Terapia fotodynamiczna komórek raka szyjki macicy w warunkach in vitro

Abstract Objectives: Photodynamic therapy is one of the modern therapeutic techniques, in which cytotoxic effects are induced by light. Currently, investigators show that it can be used to eliminate cervical cancer cells. Aim: In this study we decided to check whether the Low Level Lasers Therapy (LLLT) can induce the phototoxic changes in HeLa cells, after their photosesitization. Material and methods: The studies were performed in vitro on HeLa cervical cancer cell line. Protoporphyrin IX (PpIX) in final concentrations: 0.5, 1.0, 5.0 and 10.0μmol/l was used as photosensitizer. The cells were preincubated with specific concentrations for 6 and 18 hours. After these defined periods of time the cultures were illuminated for 8 minutes by laser light (635nm and 30mW/cm2). The cytotoxic effects were assessed by a colorimetric test XTT, 24 and 48 hours after irradiation. Results: Significantly augmented cytotoxic changes were found in HeLa cells 18 hours after preincubation and 48 hours after illumination. Moreover, biostimulating laser exposure preceded by preincubation with protoporphyrin IX caused the cytotoxic changes in cervical cancer cells. Conclusion: The obtained results allow us to assume that photodynamic therapy of cervical cancer using biostimulating laser light should be performed 18 hours after the application of protoporphyrin IX.Streszczenie Wstęp: Terapia fotodynamiczna jest jedną z nowych metod leczniczych, w których efekty cytotoksyczne są wywołane przez fotouczulacz po jego aktywacji światłem. Obecnie wskazuje się możliwość zastosowania tej metody w celu eliminacji komórek raka szyjki macicy. Cel badań: Celem niniejszych badań było sprawdzenie czy lasery niskoenergetyczne mogą zostać wykorzystane jako źródło wzbudzenia fotouczulaczy w komórkach raka szyjki macicy. Materał i metody: Badania przeprowadzono w warunkach in vitro na linii komórek raka szyjki macicy HeLa. Jako substancję fotouczulającą zastosowano protoporfirynę IX (PpIX) o końcowych stężeniach: 0,5, 1,0, 5,0 i 10,0mmol/l. Komórki preinkubowano z protoporfiryną IX przez okres 6 i 18 godzin. Po tych czasach zawartą w komórkach protoporfirynę IX aktywowano przez 8 minut światłem laserowym (635nm, 30mW/cm2). Komórki inkubowano dalsze 24 i 48 godzin a zmiany cytotoksyczne oceniano kolorymetrycznym testem XTT. Wyniki: Zaobserwowano, że najwyższe efekty cytotoksyczne w komórkach HeLa można otrzymać podczas 18 godzinnej preinkubacji oraz 48 godzinnej inkubacji po naświetlaniu. Wyniki wskazują na możliwość zastosowania laserów biostymulacyjnych w terapii fotodynamicznej komórek raka szyjki macicy. Wnioski: Uzyskane wyniki pozwalają przypuszczać, że terapię fotodynamiczną raka szyjki macicy przy pomocy laserów niskoenergetycznych należy wykonać w czasie 18 godzin od momentu aplikacji protoporfiryny IX

Nucleases isolated from Chelidonium majus L. milky sap can induce apoptosis in human cervical carcinoma HeLa cells but not in Chinese Hamster Ovary CHO cells.

Milky sap isolated from Chelidonium majus L. (Greater Celandine) serves as a rich source of various biologically active substances such as alkaloids, flavonoids and phenolic acids. Previous research showed that the activity of Ch. majus milky sap may depend also on the presence of biologically active proteins. The goal of this study was to evaluate the biological effect of two nucleases isolated from Ch. majus milk sap, CMN1 of 20 kDa and CMN2 of 36 kDa, on HeLa and CHO tumour cell lines. Both studied nucleases together with other proteins in the sap of the plant are involved in stress and defence reactions against different pathogens. After 48 h incubation of CMN1 and CMN2 only with HeLa cells, the dependence between the number of apoptotic lesions and the concentration of applied nuclease was observed. The highest proapoptotic activity was induced by 13.3 ng/ml concentration of CMN2 collected in May (62 +/- 3% HeLa cells were apoptotic). Moreover, the proportion of necrotic cells in all concentrations of the nucleases and both cell lines was relatively low (1-8 +/- 0.5%). In summary, results of this study show that purified nucleases CMN1 and CMN2 isolated from Ch. majus milky sap exhibit apoptotic activity in HeLa tumour cell line, but not in CHO cells, without inflammatory reaction

Trichrome Mallory's stain may indicate differential rates of RNA synthesis in eutopic and ectopic endometrium.

