Rajendiran Rajesh

Development of a new carbon nanotube–alginate–hydroxyapatite tricomponent composite scaffold for application in bone tissue engineering

In recent times, tricomponent scaffolds prepared from naturally occurring polysaccharides, hydroxyapatite, and reinforcing materials have been gaining increased attention in the field of bone tissue engineering. In the current work, a tricomponent scaffold with an oxidized multiwalled carbon nanotube (fMWCNT)–alginate–hydroxyapatite with the required porosity was prepared for the first time by a freeze-drying method and characterized using analytical techniques. The hydroxyapatite for the scaffold was isolated from chicken bones by thermal calcination at 800°C. The Fourier transform infrared spectra and X-ray diffraction data confirmed ionic interactions and formation of the fMWCNT–alginate–hydroxyapatite scaffold. Interconnected porosity with a pore size of 130–170 µm was evident from field emission scanning electron microscopy. The total porosity calculated using the liquid displacement method was found to be 93.85%. In vitro biocompatibility and cell proliferation on the scaffold was checked using an MG-63 cell line by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cell attachment by Hoechst stain assay. In vitro studies showed better cell proliferation, cell differentiation, and cell attachment on the prepared scaffold. These results indicate that this scaffold could be a promising candidate for bone tissue engineering.

Nerve growth factor-loaded heparinized cationic solid lipid nanoparticles for regulating membrane charge of induced pluripotent stem cells during differentiation

Nerve growth factor (NGF)-loaded heparinized cationic solid lipid nanoparticles (NGF-loaded HCSLNs) were developed using heparin-stearic acid conjugate, cacao butter, cholesterol, stearylamine (SA), and esterquat 1 (EQ 1). The effect of cationic lipids and lipid matrix composition on the particle size, particle structure, surface molecular composition, chemical structure, electrophoretic mobility, and zeta potential of HCSLNs was investigated. The effect of HCSLNs on the membrane charge of induced pluripotent stem cells (iPSCs) was also studied. The results indicated that the average diameter of HCSLNs was 90-240nm and the particle size of HCSLNs with EQ 1 was smaller than that with SA. The zeta potential and electrophoresis analysis showed that HCSLNs with SA had a positively charged potential and HCSLNs with EQ 1 had a negatively charged potential at pH7.4. The high-resolution transmission electron microscope confirmed the loading of NGF on the surface of HCSLNs. Differentiation of iPSCs using NGF-loaded HCSLNs with EQ 1 exhibited higher absolute values of the electrophoretic mobility and zeta potential than differentiation using NGF-loaded HCSLNs with SA. The immunochemical staining of neuronal nuclei revealed that NGF-loaded HCSLNs can be used for differentiation of iPSCs into neurons. NGF-loaded HCSLNs with EQ 1 had higher viability of iPSCs than NGF-loaded HCSLNs with SA. NGF-loaded HCSLNs with EQ 1 may be promising formulation to regulate the membrane charge of iPSCs during neuronal differentiation.

Chitosan/γ-poly(glutamic acid) scaffolds with surface-modified albumin, elastin and poly-l-lysine for cartilage tissue engineering

Cartilage has limited ability to self-repair due to the absence of blood vessels and nerves. The application of biomaterial scaffolds using biomimetic extracellular matrix (ECM)-related polymers has become an effective approach to production of engineered cartilage. Chitosan/γ-poly(glutamic acid) (γ-PGA) scaffolds with different mass ratios were prepared using genipin as a cross-linker and a freeze-drying method, and their surfaces were modified with elastin, human serum albumin (HSA) and poly-l-lysine (PLL). The scaffolds were formed through a complex between NH3+ of chitosan and COO- of γ-PGA, confirmed by Fourier transform infrared spectroscopy, and exhibited an interconnected porous morphology in field emission scanning electron microscopy analysis. The prepared chitosan/γ-PGA scaffolds, at a 3:1 ratio, obtained the required porosity (90%), pore size (≥100μm), mechanical strength (compressive strength>4MPa, Youngs modulus>4MPa) and biodegradation (30-60%) for articular cartilage tissue engineering applications. Surface modification of the scaffolds showed positive indications with improved activity toward cell proliferation (deoxyribonucleic acid), cell adhesion and ECM (glycoaminoglycans and type II collagen) secretion of bovine knee chondrocytes compared with unmodified scaffolds. In caspase-3 detection, elastin had a higher inhibitory effect on chondrocyte apoptosis in vitro, followed by HSA, and then PLL. We concluded that utilizing chitosan/γ-PGA scaffolds with surface active biomolecules, including elastin, HSA and PLL, can effectively promote the growth of chondrocytes, secrete ECM and improve the regenerative ability of cartilaginous tissues.

