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Cold Atmospheric Plasma Modified Electrospun Scaffolds with Embedded Microspheres for Improved Cartilage Regeneration.

Zhu W, Castro NJ, Cheng X, Keidar M, Zhang LG - PLoS ONE (2015)

Bottom Line: However, there still remains a lack of ideal biomimetic tissue scaffolds which effectively stimulate cartilage regeneration with appropriate functional properties.More importantly, our results demonstrate, for the first time, CAP and microspheres can synergistically enhance stem cell growth as well as improve chondrogenic differentiation of human marrow-derived mesenchymal stem cells (such as increased glycosaminoglycan, type II collagen, and total collagen production).By integrating CAP, sustained bioactive factor loaded microspheres, and electrospinning, we have fabricated a promising bioactive scaffold for cartilage regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Mechanical and Aerospace Engineering, The George Washington University, Washington, District of Columbia, United States of America.

ABSTRACT
Articular cartilage is prone to degeneration and possesses extremely poor self-healing capacity due to inherent low cell density and the absence of a vasculature network. Tissue engineered cartilage scaffolds show promise for cartilage repair. However, there still remains a lack of ideal biomimetic tissue scaffolds which effectively stimulate cartilage regeneration with appropriate functional properties. Therefore, the objective of this study is to develop a novel biomimetic and bioactive electrospun cartilage substitute by integrating cold atmospheric plasma (CAP) treatment with sustained growth factor delivery microspheres. Specifically, CAP was applied to a poly(ε-caprolactone) electrospun scaffold with homogeneously distributed bioactive factors (transforming growth factor-β1 and bovine serum albumin) loaded poly(lactic-co-glycolic) acid microspheres. We have shown that CAP treatment renders electrospun scaffolds more hydrophilic thus facilitating vitronectin adsorption. More importantly, our results demonstrate, for the first time, CAP and microspheres can synergistically enhance stem cell growth as well as improve chondrogenic differentiation of human marrow-derived mesenchymal stem cells (such as increased glycosaminoglycan, type II collagen, and total collagen production). Furthermore, CAP can substantially enhance 3D cell infiltration (over two-fold increase in infiltration depth after 1 day of culture) in the scaffolds. By integrating CAP, sustained bioactive factor loaded microspheres, and electrospinning, we have fabricated a promising bioactive scaffold for cartilage regeneration.

No MeSH data available.


Related in: MedlinePlus

Confocal micrographs of MSCs spread and proliferate on various scaffolds.At day 1 (A) and day 3 (B) cells penetrated to scaffolds with different cells infiltration distance for 1 day (a) and 3 days (b). Improved cell proliferation and infiltration were observed on scaffolds with CAP treatment and/or microspheres incorporation compared to PCL only control group. Data are mean ± standard error of the mean, N = 3, *p<0.05.
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pone.0134729.g009: Confocal micrographs of MSCs spread and proliferate on various scaffolds.At day 1 (A) and day 3 (B) cells penetrated to scaffolds with different cells infiltration distance for 1 day (a) and 3 days (b). Improved cell proliferation and infiltration were observed on scaffolds with CAP treatment and/or microspheres incorporation compared to PCL only control group. Data are mean ± standard error of the mean, N = 3, *p<0.05.

Mentions: Moreover, confocal microscopy was carried out on various MSC seeded scaffolds including bare scaffolds, scaffolds with CAP treatment, and scaffolds with both CAP treatment and BSA-loaded microspheres to evaluate 3D cell spreading and infiltration. As shown in Fig 9, excellent MSC adhesion and spreading was observed throughout the entirety of the CAP and microsphere embedded scaffolds’ surface with well-defined actin filaments (stained red by Rhodamine-Phalloidin) and nuclei (stained blue by DAPI) suggesting desirable biocompatibility of the scaffold groups. More importantly, CAP treated scaffolds (with and without microspheres) can substantially enhance 3D cell infiltration when compared with PCL scaffolds without CAP treatment after 1 and 3 days culture, respectively. Image analysis revealed a 2.22 and 2.07-fold increase of infiltration depth for 1 min and 3 min CAP treated scaffolds, respectively, when compared to untreated scaffolds at day 1. At day 3, the infiltration depth on scaffolds with 1 min and 3 min CAP treatment was 1.68 and 1.58 times relative to untreated scaffolds. There was no significant difference in cell infiltration depth amongst CAP treated groups with and without microspheres at day 1 and day 3.


