Supercritical Carbon Dioxide Extraction Research Articles

In vitro growth‐inhibitory effect of essential oils and supercritical carbon dioxide extracts from Cinnamomum spp. barks and fruits against food bacterial pathogens in liquid and vapor phase

AbstractFoodborne microbial diseases constitute major health concern and therefore it is necessary to preserve food products to prolong their shelf life and safety. Promising method of food preservation is application of volatile natural agents into modified atmosphere in food packaging. Spices and condiments containing volatile oils could potentially be used. Antimicrobial effect of three essential oils (EOs) and three supercritical carbon dioxide extracts (CO2 extracts) from Cinnamomum barks and fruits was evaluated using broth‐microdilution volatilization method against common foodborne pathogens, namely Bacillus cereus, Escherichia coli, Listeria monocytogenes, and Salmonella enterica Typhimurium. All samples exhibited significant inhibitory effect with minimum inhibitory concentration (MIC) values ranging from 256 to 1,024 μg/ml in both liquid and vapor phase. EO from immature fruits of Cinnamomum cassia, known as cassia buds, was the most effective agent against all tested microorganisms with MIC values ranging from 256 to 512 μg/ml in both phases. Using gas chromatography–mass spectrometry analysis, E‐cinnamaldehyde has been detected as the most abundant constituent in all samples. Subsequently, the time series headspace analysis of vapors above the mixture of growth medium and C. cassia fruits EO was performed. Decreasing content of E‐cinnamaldehyde in the headspace was gradually replaced by δ‐cadinene and γ‐muurolene. Proposed mechanism behind this is ability of E‐cinnamaldehyde to pass from the headspace back to the liquid medium. Obtained results suggest that EOs and CO2 extracts from Cinnamomum spp. could be used in food industry as natural food preservatives. In addition, broth‐microdilution volatilization method has been validated for development of food applications.

Edible oil extracted from anchovies using supercritical CO 2 : Availability of fat‐soluble vitamins and comparison with commercial oils

In this study, oil was extracted from anchovies using supercritical carbon dioxide (SC-CO2). Additionally, a comparative experiment was conducted with organic solvent-extracted anchovy oil and commercial fish oil. Omega-3 fatty acid contents were found to be 32.10 ± 0.45 to 32.15 ± 0.38% for anchovy fish oil. The results of the acid value and the peroxide value, determinants of oil stability, were 11.06 ± 0.01 mg KOH/g, and 28.92 ± 0.19 meq/kg, respectively, using SC-CO2 for extraction. In the case of fat-soluble vitamins of anchovy oil extracted by SC-CO2, vitamins A, D, and E and coenzyme Q10 were found at high percentages 22.51 ± 0.28 µg/100 g, 4.32 ± 0.10 mg/100 g, 104.67 ± 4.75 and 2.04 ± 0.20 mg/100 g, respectively, which had a higher content of these vitamins compared with that of the hexane extraction. Therefore, SC-CO2 extracted anchovy oil is expected to be used as a functional material, which could lead to economic benefits through the high valorization of anchovies. Practical applications Anchovy is a low graded fishery raw materials can be turned as a very important source of polyunsaturated fatty acids (PUFA) rich oil containing with different fat-soluble vitamins like A, D, and E, and coenzyme Q10. Supercritical carbon dioxide extraction (SC-CO2) is one of the safe and environment-friendly lipid extraction system that can be used for extracting the PUFA enriched oil from anchovy. SC-CO2 could be an efficient valorization method of anchovy that could be an important source of human-grade food oil and contribute to the food and nutritional sector.

Fast discrimination of avocado oil for different extracted methods using headspace-gas chromatography-ion mobility spectroscopy with PCA based on volatile organic compounds

Abstract To establish a method for fast discrimination of avocado oil for different extracted methods, the headspace-gas chromatography-ion mobility spectroscopy (HS-GC-IMS) combined with principal component analysis (PCA) was used to analyze non-target volatile organic compounds (VOCs). The results showed that 40 VOCs were identified, and the VOCs of the extraction method had a significant difference and had been well distinguished in PCA. The species and content of avocado oil obtained by squeeze method were more than the aqueous methods and supercritical carbon dioxide extraction methods (SC CO2). In addition, the different avocado oil had their characteristic compounds: the 2-acetylthiazole and ethyl propionate were the unique compounds in the avocado oil obtained by SC CO2. A rapid method for the determination of avocado oil obtained by different extraction methods based on HS-GC-IMS had been established, and the method was fast and simple and had a good application prospect in the prediction of avocado oil processing methods.

