3D micro-nano structured hybrid scaffolds: An investigation into the role of nanofiber coating on viability, proliferation and differentiation of seeded mesenchymal stem cells

Mohammad Kazemi Pilehrood, Amir Atashi, Hojjat Sadeghi-Aliabadi, Pertti Nousiainen, Ali Harlin

Research output: Contribution to journalArticleScientificpeer-review

5 Citations (Scopus)

Abstract

The introduction of a three dimensional scaffold providing the closest analogies to extracellular matrix (ECM) is currently a key strategy for tackling many challenges in tissue repair. Here, we present a new hybrid scaffold constructed by coating electrospun chitosan/polyethylene oxide (PEO) nanofibers on commercial BioTek polystyrene (PS) scaffold obtained from Sigma Aldrich. The viability and proliferation rate of mesenchymal stem cells (MSCs) seeded on micro-nano structured hybrid scaffold (MNHS) and commercial PS scaffolds were analyzed by MTT assay. The results of the MTT assay revealed a higher degree of viability and proliferation rate in MSCs seeded on MNHS compared with the commercial PS scaffold. DAPI images also confirmed the higher degree of attachment and viability of MSCs seeded on MNHS. Moreover, MSCs on both scaffolds differentiated to osteoblasts and adipocytes cells, as reflected by the images obtained from Alizarin Red and Oil Red-O staining. Alkaline phosphatase activity (ALP) and calcium content assays revealed that the MNHS has a higher potential for osteogenic differentiation than the commercial scaffold. To quantify the osteoblast and adipocyte gene expression, quantitative RT-PCR was carried out for MNHS, commercial scaffold and Tissue culture polystyrene (TCPS). It was found that MNHS can express a higher level of Runt-related transcription factor 2 (Runx2), osteonectin and osteocalcin in osteogenic differentiation as well as increased expression of PPAR? and UCP-1 in adipogenic differentiation. The enhancement of the attachment, viability and proliferation as well as bi-lineage differentiation may result from the biochemical and structural analogies of MNHS to native ECM. Furthermore, it was observed that biocompatible MNHS scaffold can potentially be utilized as a suitable scaffold for bone and connective tissue engineering.
Original languageEnglish
Pages (from-to)9000-9007
JournalJournal of Nanoscience and Nanotechnology
Volume16
Issue number9
DOIs
Publication statusPublished - 2016
MoE publication typeA1 Journal article-refereed

Fingerprint

Nanofibers
Mesenchymal Stromal Cells
Polystyrenes
Osteoblasts
Adipocytes
Extracellular Matrix
Osteonectin
Tissue Scaffolds
Peroxisome Proliferator-Activated Receptors
Osteocalcin
Chitosan
Tissue Engineering
Connective Tissue
Alkaline Phosphatase
Transcription Factors
Staining and Labeling
Calcium
Gene Expression
Bone and Bones
Polymerase Chain Reaction

Keywords

  • adipogenic differentiation
  • hybrid scaffold
  • mesenchymal stem cell
  • micro porosity
  • nanofiber
  • osteogenic differentiation

