RESUMO
OBJECTIVE: Bone defects or atrophy may arise as a consequence of injury, inflammation of various etiologies, and neoplastic or traumatic processes or as a result of surgical procedures. Sometimes the regeneration process of bone loss is impaired, significantly slowed down, or does not occur, e.g., in congenital defects. For the bone defect reconstruction, a piece of the removed bone from ala of ilium or bone transplantation from a decedent is used. Replacement of the autologous or allogenic source of the bone-by-bone substitute could reduce the number of surgeries and time in the pharmacological coma during the reconstruction of the bone defect. Application of mesenchymal stem cells in the reconstruction surgery may have positive influence on tissue regeneration by secretion of angiogenic factors, recruitment of other MSCs, or differentiation into osteoblasts. Materials and Methods. Mesenchymal stem cells derived from the umbilical cord (Wharton's jelly (WJ-MSC)) were cultured in GMP-grade DMEM low glucose supplemented with heparin, 10% platelet lysate, glucose, and antibiotics. In vitro WJ-MSCs were seeded on the bone substitute Bio-Oss Collagen® and cultured in the StemPro® Osteogenesis Differentiation Kit. During the culture on the 1st, 7th, 14th, and 21st day (day in vitro (DIV)), we analyzed viability (confocal microscopy) and adhesion capability (electron microscopy) of WJ-MSC on Bio-Oss scaffolds, gene expression (qPCR), and secretion of proteins (Luminex). In vivo Bio-Oss® scaffolds with WJ-MSC were transplanted to trepanation holes in the cranium to obtain their overgrowth. The computed tomography was performed 7, 14, and 21 days after surgery to assess the regeneration. RESULTS: The Bio-Oss® scaffold provides a favourable environment for WJ-MSC survival. WJ-MSCs in osteodifferentiation medium are able to attach and proliferate on Bio-Oss® scaffolds. Results obtained from qPCR and Luminex® indicate that WJ-MSCs possess the ability to differentiate into osteoblast-like cells and may induce osteoclastogenesis, angiogenesis, and mobilization of host MSCs. In animal studies, WJ-MSCs seeded on Bio-Oss® increased the scaffold integration with host bone and changed their morphology to osteoblast-like cells. CONCLUSIONS: The presented construct consisted of Bio-Oss®, the scaffold with high flexibility and plasticity, approved for clinical use with seeded immunologically privileged WJ-MSC which may be considered reconstructive therapy in bone defects.
RESUMO
Brain infarct triggers neurodegeneration that often shades spontaneous plasticity, occurring in the areas related anatomically and functionally to the infarcted structures. Neurotrophins which promote neuronal survival and plasticity, may protect neurons and enhance remodeling of the remaining circuits, leading to restoration of function. In particular, the crucial role of brain-derived neurotrophic factor (BDNF) in cortical function is well documented. Since BDNF was implicated in the mechanism of postinfarct recovery, we investigated whether focal photothrombosis in the motor cortex of adult rats modifies cortical BDNF protein levels in a time- and region-dependent fashion. In parallel, we aimed to establish, which cortical cells respond with altered BDNF expression and whether these alterations are reflected by forelimb motor skill impairment and recovery, evaluated up to 1 month postinfarct. The distribution of BDNF protein was visualized immunohistochemically and BDNF tissue levels were evaluated with enzyme-linked immunosorbent assay (ELISA). Ipsilateral to the infarct, an increase in BDNF levels occurred both in injured and neighboring regions already 24 h after photothrombosis. This increase was sustained up to postlesion day 7 in the motor cortex and reduced at 28 days. No BDNF changes were detected in homotopic regions of the contralateral cortex. The time-course of enhanced neurotrophic expression was paralleled by bilateral deficits in skilled reaching, which was the only clear and measurable motor impairment observed in the study. We conclude that the spontaneous increase of BDNF is not sufficient to protect neurons from degeneration in the lesion proximity whereas plasticity reported in the adjacent regions may be attributable to enhanced BDNF-related stimuli, which do not counteract the impairment of skilled reaching but might be, at least in part, responsible for the absence of deficits in other functional/behavioral tests.
