Characterization and cytocompatibility of 3D porous biomimetic scaffold derived from rabbit nucleus pulposus tissue in vitro

Yu Zhang,Wei Tan,Wei Tan,Wei Tan,Wei Tan,Chu-Song Zhou,Jin Sun,You Wu,Mingxin Wu,Mingxin Wu,Wei Cao

doi:10.1007/s10856-020-06480-9

Abstract

Intervertebral disc (IVD) degeneration is one of the most important causes of lower back pain. Tissue engineering provides a new method for the experimental treatment of degenerative disc diseases. This study aims to develop a natural, acellular, 3D interconnected porous scaffold derived from the extracellular matrix (ECM) of nucleus pulposus. The nucleus pulposus (NP) was decellularized by sequential detergent-nuclease methods, including physical crushing, freeze-drying and cross-linking. These 3D porous scaffolds were fabricated with a high porosity of (81.28 ± 4.10)%, an ideal pore size with appropriate mechanical properties. Rabbit bone marrow mesenchymal stem cells (rBMSCs) were seeded and cultured on the scaffolds. And the mechanical tests showed the compressive elastic modulus of the scaffolds cultured for 4 weeks reached 0.12 MPa, which was better than that of the scaffolds cultured for 2 weeks (0.07 MPa) and that of the control group (0.04 MPa). Scanning electron microscopy (SEM), histological assays, molecular biology assays revealed that the scaffolds could provide an appropriate microstructure and environment for the adhesion, proliferation, migration and secretion of seeded cells in vitro. As assays like histology, immunohistochemistry and the real-time qRT-PCR showed, NP-like tissues were preliminarily formed. In conclusion, the 3D porous scaffold derived from NP ECM is a potential biomaterial for the regeneration of NP tissues.A natural, acellular, 3D interconnected porous scaffold derived from the extracellular matrix (ECM) of nucleus pulposus was developed by sequential detergent-nuclease and freeze-drying method, which can reduce the damage of protein activity to the minimum. It is very similar to the composition and internal environment of the natural nucleus pulposus, because it derived from the natural nucleus pulposus. Scanning electron microscopy (SEM), histological assays, molecular biology assays revealed that the scaffolds could provide an appropriate microstructure and environment for the adhesion, proliferation, migration, and secretion of seeded cells in vitro.

Highlights

Intervertebral disc degeneration (IDD) can result in lower back pain and significant socioeconomic burden
It is well known that IDD is a multifactor process, including genetic, biochemical, biomechanical factors, etc [1,2,3], which can result in the changes of cell-conditioned medium [4] within the extracellular matrix (ECM) of intervertebral disc (IVD)
The Scanning electron microscopy (SEM) results showed that the pore sizes of the scaffolds range from 45 to 450 μm, very similar to the ideal size [33] required for tissue engineering

Summary

Introduction

Intervertebral disc degeneration (IDD) can result in lower back pain and significant socioeconomic burden. The traditional clinical treatment of IDD, which includes conservative treatment (drug and physical therapy) [8] and surgery (spine fusion and total disc removal) [9], is not absolutely avoided from concerns about possible comorbidities, cost-effectiveness, secondary risks and long-lasting outcomes. These therapies can only alleviate the symptoms of lower back pain, but fail to treat the underlying cause of degeneration. Can a new biomaterial be found as a substitution for the NP tissue?

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Methods

Results

Conclusion