Biofabricated Marine Hydrozoan: A Bioactive Crystalline Material Promoting Ossification of Mesenchymal Stem Cells

Abstract

This study introduces a novel three-dimensional biomatrix obtained from the marine hydrocoral Millepora dichotoma as a scaffold for hard tissue engineering. Millepora dichotoma was biofabricated under field and laboratory conditions. Three-dimensional biomatrices were made in order to convert mesenchymal stem cells (MSCs) to exemplify osteoblastic phenotype. We investigated the effect of the biomatrices on MSCs proliferation and differentiation at 2, 3, 4, 7, 10, 14, 21, 28, and 42 days. Different analyses were made: light microscopy, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), calcium incorporation to newly formed tissue (alizarin red), bone nodule formation (von Kossa), fat aggregate formation (oil red O), collagen type I immunofluorescence, DNA concentrations, alkaline phosphatase (ALP) activity, and osteocalcin concentrations. MSCs seeded on Millepora dichotoma biomatrices showed higher levels of calcium and phosphate incorporation and higher type I collagen levels than did control Porites lutea biomatrices. ALP activity revealed that MSCs seeded on M. dichotoma biomatrices are highly osteogenic compared to those on control biomatrices. The osteocalcin content of MSCs seeded on M. dichotoma remained constant up to 2 weeks before rising to surpass that of seeded P. lutea biomatrices after 28 days. Our study thus showed that M. dichotoma biomatrices enhance the differentiation of MSCs into osteoblast and hence have excellent potential as bioscaffold for hard tissue engineering.

Calcite Biohybrids as Microenvironment for Stem Cells

“…The coral exoskeleton, made of calcium carbonate in aragonite polymorph, has a structure similar to that of cancellous bone. These scaffolds have since been shown to be biocompatible, osteoconductive, and biodegradable [14][15][16]. A calcite polymorph of calcium carbonate from natural limestone was used as a bone substitute in vivo in rabbits [17].…”

Uncultured Marrow Mononuclear Cells Delivered Within Fibrin Glue Hydrogels to Porous Scaffolds Enhance Bone Regeneration Within Critical-Sized Rat Cranial Defects

“…This result was somewhat surprising, as previous work has demonstrated MSC osteogenic differentiation and ingrowth on similar scaffolds. [70][71][72] Although reports showing little to no osteogenesis on coralline HA scaffolds do exist, 73 bony ingrowth did occur in all of the tested formulations using coral.…”

Effects of Three Dimensional Microenvironment on Tumorigenicity of Fibrosarcoma in vitro

“…Previously, the POR biolattice was proven to induce spontaneous MSCs differentiation towards osteblasts. 41,50 However, the same microenvironment induced tumor promoting effects in the case of a fibrosarcomas, such as increased migration (Figure 2) and elevated tumor-related genes CCL5 and c-Myc ( Figure 5); this phenomena were not evident in the non-tumorigenic cells (Figure 7), indicating that the tumorigenic effect of the microenvironment is limited to malignant cells. Bone marrow has been described as a reservoir of Cancer Stem Cells (CSCs); [61][62][63]64 however, it remains to be established whether cancer cells reach their bone marrow niches already equipped with upgraded tumorigenic capacities, or whether their tumorgenic potential may be regulated by this unique microenvironment.…”

“…In light of our group's earlier studies in which MSCs demonstrated spontaneous differentiation towards an osteoblastic fate when cultivated on the POR biolattice, 41 we speculated that restrained proliferation could point to differentiation and hence restricted movement and tumorgenic functioning. However, this notion was dispelled by our further results: on both 2D surfaces and 3D POR scaffolds, cells grown in the presence of calcium displayed enhanced migration abilities (Figure 2a).…”