Effect of initial cell seeding density on 3D-engineered silk fibroin scaffolds for articular cartilage tissue engineering

Talukdar, Sarmistha; Nguyen, Quoc-Thang; Chen, Albert C.; Sah, Robert L.; Kundu, Subhas C.

doi:10.1016/j.biomaterials.2011.08.027

Cited by 106 publications

(72 citation statements)

References 59 publications

(53 reference statements)

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“…This observation is in line with other studies that demonstrated that differences in DNA levels and tissue formation due to different initial cell densities disappear during long-term culture (Hansen et al, 2013;Kon et al, 2010). However, other studies reported beneficial effects of increased cell densities using higher initial cell densities compared to the current study (Chen et al, 1997;Puelacher et al, 1994;Talukdar et al, 2011). Possibly, the effect of different cell densities is influenced by the initial cell density and the scaffold/hydrogel in which the chondrocytes are grown.…”

Section: Discussionsupporting

confidence: 92%

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair

Mouser

2018

ALTEX

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Summary The implantation of chondrocyte-laden hydrogels is a promising cartilage repair strategy. Chondrocytes can be spatially positioned in hydrogels and thus in defects, while current clinical cell therapies introduce chondrocytes in the defect depth. The main aim of this study was to evaluate the effect of spatial chondrocyte distribution on the reparative process. To reduce animal experiments, an ex vivo osteochondral plug model was used and evaluated. The role of the delivered and endogenous cells in the repair process was investigated. Full thickness cartilage defects were created in equine osteochondral plugs. Defects were filled with (A) chondrocytes at the bottom of the defect, covered with a cell-free hydrogel, (B) chondrocytes homogeneously encapsulated in a hydrogel, and (C, D) combinations of A and B with different cell densities. Plugs were cultured for up to 57 days, after which the cartilage and repair tissues were characterized and compared to baseline samples. Additionally, at day 21, the origin of cells in the repair tissue was evaluated. Best outcomes were obtained with conditions C and D, which resulted in well-integrated cartilage-like tissue that completely filled the defect, regardless of the initial cell density. A critical role of the spatial chondrocyte distribution in the repair process was observed. Moreover, the osteochondral plugs stimulated cartilage formation in the hydrogels when cultured in the defects. The resulting repair tissue originated from the delivered cells. These findings confirm the potential of the osteochondral plug model for the optimization of the composition of cartilage implants and for studying repair mechanisms.

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Section: Discussionsupporting

confidence: 92%

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair

Mouser

2018

ALTEX

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“…Such 3D scaffolds coated with a native ECM synthesized by bone marrow stromal cells exhibited a biomimetic bonemarrow-like microenvironment (or ''niche'') in vitro. Due to the large differences in materials, geometry, patterns of architectures, 38,39 stiffness, 40,41 and even cell seeding [42][43][44] between 2D tissue culture plastic versus 3D Col/HA scaffolds, it is unreasonable to directly compare the properties of a native ECM coated on 2D tissue culture plastic versus on 3D Col/HA scaffold in controlling the fate of MSCs. Therefore, in the present study, we only focused on comparing the behavior of MSCs maintained on a 3D scaffold made by the same materials with or without coating a native bone-marrow-derived ECM.…”

Section: Discussionmentioning

confidence: 99%

Stromal-Cell-Derived Extracellular Matrix Promotes the Proliferation and Retains the Osteogenic Differentiation Capacity of Mesenchymal Stem Cells on Three-Dimensional Scaffolds

