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Surface Entrapment of Fibronectin on Electrospun PLGA Scaffolds for Periodontal Tissue Engineering.

Campos DM, Gritsch K, Salles V, Attik GN, Grosgogeat B - Biores Open Access (2014)

Bottom Line: Suitable degradation behavior without pH variations was observed for all samples up to 28 days.All treated materials presented strong shrinkage, fiber orientation loss, and collapsed fibers.However, functionalization process using 0.01 M NaOH concentration resulted in unchanged scaffold porosity, preserved chemical composition, and similar mechanical properties compared with untreated scaffolds.

View Article: PubMed Central - PubMed

Affiliation: Laboratoire des Multimatériaux et Interfaces CNRS UMR 5615, Université Lyon 1 , Villeurbanne, France . ; UFR d'odontologie, Université Lyon 1 , Villeurbanne, France .

ABSTRACT
Nowadays, the challenge in the tissue engineering field consists in the development of biomaterials designed to regenerate ad integrum damaged tissues. Despite the current use of bioresorbable polyesters such as poly(l-lactide) (PLA), poly(d,l-lactide-co-glycolide) (PLGA), and poly-ɛ-caprolactone in soft tissue regeneration researches, their hydrophobic properties negatively influence the cell adhesion. Here, to overcome it, we have developed a fibronectin (FN)-functionalized electrospun PLGA scaffold for periodontal ligament regeneration. Functionalization of electrospun PLGA scaffolds was performed by alkaline hydrolysis (0.1 or 0.01 M NaOH). Then, hydrolyzed scaffolds were coated by simple deposition of an FN layer (10 μg/mL). FN coating was evidenced by X-ray photoelectron analysis. A decrease of contact angle and greater cell adhesion to hydrolyzed, FN-coated PLGA scaffolds were noticed. Suitable degradation behavior without pH variations was observed for all samples up to 28 days. All treated materials presented strong shrinkage, fiber orientation loss, and collapsed fibers. However, functionalization process using 0.01 M NaOH concentration resulted in unchanged scaffold porosity, preserved chemical composition, and similar mechanical properties compared with untreated scaffolds. The proposed simplified method to functionalize electrospun PLGA fibers is an efficient route to make polyester scaffolds more biocompatible and shows potential for tissue engineering.

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Related in: MedlinePlus

Biocompatibility observations of PDL cells cultured on untreated PLGA and PLGAH001FN samples. (A,B) SEM micrographs of PDL-like fibroblasts cultured on (A) untreated PLGA [scale bar (i) 100 μm and (ii) 30 μm] and (B) PLGAH001FN [scale bar (i) 100 μm and (ii) 30 μm] up to 7 days. (C,D) Confocal images of PDL cells cultured on (C) untreated PLGA surface and (D) PLGAH001FN surfaces up to 24 h; (i) and (ii) correspond to superior and inferior surfaces of samples, respectively. Scale bar (C,D) 100 μm. (E) Quantitative resazurin results of PDL cells cultured on PLGA and PLGAH001FN samples up to 24 h. Data are expressed as mean±SD (n=3). PDL, periodontal ligament.
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f6: Biocompatibility observations of PDL cells cultured on untreated PLGA and PLGAH001FN samples. (A,B) SEM micrographs of PDL-like fibroblasts cultured on (A) untreated PLGA [scale bar (i) 100 μm and (ii) 30 μm] and (B) PLGAH001FN [scale bar (i) 100 μm and (ii) 30 μm] up to 7 days. (C,D) Confocal images of PDL cells cultured on (C) untreated PLGA surface and (D) PLGAH001FN surfaces up to 24 h; (i) and (ii) correspond to superior and inferior surfaces of samples, respectively. Scale bar (C,D) 100 μm. (E) Quantitative resazurin results of PDL cells cultured on PLGA and PLGAH001FN samples up to 24 h. Data are expressed as mean±SD (n=3). PDL, periodontal ligament.

Mentions: Physical and chemical characterizations were performed to propose an optimal functionalized scaffold to PDL TE application. From these results, the PLGAH001FN group was selected to verify the capacity of alkaline hydrolysis and FN coating to improve biocompatibility properties. From SEM micrographs (Fig. 6 A,B), PDL cells were able to adhere on both sample surfaces: untreated PLGA and PLGAH001FN samples. However, PDL cells showed round-shape and unspread morphology at the untreated PLGA surface (Fig. 6A). In contrast, when cells were cultured on PLGAH001FN surface, cell adhesion structures were visualized with spread morphology and all surface was homogenously colonized by cells (data not shown). Some observed structures may be related to a novel ECM deposition (Fig. 6B). To demonstrate the capacity of cells to migrate through the scaffold, confocal images of both untreated and hydrolyzed, FN-coated superior and inferior surfaces were compared. After 24 h, PDL cells cultured on hydrolyzed, FN-coated surface were able to migrate into inferior surface (Fig. 6D-ii). Nevertheless, from quantitative results from resazurin assay, no significant difference was observed either in untreated or in hydrolyzed, FN-coated surfaces for 24 h (Fig. 6E).


