Guided osteoporotic bone regeneration with composite scaffolds of mineralized ECM/heparin membrane loaded with BMP2-related peptide

Abstract

IntroductionAt present, the treatment of osteoporotic defects poses a great challenge to clinicians, owing to the lower regeneration capacity of the osteoporotic bone as compared with the normal bone. The guided bone regeneration (GBR) technology provides a promising strategy to cure osteoporotic defects using bioactive membranes. The decellularized matrix from the small intestinal submucosa (SIS) has gained popularity for its natural microenvironment, which induces cell response.Materials and methodsIn this study, we developed heparinized mineralized SIS loaded with bone morphogenetic protein 2 (BMP2)-related peptide P28 (mSIS/P28) as a novel GBR membrane for guided osteoporotic bone regeneration. These mSIS/P28 membranes were obtained through the mineralization of SIS (mSIS), followed by P28 loading onto heparinized mSIS. The heparinized mSIS membrane was designed to improve the immobilization efficacy and facilitate controlled release of P28. P28 release from mSIS-heparin-P28 and its effects on the proliferation, viability, and osteogenic differentiation of bone marrow stromal stem cells from ovariectomized rats (rBMSCs-OVX) were investigated in vitro. Furthermore, a critical-sized OVX calvarial defect model was used to assess the bone regeneration capability of mSIS-heparin-P28 in vivo.ResultsIn vitro results showed that P28 release from mSIS-heparin-P28 occurred in a controlled manner, with a long-term release time of 40 days. Moreover, mSIS-heparin-P28 promoted cell proliferation and viability, alkaline phosphatase activity, and mRNA expression of osteogenesis-related genes in rBMSCs-OVX without the addition of extra osteogenic components. In vivo experiments revealed that mSIS-heparin-P28 dramatically stimulated osteoporotic bone regeneration.ConclusionThe heparinized mSIS loaded with P28 may serve as a potential GBR membrane for repairing osteoporotic defects.

Diverse preparation methods for small intestinal submucosa (SIS): Decellularization, components, and structure

“…Small intestinal submucosa is widely used for tissue repair and regeneration, with benefits from its ECM functional components and structure . It is considered to positively modulate cell behaviors and induce vascularization for tissue repair and remolding .…”

“…Small intestinal submucosa is widely used for tissue repair and regeneration, with benefits from its ECM functional components and structure. 3,4,29,30 It is considered to positively modulate cell behaviors and induce vascularization for tissue repair and remolding. 31,32 As a graft to be implanted into the body, most of the intrinsic cells of SIS need to be removed to avoid adverse immunogenic reactions.…”

<p>Creation of Bony Microenvironment with Extracellular Matrix Doped-Bioactive Ceramics to Enhance Osteoblast Behavior and Delivery of Aspartic Acid-Modified BMP-2 Peptides</p>

“… 28 Inspired by the natural delivery mechanisms observed in vivo, the surface of MBG/SIS scaffolds was reinforced with a heparin domain to regulate P28 release. 21 Besides, SIS was efficiently immobilized on MBG scaffolds via the pDA-assisted coating method, which further maintained the control release profile of P28. 38 A mussel-inspired immobilization strategy, pDA chemistry, would provide an easy, safe, and efficient method to deposit proteins and peptides onto polymeric scaffolds.…”

“…Alizarin Red S staining was used to observe ECM calcification as described previously. 21 In brief, 2*10 5 MC3T3-E1 cells seeded on MBG/SIS, MBG/SIS-P28 and MBG/SIS-H-P28 scaffolds (1×1×0.2 cm 3 ) were cultured in osteogenic medium for 21 d. At 14 and 21 d, cells/scaffolds were stained according to the manufacturer’s protocol for Alizarin red S staining (Beyotime Biotechnology, Nanjing, China). To quantify calcium nodule deposition further, samples were treated with 10% (v/v aqueous solution) cetylpyridinium chloride monohydrate (Sigma-Aldrich, USA) to extract the alizarin red dye.…”

“… 16 – 19 Previous studies conducted by us and other research groups have demonstrated that SIS-based biological scaffolds exhibit excellent capability for guiding bone regeneration. 20 , 21 As such, SIS was expected to be a coating modifier able to provide a bone-like matrix for MBG scaffolds.…”