Old Plant, New Possibilities: Wild Bilberry (Vaccinium myrtillus L., Ericaceae) in Topical Skin Preparation.

Bilberry represents a valuable source of antioxidant substances responsible for its application for the treatment of different conditions (such as inflammation, cardiovascular disease, cancer, diabetes, and different age-related diseases) associated with increased oxidative stress. As oxidative stress might cause skin impairments, we aim to evaluate a topical preparation containing bilberry leaves extract and bilberry seeds oil, obtained as a byproduct of the food industry. To obtain the extracts, the conventional maceration technique for leaves, and supercritical carbon dioxide extraction for seeds were employed. The chemical profile of both actives was achieved by HPLC and GC methods, revealing the presence of phenolic acids (chlorogenic being the most abundant), flavonoids (isoquercetin in the highest amount), and resveratrol in leaves extract, while in seeds oil the essential ω-3 and ω-6 fatty acids were determined in favorable ratio, almost being 1. Antioxidant potential of the wild bilberry extract and seed oil was evaluated using in vitro DPPH and FRAP assays. Finally, effects of the oil-in-water creams with mentioned wild bilberry isolates on the skin were investigated in an in vivo study conducted on healthy human volunteers, revealing the significant beneficial effects when topically applied.

Development of a decellularized porcine bone graft by supercritical carbon dioxide extraction technology for bone regeneration.

A series of novel decellularized porcine collagen bone graft (DPB) materials in a variety of shapes and sizes were developed by the supercritical carbon dioxide (SCCO2 ) extraction technique. The complete decellularization of DPB was confirmed by hematoxylin and eosin staining, 4,6-diamidino-2-phenylindole (DAPI) staining, and residual DNA analysis. The native intact collagen remained in the DPB after the SCCO2 process was confirmed by Masson trichrome staining. The physicochemical characteristics of DPB were investigated by scanning electron microscopy and x-ray diffraction. The cytotoxicity and biocompatibility tests according to ISO10993 and its efficacy for bone regeneration in osteochondral defects in rabbits were evaluated. The rabbit pyrogen test confirmed DPB was non-toxic. In vitro and in vivo biocompatibility tests of the DPB did not show any toxic or mutagenic effects. The bone regeneration potential of the DPB presented no significant histological differences compared to commercially available deproteinized bovine bone. In conclusion, DPB produced by SCCO2 exhibited similar chemical characteristics to human bone, no toxicity, good biocompatibility, and enhanced bone regeneration in rabbits comparable to that of deproteinized bovine bone. Results from this study could shed light on the potential application of the SCCO2 extraction technique to generate a native decellularized scaffold for bone tissue regeneration in human clinical trials.

Chemical Composition, Antioxidant and Cytoprotective Potentials of Carica papaya Leaf Extracts: A Comparison of Supercritical Fluid and Conventional Extraction Methods.

The leaves of Carica papaya (CP) are rich in natural antioxidants. Carica papaya has traditionally been used to treat various ailments, including skin diseases. This study aims to decipher the antioxidant effects and phytochemical content of different CP leaf extracts (CPEs) obtained using supercritical carbon dioxide (scCO2) and conventional extraction methods. The antioxidant activities of CPEs were evaluated by cell-free (1,1-diphenyl-2-picryl-hydrazyl (DPPH) and ferric-reduced antioxidative power (FRAP)) and cell-based (H2O2) assay. Both C. papaya leaf scCO2 extract with 5% ethanol (CPSCE) and C. papaya leaf scCO2 extract (CPSC) exhibited stronger DPPH radical scavenging activity than conventional extracts. In the FRAP assay, two hydrophilic extracts (C. papaya leaf ethanol extract (CPEE) and C. papaya freeze-dried leaf juice (CPFD)) showed relatively stronger reducing power compared to lipophilic extracts. Cell-based assays showed that CPFD significantly protected skin fibroblasts from H2O2-induced oxidative stress in both pre-and post-treatment. CPEE protected skin fibroblasts from oxidative stress in a dose-dependent manner while CPSCE significantly triggered the fibroblast recovery after treatment with H2O2. GC-MS analysis indicated that CPSCE had the highest α-tocopherol and squalene contents. By contrast, both CP hydrophilic extracts (CPEE and CPFD) had a higher total phenolic content (TPC) and rutin content than the lipophilic extracts. Overall, CPEs extracted using green and conventional extraction methods showed antioxidative potential in both cell-based and cell-free assays due to their lipophilic and hydrophilic antioxidants, respectively.

