Non-union fractures represent a significant clinical challenge requiring innovative therapeutic approaches. Silk fibroin (SF) scaffolds have gained recognition as advantageous biomaterials for bone tissue engineering due to their biocompatibility and mechanical characteristics. This study investigated the biocompatibility and osteoinductive potential of SF scaffolds functionalized with hydroxyapatite (HA) and loaded with platelet growth factors (PGFs) using hematopoietic stem cells (HSCs). SF scaffolds were prepared and functionalized with HA through methanol impregnation, while PGFs were obtained from platelet lysate via apheresis procedures. HSCs were cultured on different experimental groups, namely SF, SF-HA, PGF, SF-PGF, and SF-HA-PGF, assessing biocompatibility through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Live/Dead staining, and cytoskeleton analysis over 7 days. Osteoinductive properties were evaluated using Alizarin Red staining for mineral matrix deposition at 14 and 21 days. The MTT assay revealed the biocompatibility of all the experimental groups. The Live/Dead assay confirmed high cell viability, while the cytoskeleton analysis revealed well-organized actin filaments comparable to controls. Alizarin Red staining showed that PGF alone promoted early mineral matrix deposition at day 14, while SF-HA, SF-PGF, and SF-HA-PGF groups demonstrated significantly enhanced mineralization at day 21 compared with SF alone. The combination of silk fibroin scaffolds with platelet growth factors alone or with hydroxyapatite and platelet growth factors creates a biomimetic environment that supports cell viability and induces the osteogenic differentiation of hemopoietic stem cells. These findings suggest significant potential for clinical translation in treating non-union fractures and bone defects.

Concentrated growth factor (CGF) is an autologous blood-derived product widely used in regenerative medicine due to its high concentration of growth factors and platelets. In this study, the ability of primary stem cells isolated from human CGF to differentiate into adipocytes, endothelial cells, and neuronal-like cells was evaluated in vitro. CGF primary cells (CPCs) were obtained from CGF fragments and characterized after one month in culture. These cells were positive for the surface markers CD105, CD45, CD31, and CD14, and also expressed mRNA levels of the stemness markers Nanog and Oct3/4 comparable to human bone marrow mesenchymal stem cells (BMSCs). Results showed that, following appropriate differentiation protocols, CPCs, similarly to BMSCs, were able to differentiate into adipogenic, endothelial, and neuronal lineages, acquiring specific phenotypic and molecular markers. Adipogenic induction resulted in lipid accumulation and the upregulation of key genes, including PLIN2, FABP4, CD36, and FASN. Under pro-endothelial conditions, the cells exhibited increased expression of endothelial markers, eNOS, VEGFR-2, and CD31. Neuronal induction promoted the expression of β-tubulin III, Nestin, and Neurofilament. Overall, this work highlights the remarkable plasticity of CPCs and supports their potential application in multilineage regenerative therapies.

Background: Glioblastoma (GBM) is a highly aggressive tumor characterized by elevated plasticity and poor differentiation. Platelet-derived preparations such as Concentrated Growth Factors (CGF) are rich in bioactive molecules, but their effects on tumor biology remain underexplored. Methods: U87MG glioblastoma cells were cultured with a conditioned medium obtained from CGF over 14 days (CGF-CM). We analyzed cell viability, morphology, DNA integrity, migration, proliferation, and expression of astrocytic markers. Results: CGF-CM treatment induced early enhancement of cell viability, followed by decreased proliferation and reduced migration at later time points. Morphological analyses revealed astrocyte-like features. The expression of glial fibrillary acidic protein (GFAP), an astrocytic marker, and its α/δ isoform ratio increased over time, while GBM -GBM-associated markers, such as AQP-4 and S100B, were downregulated. Conclusions: Our findings demonstrate that CGF-CM modulates the phenotypic plasticity of U87MG cells and promotes differentiation toward an astroglial-like profile. These results provide a basis for future studies on the modulation of GBM aggressiveness using bioactive autologous derivatives.