Fibroblast proliferation over dialysis membrane: an experimental model for “tissue” biocompatibility evaluation

The present study reports on a biological model based on fibroblast proliferation applied to 3 different types of flat-plate dialysis membrane, in order to ascertain whether the artificial materials currently used in hemodialysis cause in vitro cellular proliferation. The study plan we followed involved plate membrane isolation from non-used dialyzers and used dialyzers, observed through scanning electron microscopy (SEM) both before and after testing with human fibroblasts by means of cell culture. Fibroblast growth was assessed by phase contrast light microscopy examination and cytometric DNA content evaluation. Our investigations proved that the artificial materials we considered interact with fibroblast cultures. Noticeable proliferative response was observed both after contact with unused material and on mediation by the protein layer absorbed on the membrane surface at the end of dialysis sessions. In this last case fibroblast proliferative activity appeared higher than that observed with unused membranes, showing that the soluble molecules entrapped in the protein layer appeared able to exert a biological activity even in in vitro tests

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Anti-Diabetic Activity of a Mineraloid Isolate, in vitro and in Genetically Diabetic Mice

International Journal for Vitamin and Nutrition Research ◽

10.1024/0300-9831/a000048 ◽

2011 ◽

Vol 81 (1) ◽

pp. 34-42 ◽

Cited By ~ 1

Author(s):

Joel Deneau ◽

Taufeeq Ahmed ◽

Roger Blotsky ◽

Krzysztof Bojanowski

Keyword(s):

Dna Microarrays ◽

Human Fibroblasts ◽

Diabetic Animal ◽

In Vitro Tests ◽

Diabetic Mice ◽

Fossil Material ◽

Expression Of Genes ◽

Enhanced Expression ◽

Genetically Diabetic Mice

Type II diabetes is a metabolic disease mediated through multiple molecular pathways. Here, we report anti-diabetic effect of a standardized isolate from a fossil material - a mineraloid leonardite - in in vitro tests and in genetically diabetic mice. The mineraloid isolate stimulated mitochondrial metabolism in human fibroblasts and this stimulation correlated with enhanced expression of genes coding for mitochondrial proteins such as ATP synthases and ribosomal protein precursors, as measured by DNA microarrays. In the diabetic animal model, consumption of the Totala isolate resulted in decreased weight gain, blood glucose, and glycated hemoglobin. To our best knowledge, this is the first description ever of a fossil material having anti-diabetic activity in pre-clinical models.

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In vitro tests for distinguishing possible immune-mediated aplastic anemia from transfusion-induced sensitization

Blood ◽

10.1182/blood.v55.2.211.211 ◽

1980 ◽

Vol 55 (2) ◽

pp. 211-215 ◽

Cited By ~ 74

Author(s):

BJ Torok-Storb ◽

C Sieff ◽

R Storb ◽

J Adamson ◽

ED Thomas

Keyword(s):

T Cells ◽

T Cell ◽

Aplastic Anemia ◽

Colony Growth ◽

Cell Depletion ◽

In Vitro Tests ◽

T Cell Depletion ◽

Immune Mediated ◽

Before And After

Abstract Forty-two patients with aplastic anemia (AA) were studied to determine whether or not transfusion-induced sensitization is responsible for the in vitro inhibition by patient lymphocytes of HLA-identical erythroid burst-forming units (BFU-E). The results indicate that lymphocytes from 12 of 34 transfused patients inhibited normal colony growth. In contrast, lymphocytes from none of the 8 untransfused patients demonstrated inhibition. These data were interpreted to mean that coculture studies would not be useful for identifying immune-mediated AA in transfused patients. Therefore, in order to identify possible immune-related AA, we assayed BFU-E from patient blood before and after T-cell depletion. In all 32 patients studied, BFU-E failed to grow from peripheral blood cells before T-cell depletion, but in 8 cases, normal- appearing BFU-E grew after T cells had been removed. Growth of patient BFU-E colonies was inhibited in 6 cases when patient T cells were added back to the culture, indicating that in these 6 patients, an “autoimmune” mechanism may have been present.