Targeted delivery of rosmarinic acid across the blood–brain barrier for neuronal rescue using polyacrylamide-chitosan-poly(lactide-co-glycolide) nanoparticles with surface cross-reacting material 197 and apolipoprotein E

Rosmarinic acid-loaded polyacrylamide-chitosan-poly(lactide-co-glycolide) nanoparticles (RA-PAAM-CH-PLGA NPs) were grafted with cross-reacting material 197 (CRM197) and apolipoprotein E (ApoE) for targeting of the blood-brain barrier (BBB) and rescuing degenerated neurons. The polymeric nanocarriers were prepared by microemulsion, solvent diffusion, grafting, and surface modification, and CRM197-ApoE-RA-PAAM-CH-PLGA NPs were used to treat human brain-microvascular endothelial cells, RWA264.7 cells, and Aβ-insulted SK-N-MC cells. Experimental results revealed that an increase in the weight percentage of PAAM decreased the particle size, zeta potential, and grafting efficiency of CRM197 and ApoE. In addition, surface DSPE-PEG(2000) could protect CRM197-ApoE-RA-PAAM-CH-PLGA NPs against uptake by RWA264.7 cells. An increase in the concentration of CRM197 and ApoE decreased the transendothelial electrical resistance and increased the ability of propidium iodide and RA to cross the BBB. The order in the viability of apoptotic SK-N-MC cells was CRM197-ApoE-RA-PAAM-CH-PLGA NPs > CRM197-RA-PAAM-CH-PLGA NPs > RA. Thus, CRM197-ApoE-RA-PAAM-CH-PLGA NPs can be a promising formulation to deliver RA to Aβ-insulted neurons in the pharmacotherapy of Alzheimers disease.

Pancreatic differentiation of induced pluripotent stem cells in activin A-grafted gelatin-poly(lactide-co-glycolide) nanoparticle scaffolds with induction of LY294002 and retinoic acid

The differentiation of induced pluripotent stem cells (iPSCs) in biomaterial scaffolds is an emerging area for biomedical applications. This study proposes, for the first time, the production of pancreatic cells from iPSCs in gelatin-poly(lactide-co-glycolide) nanoparticle (PLGA NP) scaffolds. The porosity and swelling ratio of the scaffolds decreased with increases in gelatin and PLGA NP concentrations. The adhesion efficiency of iPSCs in gelatin-PLGA NP scaffolds was found to be higher at 6.7% (w/w) PLGA NP. A 3-step induction of iPSCs was used to differentiate into pancreatic islet cells using activin A, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002), and retinoic acid (RA). The ability of iPSCs to differentiate into pancreatic islet cells in a scaffold was demonstrated by immunofluorescence staining and flow-cytometry analysis. The results indicate that the concentration of activin A, LY294002, and RA plays a decisive role in the differentiation of iPSCs into pancreatic cells. Activin A and LY294002 induce the iPSCs into endoderm and RA induces endoderm into islet cells. A maximum insulin secretion by glucose stimulation was obtained with a higher concentration (2μM) of RA. The use of activin A-grafted gelatin-PLGA NP scaffolds induced by LY294002 and RA can be a promising approach to developing pancreatic islet cells from iPSCs.

Alginate in Bone Tissue Engineering

Regenerative medicine has witnessed a paradigm shift from synthetic implants and tissue grafts to a bone tissue engineering (BTE) approach that incorporates biodegradable bioceramic composite scaffolds with biological cells. Polysaccharides, a biodegradable naturally occurring polymer can be useful either as carriers systems for active biomolecules or as cell carriers. It plays a crucial role to mimic the natural extra cellular matrix (ECM) for the development of 3D scaffold material in BTE and regenerative medicine. Alginate, a natural polysaccharide acts a biodegradable gel when crosslinked with calcium ions. Further, it encapsulates and immobilizes a variety of cells for immunoisolatory and biochemical processing applications. The bone-forming osteoblast cells can easily be attached to the alginate surface and therefore can proliferate well. Hence, alginate is widely utilized in BTE for its bioavailability, biodegradability, and ease of access.