Partner: Andrzej Chwojnowski

Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)

Recent publications
1.Wasyłeczko M., Remiszewska E., Sikorska W., Dulnik J., Chwojnowski A., Scaffolds for Cartilage Tissue Engineering from a Blend of Polyethersulfone and Polyurethane Polymers, Molecules, ISSN: 1420-3049, DOI: 10.3390/molecules28073195, Vol.28, No.7, pp.3195-1-28, 2023
Abstract:

In recent years, one of the main goals of cartilage tissue engineering has been to find appropriate scaffolds for hyaline cartilage regeneration, which could serve as a matrix for chondrocytes or stem cell cultures. The study presents three types of scaffolds obtained from a blend of polyethersulfone (PES) and polyurethane (PUR) by a combination of wet-phase inversion and salt-leaching methods. The nonwovens made of gelatin and sodium chloride (NaCl) were used as precursors of macropores. Thus, obtained membranes were characterized by a suitable structure. The top layers were perforated, with pores over 20 µm, which allows cells to enter the membrane. The use of a nonwoven made it possible to develop a three-dimensional network of interconnected macropores that is required for cell activity and mobility. Examination of wettability (contact angle, swelling ratio) showed a hydrophilic nature of scaffolds. The mechanical test showed that the scaffolds were suitable for knee joint applications (stress above 10 MPa). Next, the scaffolds underwent a degradation study in simulated body fluid (SBF). Weight loss after four weeks and changes in structure were assessed using scanning electron microscopy (SEM) and MeMoExplorer Software, a program that estimates the size of pores. The porosity measurements after degradation confirmed an increase in pore size, as expected. Hydrolysis was confirmed by Fourier-transform infrared spectroscopy (FT-IR) analysis, where the disappearance of ester bonds at about 1730 cm −1 wavelength is noticeable after degradation. The obtained results showed that the scaffolds meet the requirements for cartilage tissue engineering membranes and should undergo further testing on an animal model.

Keywords:

articular cartilage, cartilage tissue engineering, hydrolysis process, materials for scaffolds, partly degradable scaffolds, polyethersulfone–polyurethane scaffolds, polyurethane degradation, regenerative medicine, scaffold requirements, tissue engineering

Affiliations:
Wasyłeczko M.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Remiszewska E.-other affiliation
Sikorska W.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Dulnik J.-IPPT PAN
Chwojnowski A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
2.Kołakowska A., Kołbuk-Konieczny D., Chwojnowski A., Rafalski A., Gadomska-Gajadhur A., Chitosan-Based High-Intensity Modification of the Biodegradable Substitutes for Cancellous Bone, Journal of Functional Biomaterials, ISSN: 2079-4983, DOI: 10.3390/jfb14080410, Vol.14, No.8, pp.410-1-15, 2023
Abstract:

An innovative approach to treating bone defects is using synthetic bone substitutes made of biomaterials. The proposed method to obtain polylactide scaffolds using the phase inversion technique with a freeze extraction variant enables the production of substitutes with morphology similar to cancellous bone (pore size 100–400 µm, open porosity 94%). The high absorbability of the implants will enable their use as platelet-rich plasma (PRP) carriers in future medical devices. Surface modification by dipping enabled the deposition of the hydrophilic chitosan (CS) layer, maintaining good bone tissue properties and high absorbability (850% dry weight). Introducing CS increases surface roughness and causes local changes in surface free energy, promoting bone cell adhesion. Through this research, we have developed a new and original method of low-temperature modification of PLA substitutes with chitosan. This method uses non-toxic reagents that do not cause changes in the structure of the PLA matrix. The obtained bone substitutes are characterised by exceptionally high hydrophilicity and morphology similar to spongy bone. In vitro studies were performed to analyse the effect of morphology and chitosan on cellular viability. Substitutes with properties similar to those of cancellous bone and which promote bone cell growth were obtained.

Keywords:

chitosan, polylactide, bone substitute, tissue regeneration

Affiliations:
Kołakowska A.-other affiliation
Kołbuk-Konieczny D.-IPPT PAN
Chwojnowski A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Rafalski A.-other affiliation
Gadomska-Gajadhur A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
3.Wasyłeczko M., Krysiak Z.J., Łukowska E., Gruba M., Sikorska W., Kruk A., Dulnik J., Czubak J., Chwojnowski A., Three-dimensional scaffolds for bioengineering of cartilage tissue, Biocybernetics and Biomedical Engineering, ISSN: 0208-5216, DOI: 10.1016/j.bbe.2022.03.004, Vol.42, No.2, pp.494-511, 2022
Abstract:

