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Comparison of human Mesenchymal Stem Cells biocompatibility data growth on gelatin and silk fibroin scaffolds.

ABSTRACT: The data showed how gelatin hydrogel and silk fibroin scaffolds could facilitate the growth of human Mesenchymal Stem Cells (hMSC). Gelatin hydrogel and silk fibroin are biodegradable materials. Gelatin hydrogel already has many uses in the medical field, especially in tissue engineering, but silk fibroin scaffold, which is made from the cocoon of silkworm by salt leaching, its role in facilitating growth of hMSC still needs to be proven. Data was obtained by characterization of hMSC, then growing hMSC on silk fibroin scaffolds with pore sizes of ±500 ?m and ±900 ?m and on gelatin hydrogel scaffolds as control. Testing was performed by counting cell growth on days 1, 3, 5, 7 and 14 with the MTT cytotoxicity assay protocol. The morphology of hMSC that grew on gelatin and silk fibroin scaffolds was observed with a Scanning Electron Microscope (SEM) on day 3. Characterization of the hMSC showed that it fulfilled the requirements of the International Society for Cellular Therapy (ISCT). The water content of the gelatin hydrogel scaffold was higher than the silk fibroin scaffold. Biocompatibility testing showed that the gelatin hydrogel scaffold could support cell growth until day 7, then decreased on day 14 compared to the silk fibroin scaffold based on absorbance on the MTT cytotoxicity assay, while growth on silk fibroin scaffold with pore size 833 ± 147 ?m was consistently higher than on pore size 462 ± 66 ?m from day 1 to day 14. Cell binding to the silk fibroin was proven from SEM observation.

SUBMITTER: Vanawati N

PROVIDER: S-EPMC6915795 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature

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Comparison of human Mesenchymal Stem Cells biocompatibility data growth on gelatin and silk fibroin scaffolds.

Vanawati Noviana N Barlian Anggraini A Tabata Yasuhiko Y Judawisastra Hermawan H Wibowo Indra I

Data in brief 20191017

The data showed how gelatin hydrogel and silk fibroin scaffolds could facilitate the growth of human Mesenchymal Stem Cells (hMSC). Gelatin hydrogel and silk fibroin are biodegradable materials. Gelatin hydrogel already has many uses in the medical field, especially in tissue engineering, but silk fibroin scaffold, which is made from the cocoon of silkworm by salt leaching, its role in facilitating growth of hMSC still needs to be proven. Data was obtained by characterization of hMSC, then growi ...[more]

PMID: 31871963

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Project description:Three-dimensional porous scaffolds prepared from regenerated silk fibroin using either an all-aqueous process or a process involving an organic solvent, hexafluoroisopropanol (HFIP), have shown promise in cell culture and tissue engineering applications. However, their biocompatibility and in vivo degradation have not been fully established. The present study was conducted to systematically investigate how processing method (aqueous vs. organic solvent) and processing variables (silk fibroin concentration and pore size) affect the short-term (up to 2 months) and long-term (up to 1 year) in vivo behavior of the protein scaffolds in both nude and Lewis rats. The samples were analyzed by histology for scaffold morphological changes and tissue ingrowth, and by real-time RT-PCR and immunohistochemistry for immune responses. Throughout the period of implantation, all scaffolds were well tolerated by the host animals and immune responses to the implants were mild. Most scaffolds prepared from the all-aqueous process degraded to completion between 2 and 6 months, while those prepared from organic solvent (hexafluoroisopropanol (HFIP)) process persisted beyond 1 year. Due to widespread cellular invasion throughout the scaffold, the degradation of aqueous-derived scaffolds appears to be more homogeneous than that of HFIP-derived scaffolds. In general and especially for the HFIP-derived scaffolds, a higher original silk fibroin concentration (e.g. 17%) and smaller pore size (e.g. 100-200microm) resulted in lower levels of tissue ingrowth and slower degradation. These results demonstrate that the in vivo behavior of the three-dimensional silk fibroin scaffolds is related to the morphological and structural features that resulted from different scaffold preparation processes. The insights gained in this study can serve as a guide for processing scenarios to match desired morphological and structural features and degradation time with tissue-specific applications.

