Extrusion 3D-printed tricalcium phosphate-polycaprolactone biocomposites for quercetin-KCl delivery in bone tissue engineering.

Critical-sized bone defects pose a significant challenge in advanced healthcare due to limited bone tissue regenerative capacity. The complex interplay of numerous overlapping variables hinders the development of multifunctional biocomposites. Phytochemicals show promise in promoting bone growth, but their dose-dependent nature and physicochemical properties halt clinical use. To develop a comprehensive solution, a 3D-printed (3DP) extrusion-based tricalcium phosphate-polycaprolactone (TCP-PCL) scaffold is augmented with quercetin and potassium chloride (KCl). This composite material demonstrates a compressive strength of 30 MPa showing promising stability for low load-bearing applications. Quercetin release from the scaffold follows a biphasic pattern that persists for up to 28 days, driven via diffusion-mediated kinetics. The incorporation of KCl allows for tunable degradation rates of scaffolds and prevents the initial rapid release. Functionalization of scaffolds facilitates the attachment and proliferation of human fetal osteoblasts (hfOB), resulting in a 2.1-fold increase in cell viability. Treated scaffolds exhibit a 3-fold reduction in osteosarcoma (MG-63) cell viability as compared to untreated substrates. Ruptured cell morphology and decreased mitochondrial membrane potential indicate the antitumorigenic potential. Scaffolds loaded with quercetin and quercetin-KCl (Q-KCl) demonstrate 76% and 89% reduction in bacterial colonies of Staphylococcus aureus, respectively. This study provides valuable insights as a promising strategy for bone tissue engineering (BTE) in orthopedic repair.

3D Printed SiO2-Tricalcium Phosphate Scaffolds Loaded with Carvacrol Nanoparticles for Bone Tissue Engineering Application.

Bone damage resulting from trauma or aging poses challenges in clinical settings that need to be addressed using bone tissue engineering (BTE). Carvacrol (CA) possesses anti-inflammatory, anticancer, and antibacterial properties. Limited solubility and physicochemical stability restrict its biological activity, requiring a stable carrier system for delivery. Here, we investigate the utilization of a three-dimensional printed (3DP) SiO2-doped tricalcium phosphate (TCP) scaffold functionalized with carvacrol-loaded lipid nanoparticles (CA-LNPs) to improve bone health. It exhibits a negative surface charge with an entrapment efficiency of ∼97% and size ∼129 nm with polydispersity index (PDI) and zeta potential values of 0.18 and -16 mV, respectively. CA-LNPs exhibit higher and long-term release over 35 days. The CA-LNP loaded SiO2-doped TCP scaffold demonstrates improved antibacterial properties against Staphylococcus aureus and Pseudomonas aeruginosa by >90% reduction in bacterial growth. Functionalized scaffolds result in 3-fold decrease and 2-fold increase in osteosarcoma and osteoblast cell viability, respectively. These findings highlight the therapeutic potential of the CA-LNP loaded SiO2-doped TCP scaffold for bone defect treatment.

Customized 3D-printed hollow capsular device filled with norfloxacin-loaded micropellets for controlled-release delivery.

Pharmacotherapy has become more focused on the personalized treatment of patients with various diseases. This field of pharmacology and pharmacogenomics focuses on developing drug delivery systems designed to address the unique characteristics of individual patients. Three-dimensional printing technology can be used to fabricate personalized drug delivery systems with desired release properties according to patient needs. Norfloxacin (NOR)-loaded micropellets (MPs) were fabricated and filled inside a stereolithography (SLA) 3D printing technology-mediated hollow capsular device in accordance with a standard size of 09 (8.4 mm length × 2.70 mm diameter). The prepared 3D-printed hollow capsular device filled with pristine NOR and NOR-loaded MPs were characterized in terms of both in vitro and in vivo means. MPs with the particle size distribution of 1540.0 ± 26 µm showed 95.63 ± 2.0% NOR content with pellet-shaped surface morphology. The in vitro release profile showed an initial lag phase of approximately 30 min, followed by the sustained release of NOR from MPs from the 3D-printed hollow capsular device. The pharmacokinetic profile showed prolonged Tmax, AUC, and evidence of good RBA of NOR compared to pure NOR after a single oral administration in the experimental animal model. The overall results confirm the feasibility of SLA-mediated 3D printing technology for preparing customized solid oral unit dosage carriers that can be filled with pure NOR- and NOR-loaded MPs with controlled-release delivery features.

A sensitive UPLC/ESI/MS/MS method for concomitant quantification of active plant constituent combinations in rat plasma after single oral administration.

