Identification of RORγ as a favorable biomarker for colon cancer

ObjectiveTo evaluate the expression of retinoid-related orphan receptor gamma (RORγ) and its potential role in the prognosis of colon cancer.MethodsThe Cancer Genome Atlas and {"type":"entrez-geo","attrs":{"text":"GSE117606","term_id":"117606"}}GSE117606 were used to evaluate to RORγ levels in colon cancer, and real-time quantitative polymerase chain reaction was applied for validation. UALCAN and MEXPRESS were used to analyze the associations of RORγ expression with clinical parameters. The survival analysis was conducted in GEPIA.ResultsRORγ expression was significantly lower in colon tumors than in adjacent normal mucosa tissues. RORγ expression was significantly associated with tumor stage, lymph node metastasis, and liver metastasis. The area under the curve for diagnosis was 0.71. Decreased RORγ expression was positively correlated with the incidence of lymphatic invasion, microsatellite instability, the presence of residual tumor, venous invasion, and copy number variation. Overall survival was longer in patients with higher RORγ expression, especially those with microsatellite instability-high features. Methylation analysis revealed that hypermethylation of the RORγ promoter was associated with the colon cancer stage.ConclusionsRORγ downregulation could be a potential biomarker for colon cancer, especially for predicting prognosis. Decreased RORγ expression in colon tumor may be associated with promoter hypermethylation.     

Differentiation of ex vivo human breast tissue using polarization-sensitive optical coherence tomography

Successful treatment of breast cancer typically requires surgical removal of the tumor. Optical coherence tomography (OCT) has been previously developed for real-time imaging of the surgical margin. However, it can be difficult to distinguish between normal stromal tissue and cancer tissue based on scattering intensity and structure alone. Polarization-sensitive optical coherence tomography (PS-OCT) is sensitive to form birefringence of biological tissue. We report on the development of a high-speed PS-OCT system and imaging of ex vivo human breast tissue, showing enhanced contrast between healthy and cancerous tissues based upon collagen content confirmed with corresponding histology. These results demonstrate the feasibility of using PS-OCT to supplement structural OCT as a possible method for intraoperative tumor margin evaluation.OCIS codes: (110.4500) Optical coherence tomography, (170.4500) Optical coherence tomography, (170.3880) Medical and biological imaging, (260.1440) Birefringence, (110.5405) Polarimetric imaging, (170.6935) Tissue characterization     

Gene transfer to adult human lung tissue ex vivo

The potential of gene therapy for treatment of lung disease remains unrealised. Early model systems often resulted in promising efficiency of gene transfer, only to prove irreproducible in the clinic. While problems such as induction of host immune responses and duration of expression also need to be addressed, it is now widely believed that alternative, relevant models which more accurately reflect gene transfer efficiencies in human lungs are urgently required. We report here on a human lung slice culture system to assess gene transfer to adult lung epithelium. A lacZ-expressing adenovirus (AdCA35lacZ) was used as a reporter vector. A solution of AdCA35lacZ was instilled via bronchioles into resected lung tissue, a route analogous to clinical administration. Following a 1 h incubation, the tissue was inflated with a 0.4% agarose solution, instilled via the same bronchioles. Once solidified, 500 microm slices of the tissue were prepared and cultured for 4 days. beta-Galactosidase staining revealed lacZ transgene expression in bronchiolar and alveolar cells of the lung slices throughout the 4 days in culture. This system, which can also be used to study other viral and liposome vectors, could prove to be a useful alternative model for assessing gene delivery to adult human lung epithelium.     

Epidural needle with embedded optical fibers for spectroscopic differentiation of tissue: ex vivo feasibility study

Epidural injection is commonly used to provide intraoperative anesthesia, postoperative and obstetric analgesia, and to treat acute radicular pain. Identification of the epidural space is typically carried out using the loss of resistance (LOR) technique, but the usefulness of this technique is limited by false LOR and the inability to reliably detect intravascular or subarachnoid needle placement. In this study, we present a novel epidural needle that allows for the acquisition of optical reflectance spectra from tissue close to the beveled surface. This needle has optical fibers embedded in the cannula that deliver and receive light. With two spectrometers, light received from tissue is resolved across the wavelength range of 500 to 1600 nm. To determine the feasibility of optical tissue differentiation, spectra were acquired from porcine tissues during a post mortem laminectomy. The spectra were processed with an algorithm that derives estimates of the hemoglobin and lipid concentrations. The results of this study suggest that the optical epidural needle has the potential to improve the accuracy of epidural space identification.     

