A novel egg-shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineering
Abstract Background The main issue in cutaneous regeneration is to develop engineered scaffolds based on natural extracellular matrix to promote dynamics of skin progenitor cells and accelerate differentiation into mature keratinocytes. Methods In this study, nanofibrous scaffolds composed of a blen...
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| Format: | Article |
| Language: | English |
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BMC
2019-10-01
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| Series: | Journal of Biological Engineering |
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| Online Access: | http://link.springer.com/article/10.1186/s13036-019-0208-x |
| _version_ | 1818348585178103808 |
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| author | Leila Mohammadzadeh Reza Rahbarghazi Roya Salehi Mehrdad Mahkam |
| author_facet | Leila Mohammadzadeh Reza Rahbarghazi Roya Salehi Mehrdad Mahkam |
| author_sort | Leila Mohammadzadeh |
| collection | DOAJ |
| description | Abstract Background The main issue in cutaneous regeneration is to develop engineered scaffolds based on natural extracellular matrix to promote dynamics of skin progenitor cells and accelerate differentiation into mature keratinocytes. Methods In this study, nanofibrous scaffolds composed of a blend poly (ɛ-caprolactone) (PCL), silk fibroin (SF), soluble eggshell membrane (SESM), and Aloe vera (AV) gel were developed by electrospinning method and human basal cells were used to examine differentiation capacity toward keratinocyte-like cells. For this propose, cells were allocated to four distinct groups; control, PCL/SF, PCL/SF/SESM, and PCL/SF/SESM/AV. In all groups, cells were incubated with differentiation medium. Morphology, composition, hydrophilicity and mechanical features of PCL/SF, PCL/SF/SESM and PCL/SF/SESM/AV nanofibers were studied by scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy (FT-IR), water contact angle and tensile tests. To examine the orientation of basal cells to mature keratinocytes, we performed immunofluorescence analysis by monitoring cytokeratin-19. The expression of genes such as involucrin, keratin-14 and -5 was monitored by real-time PCR assay. Results PCL/SF, PCL/SF/SESM, and PCL/SF/SESM/AV had suitable physic chemical indices and biological activities to be applied as biomimetic scaffolds for the restoration cutaneous tissue. Compared to control, we found an increased basal cell proliferation at 7 and 14 days after plating on scaffolds and reach maximum levels in group PCL/SF/SESM/AV on day 14 (p < 0.05). Electron microscopy showed cell flattening, morphological adaptation. An integrated cell-to-cell connection was generated after cell seeding on scaffolds in all groups. Immunofluorescence imaging showed the ability of basal cells to synthesize cytokeratin-19 in PCL/SF, PCL/SF/SESM, and positive control cells after exposure to differentiation medium. However, these values were less in PCL/SF/SESM/AV compared to other groups. Real-time PCR analysis showed the potency of all scaffolds to induce the transcription of involucrin, keratin-14 and -5, especially involucrin in PCL/SF/SESM/AV group compared to the negative control. Conclusion Modulation of scaffolds with natural biopolymers could enable us to synthesize structures appropriate for cutaneous regeneration. |
| first_indexed | 2024-12-13T17:52:23Z |
| format | Article |
| id | doaj.art-9ab8700bdaf649b28fef2ce049e17c83 |
| institution | Directory Open Access Journal |
| issn | 1754-1611 |
| language | English |
| last_indexed | 2024-12-13T17:52:23Z |
| publishDate | 2019-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Biological Engineering |
| spelling | doaj.art-9ab8700bdaf649b28fef2ce049e17c832022-12-21T23:36:28ZengBMCJournal of Biological Engineering1754-16112019-10-0113111510.1186/s13036-019-0208-xA novel egg-shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineeringLeila Mohammadzadeh0Reza Rahbarghazi1Roya Salehi2Mehrdad Mahkam3Chemistry Department, Faculty of Science, Azarbaijan Shahid Madani UniversityStem Cell research Center, Tabriz University of Medical SciencesDrug Applied Research Center and Department of Medical Nanotechnology, Faculty of Advanced Medical Science, Tabriz University of Medical ScienceChemistry Department, Faculty of Science, Azarbaijan Shahid Madani UniversityAbstract Background The main issue in cutaneous regeneration is to develop engineered scaffolds based on natural extracellular matrix to promote dynamics of skin progenitor cells and accelerate differentiation into mature keratinocytes. Methods In this study, nanofibrous scaffolds composed of a blend poly (ɛ-caprolactone) (PCL), silk fibroin (SF), soluble eggshell membrane (SESM), and Aloe vera (AV) gel were developed by electrospinning method and human basal cells were used to examine differentiation capacity toward keratinocyte-like cells. For this propose, cells were allocated to four distinct groups; control, PCL/SF, PCL/SF/SESM, and PCL/SF/SESM/AV. In all groups, cells were incubated with differentiation medium. Morphology, composition, hydrophilicity and mechanical features of PCL/SF, PCL/SF/SESM and PCL/SF/SESM/AV nanofibers were studied by scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy (FT-IR), water contact angle and tensile tests. To examine the orientation of basal cells to mature keratinocytes, we performed immunofluorescence analysis by monitoring cytokeratin-19. The expression of genes such as involucrin, keratin-14 and -5 was monitored by real-time PCR assay. Results PCL/SF, PCL/SF/SESM, and PCL/SF/SESM/AV had suitable physic chemical indices and biological activities to be applied as biomimetic scaffolds for the restoration cutaneous tissue. Compared to control, we found an increased basal cell proliferation at 7 and 14 days after plating on scaffolds and reach maximum levels in group PCL/SF/SESM/AV on day 14 (p < 0.05). Electron microscopy showed cell flattening, morphological adaptation. An integrated cell-to-cell connection was generated after cell seeding on scaffolds in all groups. Immunofluorescence imaging showed the ability of basal cells to synthesize cytokeratin-19 in PCL/SF, PCL/SF/SESM, and positive control cells after exposure to differentiation medium. However, these values were less in PCL/SF/SESM/AV compared to other groups. Real-time PCR analysis showed the potency of all scaffolds to induce the transcription of involucrin, keratin-14 and -5, especially involucrin in PCL/SF/SESM/AV group compared to the negative control. Conclusion Modulation of scaffolds with natural biopolymers could enable us to synthesize structures appropriate for cutaneous regeneration.http://link.springer.com/article/10.1186/s13036-019-0208-xNanofibrous scaffoldsBasal cellsDifferentiationEggshell membranesAloe vera |
| spellingShingle | Leila Mohammadzadeh Reza Rahbarghazi Roya Salehi Mehrdad Mahkam A novel egg-shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineering Journal of Biological Engineering Nanofibrous scaffolds Basal cells Differentiation Eggshell membranes Aloe vera |
| title | A novel egg-shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineering |
| title_full | A novel egg-shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineering |
| title_fullStr | A novel egg-shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineering |
| title_full_unstemmed | A novel egg-shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineering |
| title_short | A novel egg-shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineering |
| title_sort | novel egg shell membrane based hybrid nanofibrous scaffold for cutaneous tissue engineering |
| topic | Nanofibrous scaffolds Basal cells Differentiation Eggshell membranes Aloe vera |
| url | http://link.springer.com/article/10.1186/s13036-019-0208-x |
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