Ultra-performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC/ESI/MS/MS) for the concomitant quantification of active plant constituents, namely quercetin and piperine, in rat plasma was developed and validated to assess pharmacokinetics after a single oral administration. Liquid-liquid extraction technique with ethyl acetate and n-hexane (1 : 1) was used, and fisetin was added as an internal standard (IS). Effective chromatographic separation of quercetin, piperine and IS was executed on a Waters Acquity BEH C18 column (50.0 mm × 2.1 mm, 1.7 µm) using formic acid both (0.1% w/v) in water (A) and acetonitrile (B) as the mobile phase in gradient mode. For detection purposes, positive electrospray ionization (ESI) mode was used with multiple reaction monitoring (MRM) mode for estimation using [M + H]+ fragment ions m/z 303.04 → 152.9 for quercetin, 286.12 → 201.04 for piperine and 287.01 → 136.93 for IS. The method was linear over the calibration range of 0.1-200 ng mL-1. The lower limit of quantification (LLOQ) of quercetin and piperine was obtained as 0.1 ng mL-1 in rat plasma, along with negligible matrix effect and acceptable stability. Furthermore, the bioanalytical method was successfully implemented to determine the pharmacokinetic profiles of quercetin-and piperine-enriched nanostructured lipid carriers (NLCs) in rat plasma after oral administration. The enhancement in the oral bioavailability of quercetin and piperine was 20.72 and 4.67 fold, respectively, compared to their native pristine dispersions. Future exploration of the concentrations of these active constituents in human plasma and organs is feasible using this sensitive, validated UPLC/ESI/MS/MS method.

3D Printed Housing Devices for Segregated Compartmental Delivery of Oral Fixed-Dose Anti-Tubercular Drugs Adopting Print and Fill Strategy.

World Health Organization (WHO) recommends the use of first-line anti-tuberculosis drugs, that is, rifampicin (RIF) and isoniazid (INH) fixed-dose combination (FDC) therapies in tuberculosis (TB) disease. The absorption of RIF from an FDC incorporates INH, and it is significantly compromised due to its reaction with INH, resulting in a severe loss of RIF under gastric stomach pH condition. Such reduction in the dose of both drugs from FDC formulations has been alleged to be one of the chief obstacles in effective TB treatment. This emphasizes a need to develop suitable cutting-edge advanced bioengineered delivery devices that can attenuate this severe problem to mitigate this chief obstacle. Therefore, we designed, prototyped, and characterized bioengineered 3D printed housing devices in the form of printed tablets adopting print and fill strategy for segregated compartmental delivery of RIF into the intestine (to avoid stomach gastric pH induced chemical degradation as alone and FDC) and INH into the stomach (no degradation observed as alone and FDC in stomach gastric pH conditions) for the desired treatment outcome against TB. Prepared 3D printed housings showed almost zero friability, enough hardness along weight variations <±3.0%. Different thermal and morphological analyses confirmed the insignificant changes in the nature of the polymer as before and after printing. The in vitro release for INH from polyvinyl alcohol mediated 3D printed housings showed almost 100% release within 2.5 h in acidic medium, whereas poly-lactic acid (PLA) mediated 3D printed housings continued to release RIF above 70% in the presence of physiological enzymes in alkaline medium for 432 h. The in vivo bioavailability assessment correlated with in vitro dissolution behavior for INH and RIF, whereas RIF did not release from 3D printed PLA housings in vivo.

Quercetin and piperine enriched nanostructured lipid carriers (NLCs) to improve apoptosis in oral squamous cellular carcinoma (FaDu cells) with improved biodistribution profile.

Oral squamous cellular carcinoma (OSCC) is considered a life-threatening disease with detection in late stages, which forces us to opt for dangerous treatment with a combination of chemotherapy and radiotherapy. Herbal components such as piperine and quercetin are derived from edible sources, proving their anticancer potential against oral cancer cells in vitro. Encapsulation into lipid matrix-mediated nanostructured lipid carriers (NLCs) can make both drugs bio-accessible. NLCs were synthesised using the high shear homogenisation method and characterised for their physicochemical properties, followed by in vitro cellular evaluation in FaDu oral cancer cells. NLCs showed negatively charged particles smaller than 180 nm with a polydispersity index (PDI) of <0.3. Both drugs were found to encapsulate sufficiently, with >85% entrapment efficiency and an improved drug release profile compared to their pristine counterparts. Differential scanning calorimetry (DSC) thermograms showed conversion into an amorphous matrix in lyophilized NLCs, which was supported by X-ray diffraction (XRD) analysis. The cytotoxicity assay showed the IC50 concentration for dual drug-loaded NLCs, which was more effective than the pure drug solution. NLCs were found to be internalised in cells in a short time with an almost 95% co-localization rate. Dual drug-loaded NLCs showed maximum depolarisation of the mitochondrial membrane along with more apoptotic changes. Improved apoptosis was confirmed in NLCs using flow cytometry. The in vivo biodistribution of Coumarin-6 labelled NLCs in rats confirmed their efficient distribution in various parts of the oral cavity through oral administration. Optimised dual drug-loaded NLCs provide a better option for delivering both drugs through a single lipid matrix against oral cancer.

Nanostructured lipid carriers as a strategy for encapsulation of active plant constituents: Formulation and in vitro physicochemical characterizations.