Composition and Antioxidant Activity, Supercritical Carbon Dioxide Extraction Extracts, and Residue after Extraction of Biologically Active Compounds from Freeze-Dried Tomato Matrix

Supercritical carbon dioxide extraction (SCE-CO2) is an attractive, green technology that is used for the recovery of biologically active compounds from plant material. The antioxidant potential of lipophilic fractions (extract obtained with SCE-CO2) and hydrophilic fractions (extracts obtained from the residue after extraction) obtained from a matrix of freeze-dried tomatoes (cvs. “Admiro” F1, “Jurgiai”, “Vilina”, “Pirmutis”, and “Skariai”) was assessed via different antioxidant activity methods. The total amount of polyphenols, carotenoids, and carotenoid isomers before and after SCE-CO2 extraction was also determined. To investigate the effect of the SCE-CO2 extract on the viability of cancer cells, rat glioblastoma C6 cells were chosen. The SCE-CO2 yielded an average of 800 mg of lipophilic fraction per 100 g of freeze-dried tomatoes. The ABTS•+ scavenging activity of the extract was 251 ± 3.4 µmol TE/g. After SCE-CO2 extraction, the DPPH•-RSA of the freeze-dried tomato matrix was 7 to 12% higher. There was a strong positive correlation (R = 0.84) between the total polyphenolics content and the DPPH•-RSA of the tomato samples. The SCE-CO2 increased the radical scavenging activity of the extraction residue, indicating that a considerable fraction of the hydrophilic compounds with particular antioxidant capacity remain unextracted from the tomato matrix. Our results reveal the cytotoxic effect of lycopene extract rich in cis-isomers (62% cis-isomers of the total lycopene content) on rat glioblastoma C6 cells. The viability of the glioblastoma C6 cells significantly decreased (−42%) at a total lycopene concentration of 2.4 µM after 24 h of incubation.

Supercritical carbon dioxide extraction of pomegranate peel-seed mixture: Yield and modelling

By-products produced during the processing of pomegranate (Punica granatum L.) juice are currently undervalued and discarded, hence, this study investigated the effects of temperature (30–60 °C) and pressure (10–40 MPa) on the supercritical carbon dioxide (scCO2) extraction of a mixture of pomegranate peels and seeds (1:1) at a fixed CO2 flow rate of 10 L/min (0.1 MPa, 25 °C). The global yield increased with temperature but decreased with pressure, with the highest global yield (11.5%) obtained at 40 MPa and 40 °C. The reaction constants (0.044–0.066 min−1) and activation energy (6.06 kJ/mol) were determined with the first-order model. The Pardo-Castaño model I correlated with the experimental data with an average absolute relative deviation (AARD) of 6.1%, with the Chrastil model correlating with the solubility data with an AARD of 2.5%. The synergistic effect of PME was demonstrated with regard to yield as it had a higher extract to seed ratio.

\u03b2-Carotene extraction from Dunaliella salina by supercritical CO2

This paper reports the results of supercritical carbon dioxide (scCO2) extraction of β-carotene from Dunaliella salina as potential alternative to conventional organic solvent extraction. In pilot-scale scCO2 experiments, the pressure, temperature, and co-solvent concentration were varied. The supercritical extraction at 500 bar, 70 °C, and 10 wt% ethanol as co-solvent yielded in the highly efficient pigment recovery of over 90%. Techno-economic assessment demonstrated higher energy consumption for the scCO2 extraction that was compensated by lower solvent costs. Thus, comparable pigment production costs to the reference extraction with n-hexane were estimated for the scCO2 process. Due to the green solvent properties of scCO2 and ethanol, this approach is highly promising for extraction of algal biomass in industrial scale.

