Fibrin membranes are widely used in regenerative medicine because they are biocompatible, biodegradable, contain growth factors, and support cell attachment. Most commonly they are produced from serum, but they can also be isolated from activated plasma. To increase the fibrinogen concentration of plasma, cryoprecipitate isolation is a possible solution. In this work, cryoprecipitate was prepared from fresh frozen plasma, isolated by plasmapheresis. The concentration of cellular elements, fibrinogen, total protein, and immunoglobulins among others was measured in different concentrations of cryoprecipitates. After activation with Ca-gluconate, fibrin membranes were produced in different thicknesses, and human mesenchymal stem cells were seeded onto the membranes. They were visualized by live-dead staining and their viability was determined by XTT. The platelet-derived growth factor AB content was quantified by ELISA. Our results showed that fibrinogen and platelet concentration can be multiplied in plasma by cryoprecipitate isolation, which affects the thickness and slightly the growth factor content of the membranes. According to live-dead staining, the thickness of the membranes does not influence cell attachment, and XTT measurement did not reveal a significant difference in cell attachment capacity either; however, a growing trend could be observed in the case of some membranes.
In this study, promising compositions of cross-linked gelatin-based scaffolds were tested in vitro and in vivo. Our aim was to utilize a solid matrix that is suitable for medical applications, and to be regulated as a medical device as a soft tissue implant. Three different cross-linkers were used in vitro, and the optimal composition was chosen for in vivo testing. The surfaces of the scaffolds were observed with SEM, and, in the case of divinyl sulfone (DVS), small cracks appeared, and the structure was rigid. With the use of poly(ethylene glycol) diglycidyl ether (PEGDE), the surface was found to be uneven, but generally, the appearance was similar in each case. The optimal scaffold contained 5 v/v % 1,4-butanediol diglycidyl ether (BDDE), and was implanted for either one month or three months in the back of BL6 mice. The explants were assessed using analytical techniques, including microscopic imaging and histological analysis, and it was found that cells, connective tissue, and extracellular matrix (ECM) were all able to successfully infiltrate the scaffolds and did not induce any inflammation. In summary, these novel implants seem to promote blood vessel formation and support the adherence of adipose tissue, as confirmed by optical microscopy and histological evaluations.