Human tissue color as viewed in high dynamic range optical spectral transmission measurements

High dynamic range optical-to-near-infrared transmission measurements for different parts of human body in the spectral range from 650 to 950 nm have been performed. Experimentally measured spectra are correlated with Monte Carlo simulations using chromaticity coordinates in CIE 1976 L*a*b* color space. Both a qualitative and a quantitative agreement have been found, paving a new way of characterizing human tissues in vivo. The newly developed experimental and computational platform for assessing tissue transmission spectra is anticipated to have a considerable impact on identifying favorable conditions for laser surgery and optical diagnostics, while providing supplementary information about tissue properties.OCIS codes: (170.6510) Spectroscopy, tissue diagnostics; (170.3660) Light propagation in tissues; (170.6935) Tissue characterization; (200.4960) Parallel processing; (300.6550) Spectroscopy, visible; (330.1730) Colorimetry     

Surgical tissue handling methods to optimize ex vivo fluorescence with the activatable optical probe γ‐glutamyl hydroxymethyl rhodamine green

Optical fluorescence imaging has been developed as an aid to intraoperative diagnosis to improve surgical and endoscopic procedures. Compared with other intraoperative imaging methods, it is lower in cost, has a high safety margin, is portable and easy to use. γ‐glutamyl hydroxymethyl rhodamine green (gGlu‐HMRG) is a recently developed activatable fluorescence probe that emits strong fluorescence in the presence of the enzyme γ‐glutamyl transpeptidase (GGT), which is overexpressed in many cancers, including ovarian cancer. Ex vivo testing is important for clinical approval of such probes. The diagnostic performance of gGlu‐HMRG in fresh excised surgical specimens has been reported; however, details of tissue handling have not been optimized. In this study, we investigated four different tissue handling procedures to optimize imaging in excised tumor specimens. The fluorescence intensity time courses after the different tissue handling methods were compared. Additionally, the fluorescence positive areas were correlated with the presence of red fluorescent protein (RFP) in an RFP positive cell line as the standard of reference for cancer location. In the ‘intact’ groups, tumors yielded quick and homogeneous activation of gGlu‐HMRG. In the ‘rinse’ and ‘cut’ groups, the fluorescence intensity of the tumor was a little lower than that in the intact group. In the ‘pressed’ groups, however, fluorescence intensity from gGlu‐HMRG was lower over the entire time course, suggesting a decrease or relocation of excreted GGT. In conclusion, we demonstrate that the method of tissue handling prior to ex vivo imaging with the activatable probe gGlu‐HMRG has a strong influence on the signal derived from the specimen. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.     

Feasibility of using multiphoton excited tissue autofluorescence for in vivo human histopathology

Rapid and direct imaging of microscopic tissue morphology and pathology can be achieved by multiphoton imaging of intrinsic tissue fluorophores and second harmonic signals. Engineering parameters for developing this technology for clinical applications include excitation levels and collection efficiencies required to obtain diagnostic quality images from different tissue types and whether these levels are mutagenic. Here we provide data on typical average powers required for high signal-to-noise in vivo tissue imaging and assess the risk potential of these irradiance levels using a mammalian cell gene mutation assay. Exposure times of ~16 milliseconds per cell to 760 nm, ~200 fs raster-scanned laser irradiation delivered through a 0.75 NA objective produced negligible mutagenicity at powers up to about 50 mW.     

Potential diagnostic value of the hematological parameters lymphocyte–monocyte ratio and hemoglobin–platelet ratio for detecting colon cancer

ObjectiveTo evaluate the efficacy of using the lymphocyte–monocyte ratio (LMR), hemoglobin–platelet ratio (HPR), and carcinoembryonic antigen (CEA) levels alone or in combination for diagnosing colon cancer.MethodsWe assessed 124 consecutive patients who were pathologically diagnosed with colon cancer and 131 patients who were diagnosed with benign colon tumors in this retrospective study. We then analyzed correlations between LMR, HPR, and clinicopathological findings. The diagnostic values of LMR, HPR, and CEA alone or in combination in colon cancer patients were evaluated by receiver operating characteristic curves.ResultsThe median LMR, HPR, and CEA values in colon cancer patients showed significant correlation with the depth of tumor invasion, lymph node metastasis, and TNM stage. Moreover, there was a significant difference in HPR between patients with tumor size ≥5 cm and those with tumor size <5 cm. Compared with LMR, HPR, or CEA alone, combinations of CEA with LMR, CEA with HPR, and HPR with LMR all had higher area under the curve values, among which the combination of all three (LMR, HPR, and CEA) had the highest area under the curve.ConclusionThe combination of LMR, HPR, and CEA may be a valuable indicator for monitoring colon cancer.     

