Bone Morphogenetic Protein‐6‐loaded Chitosan Scaffolds Enhance the Osteoblastic Characteristics of MC3T3‐E1 Cells

Abstract

Abstract:The purpose of this study is to investigate the convenience of bone morphogenetic protein-6 (BMP-6)-loaded chitosan scaffolds with preosteoblastic cells for bone tissue engineering. MC3T3-E1 cells were seeded into three different groups: chitosan scaffolds, BMP-6-loaded chitosan scaffolds, and chitosan scaffolds with free BMP-6 in culture medium. Tissue-engineered constructs were characterized by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay, scanning electron microscopy (SEM), mineralization assay (von Kossa), alkaline phosphatase (ALP) activity, and osteocalcin (OCN) assays. BMP-6-loaded chitosan scaffolds supported proliferation of the MC3T3-E1 mouse osteogenic cells in a similar pattern as the unloaded chitosan scaffolds group and as the chitosan scaffolds with free BMP-6 group. SEM images of the cell-seeded scaffolds revealed significant acceleration of extracellular matrix synthesis in BMP-6-loaded chitosan scaffolds. Both levels of ALP and OCN were higher in BMP-6-loaded chitosan scaffold group compared with the other two groups. In addition, BMP-6-loaded scaffolds showed strong staining in mineralization assays. These findings suggest that BMP-6-loaded chitosan scaffold supports cellular functions of the osteoblastic cells; therefore, this scaffold is considered as a new promising vehicle for bone tissue engineering applications.

Tissue Engineering for Tissue and Organ Regeneration

“…The presence of a small reservoir of cardiac resident progenitor cells (CPC or CSC) has been recently demonstrated in human as well as in other mammals' heart [24]. Such tissueresident cells participate in myocardial homeostasis and retain a limited regenerative capacity throughout organism lifespan [1]. All the subsets so far identified through the expression of stemness markers (c-kit+, Sca-1+, Islet-1+) demonstrated the ability to give birth to new contractile cells in vitro, while only c-kit+, Sca-1+ progenitors were shown to be Tissue Engineering for Tissue and Organ Regeneration 22 involved in post-natal cardiac tissue homeostasis in vivo [25].…”

“…The low isolation efficiency is particularly problematic regarding the use of autologous fibrin gel scaffolds in paediatric patients, as the volume of sampled blood needs to be kept to an absolute minimum. Therefore, the use of alternative precipitation methods with different chemicals has been evaluated: (1) ethanol (Kjaergard et al, 1992;Weis-Fogh, 1988), (2) ammonium sulphate alone, and (3) in combination with the cryoprecipitation method (Wolf, 1983), (4) albumin plus cryoprecipitation, and (5) polyethylene glycol (PEG) plus cryoprecipitation (Epstein et al, 1986). Heselhaus investigated each of these different precipitation methods with regard to their efficiency and their use in the development of fibrin scaffold materials for cardiovascular applications (Heselhaus, 2011):…”

“…A number of stem cells and progenitors have been so far proposed for cardiac repair, due to the inability of cardiomyocytes to proliferate after birth [1]. Among the cell sources challenged for the possibility to produce new cardiomyocytes, skeletal myoblasts have proven to be able to acquire a contractile phenotype in vitro [13].…”

“…In total, less than 50% of cardiomyocytes are renewed during a normal human life span [1]. For this reason, the topic of cardiac repair is among the major challenges for the tissue engineers worldwide.…”

Nanomaterials for cartilage tissue engineering

“…TGF-β1 could be released from the microspheres in a controlled multiphasic manner and stimulated cell proliferation and the synthesis of collagen type II. Another chondrogenic differentiation factor, bone morphogenetic protein-6 (BMP-6), was loaded onto chitosan scaffolds and increased the contents of GAG, collagen II and DNA in cultured cell-scaffold constructs [84].…”