Mallory's triple staining is a histochemical technique used mainly for analysing connective tissues and glands and other tissues. We have described the differences in the nuclear staining between eutopic and ectopic endometrium as well as endometrial hyperplasia and adenocarcinoma using the Mallory's method. The ultrastructural differences between eutopic and ectopic endometrium have been detected. In normal and hyperplastic endometrium the presence of stromal cell nuclei with an increased affinity to aniline blue has been observed. The affinity has disappeared after digestion of tissues with RNase. In cases of endometriosis, independently of cell types, the nuclei have shown affinity to orange G. Similar effects in adenocarcinoma have been noted. The ultrastructural studies have shown that in normal endometrium the stroma contained cells with euchromatic and low electron density cell nuclei. In endometriosis heterochromatic cell nuclei present both in the stroma and within glands have been detected. The results indicate that the Mallory's technique may be a useful tool for recognizing the differences between eutopic and ectopic endometrium. The affinity for aniline blue in normal and hyperplastic endometrium occurs most likely due to increased RNA synthesis. Based on Mallory's staining a similarity between hyperplasia and unchanged endometrium in contrast to similar results of the staining obtained in cases of adenocarcinoma and endometriosis may be suggested

The 42nd Symposium Chromatographic Methods of Investigating Organic Compounds : Book of abstracts

The 42nd Symposium Chromatographic Methods of Investigating Organic Compounds : Book of abstracts. June 4-7, 2019, Szczyrk, Polan

Biomimetic Multispiked Connecting Ti-Alloy Scaffold Prototype for Entirely-Cementless Resurfacing Arthroplasty Endoprostheses—Exemplary Results of Implantation of the Ca-P Surface-Modified Scaffold Prototypes in Animal Model and Osteoblast Culture Evaluation

We present here—designed, manufactured, and tested by our research team—the Ti-alloy prototype of the multispiked connecting scaffold (MSC-Scaffold) interfacing the components of resurfacing arthroplasty (RA) endoprostheses with bone. The spikes of the MSC-Scaffold prototype mimic the interdigitations of the articular subchondral bone, which is the natural biostructure interfacing the articular cartilage with the periarticular trabecular bone. To enhance the osteoinduction/osteointegration potential of the MSC-Scaffold, the attempts to modify its bone contacting surfaces by the process of electrochemical cathodic deposition of Ca-P was performed with further immersion of the MSC-Scaffold prototypes in SBF in order to transform the amorphous calcium-phosphate coating in hydroxyapatite-like (HA-like) coating. The pilot experimental study of biointegration of unmodified and Ca-P surface-modified MSC-Scaffold prototypes was conducted in an animal model (swine) and in osteoblast cell culture. On the basis of a microscope-histological method the biointegration was proven by the presence of trabeculae in the interspike spaces of the MSC-Scaffold prototype on longitudinal and cross-sections of bone-implant specimens. The percentage of trabeculae in the area between the spikes of specimen containing Ca-P surface modified scaffold prototype observed in microCT reconstructions of the explanted joints was visibly higher than in the case of unmodified MSC-Scaffold prototypes. Significantly higher Alkaline Phosphatase (ALP) activity and the cellular proliferation in the case of Ca-P-modified MSC-Scaffold pre-prototypes, in comparison with unmodified pre-prototypes, was found in osteoblast cell cultures. The obtained results of experimental implantation in an animal model and osteoblast cell culture evaluations of Ca-P surface-modified and non-modified biomimetic MSC-Scaffold prototypes for biomimetic entirely-cementless RA endoprostheses indicate the enhancement of the osteoinduction/osteointegration potential by the Ca-P surface modification of the Ti-alloy MSC-Scaffold prototype. Planned further research on the prototype of this biomimetic MSC-Scaffold for a new generation of RA endoprostheses is also given