Targeted delivery of AAV‐transduced mesenchymal stromal cells to hepatic tissue for ex vivo gene therapy

Adeno‐associated virus (AAV)‐mediated gene therapy holds great promise if challenges related to vector neutralization by pre‐existing antibodies are circumvented. The use of autologous or allogeneic cells to shield the vector might offer the possibility of successful gene transfer in such a situation. In the present study, we evaluated the feasibility of AAV‐transduced mesenchymal stromal cells (MSCs) as a vehicle for hepatic gene transfer in a murine liver injury model. In our initial studies to determine the most suitable vector, we observed that AAV1 (91%) and AAV6 (72%) serotypes are highly efficient in transducing MSCs. Subsequently, we generated a transient liver injury model to analyse the efficacy of MSCs homing to the liver, as well as their hepatic gene transfer efficiency; our data show that administration of acetaminophen (500 mg/kg) served as a cue for the homing of MSCs to the liver. Furthermore, sex‐mismatched transplantation of AAV1‐infected MSCs demonstrated a 3.5‐fold (day 7) and 2.2‐fold (day 28) higher hepatic gene transfer efficiency. To further corroborate this, we estimated the donor cell Y chromosome copies in the liver of recipient female mice. Our data revealed a 12.7‐fold increase in average genome copies of male MSCs in the livers of recipient mice with injury compared to control, 60 days after transplantation. However, in vivo administration of AAV‐transduced MSCs in the presence of neutralization antibodies (intravenous immunoglobulin, IVIG) was not beneficial. This is possibly due to the clearance of transplanted MSCs by circulating IVIG and underscores the need to develop suitable in vivo models to study such a mode of gene transfer. Copyright © 2015 John Wiley & Sons, Ltd.     

Evaluating a semi‐nested PCR to support histopathology reports of fungal rhinosinusitis in formalin‐fixed paraffin‐embedded tissue samples

BackgroundFungal rhinosinusitis (FRS) encompasses a various spectrum of diseases. Histopathology is the “reference method” for diagnosing FRS, but it cannot determine the genus and species. Moreover, in more than 50% of the histopathologically proven cases, the culture elicited no reliable results. This study was an attempt to evaluate the diagnostic efficiency of semi‐nested polymerase chain reaction (PCR) from formalin‐fixed paraffin‐embedded (FFPE) functional endoscopic sinus surgery (FESS) in FRS patients.MethodsOne hundred ten specimens were subjected to DNA extraction and histopathology examination. The amplification of the β‐globin gene by conventional PCR was used to confirm the quality of extracted DNA. The semi‐nested PCR was performed using ITS1, ITS2, and ITS4 primers during two steps. Sequencing the internal transcribed spacer region (ITS1‐5.8S‐ITS2) to identify causative agents was performed on PCR products.ResultsSixty‐four out of 110 samples were positive by histopathology evidence, of which 56 samples (87.5%) were positive by PCR. Out of 46 negative samples by histopathological methods, five samples (10.9%) yielded positive results by PCR. Sensitivity, specificity, positive predictive value, and negative predictive value of the semi‐nested PCR method were reported 87.5%, 89.2%, 92.7%, and 85.2%, respectively. The kappa factor between PCR and histopathological methods was 0.76, indicating substantial agreements between these two tests.ConclusionDue to the acceptable sensitivity and specificity of the present method, it might be used to diagnose fungal sinusitis infections along with microscopic techniques. This method is recommended to confirm the diagnose of suspected fungal sinusitis with negative histopathology results.     

Wide-field imaging of fluorescent deoxy-glucose in ex vivo malignant and normal breast tissue

Rapid in situ determination of surgical resection margins during breast cancer surgery would reduce patient time under anesthesia. We present preliminary data supporting the use of a fluorescent glucose analog (2-NBDG) as an optical contrast agent to differentiate freshly excised breast tissue containing cancerous cells from normal breast tissue. Multi-spectral images of 14 breast cancer specimens acquired before and after incubation with 2-NBDG demonstrated increased fluorescent signal in all of the malignant tissue due to increased 2-NBDG consumption. We demonstrate that 2-NBDG has potential as an optical contrast agent to differentiate cancerous from non-cancerous tissue.     

