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1

Surface functionalization of poly(L-lactide-co-glycolide) membranes with amphiphilic poly(2-oxazoline) for guided tissue regeneration and treatment of bone tissue defects

Tryba A. M., Krok-Borkowicz M., Paluszkiewicz C., Pamuła E.

Engineering of Biomaterials

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2018

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Vol. 21, no. 147

16--20

EN

The main challenge of this research was to functionalize the surface of poly(L-lactide-co-glycolide) (PLGA) membranes with amphiphilic poly(2-oxazoline) (POx) in order to change PLGA chemical state and properties. Poly(2-oxazolines) are very powerful polymers, which thanks to active pendant groups can be easily functionalized with biologically active molecules or peptides. The membranes were prepared by dissolving PLGA, POx, and poly(ethylene glycol) (PEG, 1000 Da) in methylene chloride (DCM), followed by PEG leaching. POx molecules were preferentially adsorbed at the interface PLGA-POx-PEG thanks to affinity to both hydrophilic (PEG) and hydrophobic (PLGA) chains. The properties of the membranes were characterized with Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and wettability tests. Cytocompatibility of the materials in contact with osteoblast-like MG-63 cells was studied by evaluation of cell viability (Alamar-Blue test), live/dead and phalloidin/DAPI staining. The results show that the presence of POx influenced topography of the PLGA membranes, but did not have an impact on their wettability. All membranes were fo-und cytocompatible with model osteoblasts. Presence of POx resulted in better cell adhesion as shown by microscopic studies after fluorescence staining for nuclei and cytoskeleton actin filaments. In summary, one-step phase separation process between PLGA, PEG, and POx, dissolved in DCM followed by drying and PEG leaching resulted in cytocompatible PLGA membranes with immobilised POx, which might be considered for guided tissue regeneration technique in periodontology and in bone tissue engineering.

2

Badania biopolimerów technikami spektroskopii oscylacyjnej

Paluszkiewicz C.

Przetwórstwo Tworzyw

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2011

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[R.] 17, nr 6

493-497

PL

W prezentowanej pracy pokazano możliwości zastosowania metod spektroskopii oscylacyjnej tj. fourierowskiej spektroskopii w środkowej podczerwieni (FTIR) oraz fourierowskiej spektroskopii Ramana (FT-Raman) do identyfikacji i modyfikacji biopolimerów i ich kompozytów. Analizowano biopolimery takie jak - poli(2-de-oksy-2-aminoglukoza) popularnie nazywana chitozanem (CS) oraz polikaprolakton (PCL), które stanowiły matryce do otrzymywania nanokompozytów. Jako modyfikatory stosowano: glinokrzemian warstwowy montmorylonit (K5) oraz hydroksyapatyt (HAp). Nanokompozyty na bazie biopolimerów otrzymano metodą odlewania. Badaniom metodami spektroskopii oscylacyjnej poddano zarówno materiały wyjściowe jak i produkty reakcji. Wykazano, że na podstawie przeprowadzanych analiz technikami spektroskopii oscylacyjnej można uzyskać informacje uśrednione z całego polimeru oraz z jego powierzchni. Zastosowane metody spektroskopii oscylacyjnej pozwoliły również na ocenę biozgodności i degradacji badanych materiałów.

EN

The paper presents the possibility of application of vibrational spectroscopy methods, such as Fourier Transform Infrared Spectroscopy (FTIR) and Fourier Transform Raman Spectroscopy (FT Raman) to identify and determine modification of biopolymers and their composites. Biopolymers, namely chitozan and polycaprolacton were used as matrices in the preparation of nanocomposites. Monmorylonite (K5) and hydroxyapatite (HAp) were applied as nanoparticles. The nanocomposite materials were obtained by casting method. In this study biopolymer matrices as well as modifiers and modification products were analyzed. It is shown that application of spectroscopic techniques allows to receive information from the surface and the bulk of the materials. Application of these methods permits us to characterize dispersion of nanoparticles in a polymer matrix as well as biocompatibility and degradation of the analyzed materials.

3

Diagenetic signals from ancient human remains - bioarchaeological applications

Szostek K., Stepańczak B., Szczepanek A., Kępa M., Głąb H., Jarosz P., Włodarczak P., Tunia K., Pawlyta J., Paluszkiewicz C., Tylko G.