Surface Entrapment of Fibronectin on Electrospun PLGA Scaffolds for Periodontal Tissue Engineering.

Campos DM, Gritsch K, Salles V, Attik GN, Grosgogeat B - Biores Open Access (2014)

Biocompatibility observations of PDL cells cultured on untreated PLGA and PLGAH001FN samples. (A,B) SEM micrographs of PDL-like fibroblasts cultured on (A) untreated PLGA [scale bar (i) 100 μm and (ii) 30 μm] and (B) PLGAH001FN [scale bar (i) 100 μm and (ii) 30 μm] up to 7 days. (C,D) Confocal images of PDL cells cultured on (C) untreated PLGA surface and (D) PLGAH001FN surfaces up to 24 h; (i) and (ii) correspond to superior and inferior surfaces of samples, respectively. Scale bar (C,D) 100 μm. (E) Quantitative resazurin results of PDL cells cultured on PLGA and PLGAH001FN samples up to 24 h. Data are expressed as mean±SD (n=3). PDL, periodontal ligament.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4048976&req=5

f6: Biocompatibility observations of PDL cells cultured on untreated PLGA and PLGAH001FN samples. (A,B) SEM micrographs of PDL-like fibroblasts cultured on (A) untreated PLGA [scale bar (i) 100 μm and (ii) 30 μm] and (B) PLGAH001FN [scale bar (i) 100 μm and (ii) 30 μm] up to 7 days. (C,D) Confocal images of PDL cells cultured on (C) untreated PLGA surface and (D) PLGAH001FN surfaces up to 24 h; (i) and (ii) correspond to superior and inferior surfaces of samples, respectively. Scale bar (C,D) 100 μm. (E) Quantitative resazurin results of PDL cells cultured on PLGA and PLGAH001FN samples up to 24 h. Data are expressed as mean±SD (n=3). PDL, periodontal ligament.
Mentions: Physical and chemical characterizations were performed to propose an optimal functionalized scaffold to PDL TE application. From these results, the PLGAH001FN group was selected to verify the capacity of alkaline hydrolysis and FN coating to improve biocompatibility properties. From SEM micrographs (Fig. 6 A,B), PDL cells were able to adhere on both sample surfaces: untreated PLGA and PLGAH001FN samples. However, PDL cells showed round-shape and unspread morphology at the untreated PLGA surface (Fig. 6A). In contrast, when cells were cultured on PLGAH001FN surface, cell adhesion structures were visualized with spread morphology and all surface was homogenously colonized by cells (data not shown). Some observed structures may be related to a novel ECM deposition (Fig. 6B). To demonstrate the capacity of cells to migrate through the scaffold, confocal images of both untreated and hydrolyzed, FN-coated superior and inferior surfaces were compared. After 24 h, PDL cells cultured on hydrolyzed, FN-coated surface were able to migrate into inferior surface (Fig. 6D-ii). Nevertheless, from quantitative results from resazurin assay, no significant difference was observed either in untreated or in hydrolyzed, FN-coated surfaces for 24 h (Fig. 6E).

Bottom Line: Suitable degradation behavior without pH variations was observed for all samples up to 28 days.All treated materials presented strong shrinkage, fiber orientation loss, and collapsed fibers.However, functionalization process using 0.01 M NaOH concentration resulted in unchanged scaffold porosity, preserved chemical composition, and similar mechanical properties compared with untreated scaffolds.

View Article: PubMed Central - PubMed

Affiliation: Laboratoire des Multimatériaux et Interfaces CNRS UMR 5615, Université Lyon 1 , Villeurbanne, France . ; UFR d'odontologie, Université Lyon 1 , Villeurbanne, France .

ABSTRACT
Nowadays, the challenge in the tissue engineering field consists in the development of biomaterials designed to regenerate ad integrum damaged tissues. Despite the current use of bioresorbable polyesters such as poly(l-lactide) (PLA), poly(d,l-lactide-co-glycolide) (PLGA), and poly-ɛ-caprolactone in soft tissue regeneration researches, their hydrophobic properties negatively influence the cell adhesion. Here, to overcome it, we have developed a fibronectin (FN)-functionalized electrospun PLGA scaffold for periodontal ligament regeneration. Functionalization of electrospun PLGA scaffolds was performed by alkaline hydrolysis (0.1 or 0.01 M NaOH). Then, hydrolyzed scaffolds were coated by simple deposition of an FN layer (10 μg/mL). FN coating was evidenced by X-ray photoelectron analysis. A decrease of contact angle and greater cell adhesion to hydrolyzed, FN-coated PLGA scaffolds were noticed. Suitable degradation behavior without pH variations was observed for all samples up to 28 days. All treated materials presented strong shrinkage, fiber orientation loss, and collapsed fibers. However, functionalization process using 0.01 M NaOH concentration resulted in unchanged scaffold porosity, preserved chemical composition, and similar mechanical properties compared with untreated scaffolds. The proposed simplified method to functionalize electrospun PLGA fibers is an efficient route to make polyester scaffolds more biocompatible and shows potential for tissue engineering.

No MeSH data available.


Related in: MedlinePlus