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Osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells encapsulated in Thai silk fibroin/collagen hydrogel: a pilot study in vitro

Jirun Apinun
Jirun Apinun
, 
Sittisak Honsawek
Sittisak Honsawek
, 
Somsak Kuptniratsaikul
Somsak Kuptniratsaikul
, 
Jutarat Jamkratoke
Jutarat Jamkratoke
 and 
Sorada Kanokpanont
Sorada Kanokpanont
   | Oct 24, 2019
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Article Category: Brief communication (original)
Published Online: Oct 24, 2019
Page range: 273 - 279
DOI: https://doi.org/10.1515/abm-2019-0030
Keywords
© 2018 Jirun Apinun et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Figure 1

Growth kinetics of rat mesenchymal stem cells cultured in hydrogels. Squares indicate hydrogels made of 4% silk fibroin (SF) solution only (SF hydrogel), circles indicate hydrogels made of 4% SF solution + 0.05% collagen solution (SF/0.05C hydrogel), and triangles indicate hydrogels made of 4% SF solution + 0.1% collagen solution (SF/0.1C hydrogel). Error bars are standard deviations of triplicate experiments.
Growth kinetics of rat mesenchymal stem cells cultured in hydrogels. Squares indicate hydrogels made of 4% silk fibroin (SF) solution only (SF hydrogel), circles indicate hydrogels made of 4% SF solution + 0.05% collagen solution (SF/0.05C hydrogel), and triangles indicate hydrogels made of 4% SF solution + 0.1% collagen solution (SF/0.1C hydrogel). Error bars are standard deviations of triplicate experiments.

Figure 2

DNA assay of rat mesenchymal stem cells cultured in hydrogels in osteogenic induction medium. Squares indicate hydrogels made of 4% silk fibroin (SF) solution only (SF hydrogel), circles indicate hydrogels made of 4% SF solution + 0.05% collagen solution (SF/0.05C hydrogel), and triangles indicate hydrogels made of 4% SF solution + 0.1% collagen solution (SF/0.1C hydrogel). *P < 0.05 (one-way analysis of variance with a post hoc least significant difference test). Error bars are standard deviations of triplicate experiments.
DNA assay of rat mesenchymal stem cells cultured in hydrogels in osteogenic induction medium. Squares indicate hydrogels made of 4% silk fibroin (SF) solution only (SF hydrogel), circles indicate hydrogels made of 4% SF solution + 0.05% collagen solution (SF/0.05C hydrogel), and triangles indicate hydrogels made of 4% SF solution + 0.1% collagen solution (SF/0.1C hydrogel). *P < 0.05 (one-way analysis of variance with a post hoc least significant difference test). Error bars are standard deviations of triplicate experiments.

Figure 3

Alkaline phosphatase (ALP) activity of rat mesenchymal stem cells cultured on hydrogels in osteogenic induction medium. Units indicate mM p-nitrophenol produced per min per cell. Squares indicate hydrogels made of 4% silk fibroin (SF) solution only (SF hydrogel), circles indicate hydrogels made of 4% SF solution + 0.05% collagen solution (SF/0.05C hydrogel), and triangles indicate hydrogels made of 4% SF solution + 0.1% collagen solution (SF/0.1C hydrogel). Error bars are standard deviations of triplicate experiments.
Alkaline phosphatase (ALP) activity of rat mesenchymal stem cells cultured on hydrogels in osteogenic induction medium. Units indicate mM p-nitrophenol produced per min per cell. Squares indicate hydrogels made of 4% silk fibroin (SF) solution only (SF hydrogel), circles indicate hydrogels made of 4% SF solution + 0.05% collagen solution (SF/0.05C hydrogel), and triangles indicate hydrogels made of 4% SF solution + 0.1% collagen solution (SF/0.1C hydrogel). Error bars are standard deviations of triplicate experiments.

Figure 4

Calcium content of rat mesenchymal stem cells cultured on hydrogels in osteogenic induction medium. Squares indicate hydrogels made of 4% silk fibroin (SF) solution only (SF hydrogel), circles indicate hydrogels made of 4% SF solution + 0.05% collagen solution (SF/0.05C hydrogel), and triangles indicate hydrogels made of 4% SF solution + 0.1% collagen solution (SF/0.1C hydrogel). *P < 0.05 (one-way analysis of variance with a post hoc least significant difference test). Error bars are standard deviations of triplicate experiments.
Calcium content of rat mesenchymal stem cells cultured on hydrogels in osteogenic induction medium. Squares indicate hydrogels made of 4% silk fibroin (SF) solution only (SF hydrogel), circles indicate hydrogels made of 4% SF solution + 0.05% collagen solution (SF/0.05C hydrogel), and triangles indicate hydrogels made of 4% SF solution + 0.1% collagen solution (SF/0.1C hydrogel). *P < 0.05 (one-way analysis of variance with a post hoc least significant difference test). Error bars are standard deviations of triplicate experiments.

Figure 5

SEM images of rat MSCs cultured in hydrogels under different magnifications. SF (hydrogel made of 4% silk fibroin (SF) solution only, SF/0.05C (hydrogel made of 4% SF solution + 0.05% collagen solution), and SF/0.1C (hydrogel made of 4% SF solution + 0.1% collagen solution).
SEM images of rat MSCs cultured in hydrogels under different magnifications. SF (hydrogel made of 4% silk fibroin (SF) solution only, SF/0.05C (hydrogel made of 4% SF solution + 0.05% collagen solution), and SF/0.1C (hydrogel made of 4% SF solution + 0.1% collagen solution).

Elemental analysis result by EDS

HydrogelsC (% wt)O (% wt)Ca (% wt)P (% wt)Ca/P
SF45.03 ± 5.5637.61 ± 3.7513.99 ± 1.684.03 ± 1.814.28 ± 2.38
SF/0.05C39.36 ± 2.9633.99 ± 10.5619.27 ± 7.137.38 ± 3.372.91 ± 1.09
SF/0.1C45.92 ± 10.0038.96 ± 3.1110.86 ± 6.044.27 ± 4.373.61 ± 1.75
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