Mineralogia

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2011

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Vol. 42, iss. 2/3

93--112

EN

This preliminary study examines the potential effects of diagenetic processes on the oxygen-isotope ratios of bone and tooth phosphate (δ18O) from skeletal material of individuals representing the Corded Ware Culture (2500–2400 BC) discovered in Malżyce (Southern Poland). Intra-individual variability of Ca/P, CI, C/P, collagen content (%) and oxygen isotopes was observed through analysis of enamel, dentin and postcranial bones. Using a variety of analytical techniques, it was found that, despite the lack of differences in soil acidity, not all the parts of a skeleton on a given site had been equally exposed to diagenetic post mortem changes. In a few cases, qualitative changes in the FTIR spectrum of analysed bones were observed. The data suggest that apart from quantitative analyses, i.e., the calculation of Ca/P, CI, C/P and collagen content, qualitative analyses such as examination of the absorbance line are recommended. The degree to which a sample is, contaminated on the basis of any additional, non-biogenic peaks, deemed to be contaminated should also be specified.

4

Vibrational spectroscopy investigation of montmorillonite-chitosane nanocomposite materials

Paluszkiewicz C., Stodolak E.

Engineering of Biomaterials

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2011

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Vol. 14, no. 109-111 spec. iss.

44

EN

Biomaterials basing on natural polysaccharides, i.e. hiauronic acid, alginate, chitosane are an alternative for already applied bioresorbable synthetic materials basing on synthetic polyhydroxy acids. Their main advantages are good accessibility, low cost, easy forming and high biocompatibility. Additionally , they are a perfect matrix for bioactive nanoparticles i.e. hydroxy apatite (HAp), tricalcium phosphate (TCP) and silica (SiO2). The work presents results of research on nanocomposite consisting of chitosane matrix (CS) modified with a nanofiller, which was natural montmorillonite (MMT). Nanocomposite foils were produced by the casting method. In order to induce better biocompatibility, the surface of the CS/MMT composite was neutralized (bath in NaOH solution). The nanocomposite foils were subjected to a bioactivity test by incubation in SBF at 37oC for 7 days. It was observed that the CS/ MMT material surface showed a local supersaturation, which was a result of apatite nucleation. The CS/ MMT nanocomposites were investigated using FT-IR ( Fourier Transform Infrared Spectroscopy) and Fourier Raman Spectroscopy. FTIR measurements o f the samples were carried out on the transmission and reflection modes. The FTIR microscopy spectra were collected using BioRad Excalibur with ATR attachment as well as microscope UMA500 equipped with MCT detector. Spectra were measure at 4 cm -1 resolution in the region from 4 000 cm -1 to 600 cm -1 . FT-Raman spectra were obtained using a FTS 6000 Bio-Rad spectrometer with Ge detector. The samples were excited with a Nd-YAG laser (1064nm). Additional all materials in all steps experiments were observed under Scanning Electron Microscopy (Nova Nano SEM). Vibrational spectroscopy methods (FT Raman and FTIR) can be used for investigation of nanocomposite foils basing on biopolymers. High sensitivity the applied spectroscopy techniques show that in the result of the neutralization of CS/ MMT foil (via incubation in NaOH solution the biopolymer chain breaks. This phenomena is visible by intensity ratio between COC/COH bands. Increase of reactivity of chitosane chain lead to entrapment of PO4+3-, which is the origin of the apatite forms nucleation process. Chemical treatment of the nanocomposite foils, i.e. NaOH washes in fluences their chemical structure and microstructure. Neutralisation of the foils is the first processing stage which precedes the potential use o fCS/ MMT foils in biomedical applications. The materials show a tendency to apatite crystallisation which may support regeneration of damaged bone tissue. The applied spectroscopic methods allowed to observe changes in the whole volume of the sample. Individual ATR measurements taken at various spectral ranges and penetration depths allow to observe subtle changes in the polymer matrix caused by chemical treatment (NaOH and SBF incubation). Results of the investigations indicate that in the CS/ MMT systems new chemical bonds and related to them vibrations appear. Quantity and quality of the interact ions is related to characteristics of the nanoparticle and the presence of forming apatite structures.

5

The soft-tissue response to implants of chitosan foils

Paluszkiewicz C., Menaszek E., Żołnierek M.

Engineering of Biomaterials

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2011

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Vol. 14, no. 109-111 spec. iss.