The cartilage tissue is neither supplied with blood nor innervated, so it cannot heal by itself. Thus, its reconstruction is highly challenging and requires external support. Cartilage diseases are becoming more common due to the aging population and obesity. Among young people, it is usually a post-traumatic complication. Slight cartilage damage leads to the spontaneous formation of fibrous tissue, not resistant to abrasion and stress, resulting in cartilage degradation and the progression of the disease. For these reasons, cartilage regeneration requires further research, including use of new type of biomaterials for scaffolds. This paper shows cartilage characteristics within its most frequent problems and treatment strategies, including a promising method that combines scaffolds and human cells. Structure and material requirements, manufacturing methods, and commercially available scaffolds were described. Also, the comparison of poly(L-lactide) (PLLA) and polyethersulfone (PES) 3D membranes obtained by a phase inversion method using nonwovens as a pore-forming additives were reported. The scaffolds' structure and the growth ability of human chondrocytes were compared. Scaffolds' structure, cells morphology, and protein presence in the membranes were examined with a scanning electron microscope. The metabolic activity of cells was tested with the MTT assay. The structure of the scaffolds and the growth capacity of human chondrocytes were compared. Obtained results showed higher cell activity and protein content for PES scaffolds than for PLLA. The PES membrane had better mechanical properties (e.g. ripping), greater chondrocytes proliferation, and thus a better secretion of proteins which build up the cartilage structure.

Keywords:

3D-scaffolds, membrane structure, polyethersulfone, poly(L-lactide), chondrocyte culture, cartilage regeneration

Affiliations:
Wasyłeczko M.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Krysiak Z.J.-other affiliation
Łukowska E.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Gruba M.-Gruca Orthopedic and Trauma Teaching Hospital, Centre of Postgraduate Medical Education (PL)
Sikorska W.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Kruk A.-Warsaw University of Technology (PL)
Dulnik J.-IPPT PAN
Czubak J.-Gruca Orthopedic and Trauma Teaching Hospital, Centre of Postgraduate Medical Education (PL)
Chwojnowski A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
4.Wasyleczko M., Sikorska W., Przytulska M., Dulnik J., Chwojnowski A., Polyester membranes as 3D scaffolds for cell culture, Desalination and Water Treatment, ISSN: 1944-3994, DOI: 10.5004/dwt.2021.26658, Vol.214, pp.181-193, 2021
Abstract:

The study presents two types of three-dimensional membranes made of the biodegradable copolymer. They were obtained by the wet-phase inversion method using different solvent and pore precursors. In one case, a nonwoven made of gelatin and polyvinylpyrrolidone (PVP) as precursors of macropores and small pores, respectively, were used. In the second case, PVP nonwovens and Pluronic were used properly for macro- and micro-pores. As the material, a biodegradable poly(L-lactide-co-ε-caprolactone) is composed of 30% ε-caprolactone and 70% poly(L-lactic acid) was used. Depending on the pore precursors, different membrane structures were obtained. The morphology of pores was studied using the MeMoExplorer™, an advanced software designed for computer analysis of the scanning electron microscopy images. The scaffolds were degraded in phosphate-buffered saline and Hank’s balanced salt solutions at 37°C. Moreover, the porosity of the membranes before and after hydrolysis was calculated.

Keywords:

3D scaffolds, poly(L-lactide-co-ε-caprolactone), porosity of membrane, phase inversion method, degradation of scaffolds

Affiliations:
Wasyleczko M.-other affiliation
Sikorska W.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Przytulska M.-other affiliation
Dulnik J.-IPPT PAN
Chwojnowski A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
5.Gadomska‐Gajadhur A., Kruk A., Ruśkowski P., Sajkiewicz P., Dulnik J., Chwojnowski A., Original method of imprinting pores in scaffolds for tissue engineering, Polymers for Advanced Technologies, ISSN: 1042-7147, DOI: 10.1002/pat.5091, pp.1-13, 2020
Abstract:

Results of the preparation of biodegradable porous scaffolds using an original modification of a wet phase inversion method were presented. Influence of gelatin non‐woven as a non‐classic pore precursor and polyvinylpyrrolidone, Pluronic as classic pore precursors on the structure of obtained scaffolds was analyzed. It was shown that the addition of gelatin non‐wovens enables the preparation of scaffolds, which allow for the growth of cells (size, distribution, and shape of pores). Mechanical properties of the obtained cell scaffolds were determined. The influence of pore precursors on mass absorption of scaffolds against isopropanol and plasma was investigated. Interaction of scaffolds with a T‐lymphocyte line (Jurkat) and with fibroblasts (L929) was investigated. Obtained scaffolds are not cytotoxic and can be used as implants, for example, the regeneration of cartilage tissue.