Supercritical carbon dioxide extraction of oil and minor lipid compounds of cake byproduct from Brazil nut (Bertholletia excelsa) beverage production

Supercritical carbon dioxide extraction (SC-CO2) was performed to recover the lipid fraction of the cake by-product from Brazil nut beverage (BNM) production. Extraction kinetics and the effect of pressure and temperature on the yield, oil recovery, fatty acid (FA) profile and minor lipid compounds were evaluated. The modeling of the kinetic assays indicated that most of the oil is readily accessible and the extraction velocity is dominated by the oil solubility. High yield with c.a. 100% oil recovery was achieved at 400 bar and 60 °C. While the FA profile of extracts did not present relevant variations with the extraction conditions, higher content of squalene and β-sitosterol (590.8 and 283.9 mg/g extract) were obtained at the lowest pressure and highest temperature assessed (200 bar and 60 °C). Therefore, SC-CO2 extraction is a suitable technique to recover the oil and other minor lipid bioactive compounds of the cake by-product from BNM production, with a potential application in foods, cosmetics or nutraceuticals.

Supercritical extraction of biomass as an effective pretreatment step for the char yield control in pyrolysis

This work demonstrates that the coupling of supercritical carbon dioxide extraction with pyrolysis is an effective method for the removal of extractives from forestry residues and generation of solid char with different properties from the remaining solid wood fractions. Extraction of the needles and stumps shows greater yields of resin acids, terpenes, steroids and other derivatives than that of pinewood bark, cones and branches. The char yields of both non-treated and scCO2 extracted wood fractions varied from approximately 17.5 to 38.5 wt. % on dry basis at fast heating rates. The catalytic effect of extractives is significant on the yields and morphology of solid chars in fast pyrolysis and less pronounced at slow heating rates. These results are promising as they show that both the composition and location of extractives inclusions in the interior of wood particle can affect the morphology of char samples. Moreover, the impact of alkali metals on the wood devolatilization appears to be less compared to the lignocellulosic composition in slow pyrolysis. These results demonstrate that supercritical carbon dioxide extraction can be integrated in biorefinery as a pretreatment step to control the properties of pyrolysis products by varying the heating rate.

Solubility of catechin and epicatechin from Arachis Hypogea skins wastes by using supercritical carbon dioxide-ethanol and its optimization

This research aims to optimize a supercritical carbon dioxide extraction assisted by co-solvent ethanol for the recovery of catechin and epicatechin from peanut (Arachis Hypogea) skin. The second objective was to determine the solubility of catechin and epicatechin by semi-empirical models. The operating conditions were: pressure of 10 to 30 MPa; temperature of 313 to 343 K; and ethanol rate of 0.075 to 0.225 mL/min. Response surface methodology (RSM) was applied to determine the optimum condition to recover the catechin and epicatechin. The optimum condition was found to be 21.86 MPa, 332.23 K, and 0.17 mL/min with 752.03 µg/g of catechin and 3399.84 µg/g of epicatechin. Putra model successfully fitted the solubility of catechin and epicatechin in range of 1.21 E-5 g/L and 6.02 E-5 g/L with 3.11% of AARD (%). Application of RSM was succesful to optimize the conditions with high quantity of catechin and epicatechin.

Process optimization for the supercritical carbon dioxide extraction of Foeniculum vulgare Mill. seeds aromatic extract with respect to yield and trans‐anethole contents using Box‐Behnken design

AbstractFennel (seeds of Foeniculum vulgare Mill.) is the widely used aromatic and medicinal plant throughout the world. In this research, supercritical carbon dioxide (SC‐CO2) extraction was carried out to extract the aromatic extract from F vulgare seeds under various operating conditions, and aromatic extract samples were characterized for properties such as yield and trans‐anethole content. A three‐level Box‐Behnken design from response surface method was applied to optimize the extraction parameters such as pressure, temperature, and extraction time with an aim to obtain high yield and rich trans‐anethole content. The optimum extraction conditions were found to be 177 bar, 40°C and 68 min where the predicted oil yield and trans‐anethole contents were found to be 3.798% and 95 mg/g, respectively. Under the optimum conditions, the experimental oil yield was 3.85%, and the content of trans‐anethole was 94 mg/g. The results were very close to the values predicted by the model. At the same time, the chemical compositions of the oils from two different extraction methods, hydrodistillation and SC‐CO2, were compared. The chemical constituents of oils from two different extraction methods found to be the same, and they showed slight difference in terms of the quantity of the main chemicals. Antibacterial and antioxidant effects of the oils obtained by the two extraction methods were compared, and they showed similar effects.

