Boosting in vitro cartilage tissue engineering through the fabrication of polycaprolactone-gelatin 3D scaffolds with specific depth-dependent fiber alignments and mechanical stimulation

Semitela, Ângela; Girão, André F.; Fernandes, Carla; Ramalho, Gonçalo; Pinto, Susana; Completo, António; Marques, Paula A.A.P.

doi:10.1016/j.jmbbm.2021.104373

Cited by 15 publications

(26 citation statements)

References 63 publications

(96 reference statements)

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“…Although the small pore sizes of the electrospun meshes were insufficient to ensure cell migration through the layers, they were sufficient to assess the survivability of chondrocytes exposed to electrospraying and integration on multilayer constructions, as previously demonstrated. 27,30,31…”

Section: Discussionmentioning

confidence: 99%

Multi-layered electrospinning and electrospraying approach: Effect of polymeric supplements on chondrocyte suspension

et al. 2021

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Articular cartilage was expected to be one of the first tissues to be successfully engineered, but replicating the complex fibril architecture and the cellular distribution of the native cartilage has proven difficult. While electrospinning has been widely used to reproduce the depth-dependent fibre architecture in 3D scaffolds, the chondrocyte-controlled distribution remains an unsolved problem. To incorporate cells homogeneously through the depth of scaffolds, a combination of polymer electrospinning and cell seeding is necessary. A multi-layer approach alternating between polymer electrospinning with chondrocyte electrospraying can be a solution. Still, the success of this process is related to the survival rate of the electrosprayed chondrocytes embedded within the electrospun mesh. In this regard, the present study investigated the impact of the multi-layered process and the supplementation of the electrospray chondrocyte suspension with different concentrations of Gelatin and Alginate on the viability of electrosprayed chondrocytes embedded within a Polycaprolactone/Gelatin electrospun mesh and on the mechanical properties of the resulting meshes. The addition of Gelatin in the chondrocyte suspension did not increase significantly ( p > 0.05) the percentage of viable electrosprayed chondrocytes (25%), while 3 wt% Alginate addition led to a significant ( p < 0.05) increase in chondrocyte viability (50%) relative to the case without polymer supplement (15%). Furthermore, the addition of both polymer supplements increased the mechanical properties of the multi-layer construct. These findings imply that this multi-layered approach can be applied to cartilage TE allowing for automated chondrocyte integration during scaffolds creation.

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Section: Discussionmentioning

confidence: 99%

Multi-layered electrospinning and electrospraying approach: Effect of polymeric supplements on chondrocyte suspension

et al. 2021

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“…The rotational stacking only needs to prepare the same 2D aligned building blocks, which make it suitable for engineering PMA with unrepeatable interlayer angles, such as that of the myocardium. [14,103] Like the bundling, the stacking process is highly manual, as a result, it [24,59,95] • Peripheral nerve [11,62,63] • Tendon and ligament [67][68][69] • Tendon and ligament [12,[60][61]64,[90][91][128][129][130][131][132] • Myocardium [14] • Myocardium [14,19,103,133] • Corneal stroma [70,94,99,100] • Articular cartilage [46,[72][73][74][75][76]92] • Annulus fibrosus [77][78][79][80]96,97] • Smooth muscle [81,82] • Skeletal muscle [10,…”

Section: Modular Assembly Methodsmentioning

confidence: 99%

“…Various biomaterials [10,[65][66][67][68][69][70][71][72][73][74][75][76][77][78][79][80][81][82][83][84] • Applicable to various biomaterials • Ease of use • Limited mechanical property Electrocompaction Natural collagen [85][86][87][88][89][90][91] • Providing friendly environment • Only applicable for collagen fibers • Only applied to collagen-rich tissues Wetspinning Gel, [92] PCL/SF [93] • Simple and low cost • Easy to cell encapsulation • Low resolution…”

Section: Electrospinningmentioning

confidence: 99%

“…Subsequently, various trilayered scaffolds with varying-aligned fiber or pore, prepared from natural or synthetic polymers, have been orderstacked for partially or fully mimicking the collagen fiber-orientated characteristics of the superficial, middle, and deep zones of native articular cartilage, aiming to direct growth or differentiation of chondrocytes or MSCs to, respectively, generate engineered cartilages that reproduce the stratified architecture and mechanical behavior of the native analogs. [46,[73][74][75] For instance, a porous multilayer scaffold was assembled from a paralleloriented, an isotropic and an orthogonal-oriented PLA layer with or without functionalized cellulose nanocrystals, which can direct the morphology, orientation and ECM deposition of seeded chondrocytes in a zone-dependent way, and promote the formation of chondral tissue with features analogous to the natural counterparts (Figure 3E). [46] It is important to point out that some scaffolds are composed of three layers, but they only can mimic the collagen morphology in the superficial and middle zones.…”