Keywords:

cell cultures, cytotoxic, fibroblasts, imprinted scaffolds

Affiliations:
Gadomska‐Gajadhur A.-other affiliation
Kruk A.-Warsaw University of Technology (PL)
Ruśkowski P.-Warsaw University of Technology (PL)
Sajkiewicz P.-IPPT PAN
Dulnik J.-IPPT PAN
Chwojnowski A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
6.Gadomska‐Gajadhur A., Kruk A., Dulnik J., Chwojnowski A., New polyester biodegradable scaffolds for chondrocyte culturing: preparation, properties, and biological activity, JOURNAL OF APPLIED POLYMER SCIENCE, ISSN: 0021-8995, DOI: 10.1002/app.50089, pp.e50089-1-14, 2020
Abstract:

An innovative modification of the wet inversion phase method, consisting in the use of a polymer nano‐nonwoven as a nonclassic pore precursor. Mechanical properties of the obtained scaffolds were determined, their hydrophilic properties (serum absorbability) were tested, and the content of residues of materials used in the scaffold preparation was determined. Nontoxicity of the developed scaffolds toward T lymphocyte cells was proved. Cultures of primary chondrocytes were obtained successfully. It was proved that an addition of a polymer nano‐nonwoven changes the properties of the scaffolds favorably in respect of their subsequent application in tissue engineering.

Keywords:

cartilage regeneration, chondrocytes, nano-nonwoven, polyvinylpyrrolidone, T lymphocytes

Affiliations:
Gadomska‐Gajadhur A.-other affiliation
Kruk A.-Warsaw University of Technology (PL)
Dulnik J.-IPPT PAN
Chwojnowski A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
7.Lewińska D., Chwojnowski A., Wojciechowski C., Kupikowska-Stobba B., Grzeczkowicz M., Weryński A., Electrostatic droplet generator with 3-coaxial-nozzle head for microencapsulation of living cells in hydrogel covered by synthetic polymer membranes, Separation Science and Technology, ISSN: 1520-5754, DOI: 10.1080/01496395.2011.617350, Vol.47, No.3, pp.463-469, 2012
8.Płończak M., Czubak J., Chwojnowski A., Kupikowska-Stobba B., Culture of human autologous chondrocytes on polysulphonic membrane - preliminary studies, Biocybernetics and Biomedical Engineering, ISSN: 0208-5216, DOI: 10.1016/s0208-5216(12)70042-6, Vol.32, No.3, pp.63-67, 2012
9.Chwojnowski A., Wojciechowski C., Lewińska D., Łukowska E., Nowak J., Kupikowska-Stobba B., Grzeczkowicz M., Studies on the structure of semipermeable membranes by means of SEM. Problems and potential sources of errors, Biocybernetics and Biomedical Engineering, ISSN: 0208-5216, DOI: 10.1016/s0208-5216(12)70032-3, Vol.32, No.1, pp.51-64, 2012
10.Kupikowska-Stobba B., Lewińska D., Dudziński K., Jankowska-Śliwińska J., Grzeczkowicz M., Wojciechowski C., Chwojnowski A., Influence of changes in composition of the membrane-forming solution on the structure of alginate-polyethersulfone microcapsules, Biocybernetics and Biomedical Engineering, ISSN: 0208-5216, Vol.29, No.3, pp.61-69, 2009

List of chapters in recent monographs
1.
701
Chwojnowski A., Wojciechowski C., Lewińska D., Kupikowska-Stobba B., Grzeczkowicz M., Dudziński K., Membrany i procesy membranowe w ochronie środowiska. Monografie Komitetu Inżynierii Środowiska PAN, rozdział: Badanie budowy membran półprzepuszczalnych za pomocą SEM. Problemy i potencjalne źródła błędów, Komitet Inżynierii Środowiska PAN, 65, pp.109-120, 2010
2.
702
Kupikowska-Stobba B., Dudziński K., Chwojnowski A., Gutowska M., Sabalińska S., Lewińska D., Płończak M., Czubak J., Membrany i procesy membranowe w ochronie środowiska. Monografie Komitetu Inżynierii Środowiska PAN , rozdział: Wykorzystanie membran półprzepuszczalnych do hodowli chondrocytów króliczych, Komitet Inżynierii Środowiska PAN , 65, pp.341-352, 2010

Conference papers
1.Chwojnowski A., Kruk A., Wojciechowski C., Łukowska E., Dulnik J., Sajkiewicz P., The dependence of the membrane structure on the non-woven forming the macropores in the 3D scaffolds preparation, Desalination and Water Treatment, ISSN: 1944-3994, DOI: 10.5004/dwt.2017.11394, Vol.64, pp.324-331, 2017
Abstract:

Three types of membrane structures with wide pores were compared in this study. One of the membranes was obtained from polyethersulfone using cellulose fibers as the macropore precursors. Two of the fibers were obtained from poly(L-lactide). As the macropore precursors olyvinylpyrrolidone (1.2 MDa) and pork gelatin non-woven were used, the influence of non-woven fibers on the structure of membranes was shown. Necessity of specific membrane structure application was explained. The hoice of polymers and co-polymers with a range of biodegradation times can determine the scaffold type suitable for the age of a patient.