Local aldosterone synthesis in the large intestine of mouse: An ex vivo incubation study

ObjectiveTo investigate the regulation of local aldosterone synthesis by physiological stimulants in the murine gut.MethodsMale mice were fed for 14 days with normal, high (1.6%) or low (0.01%) sodium diets. Tissue liver receptor homolog-1 and aldosterone in the colon and caecum were detected using an enzyme-linked immunosorbent assay (ELISA). Released corticosterone and aldosterone in tissue incubation experiments after stimulation with angiotensin II (Ang II) and dibutyryl-cAMP (DBA; the second messenger of adrenocorticotropic hormone) were assayed using an ELISA. Tissue aldosterone synthase (CYP11B2) protein levels were measured using an ELISA and Western blots.ResultsIn incubated colon tissues, aldosterone synthase levels were increased by a low-sodium diet; and by Ang II and DBA in the normal diet group. Release of aldosterone into the incubation buffer was increased from the colon by a low-sodium diet and decreased by a high-sodium diet in parallel with changes in aldosterone synthase levels. In mice fed a normal diet, colon incubation with both Ang II and DBA increased the release of aldosterone as well as its precursor corticosterone.ConclusionLocal aldosterone synthesis in the large intestine is stimulated by a low-sodium diet, dibutyryl-cAMP and Ang II similar to the adrenal glands.     

C18 hydroxy fatty acids as markers of lipid peroxidation ex vivo and in vivo

Different C18 monohydroxy fatty acids (OHFAs) were evaluated for their usefulness as markers of plasma lipid peroxidation (unsaturated fatty acid oxidation) ex vivo and in vivo. First, plasma samples (n=5) were exposed for 3?h to different radical fluxes ex vivo. The formation of OHFAs was assessed by using varying concentrations of Cu²⁺ions and AAPH (2,2′‐azobis(2‐amidinopropane) hydrochloride) as radical flux initiators. Secondly, a cross‐sectional study was carried out in 47 middle‐aged men. In this study, plasma concentrations of different in vivo OHFAs were compared with other indices of lipid peroxidation. Under mild oxidation conditions (heparin plasma containing 4.2 or 8.3?mM AAPH), concentrations of all the measured OHFAs (8, 9, 10, 11, 12, 13, 15 and 16‐OH acids) increased in an identical manner, but under highly oxidative conditions (heparin plasma containing 83?mM AAPH or 4.2 to 8.3?mM CuSO₄) mainly 9 and 13‐OHFAs were formed. In the cross‐sectional study, plasma 11 and 13‐OHFA levels were associated statistically significantly with plasma free F_2α‐isoprostanes, recognized index of in vivo lipid peroxidation (r=0.305, p=0.037 and r=0.308, p=0.035, respectively). In addition, 16‐OHFA levels correlated with the ratio of electronegatively charged LDL to total LDL (r=0.335, p=0.021). With respect to the other OHFAs, 15‐OHFA had no correlation with either other OHFAs or the reference substances used. In addition, occasionally there were contamination problems in the assessment of 12‐OHFA. It is concluded that all of the measured C18 OHFAs can be used as indicators of plasma lipid peroxidation under mild oxidation conditions, though the 12 and 15‐OHFAs may need to be used with some caution. Under high oxidation conditions, 9‐and 13‐OHFAs seem to be the most useful indices because of their high formation capacity.     

