Preparation and in vitro characterization of histidine trimethyl chitosan conjugated nanocomplex incorporated into injectable thermosensitive hydrogels for localized gene delivery

Abstract

Localized and sustained delivery of DNA using biomaterial scaffolds would increase the applicability of gene therapy in cancer treatment and tissue engineering. The most promising approach is the application of hydrogel scaffolds to encapsulate and deliver DNA in the form of nanocomplex to the target sites. In the present work, histidine conjugated trimethylated chitosan (HTMC) was synthesized and nanocomplexes fabricated with pDNA at different N/P ratios. The zeta potential and size of the HTMC/pDNA nanoparticles were determined using dynamic light scattering technique and the results were confirmed by scanning electron microscopy (SEM). The morphology of the nanoparticles was found spherical in shape having core-shell nanostructure. Based on gel retardation assay, polyplex dissociation following incubation with heparin, protection against DNase Ι, cytotoxicity and transfection experiments, HTMC/pDNA at N/P 15 was selected as an optimized formulation and loaded into CTS/PF127 and HA/PF127 hydrogel. The optimized HTMC/pDNA nanocomplex was homogenously distributed in the CTS/PF127 hydrogel. Transfection experiments were carried out on HEK293T cell lines and the results revealed that HTMC/pDNA complexes are stable and active inside the hydrogel, however, the transfection efficacy was decreased after incorporation into the hydrogel.

A novel histidine functionalized chitosan nanoformulation: Synthesis, characterization and its bioactivity in tomato plant

“…These nanoformulations have been used to enhance plant immunity against plant diseases and growth for getting an adequate yield. However, limited literature is available where chitosan histidine nanoparticles have been developed without TPP for metal ion absorption [9][10][11][12], drug delivery [13], gene delivery [14][15][16], cancer therapy [17] , and wound healing [18]. To the best of current knowledge, chitosan histidine nanoformulations have not undergone evaluation for their impact on physiological and biochemical responses in plants.…”

“…Furthermore, the versatility of these formulations extends to drug delivery applications [13]. Additionally, research has explored their utility in gene delivery [14][15][16]. Moreover, the chitosan-based formulations have shown promise in cancer therapy [17], and in the realm of wound healing [18].…”

Chitosan: A Potential Biopolymer in Drug Delivery and Biomedical Applications

“…On the other hand, the nanoparticles were uniformly distributed in the chitosan/PF127 system. The platform facilitated the delivery of DNA to HEK293T cells, but a lower transfection efficiency was observed (compared to just nanoparticles), possibly owing to the additional barrier brought by the hydrogel for the diffusion of nanoparticles before entering the targeted cells [240].…”

Intelligent DNA-Based Hydrogels for Bioanalysis and Therapeutics

“…DNA-based hydrogels not only utilize the skeleton structure of the hydrogel but also retain the biological function of DNA, realizing a perfect fusion of the structure and function of the hydrogel material. It has broad application prospects in the fields of biosensors, molecular diagnosis, drug and gene delivery, and disease treatment …”

“…11 DNA-based hydrogels not only utilize the skeleton structure of the hydrogel but also retain the biological function of DNA, realizing a perfect fusion of the structure and function of the hydrogel material. It has broad application prospects in the fields of biosensors, 12 molecular diagnosis, 13 drug and gene delivery, 14 and disease treatment. 15 According to the type of hydrogel cross-linking and components, DNA-based hydrogels can be divided into pure DNA-based hydrogels and hybrid DNA-based hydrogels.…”