Keywords:

Polysulfone membrane, Polyester membranes, Membrane structures, Biodegradable membranes, 3D scaffold

Affiliations:
Chwojnowski A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Kruk A.-Warsaw University of Technology (PL)
Wojciechowski C.-other affiliation
Łukowska E.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Dulnik J.-IPPT PAN
Sajkiewicz P.-IPPT PAN
2.Kruk A., Gadomska-Gajadhur A., Ruśkowski P., Chwojnowski A., Dulnik J., Synoradzki L., Preparation of biodegradable semi-permeable membranes as 3D scaffolds for cell cultures, Desalination and Water Treatment, ISSN: 1944-3994, DOI: 10.5004/dwt.2017.11415, Vol.64, pp.317-323, 2017
Abstract:

Results of the preparation of semi-permeable membranes made of biodegradable polymers membranes were presented. Among known polyesters, polylactide was selected for research. The membranes were obtained using wet phase inversion method. The influence of polyvinylpyrrolidone and polymeric nano-non-wovens as pores precursors on the structure of obtained membranes was analysed. It was shown, that utilisation of polymeric nano-non-wovens enabled preparation of semi-permeable membranes, which could be used as wide-pore 3D-type cellular scaffolds.

Keywords:

Biodegradable polymers membranes, Biodegradable polyesters, Porous three-dimensional scaffolds, Inversion phase method

Affiliations:
Kruk A.-Warsaw University of Technology (PL)
Gadomska-Gajadhur A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Ruśkowski P.-Warsaw University of Technology (PL)
Chwojnowski A.-Nałęcz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences (PL)
Dulnik J.-IPPT PAN
Synoradzki L.-Warsaw University of Technology (PL)

Conference abstracts
1.Chwojnowski A., Kruk A., Wojciechowski C., Łukowska E., Dulnik J., Sajkiewicz P., The dependence of the membrane structure on the non-woven forming the macropores in the 3D scaffolds preparation, MEMPEP 2016, XI-th Conference on Membranes and Membrane Processes in Environmental Protection, 2016-06-15/06-18, Zakopane (PL), pp.23, 2016
2.Kruk A., Gadomska-Gajadhur A., Ruśkowski P., Chwojnowski A., Dulnik J., Synoradzki L., Preparation of biodegradable semi-permeable membranes as 3D scaffolds for cell cultures, MEMPEP 2016, XI-th Conference on Membranes and Membrane Processes in Environmental Protection, 2016-06-15/06-18, Zakopane (PL), pp.24, 2016
3.Kupikowska B., Dudziński K., Chwojnowski A., Gutowska M., Sabalińska S., Lewińska D., Płończak M., Czubak J., Wykorzystanie membran półprzepuszczalnych do hodowli chondrocytów króliczych, MEMPEP 2010, VIII Krajowa Konferencja Membrany i procesy membranowe w ochronie środowiska, 2010-06-09/06-12, Zakopane (PL), pp.341, 2010
4.Chwojnowski A., Wojciechowski C., Lewińska D., Kupikowska B., Grzeczkowicz M., Dudziński K., Badanie budowy membran półprzepuszczalnych za pomocą SEM. Problemy i potencjalne źródła błędów, MEMPEP 2010, VIII Krajowa Konferencja Membrany i procesy membranowe w ochronie środowiska, 2010-06-09/06-12, Zakopane (PL), pp.109, 2010

Patents
Filing No./Date
Filing Publication
Autor(s)
Title
Protection Area, Applicant Name
Patent Number
Date of Grant
pdf
414353
2015-10-13
BUP 09/2017
2017-04-24
Chwojnowski A., Łukowska E., Wojciechowski C., Gadomska-Gajadhur A., Kruk A., Ruśkowski P., Synoradzki L., Denis P., Dulnik J., Sajkiewicz P.
Sposób otrzymywania szerokoporowatego, poliestrowego rusztowania komórkowego
PL, Instytut Biocybernetyki i Inżynierii Biomedycznej im. Macieja Nałęcza PAN, Politechnika Warszawska, Instytut Podstawowych Problemów Techniki PAN
228884
WUP 05/2018
2018-05-30



402004
2012-12-11
BUP 13/2014
2014-06-23
Lewińska D., Kupikowska-Stobba B., Chwojnowski A., Grzeczkowicz M., Łukowska E.
Sposób oznaczania stężenia komórek
PL, Instytut Biocybernetyki i Inżynierii Biomedycznej PAN
223717
WUP 10/2016
2016-10-31