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Stepwise Glucoheptoamidation of Poly(Amidoamine) Dendrimer G3 to Tune Physicochemical Properties of the Potential Drug Carrier: In Vitro Tests for Cytisine Conjugates

Pharmaceutics ◽

10.3390/pharmaceutics12050473 ◽

2020 ◽

Vol 12 (5) ◽

pp. 473 ◽

Cited By ~ 1

Author(s):

Anna Czerniecka-Kubicka ◽

Piotr Tutka ◽

Marek Pyda ◽

Małgorzata Walczak ◽

Łukasz Uram ◽

...

Keyword(s):

Differential Scanning Calorimetry ◽

Physicochemical Properties ◽

Drug Carrier ◽

Human Fibroblasts ◽

Pamam Dendrimer ◽

In Vitro Tests ◽

Scanning Calorimetry ◽

Ζ Potential ◽

Cell Accumulation

Third-generation poly(amidoamine) dendrimer (PAMAM) was modified by stepwise primary amine group amidation with d-glucoheptono-1,4-lactone. The physicochemical properties of the conjugates—size, ζ potential in lysosomal pH 5 and in neutral aqueous solutions, as well as intramolecular dynamics by differential scanning calorimetry—were determined. Internalization and toxicity of the conjugates against normal human fibroblasts BJ were monitored in vitro in order to select an appropriate carrier for a drug delivery system. It was found that initial glucoheptoamidation (up to 1/3 of amine groups of neat dendrimers available) resulted in increase of conjugate size and ζ potential. Native or low substituted dendrimer conjugates accumulated efficiently in fibroblast cells at nontoxic 1 µM concentration. Further substitution of dendrimer caused consistent decrease of size and ζ potential, cell accumulation, and toxicity. All dendrimers are amorphous at 36.6 °C as determined by differential scanning calorimetry (DSC). The optimized dendrimer, half-filled with glucoheptoamide substituents, was applied as carrier bearing two covalently attached cytisine molecules: a rigid and hydrophobic alkaloid. The conjugate with 2 cytisine and 16 glucoheptoamide substituents showed fast accumulation and no toxicity up to 200 µM concentration. The half-glucoheptoamidated PAMAM dendrimer was selected as a promising anticancer drug carrier for further applications.

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Therapeutic effectiveness of frozen platelet concentrates for transfusion

Blood ◽

10.1182/blood.v57.2.243.243 ◽

1981 ◽

Vol 57 (2) ◽

pp. 243-249 ◽

Cited By ~ 28

Author(s):

HM Lazarus ◽

EA Kaniecki-Green ◽

SE Warm ◽

M Aikawa ◽

RH Herzig

Keyword(s):

Vapor Phase ◽

Platelet Concentrate ◽

In Vitro Tests ◽

Platelet Concentrates ◽

Emergency Situations ◽

Ultrastructural Studies ◽

Significant Difference ◽

Before And After ◽

The Difference

Abstract Six patients received platelet concentrate transfusions from their HLA- identical siblings. Platelet concentrates were administered either fresh, or after being frozen in 10% dimethylsulfoxide, at a slow controlled rate (1 degree C/min) or rapidly (approximately 8 degrees C/min) in the vapor-phase of a liquid nitrogen refrigerator. The median freeze-thaw loss was 13.5%. The mean 1-hr and 20-hr corrected increments in platelet count were calculated for fresh platelet concentrates transfused before and after transfusion with controlled- rate frozen and vapor-phase frozen platelet concentrates. There was no significant difference among the first and second transfusion of fresh platelet concentrates, nor was the difference observed between fresh and controlled-rate frozen platelet concentrates significant. The difference between fresh and vapor-phase frozen platelet concentrates, and between controlled-rate frozen and vapor-phase frozen platelet concentrates were highly significant (p < 0.01). In vitro tests of aggregation using ristocetin and platelet ultrastructural studies paralleled the transfusion experience. Our results indicate that HLA- identical platelet concentrates can be successfully frozen and thawed for transfusion if a slow, controlled rate of freezing is employed. The use of HLA-identical frozen platelet concentrates may be important in emergency situations for the refractory patient and potentially for the establishment of a platelet concentrate bank.