73

EN

The objective of our in vivo study was to investigate the biocompatibility of polymer foils enriched with chitosan. Methods: three types of chitosan: non-modified(jp), and modified with montmorillonite K5 and montmorillonite K10 (p2-p3) were prepared. The in vivo studies were carried out using the rat soft tissues as a model. Small pieces of foils (2mm x 4mm)used in the experiment were sterilized by UV radiation. Next they were implanted under sterile conditions into the gluteus muscle of adult male outbred Wistar rats. Each animal received two implants: the modified foil into the right muscle, and the unmodifiedfoil used as a control into the left one. All procedures were conducted in sterile conditions and under anaesthesia. Animals were anesthetized with intraperitoneal injection of xylazine and ketamine (Biowet Puławy, Poland). Skin in the site of surgery was shaved and disinfected, and a small incision was made in the gluteus muscle. Equal pieces of foils were inserted into the such created pouch. The muscle and skin wounds were closed with 5/0 PDSII (polydioxanone) monofilament absorbable sutures (Ethicon Ltd., UK). All animals survived the surgery. No wound healing complications were observed after the surgery. The animals were maintained under standard conditions with free access to food and water. After 7, and 30 days from the surgery, at each time point 12animals were sacrificed. Tissue specimens containing the implanted materials were excised and immediately frozen in liquid nitrogen. Next they were cut into 8μm thick slides in a Shandon cryostat (Thermo-Scientific, UK) at -22°C. Obtained slides were investigated through histological and histochemical methods to estimate the intensity of inflammation, production of collagen, and metabolic activity of connective and muscle tissues surrounding implant. In order to estimate the effect of the implants on the metabolic state of surrounding tissues, activities of the marker metabolic enzymes: cytochrome c oxidase, and NADH dehydrogenase were examined. The activity of acid phosphatase was used to assess the extent of inflammation around the implants. The presence and thickness of fibrous capsule around the implants were estimated on slides stained by van Gieson’s method. Results: Differences between non-modified and modified with chitosan materials were not manifested in short, 7- and 30 days series. The activities of mitochondrial oxidative enzymes, cytochrome c oxidase and NADH dehydrogenase, in muscle fibres in close proximity to the implants were slightly lower than in those further away. Probably one month was too short period to obtain the whole recovery after the surgery. At the same time the process of regeneration seemed to be intense: numerous regenerating muscle fibres infiltrated the granulation tissue around the implanted foils were observed The inflammation respone was visibly lower in 30days series compared to7days ones what indicate that the inflammation was evoked in higher degree by surgery than by the presence of the implanted materials. The fibrous capsule around foils was thin or not present at all – there were places where pieces of foils were in direct contact with the muscle tissue. After one month experiment there were no signs of degradation of materials. The experiment has been continued to compare abilities of biomaterials in long term series. Conclusion: The regeneration and enzymic activity of muscle tissue together with the lack of continuous fibrous capsule suggest that the materials used in our study are biocompatibile and are suitable for the treatment of tissue injury.

6

Zastosowanie technik FTIR w badaniach nanokompozytów polimerowo-ceramicznych

Paluszkiewicz C., Stodolak E., Jeleń P.

Kompozyty

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2010

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R. 10, nr 1

64-69

PL

W pracy zbadano nanokompozytowe tworzywa polimerowo-ceramiczne, w których jako osnowę zastosowano biopolimer naturalny należący do grupy polisacharydów (chitosan, CS). Jako nanonapełanicz wykorzystano komercyjny glinokrzemian warstwowy - montmorylonit w dwóch odmianach: MMT-K5 i MMT-K10. Nanometryczne cząstki ceramiczne scharakteryzowano ze względu na powierzchnię rozwinięcia (BET), wielkość nanocząstek w roztworze rozpuszczalnika (DLS) i morfologię (TEM). Materiały wyjściowe: chitozan, MMT K5 i K10 poddano również analizie fourierowskiej spektroskopii w podczerwieni (FTIR) ze względu na późniejszy zakres badań otrzymanych na ich bazie tworzyw. Materiały nanokompozytowe otrzymano techniką odlewania, stosując kilkustopniową metodę dyspersji nanocząstek w matrycy. Udział nanonapełaniacza był stały i nie przekraczał 3% wag. MMT. Folie nanokompozytowe (CS/MMT-K5 i CS/MMT-K10) poddano badaniom FTIR. Materiałem odniesienia była niemodyfikowana folia CS. Widma badanych materiałów rejestrowano w zakresie środkowej podczerwieni techniką transmisyjną i refleksyjną techniką całkowitego wewnętrznego odbicia (ATR). Wykorzystując techniki FTIR, przedstawiono oddziaływania na granicy faz: nanocząstka a matryca biopolimerowa. Wykazano, że dzięki zastosowanym technikom FTIR można otrzymać informacje na temat rozłożenia nanonapełniacza w osnowie, jednorodności folii nanokompozytowej, skłonności nanonapełniacza do sedymentacji w trakcie procesu suszenia. Dobrane techniki badawcze pozwoliły na otrzymanie informacji uśrednionych (transmisja) oraz informacji z powierzchni tworzywa przy różnej głębokości penetracji (ATR z kryształem germanowym i diamentowym) i wielkości badanej powierzchni.

