Chitosan is one of the most well-known and characterized materials applied in tissue engineering. Due to its unique chemical, biological and physical properties chitosan is frequently used as the main component in a variety of biomaterials such as membranes, scaffolds, drug carriers, hydrogels and, lastly, as a component of bio-ink dedicated to medical applications. Chitosan’s chemical structure and presence of active chemical groups allow for modification for tailoring material to meet specific requirements according to intended use such as adequate endurance, mechanical properties or biodegradability time. Chitosan can be blended with natural (gelatin, hyaluronic acid, collagen, silk, alginate, agarose, starch, cellulose, carbon nanotubes, natural rubber latex, κ-carrageenan) and synthetic (PVA, PEO, PVP, PNIPPAm PCL, PLA, PLLA, PAA) polymers as well as with other promising materials such as aloe vera, silica, MMt and many more. Chitosan has several derivates: carboxymethylated, acylated, quaternary ammonium, thiolated, and grafted chitosan. Its versatility and comprehensiveness are confirming by further chitosan utilization as a leading constituent of innovative bio-inks applied for tissue engineering. This review examines all the aspects described above, as well as is focusing on a novel application of chitosan and its modifications, including the 3D bioprinting technique which shows great potential among other techniques applied to biomaterials fabrication.

3D printing is being used more extensively in modern biomedicine. One of the problems is selecting a proper crosslinking method of bioprinted material. Amongst currently used techniques we can distinguish: physical crosslinking (e.g. Ca²⁺ and Sr²⁺) and chemical crosslinking–the UV light crosslinking causing the biggest discussion. UV radiation is selectively absorbed by DNA, mainly in the UV-B region but also (to some extent) in UV-A and UV-C regions. DNA excitement results in typical photoproducts. The amount of strand breaks may vary depending on the period of exposition, it can also differ when cells undergo incubation after radiation. The aim of this study was to show whether and how the time of irradiation with 405 nm and 365 nm wavelengths affect DNA damage in cell lines and micro-organs (pancreatic islets). The degree of DNA damage caused by different wavelengths of radiation (405 nm and 365 nm) was evaluated by a comet assay. The test was performed on fibroblasts, alpha cells, beta cells and porcine pancreatic islets after 24 hours incubation period. Samples without radiation treatment were selected as a control group. Results analysis consisted of determining the percent of cells with damaged DNA and the tail intensity evaluation. The degree of DNA damage in pancreatic islets after exposure to 405 nm wavelength oscillated between 2% and 6% depending on the tested time period (10 – 300 seconds). However, treating islets using 365 nm wavelength resulted in damage up to 50%. This clearly shows significantly less damage when using 405 nm wavelength. Similar results were obtained for the tested cell lines. Crosslinking with 405 nm is better for pancreatic islets than crosslinking with 365 nm UV light.

Background: 3D bioprinting is the future of constructing functional organs. Creating a bioactive scaffold with pancreatic islets presents many challenges. The aim of this paper is to assess how the 3D bioprinting process affects islet viability. Methods: The BioX 3D printer (Cellink), 600 μm inner diameter nozzles, and 3% (w/v) alginate cell carrier solution were used with rat, porcine, and human pancreatic islets. Islets were divided into a control group (culture medium) and 6 experimental groups (each subjected to specific pressure between 15 and 100 kPa). FDA/PI staining was performed to assess the viability of islets. Analogous studies were carried out on α-cells, β-cells, fibroblasts, and endothelial cells. Results: Viability of human pancreatic islets was as follows: 92% for alginate-based control and 94%, 90%, 74%, 48%, 61%, and 59% for 15, 25, 30, 50, 75, and 100 kPa, respectively. Statistically significant differences were observed between control and 50, 75, and 100 kPa, respectively. Similar observations were made for porcine and rat islets. Conclusions: Optimal pressure during 3D bioprinting with pancreatic islets by the extrusion method should be lower than 30 kPa while using 3% (w/v) alginate as a carrier.

Pathogenesis of primary sclerosing cholangitis (PSC) may involve impaired bile acid (BA) homeostasis. We analyzed expressions of factors mediating enterohepatic circulation of BA using ileal and colonic (ascending and sigmoid) biopsies obtained from patients with PSC with and without ulcerative colitis (UC) and explanted PSC livers. Two-fold increase of BA-activated farnesoid X receptor (FXR) protein levels were seen in ascending and sigmoid colon of PSC patients with correspondingly decreased apical sodium-dependent BA transporter (ASBT) gene expression. This was associated with increased OSTβ protein levels in each part of analyzed gut. An intestinal fibroblast growth factor (FGF19) protein expression was significantly enhanced in ascending colon. Despite increased hepatic nuclear receptors (FXR, CAR, SHP), and FGF19, neither CYP7A1 suppression nor CYP3A4 induction were observed. The lack of negative regulation of BA synthesis may be accountable for lower levels of cholesterol observed in PSC in comparison to primary biliary cholangitis (PBC). In conclusion, chronic cholestasis in PSC induces adaptive changes in expression of BA transporters and FXR in the intestine. However hepatic impairment of expected in chronic cholestasis downregulation of CYP7A1 and upregulation of CYP3A4 may promote BA-induced liver injury in PSC.

Type 1 diabetes (T1D) is characterized by the destruction of over 90% of the β-cells. C-peptide is a parameter for evaluating T1D. Streptozotocin (STZ) is a standard method of inducing diabetes in animals. Eight protocols describe the administration of STZ in mice; C-peptide levels are not taken into account. The aim of the study is to determine whether the STZ protocol for the induction of beta-cell mass destruction allows for the development of a stable in vivo mouse model for research into new transplant procedures in the treatment of type 1 diabetes. Materials and methods: Forty BALB/c mice were used. The animals were divided into nine groups according to the STZ dose and a control group. The STZ doses were between 140 and 400 mg/kg of body weight. C-peptide was taken before and 2, 7, 9, 12, 14, and 21 days after STZ. Immunohistochemistry was performed. The area of the islet and insulin-/glucagon-expressing tissues was calculated. Results: Mice who received 140, 160, 2 × 100, 200, and 250 mg of STZ did not show changes in mean fasting C-peptide in comparison to the control group and to day 0. All animals with doses of 300 and 400 mg of STZ died during the experiment. The area of the islets did not show any differences between the control and STZ-treated mice in groups below 300 mg. The reduction of insulin-positive areas in STZ mice did not exceed 50%. Conclusions: Streptozotocin is not an appropriate method of inducing a diabetes model for further research on transplantation treatments of type 1 diabetes, having caused the destruction of more than 90% of the β-cell mass in BALB/c mice.

Tissue engineering is a widely developing scientific field, which combines technological solutions with the biology of the living organism. Regenerative medicine that uses tools of tissue engineering offers alternative means of therapy enhancing damaged tissues or organs. One of the development directions of contemporary chemical engineering is the scientific description of novel technologies that will enable production of porous structures – with high utility for biomedical engineering. 3D printing is one of the most popular methods used to produce scaffolds for cell culture. Nowadays a research team, in which authors are currently working, is dealing with the problem of manufacturing 3D constructs that play the role of artificial organ, obtained via 3D bioprinting. In the current article we present the possibilities and limitations of 3D bioprinting method in the context of possible application of manufactured structures as fully functional organs.