Section: Articular Cartilagementioning

confidence: 99%

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Engineering Complex Anisotropic Scaffolds beyond Simply Uniaxial Alignment for Tissue Engineering

Xing

Liu

Xu³

et al. 2021

Adv Funct Materials

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Anisotropic microarchitectures arising from an aligned organization of threadlike extracellular matrix (ECM) components or cells are ubiquitous in the human body, such as skeletal muscle, corneal stroma, and meniscus, for executing tissue-specific physiological functions. It is widely recognized that tissue engineering, whereby growing the implanted or endogenous cells in anisotropic scaffolds with geometrical resemblance to the ECM of targeted tissues, represents a promising solution for the structural and functional restoration of these anisotropic tissues. However, remarkable challenges remain in recapitulating the anisotropic complexities of native tissues beyond simply uniaxial alignment. Through unremitting endeavors over the past decade, some innovative bioengineering approaches are developed to tackle these challenges. This review focuses on the recent progress in modular assembly and 3D printing techniques exploited to construct complex anisotropic scaffolds with a key highlight on their accessibility and features for different types of anisotropies, based on understanding the whole picture of anisotropies beyond simply uniaxial alignment in native tissues, which are geometrically divided into three categories. Finally, the applications of these complex scaffolds in anisotropic tissue engineering, either in vitro modeling or in vivo regeneration, are explored.

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“…Then, MC3T3‐E1 cells were seeded onto the scaffolds at an initial density of 10000 cells per scaffold, followed by an incubation at 37 °C for 2 h. Afterward, fresh medium was added, and cell‐seeded scaffolds were cultured for 21 d, with two medium chances for a week. After a preculture period of 7 d, four cell‐laden scaffolds of each composition were subjected to dynamic compression on a designed bioreactor: [ 62 ] 2 h of sinusoidal compression (0.5 Hz, 0–10 % strain) and 4 h of rest, four times a day for 14 d. Four static controls (without any compression) were placed and cultured in the incubator for the remaining 14 d. The culture medium was refreshed two times a week.…”

Section: Methodsmentioning

confidence: 99%

Biomimetic Graphene/Spongin Scaffolds for Improved Osteoblasts Bioactivity via Dynamic Mechanical Stimulation

Semitela

Carvalho

Fernandes

et al. 2021

Macromolecular Bioscience

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Biomimetics offers excellent prospects for design a novel generation of improved biomaterials. Here the controlled integration of graphene oxide (GO) derivatives with a 3D marine spongin (MS) network is explored to nanoengineer novel smart bio‐based constructs for bone tissue engineering. The results point out that 3D MS surfaces can be homogeneously coated by layer‐by‐layer (LbL) assembly of oppositely charged polyethyleneimine (PEI) and GO. Notably, the GOPEI@MS bionanocomposites present a high structural and mechanical stability under compression tests in wet conditions (shape memory). Dynamic mechanically (2 h of sinusoidal compression cyclic interval (0.5 Hz, 0–10% strain)/14 d) stimulates GOPEI@MS seeded with osteoblast (MC3T3‐E1), shows a significant improvement in bioactivity, with cell proliferation being two times higher than under static conditions. Besides, the dynamic assays show that GOPEI@MS bionanocomposites are able to act as mechanical stimulus‐responsive scaffolds able to resemble physiological bone extracellular matrix (ECM) requirements by strongly triggering mineralization of the bone matrix. These results prove that the environment created by the system cell‐GOPEI@MS is suitable for controlling the mechanisms regulating mechanical stimulation‐induced cell proliferation for potential in vivo experimentation.

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Boosting in vitro cartilage tissue engineering through the fabrication of polycaprolactone-gelatin 3D scaffolds with specific depth-dependent fiber alignments and mechanical stimulation

Cited by 15 publications

References 63 publications

Multi-layered electrospinning and electrospraying approach: Effect of polymeric supplements on chondrocyte suspension

Multi-layered electrospinning and electrospraying approach: Effect of polymeric supplements on chondrocyte suspension

Engineering Complex Anisotropic Scaffolds beyond Simply Uniaxial Alignment for Tissue Engineering

Biomimetic Graphene/Spongin Scaffolds for Improved Osteoblasts Bioactivity via Dynamic Mechanical Stimulation

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