Cold Atmospheric Plasma Modified Electrospun Scaffolds with Embedded Microspheres for Improved Cartilage Regeneration.

Zhu W, Castro NJ, Cheng X, Keidar M, Zhang LG - PLoS ONE (2015)

Confocal micrographs of MSCs spread and proliferate on various scaffolds.At day 1 (A) and day 3 (B) cells penetrated to scaffolds with different cells infiltration distance for 1 day (a) and 3 days (b). Improved cell proliferation and infiltration were observed on scaffolds with CAP treatment and/or microspheres incorporation compared to PCL only control group. Data are mean ± standard error of the mean, N = 3, *p<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4519315&req=5

pone.0134729.g009: Confocal micrographs of MSCs spread and proliferate on various scaffolds.At day 1 (A) and day 3 (B) cells penetrated to scaffolds with different cells infiltration distance for 1 day (a) and 3 days (b). Improved cell proliferation and infiltration were observed on scaffolds with CAP treatment and/or microspheres incorporation compared to PCL only control group. Data are mean ± standard error of the mean, N = 3, *p<0.05.
Mentions: Moreover, confocal microscopy was carried out on various MSC seeded scaffolds including bare scaffolds, scaffolds with CAP treatment, and scaffolds with both CAP treatment and BSA-loaded microspheres to evaluate 3D cell spreading and infiltration. As shown in Fig 9, excellent MSC adhesion and spreading was observed throughout the entirety of the CAP and microsphere embedded scaffolds’ surface with well-defined actin filaments (stained red by Rhodamine-Phalloidin) and nuclei (stained blue by DAPI) suggesting desirable biocompatibility of the scaffold groups. More importantly, CAP treated scaffolds (with and without microspheres) can substantially enhance 3D cell infiltration when compared with PCL scaffolds without CAP treatment after 1 and 3 days culture, respectively. Image analysis revealed a 2.22 and 2.07-fold increase of infiltration depth for 1 min and 3 min CAP treated scaffolds, respectively, when compared to untreated scaffolds at day 1. At day 3, the infiltration depth on scaffolds with 1 min and 3 min CAP treatment was 1.68 and 1.58 times relative to untreated scaffolds. There was no significant difference in cell infiltration depth amongst CAP treated groups with and without microspheres at day 1 and day 3.

Bottom Line: However, there still remains a lack of ideal biomimetic tissue scaffolds which effectively stimulate cartilage regeneration with appropriate functional properties.More importantly, our results demonstrate, for the first time, CAP and microspheres can synergistically enhance stem cell growth as well as improve chondrogenic differentiation of human marrow-derived mesenchymal stem cells (such as increased glycosaminoglycan, type II collagen, and total collagen production).By integrating CAP, sustained bioactive factor loaded microspheres, and electrospinning, we have fabricated a promising bioactive scaffold for cartilage regeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Mechanical and Aerospace Engineering, The George Washington University, Washington, District of Columbia, United States of America.

ABSTRACT
Articular cartilage is prone to degeneration and possesses extremely poor self-healing capacity due to inherent low cell density and the absence of a vasculature network. Tissue engineered cartilage scaffolds show promise for cartilage repair. However, there still remains a lack of ideal biomimetic tissue scaffolds which effectively stimulate cartilage regeneration with appropriate functional properties. Therefore, the objective of this study is to develop a novel biomimetic and bioactive electrospun cartilage substitute by integrating cold atmospheric plasma (CAP) treatment with sustained growth factor delivery microspheres. Specifically, CAP was applied to a poly(ε-caprolactone) electrospun scaffold with homogeneously distributed bioactive factors (transforming growth factor-β1 and bovine serum albumin) loaded poly(lactic-co-glycolic) acid microspheres. We have shown that CAP treatment renders electrospun scaffolds more hydrophilic thus facilitating vitronectin adsorption. More importantly, our results demonstrate, for the first time, CAP and microspheres can synergistically enhance stem cell growth as well as improve chondrogenic differentiation of human marrow-derived mesenchymal stem cells (such as increased glycosaminoglycan, type II collagen, and total collagen production). Furthermore, CAP can substantially enhance 3D cell infiltration (over two-fold increase in infiltration depth after 1 day of culture) in the scaffolds. By integrating CAP, sustained bioactive factor loaded microspheres, and electrospinning, we have fabricated a promising bioactive scaffold for cartilage regeneration.

No MeSH data available.


Related in: MedlinePlus