EN

The nanocomposites from renewable sources are considered to be a group of perspective materials. The main advantage of the nanocomposite materials based on natural biopolymers such as polysaccharides e.g. alginates or chitosane is their simple utilization and safe products of degradation. The additional advantage of nanocomposites based on chitosane and montmorylonite (MMT) is availability and low cost of the component materials. However, successful preparation of the nanocomposites still encounters many problems mainly related to proper dispersion of nano-fillers. High surface area, and high free surface energy are the cause of agglomeration of the nanoparticles, which then lose ability to create first-, and second-order interactions with a polymer matrix. In the result, the nano-filler becomes submicron or micron phase which because of its low fraction, not exceeding 10% (volume or weight), act as a defect weakening the polymer matrix. The situation between nanofillier and polymer matrix can be observed used the most popular measurement technique: thermal analyses (DSC, TG, DTA), microscopic methods such as: AFM, SEM, TEM, and spectroscopic methods; FTIR, Raman, XPS. In order to follow the structural changes in the nanocomposites materials a variety of investigative methods suitable for verification of complex phenomena taking place in the materials is utilized. The spectroscopy techniques FTIR can be relatively easily applied both to powder and foil samples on nanocomposite materials. Obtained result can be averaged in the case of transmission techniques or it may describe an area of a sample in the case of ATR technique. FTIR techniques enables better observation and interpretation of subtle changes taking place in the polymer matrix than XRD and SEM/EDS methods, especially when the investigated material is a nanocomposite polymer foil. In the work efforts of production of a series of nanocomposites based on natural chitosane and nano-clay (MMT) were taken. The nanocomposites materials was obtaining by casting method. Mechanical and ultrasound stirrer was used to better homogenization between nanoclay particles and biopolymer matrix. Two types of MMT (MMT-K5 and MMT-K10) with different particle size (DLS) and specific surface area (BET) were used. The nanoparticles morphology was examined with transmission electron microscopy (TEM). In each nanocomposite, the nano-filler amount was 3% wt. Dispersion of the nano-filler within the polymer matrix was investigated with the use of FTIR method (transmission mode, FTIR microscopy). FTIR measurements of the samples were carried out on the transmission mode in the region from 400 to 4000 cm-1 and ATR method in the region from 700 to 4000 cm-1. For the transmission mode powder samples were prepared in the form of potassium bromide (KBr), whereas the foils of the polymer samples were placed directly in the spectrometer. In the ATR method which allow to analyze the deep of penetration of the samples two different crystals were used: german and diament. In this way were observed how the clay (MMT-K5 or MMT-K10) influenced on the chemical structure of biopolymer chain.

7

Vibrational spectroscopy investigation of montmorillonite - chitosane nanocomposite materials

Paluszkiewicz C., Stodolak E.

Engineering of Biomaterials

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2010

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Vol. 13, no. 99-101

128

EN

Biomaterials basing on natural polysaccharides, i.e. hiauronic acid, alginate, chitosane are an alternative for already applied bioresorbable synthetic materials basing on synthetic polyhydroxyacids. Their main advantages are good accessibility, low cost, easy forming and high biocompatibility. Additionally, they are a perfect matrix for bioactive nanoparticles i.e. hydroxyapatite (HAp), tricalcium phosphate (TCP) and silica (SiO2). The work presents results of research on nanocomposite consisting of chitosane matrix (CS) modified with a nanofiller, which was natural montmorillonite (MMT). Nanocomposite foils were produced by the casting method. In order to induce better biocompatibility, the surface of the CS/MMT composite was neutralized (bath in NaOH solution). The nanocomposite foils were subjected to a bioactivity test by incubation in SBF at 37oC for 7 days. It was observed that the CS/MMT material surface showed a local supersaturation, which was a result of apatite nucleation. The CS/MMT nanocomposites were investigated using FT-IR (Fourier Transform Infrared Spectroscopy) and Fourier Raman Spectroscopy. FTIR measurements of the samples were carried out on the transmission and reflection modes. The FTIR microscopy spectra were collected using Bio-Rad Excalibur with ATR attachment as well as microscope UMA500 equipped with MCT detector. Spectra were measured at 4 cm-1 resolution in the region from 4000 cm-1 to 600 cm-1. FT-Raman spectra were obtained using a FTS6000 Bio-Rad spectrometer with Ge detector. The samples were excited with a Nd-YAG laser (1064nm). Additional all materials in all steps experiments were observed under Scanning Electron Microscopy (Nova NanoSEM). Vibrational spectroscopy methods (FT Raman and FTIR) can be used for investigation of nanocomposite foils basing on biopolymers. High sensitivity the applied spectroscopy techniques show that in the result of the neutralization of CS/MMT foil (via incubation in NaOH solution) the biopolymer chain breaks. This phenomena is visible by intensity ratio between COC/ COH bands. Increase of reactivity of chitosane chain lead to entrapment of PO43-, which is the origin of the apatite forms nucleation process. Chemical treatment of the nanocomposite foils, i.e. NaOH washes influences their chemical structure and microstructure. Neutralisation of the foils is the first processing stage which precedes the potential use of CS/MMT foils in biomedical applications. The materials show a tendency to apatite crystallisation which may support regeneration of damaged bone tissue. The applied spectroscopic methods allowed to observe changes in the whole volume of the sample. Individual ATR measurements taken at various spectral ranges and penetration depths allow to observe subtle changes in the polymer matrix caused by chemical treatment (NaOH and SBF incubation). Results of the investigations indicate that in the CS/ MMT systems new chemical bonds and related to them vibrations appear. Quantity and quality of the interactions is related to characteristics of the nanoparticle and the presence of forming apatite structures.