C18 hydroxy fatty acids as markers of lipid peroxidation ex vivo and in vivo

Different C18 monohydroxy fatty acids (OHFAs) were evaluated for their usefulness as markers of plasma lipid peroxidation (unsaturated fatty acid oxidation) ex vivo and in vivo. First, plasma samples (n = 5) were exposed for 3 h to different radical fluxes ex vivo. The formation of OHFAs was assessed by using varying concentrations of Cu2+ ions and AAPH (2,2'-azobis(2-amidinopropane) hydrochloride) as radical flux initiators. Secondly, a cross-sectional study was carried out in 47 middle-aged men. In this study, plasma concentrations of different in vivo OHFAs were compared with other indices of lipid peroxidation. Under mild oxidation conditions (heparin plasma containing 4.2 or 8.3 mM AAPH), concentrations of all the measured OHFAs (8, 9, 10, 11, 12, 13, 15 and 16-OH acids) increased in an identical manner, but under highly oxidative conditions (heparin plasma containing 83 mM AAPH or 4.2 to 8.3 mM CuSO4) mainly 9 and 13-OHFAs were formed. In the cross-sectional study, plasma 11 and 13-OHFA levels were associated statistically significantly with plasma free F2alpha-isoprostanes, recognized index of in vivo lipid peroxidation (r = 0.305, p = 0.037 and r = 0.308, p = 0.035, respectively). In addition, 16-OHFA levels correlated with the ratio of electronegatively charged LDL to total LDL (r = 0.335, p = 0.021). With respect to the other OHFAs, 15-OHFA had no correlation with either other OHFAs or the reference substances used. In addition, occasionally there were contamination problems in the assessment of 12-OHFA. It is concluded that all of the measured C18 OHFAs can be used as indicators of plasma lipid peroxidation under mild oxidation conditions, though the 12 and 15-OHFAs may need to be used with some caution. Under high oxidation conditions, 9-and 13-OHFAs seem to be the most useful indices because of their high formation capacity.     

INHBA promotes the proliferation,migration and invasion of colon cancer cells through the upregulation of VCAN

ObjectiveColon cancer has high morbidity and mortality rates, and proliferation, invasion and migration play an important role in colon cancer progression. Here, the effects of inhibin subunit beta A (INHBA) on cell proliferation, invasion and migration were investigated.MethodsThe UALCAN database was used to assess INHBA expression in colon cancer tissues and predict the survival of patients with high and low INHBA expression. The relevant proteins were detected by RT-qPCR and western blot. Cell transfection was performed to overexpress or inhibit INHBA and versican (VCAN). The high correlation between INHBA and VCAN found through LinkedOmics and StarBase databases was verified by immunoprecipitation assays. Cell proliferation was detected by cell counting kit-8 and colony formation assays. Wound healing and Transwell assays were used to assess migration and invasion.ResultsINHBA expression was upregulated in colon cancer tissues and cells. INHBA inhibition impaired the proliferation, migration and invasion of these cells. In addition, we confirmed the correlation between INHBA and VCAN in colon cancer cells. Finally, we found that INHBA interference inhibited the aggressive behavior of colon cancer cells by downregulating VCAN.ConclusionINHBA promotes the proliferation, migration and invasion of colon cancer cells through the upregulation of VCAN.     

Effects of tissue heterogeneity on the optical estimate of breast density

Breast density is a recognized strong and independent risk factor for developing breast cancer. At present, breast density is assessed based on the radiological appearance of breast tissue, thus relying on the use of ionizing radiation. We have previously obtained encouraging preliminary results with our portable instrument for time domain optical mammography performed at 7 wavelengths (635–1060 nm). In that case, information was averaged over four images (cranio-caudal and oblique views of both breasts) available for each subject. In the present work, we tested the effectiveness of just one or few point measurements, to investigate if tissue heterogeneity significantly affects the correlation between optically derived parameters and mammographic density. Data show that parameters estimated through a single optical measurement correlate strongly with mammographic density estimated by using BIRADS categories. A central position is optimal for the measurement, but its exact location is not critical.OCIS codes: (170.6510) Spectroscopy, tissue diagnostics; (170.6935) Tissue characterization; (170.3830) Mammography; (170.5280) Photon migration     

Linking ground-based to satellite-derived phenological metrics in support of habitat assessment

Changes in the timing of plant phenology are important indicators of inter-annual climatic variations and are a critical driver of food availability and habitat use for a range of species. A number of remote sensing techniques have recently been developed to observe vegetation cycles throughout the year, including the use of inexpensive visible spectrum digital cameras at the stand level and the use of high temporal frequency Advanced Very High Resolution Radiometer National Oceanic and Atmospheric Administration (AVHRR NOAA) and MODerate resolution Imaging Spectroradiometer (MODIS) imagery at a satellite scale. A fundamental challenge with using satellite data to track plant phenology, however, is the trade-off between the level of spatial detail and the revisit time provided by the sensor, and the ability to verify the interpretation of phenological activity. One way to address this challenge is to integrate remotely sensed observations obtained at different spatial and temporal scales to provide information that contains both high temporal density and fine spatial resolution observations. In this article, we compare measures of vegetation phenology observed from a network of ground-based cameras with satellite-derived measures of greenness derived from a fused broad (MODIS) and fine spatial (Landsat) scale satellite data set. We derive and compare three key indicators of phenological activity including the start date of green-up, start date of senescence and length of growing season from both a ground-based camera network and 30 m spatial resolution synthetic Landsat scenes. Results indicate that although field-based estimates, generally, predicted an earlier start and end of the vegetation season than the fused satellite observations, highly significant relationships were found for the prediction of the start (R ²?=?0.65), end (R ²?=?0.72) and length (R ²?=?0.70) of the growing season across all sites. We conclude that some predictable bias exists however unlike visual field measures of the collected data represent both a spectral and a visual archive for later use.     