Project description:Incorporation of nanohydroxyapatite (nHAP) within a chitosan (CS)/silk fibroin (SF) nanofibrous membrane scaffold (NMS) may provide a favorable microenvironment that more closely mimics the natural bone tissue physiology and facilitates enhanced osteogensis of the implanted cell population. In this study, we prepared pristine CS/SF NMS, composite CS/SF/nHAP NMS containing intrafibrillar nHAP by in situ blending of 10% or 30% nHAP before the electrospinning step, and composite CS/SF/nHAP NMS containing extrafibrillar nHAP by depositing 30% nHAP through alternative soaking surface mineralization. We investigated the effect of the incorporation of HAP nanoparticles on the physicochemical properties of pristine and composite NMS. We confirmed the presence of ~30 nm nHAP in the composite nanofibrous membranes by thermogravimetry analysis (TGA), X-ray diffraction (XRD), and scanning electron microscopy (SEM), either embedded in or exposed on the nanofiber. Nonetheless, the alternative soaking surface mineralization method drastically influenced the mechanical properties of the NMS with 88% and 94% drop in Young's modulus and ultimate maximum stress. Using in vitro cell culture experiments, we investigated the effects of nHAP content and location on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs). The proliferation of hMSCs showed no significant difference among pristine and composite NMS. However, the extent of osteogenic differentiation of hMSCs was found to be positively correlated with the content of nHAP in the NMS, while its location within the nanofiber played a less significant role. In vivo experiments were carried out with hMSCs seeded in CS/SF/30%nHAP NMS prepared by in situ blending and subcutaneous implantation in nude mice. Micro-computed tomography images as well as histological and immunohistochemical analysis of the retrieved hMSCs/NMS construct 1 and 2 months postimplantation indicated that NMS had the potential for bone regeneration and can be suggested as a promising scaffold for bone tissue engineering.

Project description:The dissolution of Bombyx mori silk fibroin (SF) films in formic acid (FA) for the preparation of electrospinning dopes is widely exploited to produce electrospun SF scaffolds. The SILKBridge® nerve conduit is an example of medical device having in its wall structure an electrospun component produced from an FA spinning dope. Though highly volatile, residual FA remains trapped into the bulk of the SF nanofibers. The purpose of this work is to investigate the type and strength of the interaction between FA and SF in electrospun mats, to quantify its amount and to evaluate its possible toxicological impact on human health. The presence of residual FA in SF mats was detected by FTIR and Raman spectroscopy (new carbonyl peak at about 1,725 cm-1) and by solid state NMR, which revealed a new carbonyl signal at about 164.3 ppm, attributed to FA by isotopic 13C substitution. Changes occurred also in the spectral ranges of hydroxylated amino acids (Ser and Thr), demonstrating that FA interacted with SF by forming formyl esters. The total amount of FA was determined by HS-GC/MS analysis and accounted for 247 ± 20 μmol/g. The greatest part was present as formyl ester, a small part (about 3%) as free FA. Approximately 17% of the 1,500 μmol/g of hydroxy amino acids (Ser and Thr) theoretically available were involved in the formation of formyl esters. Treatment with alkali (Na2CO3) succeeded to remove the greatest part of FA, but not all. Alkali-treated electrospun SF mats underwent morphological, physical, and mechanical changes. The average diameter of the fibers increased from about 440 nm to about 480 nm, the mat shrunk, became stiffer (the modulus increased from about 5.5 MPa to about 7 MPa), and lost elasticity (the strain decreased from about 1 mm/mm to about 0.8 mm/mm). Biocompatibility studies with human adult dermal fibroblasts did not show significant difference in cell proliferation (313 ± 18 and 309 ± 23 cells/mm2 for untreated and alkali-treated SF mat, respectively) and metabolic activity. An in-depth evaluation of the possible toxicological impact of residual FA was made using the SILKBridge® nerve conduit as case study, following the provisions of the ISO 10993-1 standard. The Potential Patient Daily Intake, calculated from the total amount of FA determined by HS-GC/MS, was 2.4 mg/day and the Tolerable Exposure level was set to 35.4 mg/day. This allowed to obtain a value of the Margin of Safety of 15, indicating that the amount of FA left on SF mats after electrospinning does not raise concerns for human health.

Dataset Information

Comparison of human Mesenchymal Stem Cells biocompatibility data growth on gelatin and silk fibroin scaffolds.

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Comparison of human Mesenchymal Stem Cells biocompatibility data growth on gelatin and silk fibroin scaffolds.

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