Supercritical carbon dioxide extraction of plant phytochemicals for biological and environmental applications – A review

Recently, supercritical fluid CO2 extraction (SFE) has emerged as a promising and pervasive technology over conventional extraction techniques for various applications, especially for bioactive compounds extraction and environmental pollutants removal. In this context, temperature and pressure regulate the solvent density and thereby effects the yield, selectivity, and biological/therapeutic properties of the extracted components. However, the nature of plant matrices primarily determines the extraction mechanism based on either density or vapor pressure. The present review aims to cover the recent research and developments of SFE technique in the extraction of bioactive plant phytochemicals with high antioxidant, antibacterial, antimalarial, and anti-inflammatory activities, influencing parameters, process conditions, the investigations for improving the yield and selectivity. In another portion of this review focuses on the ecotoxicology and toxic metal recovery applications. Nonpolar properties of Sc-CO2 create strong solvent strength via distinct intermolecular interaction forces with micro-pollutants and toxic metal complexes. This results in efficient removal of these contaminants and makes SFE technology as a superior alternative for conventional solvent-based treatment methods. Moreover, a compelling assessment on the therapeutic, functional, and solvent properties of SFE is rarely focused, and hence this review would add significant value to the SFE based research studies. Furthermore, we mention the limitations and potential of future perspectives related to SFE applications.

A continuous flow process for biaryls based on sequential Suzuki–Miyaura coupling and supercritical carbon dioxide extraction

A continuous flow process for the production of biaryls based on the seamless coupling of a packed-bed reactor (synthesis module) and a rapid supercritical CO2 extraction system (extraction module) is reported.

AN OVERVIEW OF OIL BASED DRILL CUTTINGS WASTE ENVIRONMENTAL EFFECT AND DISPOSAL TREATMENTS

The drilling fluid is essential for drilling operations in the oil and gas industry. It is a complex mixture of organic and inorganic materials blended together to achieve desired multi-functions. There are several types of drilling fluid commonly in use, oil-based drilling fluid considered the most hazardous and harmful for the surrounding environment because of contaminated waste produced, normally consisting of diesel, oil, asphalt, polymers, crude oil and surfactants. Produced drill cuttings are contaminated by hydrocarbons, and other chemical additives. The produced drill cuttings are hazardous for environment as they contain polycyclic aromatic hydrocarbons and undesirable heavy metals. Presently, various oil and gas operator companies in Yemen and other parts of the world dispose the drilling wastes in excavated pits or landfill sites. It is not acceptable to deal with such hazardous substances in this manner. Other operator companies use thermal desorption technique as a separation method. This method still has many disadvantages. Therefore, it is vital to search for sufficient and efficient technologies and processes to treat oil-based drilling wastes as the existing methods in use cannot be considered as a viable solution to eliminate the environmental damaged caused by contaminated drill cuttings. This paper review three existing experimental methods of extraction that have significant total organic carbon removal efficiency and highlighted factors that effect on the extraction efficiency. They are; supercritical water oxidation, superheated steam extraction, and supercritical carbon dioxide extraction. The stated experimental studies -have shown promising results and efficiency in extraction of carbon content and are considered environmentally friendly.

Advance on the preparation, physiological function and nanocarriers of resveratrol

Resveratrol is a natural compound that has many beneficial physiological functions. In recent years, the research on resveratrol has gradually become a hotspot. This article summarized the literature reports all over the world, classified and concluded the extraction technology, function and nano-delivery carriers of resveratrol. Solvent extraction, microwave-and ultrasonic-assisted extraction, supercritical carbon dioxide extraction and enzymatic hydrolysis method were respectively discussed in detail. Then the antioxidant property, antibacterial activity, antiumor effect and the prevention of cardiovascular disease of resveratrol were also discussed. Finally, the resveratrol-loaded nanocarriers, such as nanoparticles, nanoemulsions, solid lipid particles and nanosuspensions were summarized. And the future research direction of resveratrol were prospected, in order to provide references and orientations for the further development and research of resveratrol.

Fucoxanthin, A Xanthophyll from Macro- and Microalgae: Extraction Techniques, Bioactivities and Their Potential Application in Nutra- and Cosmeceutical Industries