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In vitro tests for distinguishing possible immune-mediated aplastic anemia from transfusion-induced sensitization

Blood ◽

10.1182/blood.v55.2.211.bloodjournal552211 ◽

1980 ◽

Vol 55 (2) ◽

pp. 211-215

Author(s):

BJ Torok-Storb ◽

C Sieff ◽

R Storb ◽

J Adamson ◽

ED Thomas

Keyword(s):

T Cells ◽

T Cell ◽

Aplastic Anemia ◽

Colony Growth ◽

Cell Depletion ◽

In Vitro Tests ◽

T Cell Depletion ◽

Immune Mediated ◽

Before And After

Forty-two patients with aplastic anemia (AA) were studied to determine whether or not transfusion-induced sensitization is responsible for the in vitro inhibition by patient lymphocytes of HLA-identical erythroid burst-forming units (BFU-E). The results indicate that lymphocytes from 12 of 34 transfused patients inhibited normal colony growth. In contrast, lymphocytes from none of the 8 untransfused patients demonstrated inhibition. These data were interpreted to mean that coculture studies would not be useful for identifying immune-mediated AA in transfused patients. Therefore, in order to identify possible immune-related AA, we assayed BFU-E from patient blood before and after T-cell depletion. In all 32 patients studied, BFU-E failed to grow from peripheral blood cells before T-cell depletion, but in 8 cases, normal- appearing BFU-E grew after T cells had been removed. Growth of patient BFU-E colonies was inhibited in 6 cases when patient T cells were added back to the culture, indicating that in these 6 patients, an “autoimmune” mechanism may have been present.

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The effect of Gallium Aluminium Arsenide laser on fibroblast activity: An in vitro dosimetry study

South African Journal of Physiotherapy ◽

10.4102/sajp.v54i1.600 ◽

1998 ◽

Vol 54 (1) ◽

pp. 18-22

Author(s):

Susan Mars ◽

Anil Chuturgoon ◽

Dhamarai Pillay ◽

Maurice Mars

Keyword(s):

Laser Treatment ◽

Human Fibroblasts ◽

Optimal Dose ◽

Fibroblast Proliferation ◽

Continuous Output ◽

Cleavage Assay ◽

Low Intensity Laser Therapy ◽

Gallium Aluminium Arsenide

The effect of different doses of low intensity laser therapy (L.I.L.T.) on human fibroblasts was investigated to determine the optimal dose required to stimulate fibroblast proliferation. Human fibroblasts were cultured in vitro and irradiated with different energy densities of 83Onm continuous output infra-red laser using a Gallium Aluminium Arsenide laser. The fibroblasts were irradiated on three consecutive days at energy densities, ranging from 0.2 to 5 J.cm2, delivered at an average radiant power of 30 mW, and at a constant distance of lcm from the fibroblasts. Fibroblast activity was assessed on the fourth day using a calorimetric MTT (tetrazolium) cleavage assay. There was a significant increase in fibroblast proliferation at laser treatment energy densities of 0.4 J.cm2 and 5 J.cm2. Difficulties associated with in vivo and in vitro studies of the effect of laser treatment are discussed.

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miRNA-148a Enhances the Treatment Response of Patients with Rectal Cancer to Chemoradiation and Promotes Apoptosis by Directly Targeting c-Met

Biomedicines ◽

10.3390/biomedicines9101371 ◽

2021 ◽

Vol 9 (10) ◽

pp. 1371

Author(s):

Chun-Ming Huang ◽

Ming-Yii Huang ◽

Yen-Cheng Chen ◽

Po-Jung Chen ◽

Wei-Chih Su ◽

...