Cite this

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title = "3D micro-nano structured hybrid scaffolds: An investigation into the role of nanofiber coating on viability, proliferation and differentiation of seeded mesenchymal stem cells",
abstract = "The introduction of a three dimensional scaffold providing the closest analogies to extracellular matrix (ECM) is currently a key strategy for tackling many challenges in tissue repair. Here, we present a new hybrid scaffold constructed by coating electrospun chitosan/polyethylene oxide (PEO) nanofibers on commercial BioTek polystyrene (PS) scaffold obtained from Sigma Aldrich. The viability and proliferation rate of mesenchymal stem cells (MSCs) seeded on micro-nano structured hybrid scaffold (MNHS) and commercial PS scaffolds were analyzed by MTT assay. The results of the MTT assay revealed a higher degree of viability and proliferation rate in MSCs seeded on MNHS compared with the commercial PS scaffold. DAPI images also confirmed the higher degree of attachment and viability of MSCs seeded on MNHS. Moreover, MSCs on both scaffolds differentiated to osteoblasts and adipocytes cells, as reflected by the images obtained from Alizarin Red and Oil Red-O staining. Alkaline phosphatase activity (ALP) and calcium content assays revealed that the MNHS has a higher potential for osteogenic differentiation than the commercial scaffold. To quantify the osteoblast and adipocyte gene expression, quantitative RT-PCR was carried out for MNHS, commercial scaffold and Tissue culture polystyrene (TCPS). It was found that MNHS can express a higher level of Runt-related transcription factor 2 (Runx2), osteonectin and osteocalcin in osteogenic differentiation as well as increased expression of PPAR? and UCP-1 in adipogenic differentiation. The enhancement of the attachment, viability and proliferation as well as bi-lineage differentiation may result from the biochemical and structural analogies of MNHS to native ECM. Furthermore, it was observed that biocompatible MNHS scaffold can potentially be utilized as a suitable scaffold for bone and connective tissue engineering.",
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3D micro-nano structured hybrid scaffolds: An investigation into the role of nanofiber coating on viability, proliferation and differentiation of seeded mesenchymal stem cells. / Pilehrood, Mohammad Kazemi; Atashi, Amir; Sadeghi-Aliabadi, Hojjat; Nousiainen, Pertti; Harlin, Ali.

In: Journal of Nanoscience and Nanotechnology, Vol. 16, No. 9, 2016, p. 9000-9007.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Pilehrood, Mohammad Kazemi

AU - Atashi, Amir

AU - Sadeghi-Aliabadi, Hojjat

AU - Nousiainen, Pertti

AU - Harlin, Ali

PY - 2016

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AB - The introduction of a three dimensional scaffold providing the closest analogies to extracellular matrix (ECM) is currently a key strategy for tackling many challenges in tissue repair. Here, we present a new hybrid scaffold constructed by coating electrospun chitosan/polyethylene oxide (PEO) nanofibers on commercial BioTek polystyrene (PS) scaffold obtained from Sigma Aldrich. The viability and proliferation rate of mesenchymal stem cells (MSCs) seeded on micro-nano structured hybrid scaffold (MNHS) and commercial PS scaffolds were analyzed by MTT assay. The results of the MTT assay revealed a higher degree of viability and proliferation rate in MSCs seeded on MNHS compared with the commercial PS scaffold. DAPI images also confirmed the higher degree of attachment and viability of MSCs seeded on MNHS. Moreover, MSCs on both scaffolds differentiated to osteoblasts and adipocytes cells, as reflected by the images obtained from Alizarin Red and Oil Red-O staining. Alkaline phosphatase activity (ALP) and calcium content assays revealed that the MNHS has a higher potential for osteogenic differentiation than the commercial scaffold. To quantify the osteoblast and adipocyte gene expression, quantitative RT-PCR was carried out for MNHS, commercial scaffold and Tissue culture polystyrene (TCPS). It was found that MNHS can express a higher level of Runt-related transcription factor 2 (Runx2), osteonectin and osteocalcin in osteogenic differentiation as well as increased expression of PPAR? and UCP-1 in adipogenic differentiation. The enhancement of the attachment, viability and proliferation as well as bi-lineage differentiation may result from the biochemical and structural analogies of MNHS to native ECM. Furthermore, it was observed that biocompatible MNHS scaffold can potentially be utilized as a suitable scaffold for bone and connective tissue engineering.

KW - adipogenic differentiation

KW - hybrid scaffold

KW - mesenchymal stem cell

KW - micro porosity

KW - nanofiber

KW - osteogenic differentiation

U2 - 10.1166/jnn.2016.12740

DO - 10.1166/jnn.2016.12740

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EP - 9007

JO - Journal of Nanoscience and Nanotechnology

JF - Journal of Nanoscience and Nanotechnology

SN - 1533-4880

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