Some works of literature reported that fucoxanthin has diverse potential benefits for human health. Thus, this review would explain the sources of fucoxanthin, extraction techniques, bioactivities, and its potential application in Nutra- and cosmeceutical industries. Brown algae, such asPadina australis,Undaria pinnatifida; and the microalgae, such asChaetoceros gracilis,Phaeodactylum tricornutumwere sources of fucoxanthin. Then, the chemical structure of this xanthophyll is unique and that confers its biological activities. And then, the extraction process of fucoxanthin from macro- and microalgae is more safe, accessible, and economic, although this xanthophyll can be synthesized chemically. Generally, there are two techniques for the extraction of fucoxanthin, namely liquid solvent (conventional), and supercritical carbon dioxide (non-conventional) extractions. Furthermore, there are some bioactivities of fucoxanthin, including its activities of anticancer, antidiabetic, antiobesity, antioxidant; protective effects of skin, bone, and eyes. Based onin vivoassay of the animal, it has no adverse effects of fucoxanthin supplementation. Therefore, this xanthophyll might be applied in both the Nutra- and cosmeceutical industries. In the future, fucoxanthin and its derivatives would be important for human health, contributing to the beauty industry, and playing an important in the prevention of cancer and the disease related to lifestyle.

Extract of Ganoderma sinensis spores induces cell cycle arrest of hepatoma cell via endoplasmic reticulum stress

Context Ganoderma sinensis Zhao, Xu et Zhang (Ganodermataceae) has been used for the prevention or treatment of a variety of diseases, including cancer. Objective We investigated the antitumor activity and mechanism of an extract from G. sinensis against hepatocellular carcinoma. Materials and methods A G. sinensis extract (GSE) was obtained from sporoderm-broken G. sinensis spores by supercritical fluid carbon dioxide extraction. Hepatoma cells, HepG2 cells, were treated with emulsified sample of GSE at 12.5, 25, 50, 100 and 150 μg/mL for 24 h. The Alamar Blue assay was used to examine growth inhibitory effects. Changes in cell structure and morphology were assessed via transmission electron microscopy and confocal laser scanning microscope. Cell cycle distribution was analysed by flow cytometry. Results GSE suppressed the proliferation of HepG2 cells (IC50=70.14 μg/mL). Extensive cytoplasmic vacuolation originating from dilation of the endoplasmic reticulum (ER) was shown in GSE-treated HepG2 cells. GSE treatment also upregulated the expression of ER stress-related proteins in HepG2 cells. Cells tended to be arrested at the G2/M cell cycle stage after GSE treatment (30.8 ± 1.4% and 42.2 ± 2.6% at GSE with 50 μg/mL and 100 μg/mL vs. 21.03 ± 1.10%, control). Pre-treatment with salubrinal, an inhibitor of ER stress, effectively attenuated cell cycle arrest induced by GSE. Discussion and conclusions Our findings provide new evidence that GSE suppresses growth of cancer cells in vitro through activating the ER stress pathway. The GSE may be clinically applied in the prevention and/or treatment of cancer.

A novel 3D histotypic cartilage construct engineered by supercritical carbon dioxide decellularized porcine nasal cartilage graft and chondrocytes exhibited chondrogenic capability in vitro

Augmentative and reconstructive rhinoplasty surgical procedures use autologous tissue grafts or synthetic grafts to repair the nasal defect and aesthetic reconstruction. Donor site trauma and morbidity are common in autologous grafts. The desperate need for the production of grafted 3D cartilage tissues as rhinoplasty grafts without the adverse effect is the need of the hour. In the present study, we developed a bioactive 3D histotypic construct engineered with the various ratio of adipose-derived stem cells (ADSC) and chondrocytes together with decellularized porcine nasal cartilage graft (dPNCG). We decellularized porcine nasal cartilage using supercritical carbon dioxide (SCCO2) extraction technology. dPNCG was characterized by H&E, DAPI, alcian blue staining, scanning electron microscopy and residual DNA content, which demonstrated complete decellularization. 3D histotypic constructs were engineered using dPNCG, rat ADSC and chondrocytes with different percentage of cells and cultured for 21 days. dPNCG together with 100% chondrocytes produced a solid mass of 3D histotypic cartilage with significant production of glycosaminoglycans. H&E and alcian blue staining showed an intact mass, with cartilage granules bound to one another by extracellular matrix and proteoglycan, to form a 3D structure. Besides, the expression of chondrogenic markers, type II collagen, aggrecan and SOX-9 were elevated indicating chondrocytes cultured on dPNCG substrate facilitates the synthesis of type II collagen along with extracellular matrix to produce 3D histotypic cartilage. To conclude, dPNCG is an excellent substrate scaffold that might offer a suitable environment for chondrocytes to produce 3D histotypic cartilage. This engineered 3D construct might serve as a promising future candidate for cartilage tissue engineering in rhinoplasty.