Antebi

Zhang

Wang

et al. 2015

Tissue Engineering Part C: Methods

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To date, expansion of bone-marrow-derived mesenchymal stem cells (MSCs) is typically carried out on twodimensional (2D) tissue culture plastic. Since this 2D substratum is very different from the physiological situation, MSCs gradually lose their unique multipotent properties during expansion. Recently, the role of the extracellular matrix (ECM) microenvironment (''niche'') in facilitating and regulating stem cell behavior in vivo has been elucidated. As a result, investigators have shifted their efforts toward developing threedimensional (3D) scaffolds capable of functioning like the native tissue ECM. In this study, we demonstrated that stromal-cell-derived ECM, formed within a collagen/hydroxyapatite (Col/HA) scaffold to mimic the bone marrow ''niche,'' promoted MSC proliferation and preserved their differentiation capacity. The ECM was synthesized by MSCs to reconstitute the tissue-specific 3D microenvironment in vitro. Following deposition of the ECM inside Col/HA scaffold, the construct was decellularized and reseeded with MSCs to study their behavior. The data showed that MSCs cultured on the ECM-Col/HA scaffolds grew significantly faster than the cells from the same batch cultured on the regular Col/HA scaffolds. In addition, MSCs cultured on the ECMCol/HA scaffolds retained their ''stemness'' and osteogenic differentiation capacity better than MSCs cultured on regular Col/HA scaffolds. When ECM-Col/HA scaffolds were implanted into immunocompromised mice, with or without loading MSCs, it was found that those scaffolds formed less bone as compared with regular Col/ HA scaffolds (i.e., without ECM), in both cases of with or without loading MSCs. The in vivo study further confirmed that the ECM-Col/HA scaffold was a suitable mimic of the bone marrow ''niche.'' This novel 3D stromal-cell-derived ECM system has the potential to be developed into a biomedical platform for regenerative medicine applications.

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“…Cell seeding density, as well as different concentrations of dexamethasone, would affect the results of osteogenic differentiation. 42 The differentiation potential was compared inducing different dexamethasone amounts through measuring various pathways of differentiation utilizing biochemical cues as markers. ALP is an early-stage marker of osteogenic differentiation, and its expression may affect the progression.…”

Section: Discussionmentioning

confidence: 99%

“…MC3T3-E1 cell seeding density, as well as different concentrations of dexamethasone, would affect the results of osteogenic differentiation. 42 In order to compare the differentiation potential, the MC3T3-E1 cells were induced at different concentrations of dexamethasone and cultured at varying cell seeding density to determine the osteogenic differentiation pathways. The degree of ECM mineralization was measured by staining with Alizarin Red and the respective bright field images of stained cells are shown in Figure S3.…”

Section: Osteogenic Differentiationmentioning

confidence: 99%

Investigation of silk fibroin nanoparticle-decorated poly(L-lactic acid) composite scaffolds for osteoblast growth and differentiation

Chen¹,

Kankala²,

Chen³

et al. 2017

IJN

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Attempts to reflect the physiology of organs is quite an intricacy during the tissue engineering process. An ideal scaffold and its surface topography can address and manipulate the cell behavior during the regeneration of targeted tissue, affecting the cell growth and differentiation significantly. Herein, silk fibroin (SF) nanoparticles were incorporated into poly( l -lactic acid) (PLLA) to prepare composite scaffolds via phase-inversion technique using supercritical carbon dioxide (SC-CO 2 ). The SF nanoparticle core increased the surface roughness and hydrophilicity of the PLLA scaffolds, leading to a high affinity for albumin attachment. The in vitro cytotoxicity test of SF/PLLA scaffolds in L929 mouse fibroblast cells indicated good biocompatibility. Then, the in vitro interplay between mouse preosteoblast cell (MC3T3-E1) and various topological structures and biochemical cues were evaluated. The cell adhesion, proliferation, osteogenic differentiation and their relationship with the structures as well as SF content were explored. The SF/PLLA weight ratio (2:8) significantly affected the MC3T3-E1 cells by improving the expression of key players in the regulation of bone formation, ie, alkaline phosphatase (ALP), osteocalcin (OC) and collagen 1 (COL-1). These results suggest not only the importance of surface topography and biochemical cues but also the potential of applying SF/PLLA composite scaffolds as biomaterials in bone tissue engineering.

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Effect of initial cell seeding density on 3D-engineered silk fibroin scaffolds for articular cartilage tissue engineering

Cited by 106 publications

References 59 publications

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair

Ex vivo model unravelling cell distribution effect in hydrogels for cartilage repair

Stromal-Cell-Derived Extracellular Matrix Promotes the Proliferation and Retains the Osteogenic Differentiation Capacity of Mesenchymal Stem Cells on Three-Dimensional Scaffolds

Investigation of silk fibroin nanoparticle-decorated poly(L-lactic acid) composite scaffolds for osteoblast growth and differentiation

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