In vitro and in vivo biocompatibility study on acellular sheep periosteum for guided bone regeneration

Abstract

This study addresses the fabrication of an extracellular matrix material of the acellular sheep periosteum and the systematic evaluation of its biocompatibility to explore its potential application in guided bone regeneration. Sheep periosteum was harvested and decellularized by a combined decellularization protocol. The effectiveness of cell removal was proved and residual α-Gal antigen was also quantitatively detected. Then, mouse MC3T3-E1 cells were seeded onto the acellular periosteum. A scanning electron microscope (SEM) was used to record the whole process of cell adhesion. The CCK-8 assay suggested that the acellular periosteum not only had zero toxic effect on pre-osteoblasts, but played a positive role in cell proliferation. We also tested whether the acellular periosteum possesses favorable osteogenesis induction activity using an alkaline phosphatase (ALP) assay and a quantitative real-time PCR (Col I, Runx2, OCN) assay. An in vivo study of a subcutaneous implantation test using Sprague Dawley (SD) rats was performed to detect the changes in IL-2, IFN-γ and IL-4 in serum and elucidate the host’s local response to acellular periosteum through hematoxylin and eosin (HE) and immunohistochemical staining. The results show that acellular sheep periosteum did not elicit a severe immunogenic response via the Th1 pathway, unlike fresh sheep periosteum. In conclusion, acellular sheep periosteum possesses favorable biocompatibility to be employed for guided bone regeneration.

Fabrication and evaluation of an optimized xenogenic decellularized costal cartilage graft: preclinical studies of a novel biocompatible prosthesis for rhinoplasty

“…Regarding biocompatibility and biosecurity, this study conducted in vitro tests, including cell affinity, DNA released in culture medium [ 45 ] and CCK-8 assay [ 46 ], as well as in vivo tests, including inflammatory factor ELISA and neutrophil immunohistochemical staining [ 47 ]. The possible reasons for immune rejection and cytotoxicity of decellularized ECM materials mainly include residual cell components and toxic substances remaining in the process of decellularization and sterilization [ 20 , 48 , 49 ].…”

Resveratrol benefits the lineage commitment of bone marrow mesenchymal stem cells into osteoblasts via miR‐320c by targeting Runx2

“…Runx2 is a crucial transcriptional regulator of bone formation and plays a crucial role in modulating the differentiation of BMSCs (Colbath, Kisiday, Phillips, & Goodrich, 2019; He et al., 2019; Wang et al., 2019). Previous studies showed that Runx2 was the target of many miRNAs, including miR‐23a, miR‐30c, miR‐204, and miR‐143‐5p (Hamam et al., 2014; Zhan, Liu, & Wang, 2018; Zhang et al., 2011).…”

Resveratrol Induces the Osteogenic Differentiation via MiR-320c by Targeting Runx2 and Is Involved in the Adipogenic differentiation of BMSCs

“…Runx2 is a crucial transcriptional regulator of bone formation and plays an important role in modulating the differentiation of mesenchymal stem cells [36][37][38]. Previous studies showed that Runx2 was the target of many miRNAs, such as miR-23a, miR-30c, miR-204 and miR-143-5p [39][40][41].…”