Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Seeded on Melt Based Chitosan Scaffolds for Bone Tissue Engineering Applications

Abstract

The purpose of this study was to evaluate the growth patterns and osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) when seeded onto new biodegradable chitosan/polyester scaffolds. Scaffolds were obtained by melt blending chitosan with poly(butylene succinate) in a proportion of 50% (wt) each and further used to produce a fiber mesh scaffold. hBMSCs were seeded on those structures and cultured for 3 weeks under osteogenic conditions. Cells were able to reduce MTS and demonstrated increasing metabolic rates over time. SEM observations showed cell colonization at the surface as well as within the scaffolds. The presence of mineralized extracellular matrix (ECM) was successfully demonstrated by peaks corresponding to calcium and phosphorus elements detected in the EDS analysis. A further confirmation was obtained when carbonate and phosphate group peaks were identified in Fourier Transformed Infrared (FTIR) spectra. Moreover, by reverse transcriptase (RT)-PCR analysis, it was observed the expression of osteogenic gene markers, namely, Runt related transcription factor 2 (Runx2), type 1 collagen, bone sialoprotein (BSP), and osteocalcin. Chitosan-PBS (Ch-PBS) biodegradable scaffolds support the proliferation and osteogenic differentiation of hBMSCs cultured at their surface in vitro, enabling future in vivo testing for the development of bone tissue engineering therapies.

DHEA promotes osteoblast differentiation by regulating the expression of osteoblast-related genes and Foxp3⁺ regulatory T cells

“…MSCs were cultured as previously described (15). Mice were anesthetized with 10% chloral hydrate and immersed in 75% ethanol for 10 min.…”

Synergistic intrafibrillar/extrafibrillar mineralization of collagen scaffolds based on a biomimetic strategy to promote the regeneration of bone defects

“…With the development of bone tissue engineering, scaffolds fabricated using different biomaterials, such as bioactive glasses, polylactic acid, collagen, chitosan, and hydroxyapatite (HA), have been explored for bone regeneration due to their biocompatibility, sufficient supply, and no secondary damage for patients. [1][2][3][4][5][6][7] Among these biomaterials, type 1 9 To date, mixing collagen with minerals and mineralizing collagen have been used to modify collagen scaffolds for simulating the composition and microstructure of bone tissue to the greatest extent possible. [10][11][12][13][14][15][16][17] Because HA is a primary mineral component of natural bone and possesses outstanding bioactivity, osteoconductivity, biocompatibility with bone cells, good mechanical properties, and a slow degradation rate in vivo, it has often been the first choice for modifying collagen scaffolds.…”

Response of human mesenchymal stem cells to intrafibrillar nanohydroxyapatite content and extrafibrillar nanohydroxyapatite in biomimetic chitosan/silk fibroin/nanohydroxyapatite nanofibrous membrane scaffolds

“…It is reported to be a useful material for migration and differentiation of bone marrow mesenchymal stem cells. 10 Though it is good for differentiation, poor processability and inferior mechanical properties hinder the applicability of CS alone in tissue engineering. To solve this problem, CS can be combined with other polymeric biomaterials such as alginate, gelatin, hyaluronic acid, and SF to develop better scaffolds for tissue engineering by retaining its osteogenic activities, but with improved mechanical properties.…”