for introducing multiple gradients to hydrogels

Ding

Lü

et al. 2020

Protein Cell

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Gradient biomaterials are considered as preferable matrices for tissue engineering due to better simulation of native tissues. The introduction of gradient cues usually needs special equipment and complex process but is only effective to limited biomaterials. Incorporation of multiple gradients in the hydrogels remains challenges. Here, betasheet rich silk nanofibers (BSNF) were used as building blocks to introduce multiple gradients into different hydrogel systems through the joint action of crosslinking and electric field. The blocks migrated to the anode along the electric field and gradually stagnated due to the solution-hydrogel transition of the systems, finally achieving gradient distribution of the blocks in the formed hydrogels. The gradient distribution of the blocks could be tuned easily through changing different factors such as solution viscosity, which resulted in highly tunable gradient of mechanical cues. The blocks were also aligned under the electric field, endowing orientation gradient simultaneously. Different cargos could be loaded on the blocks and form gradient cues through the same crosslinking-electric field strategy. The building blocks could be introduced to various hydrogels such as Gelatin and NIPAM, indicating the universality. Complex niches with multiple gradient cues could be achieved through the strategy. Silk-based hydrogels with suitable mechanical gradients were fabricated to control the osteogenesis and chondrogenesis. Chondrogenic-osteogenic gradient transition was obtained, which stimulated the ectopic osteochondral tissue regeneration in vivo. The versatility and highly controllability of the strategy as well as multifunction of the building blocks reveal the applicability in complex tissue engineering and various interfacial tissues.

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“…For example, Dai, Liu, Ma, Wang, and Gao (2016) used scaffold-free fibrin to induce the regeneration of full-thickness cartilage defects. Silk fibroin (Ribeiro et al, 2018) or bioprinted polycaprolactone (Theodoridis et al, 2019) scaffolds have also been explored for cartilage repair. Unfortunately, no specific hydrogel material has accurately reproduced the tissue properties of cartilage.…”

Section: Introductionmentioning

confidence: 99%

Optimization of photocrosslinked gelatin/hyaluronic acid hybrid scaffold for the repair of cartilage defect

Lin

Beck

Shimomura

et al. 2019

J Tissue Eng Regen Med

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Summary There is no therapy currently available for fully repair of articular cartilage lesion. Our laboratory has recently developed a visible light-activatable methacrylated gelatin (mGL) hydrogel with the potential for cartilage regeneration. In this study, we further optimized mGL scaffolds by supplementing methacrylated hyaluronic acid (mHA), which has been shown to stimulate chondrogenesis via activation of critical cellular signaling pathways. We hypothesized that the introduction of an optimal ratio of mHA would enhance the biological properties of mGL scaffolds and augment chondrogenesis of human bone marrow-derived mesenchymal stem cells (hBMSCs). To test this hypothesis, hybrid scaffolds consisting of mGL and mHA at different weight ratios were fabricated with hBMSCs encapsulated at 20×106 cells/mL and maintained in a chondrogenesis-promoting medium. The chondrogenenic differentiation of hBMSCs within different scaffolds was estimated after 8 weeks of culture. Our results showed that mGL/mHA at a 9:1 (%, w/v) ratio resulted in the lowest hBMSC hypertrophy and highest glycosaminoglycan production, with slightly increased volume of the entire construct. The applicability of this optimally designed mGL/mHA hybrid scaffold for cartilage repair was then examined in vivo. A full-thickness cylindrical osteochondral defect was surgically created in the rabbit femoral condyle and a 3-dimensional cell-biomaterial construct was fabricated by in situ photo-crosslinking to fully fill the lesion site. The results showed that implantation of the mGL/mHA (9:1) construct resulted in both cartilage and subchondral bone regeneration after 12 weeks, supporting its use as a promising scaffold for repair and resurfacing of articular cartilage defects in the clinical setting.

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Combinatory approach for developing silk fibroin scaffolds for cartilage regeneration

Cited by 101 publications

References 62 publications

Macroporous nanofibrous vascular scaffold with improved biodegradability and smooth muscle cells infiltration prepared by dual phase separation technique

Macroporous nanofibrous vascular scaffold with improved biodegradability and smooth muscle cells infiltration prepared by dual phase separation technique

Electric field-driven building blocks for introducing multiple gradients to hydrogels

Optimization of photocrosslinked gelatin/hyaluronic acid hybrid scaffold for the repair of cartilage defect

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