Active plant constituents obtained from edible sources have manifested their pharmacological potential as a therapy against several diseases. But the lack of their desired physicochemical properties such as solubility, permeability ultimately leads to poor bioavailability. Two potent active plant constituents namely, quercetin and piperine having a problem with either solubility or permeability or both, and hence require an advanced lipid-mediated separate formulation system to improve their aforementioned concerns. Concerning advancement in nanoformulations, lipid-based nano-carriers systems have created their mark as a novel drug delivery system. Therefore, an advanced formulation like nanostructured lipid carriers (NLCs) has been formulated individually for both the active plant constituents/drugs through the solvent evaporation technique using high shear homogenization method followed by sonication. Compritol® 888 ATO, a solid lipid, and squalene as liquid lipid was used in their optimized ratios to formulate individual NLCs. Blank and individual drugs loaded NLCs were further characterized for their in vitro physicochemical properties. NLCs showed a negative surface charge with an average particle size below 200 nm. Electron microscopy images showed an anomalous structure of both the formulated NLCs with higher % drug encapsulation efficiency (DEE) with the desired in vitro drug release profile. In the case of quercetin-NLCs, 93.18 ± 5.5 % DEE was observed followed by drug release up to 45.0 ± 1.3 % within 12 h, while piperine-NLCs showed 91.80 ± 2.51 % DEE and drug release up to 38 ± 5.2 % at the same time. XRD and DSC plots showed the conversion of both the drugs into an amorphous structure encapsulated in a lyophilized NLCs matrix. Finally, the safety profile for formulated NLCs was confirmed by haemolysis assay. Hence, the developed active plant constituents enriched NLCs can further be delivered separately and/or in combination, and also may further be evaluated both in vitro and in vivo means.

Extruded filaments derived 3D printed medicated skin patch to mitigate destructive pulmonary tuberculosis: design to delivery.

Background: Quercetin in combination with polyvinylpyrrolidone (PVP) was found to limit the spreading of necrosis to unaffected tissues in tuberculosis-infected mice. Therefore, we hypothesized that 3D printed medicated skin patch incorporated with a quercetin-PVP combination would provide an appropriate therapeutic drug concentration with desired sustained release profile.Research design and methods: We fabricated quercetin-PVP 40 extruded-filaments by hot-melt extrusion (HME) technique along with Eudragit® RSPO and tri-ethyl citrate and further printed it to make medicated skin patches using fused deposition modeling (FDM) based 3D Printing technology. Various characterizations were performed to optimize the 3D-printed patch formulation.Results: Patch formulation has been optimized for several characterization parameters and was further assessed using SEM, DSC, and XRD studies to confirm the conversion of crystalline quercetin into an amorphous form. Finally, the pharmacokinetic profile of an optimized patch was studied in rats showing prolonged Tmax, lowered Cmax, and reduced fluctuations in plasma concentrations till 18 days with single skin application of 3D-printed medicated patch.Conclusion: Overall data confirmed the feasibility of developing 3D printed medicated skin patches to provide plasma levels for continued 18 days in rats after a single application.

Analytical method development and validation of reverse-phase high-performance liquid chromatography (RP-HPLC) method for simultaneous quantifications of quercetin and piperine in dual-drug loaded nanostructured lipid carriers.

Quercetin and piperine are often used as an add-on therapy for various diseases, however both drug exhibits poor aqueous solubility and photosensitivity issue. Therefore, the aim of the present study is to improve the pharmaceutical challenges by incorporating both the drugs in nanostructured lipid carriers (NLCs) and to develop a sensitive, selective, accurate and precise reverse-phase high performance liquid chromatography (RP-HPLC) method for the simultaneous analysis of both drugs in NLCs. Effective chromatographic separation of quercetin and piperine was achieved on Hypersil gold C-18 column and mobile phase consisting of a mixture of acetonitrile and HPLC grade water (pH 2.6, adjusted with 2%v/v glacial acetic acid) in an isocratic elution mode. The flow rate of the mobile phase was 1 mL/min, column temperature at 35 ± 0.2 °C and the injection volume was 20 µL. The retention time for quercetin and piperine were found to be at 2.80 min and 10.36 min, respectively and detected at an isobestic wavelength of 346 nm using a photodiode array (PDA) detector. The method was found to be specific for the simultaneous analysis of quercetin and piperine in presence of NLCs matrix, accurate (>90%) and precise (%RSD < 2%). The validated RP-HPLC method effectively utilised to determine the percentage drug entrapment efficiency cum percentage drug loading of quercetin and piperine in NLCs enriched formulations along with the secondary estimation of in vitro cumulative percentage drug release study. The results were found to be reliable, hence the validated RP-HPLC method could be further used for the simultaneous detection and quantification of both these drugs in other lipid-based nano-formulations such as solid-lipid nanoparticles, polymer-lipid hybrid nanoparticles, lipid drug conjugates, etc. in in vitro and in vivo.