Keyword(s):

Rectal Cancer ◽

Cellular Proliferation ◽

Locally Advanced ◽

Neoadjuvant Chemoradiotherapy ◽

Complete Response ◽

Advanced Rectal Cancer ◽

Locally Advanced Rectal Cancer ◽

Before And After

Patients with locally advanced rectal cancer (LARC) who achieve a pathological complete response (pCR) to neoadjuvant chemoradiotherapy (NACRT) have an excellent prognosis, but only approximately 30% of patients achieve pCR. Therefore, identifying predictors of pCR is imperative. We employed a microRNA (miRNA) microarray to compare the miRNA profiles of patients with LARC who achieved pCR (pCR group, n = 5) with those who did not (non-pCR group, n = 5). The validation set confirmed that miRNA-148a was overexpressed in the pCR group (n = 11) compared with the non-pCR group (n = 40). Cell proliferation and clonogenic assays revealed that miRNA-148a overexpression radio-sensitized cancer cells and inhibited cellular proliferation, before and after irradiation (p < 0.01). Apoptosis assays demonstrated that miRNA-148a enhanced apoptosis before and after irradiation. Reporter assays revealed that c-Met was the direct target gene of miRNA-148a. An in vivo study indicated that miRNA-148a enhanced the irradiation-induced suppression of xenograft tumor growth (p < 0.01). miRNA-148a may be a biomarker of pCR following NACRT and can promote apoptosis and inhibit proliferation in CRC cells by directly targeting c-Met in vitro and enhancing tumor response to irradiation in vivo.

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Assessment of Human Gingival Fibroblast Proliferation After Laser Stimulation In Vitro Using Different Laser Types and Wavelengths (1064, 980, 635, 450 and 405nm) – Preliminary Report

10.20944/preprints202010.0600.v1 ◽

2020 ◽

Author(s):

Barbara Sterczała ◽

Kinga Grzech-Leśniak ◽

Olga Michel ◽

Witold Trzeciakowski ◽

Kamil Jurczyszyn

Keyword(s):

Energy Density ◽

Preliminary Report ◽

Human Fibroblasts ◽

Mitochondrial Activity ◽

Human Gingival Fibroblast ◽

Gingival Fibroblast ◽

Fibroblast Proliferation ◽

Gingival Fibroblasts ◽

Healthy Donors

Purpose: to assess the effect of photobiomodulation (PBM) on human gingival fibroblast proliferation. Methods: The study was conducted using the primary cell cultures of human fibroblasts collected from systemically healthy donors. Three different laser types: Nd:YAG (1064nm), infrared diode laser (980nm) and prototype led laser emitting 405, 450 and 635nm were used to irradiate fibroblasts. Thanks to the patented structure of that laser, it was possible to irradiate fibroblasts with a beam combining two or three wavelengths. The energy density was 3 J/cm², 25 J/cm², 64 J/cm². The viability and proliferation of cells were determined using the MTT test conducted 24, 48 and 72 hours after laser irradiation. Results: The highest percentage of mitochondrial activity (MA=122.1%) was observed in the group irradiated with the 635nm laser, with an energy density of 64 J/cm² after 48 hours. The lowest percentage of MA (94.0%) was observed in the group simultaneously irradiated with three wavelengths (405 + 450 + 635 nm). The use of the 405nm laser at 25 J/cm² gave similar results to the 635 nm laser. Conclusions: The application of the 635nm and 405nm irradiation caused a statistically significant increase in the proliferation of gingival fibroblasts.

