Do nutraceuticals boost neuronal network formation in vitro? MAP2-postive neurons are stained in red and GFAP-positive astrocytes in green. Courtesy of Federica Ruggeri.

In particular, we investigate the neuroprotective effects of human milk components for preterm infants, with a specific focus on human milk oligosaccharides (HMOS). By taking a bench-to-bedside approach, we study the therapeutic effects of HMOS in animal models for encephalopathy of prematurity and in preterm infants we aim to link neuroprotective human milk components to neurodevelopmental outcome.

^{A DHA-supplemented diet reduced lesion size and neuroinflammation specifically in males. In both sexes, brain n-3 fatty acids were increased after receiving the experimental diet. The experimental diet also improved novel object recognition, but no significant effects on motor performance were observed. Current data indicates that early life DHA supplementation may provide neuroprotection after perinatal HI.}

Another important component of human milk, docosahexaenoic acid (DHA) is an important building block for the brain. DHA serves an important role in axonal outgrowth and synaptogenesis of neurons, particularly during the first 1000 days of life. Therefore, we investigate the therapeutic effects of DHA on neonatal brain injury, as sole therapy, and also in combination with regenerative therapies with the aim to provide building blocks to repair the injured brain (see also the research line Regenerative strategies for the injured neonatal brain).

^{The effects of (milk-derived) nutrients and bioactive compounds on important processes for brain development that unfold during the neonatal period. HMOs = human milk oligosaccharides, MSCs = mesenchymal stem cells. ©Drawing by Nynke van de Haar, copyright permission has been obtained. Courtesy of Myrna Brandt.}

In addition to nutritional building blocks, milk contains components that play an active role in the regulation of the development of the new-born. One of these bioactive components are cell-derived extracellular vesicles (EVs), which are vehicles for cell-cell communication. Milk EVs are interesting candidates for therapeutic application as they have anti-inflammatory and regenerative capacities and have the potential to cross the blood brain barrier. In this ZonWM OPEN funded project, we aim to decipher the anti-inflammatory and regenerative effects of milk EVs on the injured preterm brain and investigate their therapeutic potential. Taking a bench-to-bedside approach, we will investigate in vitro functional effects and targeting of milk EVs to different neural cell types and connect these findings to the milk EV cargo analyzed by multi-omics approaches in order to unravel the molecular mechanisms of milk EV communication to neural cells. In vivo, the migratory capacity of milk EVs and therapeutic efficacy of intranasal milk EV administration will be determined in a preclinical mouse model for preterm brain injury. In the clinic, we will assess the feasibility ofintranasal administration of mother's own EV-containing milk plasma in preterm infants with brain injury (post-hemorrhagic ventricular dilatation) and investigate the effects on neuroinflammatory and neuroregenerative markers in cerebrospinal fluid. We expect these mechanistic, efficacy, and feasibility studies on milk EV-neural cell communication to deliver a protocol for the first-in-human efficacy trial to define the effects of intranasal milk EV therapy on preterm brain injury.

^{Overview of the postulated intranasal routes of milk EV delivery to the brain, neural target cells, and EV cargo that could promote neural cells to protect the brain from EoP in preterm infants.}