8

Badanie procesu degradacji kompozytów z polimerów resorbowalnych w warunkach in vitro

Chłopek J., Morawska A., Umańska L., Paluszkiewicz C.

Inżynieria Biomateriałów

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2004

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R. 7, nr 38-42

132-136

9

Fotochemiczna modyfikacja polisulfonu

Kowal J., Czajkowska B., Błażewicz M., Paluszkiewicz C.

Inżynieria Biomateriałów

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2003

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R. 6, nr 30-33

104-107

PL

Badano zmiany związane z fotochemiczną modyfikacją polisulfonu w oparciu o analizę spektralną naświetlanych próbek. Stwierdzono tworzenie połączeń karbonylowych grup hydroksylowych, a także obniżenie ilości grup eterowych i sulfonowych, które związane jest z degradacją polimeru. Obserwowany wzrost absorbancji w zakresie długofalowej częstości zakresu UV (?>290 nm) przypisano tworzeniu się sprzężonych struktur polifenylowych.

EN

The changes resulting from the photochemical modification of polysulfone were examined with the use of spectroscopic analysis of irradiated samples. The formation of carbonyl and hydroxyl groups in irradiated films and the reduction of the maount of ether and sulfone groups, connected with the degradation of the polymer were observed. The increase of the long wave UV absorption (?>290 nm) was attributed to the formation of polyphenyl conjugated structures.

10

Zastosowanie metody zol-żel w praktyce dentystycznej

Matraszek H., Stoch A., Paluszkiewicz C., Brożek A., Długoń E.

Inżynieria Biomateriałów

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2002

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R. 5, nr 23-25

72-74

11

Badania kompozytów ceramiczno węglowych metodą mikrospektroskopii FTIR

Paluszkiewicz C., Błażewicz M., Szaraniec B., Chłopek J.

Inżynieria Biomateriałów

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2001

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R. 4, nr 17-19

81-82

12

Mechanizm wiązania mas fosforanowych w świetle badań spektroskopowych w podczerwieni

Pucka G., Dobosz S. M., Paluszkiewicz C.

Krzepnięcie Metali i Stopów

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2000

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R. 2, nr 44

521--526

PL

W pracy przedstawiono badania nad opracowaniem mechanizmu wiązania samoutwardzalnych mas fosforanowych W badaniach wykorzystano technikę rozproszonego odbicia DRS. Badaniom poddano składniki masy (piasek kwarcowy wzorcowy, spoiwo Glifos-C, utwardzacz UP-2) i układ wiążący masy w temperaturze otoczenia i po obróbce cieplnej w temperaturze 950 ºC. Stwierdzono, iż badania wykonane zastosowaną techniką bardzo dokładnie odwzorowują zmiany zachodzące na powierzchni ziaren ceramicznej osnowy masy fosforanowej.

EN

In this work the mechanism self-hard bonding phosphate masses has been shown. In the examinations the DRS method has been appalled. Components of moulding sand (quartz sand, Glifos-C binder, UP-2 hardener) and hardening system at room temperature and after heat treatment 950 ºC have been examined. Appalled in this work research technique to the examination of moulding masses show accuracy changing on the particle surface of phosphate mass.