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Increased stem cells delivered using a silk gel/scaffold complex for enhanced bone regeneration.

ABSTRACT: The low in vivo survival rate of scaffold-seeded cells is still a challenge in stem cell-based bone regeneration. This study seeks to use a silk hydrogel to deliver more stem cells into a bone defect area and prolong the viability of these cells after implantation. Rat bone marrow stem cells were mingled with silk hydrogels at the concentrations of 1.0?×?105/mL, 1.0?×?106/mL and 1.0?×?107/mL before gelation, added dropwise to a silk scaffold and applied to a rat calvarial defect. A cell tracing experiment was included to observe the preservation of cell viability and function. The results show that the hydrogel with 1.0?×?107/mL stem cells exhibited the best osteogenic effect both in vitro and in vivo. The cell-tracing experiment shows that cells in the 1.0?×?107 group still survive and actively participate in new bone formation 8 weeks after implantation. The strategy of pre-mingling stem cells with the hydrogel had the effect of delivering more stem cells for bone engineering while preserving the viability and functions of these cells in vivo.

SUBMITTER: Ding X

PROVIDER: S-EPMC5438390 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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Increased stem cells delivered using a silk gel/scaffold complex for enhanced bone regeneration.

Ding Xun X Yang Guangzheng G Zhang Wenjie W Li Guanglong G Lin Shuxian S Kaplan David L DL Jiang Xinquan X

Scientific reports 20170519 1

The low in vivo survival rate of scaffold-seeded cells is still a challenge in stem cell-based bone regeneration. This study seeks to use a silk hydrogel to deliver more stem cells into a bone defect area and prolong the viability of these cells after implantation. Rat bone marrow stem cells were mingled with silk hydrogels at the concentrations of 1.0 × 105/mL, 1.0 × 106/mL and 1.0 × 107/mL before gelation, added dropwise to a silk scaffold and applied to a rat ...[more]

PMID: 28526887

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Project description:BackgroundThe invasive and complicated procedures involving the use of traditional stem cells limit their application in bone tissue engineering. Cell-free, tissue-engineered bones often have complex scaffold structures and are usually engineered using several growth factors (GFs), thus leading to costly and difficult preparations. Urine-derived stem cells (USCs), a type of autologous stem cell isolated noninvasively and with minimum cost, are expected to solve the typical problems of using traditional stem cells to engineer bones. In this study, a graphene oxide (GO)-modified silk fibroin (SF)/nanohydroxyapatite (nHA) scaffold loaded with USCs was developed for immunomodulation and bone regeneration.MethodsThe SF/nHA scaffolds were prepared via lyophilization and cross-linked with GO using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxy succinimide (NHS). Scaffolds containing various concentrations of GO were characterized using scanning electron microscopy (SEM), the elastic modulus test, Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectrometer (XPS). Examinations of cell adhesion, proliferation, viability, morphology, alkaline phosphatase activity, and osteogenesis-related gene expression were performed to compare the osteogenesis-related biological behaviors of USCs cultured on the scaffolds. The effect of USC-laden scaffolds on the differentiation of macrophages was tested using ELISA, qRT-PCR, and immunofluorescence staining. Subcutaneous implantations in rats were performed to evaluate the inflammatory response of the USC-laden scaffolds after implantation. The scaffolds loaded with USCs were implanted into a cranial defect model in rats to repair bone defects. Micro-computed tomography (μCT) analyses and histological evaluation were performed to evaluate the bone repair effects.ResultsGO modification enhanced the mechanical properties of the scaffolds. Scaffolds containing less than 0.5% GO had good biocompatibility and promoted USC proliferation and osteogenesis. The scaffolds loaded with USCs induced the M2-type differentiation and inhibited the M1-type differentiation of macrophages. The USC-laden scaffolds containing 0.1% GO exhibited the best capacity for promoting the M2-type differentiation of macrophages and accelerating bone regeneration and almost bridged the site of the rat cranial defects at 12 weeks after surgery.ConclusionsThis composite system has the capacity for immunomodulation and the promotion of bone regeneration and shows promising potential for clinical applications of USC-based, tissue-engineered bones.

Project description:BackgroundWith the development of tissue engineering, enhanced tendon regeneration could be achieved by exploiting suitable cell types and biomaterials. The accessibility, robust cell amplification ability, superior tendon differentiation potential, and immunomodulatory effects of human periodontal ligament stem cells (hPDLSCs) indicate their potential as ideal seed cells for tendon tissue engineering. Nevertheless, there are currently no reports of using PDLSCs as seed cells. Previous studies have confirmed the potential of silk scaffold for tendon tissue engineering. However, the biomimetic silk scaffold with tendon extracellular matrix (ECM)-like structure has not been systematically studied for in situ tendon regeneration. Therefore, this study aims to evaluate the effects of hPDLSCs and biomimetic silk scaffold on in situ tendon regeneration.MethodsHuman PDLSCs were isolated from extracted wisdom teeth. The differentiation potential of hPDLSCs towards osteo-, chondro-, and adipo-lineage was examined by cultured in different inducing media. Aligned and random silk scaffolds were fabricated by the controlled directional freezing technique. Scaffolds were characterized including surface structure, water contact angle, swelling ratio, degradation speed and mechanical properties. The biocompatibility of silk scaffolds was evaluated by live/dead staining, SEM observation, cell proliferation determination and immunofluorescent staining of deposited collagen type I. Subsequently, hPDLSCs were seeded on the aligned silk scaffold and transplanted into the ruptured rat Achilles tendon. Scaffolds without cells served as control groups. After 4 weeks, histology evaluation was carried out and macrophage polarization was examined to check the repair effects and immunomodulatory effects.ResultsHuman PDLSCs were successfully isolated, and their multi-differentiation potential was confirmed. Compared with random scaffold, aligned silk scaffold had more elongated and aligned pores and promoted the proliferation and ordered arrangement of hPDLSCs. After implantation into rat Achilles tendon defect, hPDLSCs seeded aligned silk scaffold enhanced tendon repair with more tendon-like tissue formation after 4 weeks, as compared to the scaffold-only groups. Higher expression of CD206 and lower expression of iNOS, IL-1β and TNF-α were found in the hPDLSCs seeded aligned silk scaffold group, which revealed its modulation effect of macrophage polarization from M1 to M2 phenotype.ConclusionsIn summary, this study demonstrates the efficacy of hPDLSCs as seed cells and aligned silk scaffold as a tendon-mimetic scaffold for enhanced tendon tissue engineering, which may have broad implications for future tendon tissue engineering and regenerative medicine researches.

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Increased stem cells delivered using a silk gel/scaffold complex for enhanced bone regeneration.

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Increased stem cells delivered using a silk gel/scaffold complex for enhanced bone regeneration.

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