Layer-by-layer paper-stacking nanofibrous membranes to deliver adipose-derived stem cells for bone regeneration

Abstract

Bone tissue engineering through seeding of stem cells in three-dimensional scaffolds has greatly improved bone regeneration technology, which historically has been a constant challenge. In this study, we researched the use of adipose-derived stem cell (ADSC)-laden layer-by-layer paper-stacking polycaprolactone/gelatin electrospinning nanofibrous membranes for bone regeneration. Using this novel paper-stacking method makes oxygen distribution, nutrition, and waste transportation work more efficiently. ADSCs can also secrete multiple growth factors required for osteogenesis. After the characterization of ADSC surface markers CD29, CD90, and CD49d using flow cytometry, we seeded ADSCs on the membranes and found cells differentiated, with significant expression of the osteogenic-related proteins osteopontin, osteocalcin, and osteoprotegerin. During 4 weeks in vitro, the ADSCs cultured on the paper-stacking membranes in the osteogenic medium exhibited the highest osteogenic-related gene expressions. In vivo, the paper-stacking scaffolds were implanted into the rat calvarial defects (5 mm diameter, one defect per parietal bone) for 12 weeks. Investigating with microcomputer tomography, the ADSC-laden paper-stacking membranes showed the most significant bone reconstruction, and from a morphological perspective, this group occupied 90% of the surface area of the defect, produced the highest bone regeneration volume, and showed the highest bone mineral density of 823.06 mg/cm 3 . From hematoxylin and eosin and Masson staining, the new bone tissue was most evident in the ADSC-laden scaffold group. Using quantitative polymerase chain reaction analysis from collected tissues, we found that the ADSC-laden paper-stacking membrane group presented the highest osteogenic-related gene expressions of osteocalcin, osteopontin, osteoprotegerin, bone sialoprotein, runt-related transcription factor 2, and osterix (two to three times higher than the control group, and 1.5 times higher than the paper-stacking membrane group in all the genes). It is proposed that ADSC-laden layer-by-layer paper-stacking scaffolds could be used as a way of promoting bone defect treatment.

A Bioactive Cartilage Graft of IGF1-Transduced Adipose Mesenchymal Stem Cells Embedded in an Alginate/Bovine Cartilage Matrix Tridimensional Scaffold

“…As shown by FACS analysis, a homogenous adherent cell population was isolated by collagenase I digestion (Figure 1(c)); the isolated cells express mesenchymal antigens CD73, CD90, and CD105 and also were negative for HSC markers CD34, CD45, and CD14 (Figure 1(d)). Similar to previous studies, the expression profile of surface markers correlates with cell-based isolation with collagenase I digestion [27, 28]. Nevertheless, we found a discrepancy regarding CD90 expression, which according to Maddox et al, its expression in ADSCs is negative.…”

Biomineralization Guided by Paper Templates

“…The deposition of calcium phosphate in the 3D paper scaffolds described here are the first functional examples of template-guided mineralization by osteoblasts in a 3D system. Although in a previous report adipose-derived stem cells (ADSC) that were grown in polycaprolactone/gelatin electrospun nanofibrous membranes demonstrated regeneration of bone in a calvarial defect in a mouse model 42 , achieving mineralization by osteoblasts is a greater challenge due to their short life-span and possibility for de-differentiation.…”

Fabrication of multilayered electrospun poly(lactic‐co‐glycolic acid)/polyvinyl pyrrolidone + poly(ethylene oxide) scaffolds and biocompatibility evaluation

“…The multilayer electrospinning technology adopted in our study was demonstrated to be capable of fabricating scaffolds meeting TE standards. The multilayer electrospinning method can further complement the advantages of the biomaterials with different functions, producing scaffolds with better flexibility, functionality, and plasticity 22,23 . In this study, we applied three materials, namely, PLGA, PVP, and PEO, and combined the benefits of the three to compensate for the limitations of each individual material, thus maximizing their respective advantages.…”