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Assessment of Human Gingival Fibroblast Proliferation after Laser Stimulation In Vitro Using Different Laser Types and Wavelengths (1064, 980, 635, 450, and 405 nm)—Preliminary Report

Journal of Personalized Medicine ◽

10.3390/jpm11020098 ◽

2021 ◽

Vol 11 (2) ◽

pp. 98

Author(s):

Barbara Sterczała ◽

Kinga Grzech-Leśniak ◽

Olga Michel ◽

Witold Trzeciakowski ◽

Marzena Dominiak ◽

...

Keyword(s):

Energy Density ◽

Human Fibroblasts ◽

Mitochondrial Activity ◽

Human Gingival Fibroblast ◽

Gingival Fibroblast ◽

Fibroblast Proliferation ◽

Gingival Fibroblasts ◽

Healthy Donors ◽

Thiazolyl Blue Tetrazolium

Purpose: to assess the effect of photobiomodulation (PBM) on human gingival fibroblast proliferation. Methods: The study was conducted using the primary cell cultures of human fibroblasts collected from systemically healthy donors. Three different laser types, Nd:YAG (1064 nm), infrared diode laser (980 nm), and prototype led laser emitting 405, 450, and 635 nm were used to irradiate the fibroblasts. Due to the patented structure of that laser, it was possible to irradiate fibroblasts with a beam combining two or three wavelengths. The energy density was 3 J/cm2, 25 J/cm2, 64 J/cm2. The viability and proliferation of cells were determined using the (Thiazolyl Blue Tetrazolium Blue) (MTT) test conducted 24, 48, and 72 h after laser irradiation. Results: The highest percentage of mitochondrial activity (MA = 122.1%) was observed in the group irradiated with the 635 nm laser, with an energy density of 64 J/cm2 after 48 h. The lowest percentage of MA (94.0%) was observed in the group simultaneously irradiated with three wavelengths (405 + 450 + 635 nm). The use of the 405 nm laser at 25 J/cm2 gave similar results to the 635 nm laser. Conclusions: The application of the 635 nm and 405 nm irradiation caused a statistically significant increase in the proliferation of gingival fibroblasts.

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Therapeutic effectiveness of frozen platelet concentrates for transfusion

Blood ◽

10.1182/blood.v57.2.243.bloodjournal572243 ◽

1981 ◽

Vol 57 (2) ◽

pp. 243-249

Author(s):

HM Lazarus ◽

EA Kaniecki-Green ◽

SE Warm ◽

M Aikawa ◽

RH Herzig

Keyword(s):

Vapor Phase ◽

Platelet Concentrate ◽

In Vitro Tests ◽

Platelet Concentrates ◽

Emergency Situations ◽

Ultrastructural Studies ◽

Significant Difference ◽

Before And After ◽

The Difference

Six patients received platelet concentrate transfusions from their HLA- identical siblings. Platelet concentrates were administered either fresh, or after being frozen in 10% dimethylsulfoxide, at a slow controlled rate (1 degree C/min) or rapidly (approximately 8 degrees C/min) in the vapor-phase of a liquid nitrogen refrigerator. The median freeze-thaw loss was 13.5%. The mean 1-hr and 20-hr corrected increments in platelet count were calculated for fresh platelet concentrates transfused before and after transfusion with controlled- rate frozen and vapor-phase frozen platelet concentrates. There was no significant difference among the first and second transfusion of fresh platelet concentrates, nor was the difference observed between fresh and controlled-rate frozen platelet concentrates significant. The difference between fresh and vapor-phase frozen platelet concentrates, and between controlled-rate frozen and vapor-phase frozen platelet concentrates were highly significant (p < 0.01). In vitro tests of aggregation using ristocetin and platelet ultrastructural studies paralleled the transfusion experience. Our results indicate that HLA- identical platelet concentrates can be successfully frozen and thawed for transfusion if a slow, controlled rate of freezing is employed. The use of HLA-identical frozen platelet concentrates may be important in emergency situations for the refractory patient and potentially for the establishment of a platelet concentrate bank.

Download Full-text