Prostate cancer detection using combined auto-fluorescence and light reflectance spectroscopy: ex vivo study of human prostates

This study was conducted to evaluate the capability of detecting prostate cancer (PCa) using auto-fluorescence lifetime spectroscopy (AFLS) and light reflectance spectroscopy (LRS). AFLS used excitation at 447 nm with four emission wavelengths (532, 562, 632, and 684 nm), where their lifetimes and weights were analyzed using a double exponent model. LRS was measured between 500 and 840 nm and analyzed by a quantitative model to determine hemoglobin concentrations and light scattering. Both AFLS and LRS were taken on n = 724 distinct locations from both prostate capsular (n_c = 185) and parenchymal (n_p = 539) tissues, including PCa tissue, benign peripheral zone tissue and benign prostatic hyperplasia (BPH), of fresh ex vivo radical prostatectomy specimens from 37 patients with high volume, intermediate-to-high-grade PCa (Gleason score, GS ≥7). AFLS and LRS parameters from parenchymal tissues were analyzed for statistical testing and classification. A feature selection algorithm based on multinomial logistic regression was implemented to identify critical parameters in order to classify high-grade PCa tissue. The regression model was in turn used to classify PCa tissue at the individual aggressive level of GS = 7,8,9. Receiver operating characteristic curves were generated and used to determine classification accuracy for each tissue type. We show that our dual-modal technique resulted in accuracies of 87.9%, 90.1%, and 85.1% for PCa classification at GS = 7, 8, 9 within parenchymal tissues, and up to 91.1%, 91.9%, and 94.3% if capsular tissues were included for detection. Possible biochemical and physiological mechanisms causing signal differences in AFLS and LRS between PCa and benign tissues were also discussed.OCIS codes: (170.1610) Clinical applications, (170.3650) Lifetime-based sensing, (170.6935) Tissue characterization, (170.4580) Optical diagnostics for medicine, (170.6510) Spectroscopy, tissue diagnostics     

The response of gingiva monolayer,spheroid, and ex vivo tissue cultures to collagen membranes and bone substitute

Collagen membranes and bone substitute are popular biomaterials in guided tissue regeneration for treatment of traumatized or diseased periodontal tissue. Development of these biomaterials starts in monolayer cell culture, failing to reflect in vivo tissue organization. Spheroid cultures potentially mimic in vivo tissues in structure and functionality. This study aims to compare gingiva cell (GC) monolayers and spheroids to ex vivo gingiva. Human GC monolayers, spheroids and gingiva ex vivo tissues were cultured on plastic surfaces, collagen membranes or bone substitute. Hematoxylin–eosin (HE) staining, immunohistochemistry for KI67 and caspase 3 (CASP3), resazurin‐based toxicity assays, quantitative polymerase chain reaction for collagen I (COL1A1), vascular endothelial growth factor (VEGF), angiogenin (ANG), interleukin (IL)6 and IL8 and ELISA for COL1A1, VEGF, ANG, IL6 and IL8 were performed in all cultures. Morphology was different in all culture set‐ups. Staining of KI67 was positive in monolayers and staining of CASP3 was positive in spheroids. All culture set‐ups were viable. COL1A1 production was modulated in monolayers and ex vivo tissues at mRNA levels, VEGF in monolayers and ex vivo tissues at mRNA levels and in spheroids at protein levels, ANG in spheroids at mRNA levels and in monolayers and spheroids at protein levels, IL6 in monolayers and spheroids at mRNA levels and in spheroids and ex vivo tissues at protein levels and IL8 in monolayers and ex vivo tissues at mRNA levels. Modulations were surface‐dependent. In conclusion, each culture model is structurally and functionally different. Neither GC monolayers nor spheroids mimicked gingiva ex vivo tissue in all measured aspects.