Bio-Membrane-Based Nanofiber Scaffolds: Targeted and Controlled Carriers for Drug Delivery-An Experimental In Vivo...

Cell population and vascular vessel distribution analysis in membrane-based scaffolds for tissue engineering is crucial. Biomimetic nanostructured membranes of methyl methacrylate/hydroxyethyl methacrylate and methyl acrylate/hydroxyethyl acrylate (MMA)1-co-(HEMA)1/(MA)3-co-(HEA)2 loaded with 5% wt SiO2-nanoparticles (Si-M) were doped with zinc (Zn-M) or doxycycline (Dox-M). Critical bone defects were effectuated on six New Zealand-bred rabbit skulls and then they were covered with the membrane-based scaffolds. After six weeks, bone cell population in terms of osteoblasts, osteoclasts, osteocytes, fibroblasts, and M1 and M2 macrophages and vasculature was determined. The areas of interest were the space above (over) and below (under) the membrane, apart from the interior (inner) compartment. All membranes showed that vasculature and most cell types were more abundant under the membrane than in the inner or above regions. Quantitatively, osteoblast density increased by approximately 35% in Zn-M and 25% in Si-M compared with Dox-M. Osteoclast counts decreased by about 78% in Dox-M, indicating strong inhibition of bone resorption. Vascular structures were nearly twofold more frequent under the membranes, particularly in Si-M, while fibroblast presence remained moderate and evenly distributed. The M1/M2 macrophage ratio was higher in Zn-M, reflecting a transient pro-inflammatory state, whereas Dox-M favored an anti-inflammatory, pro-regenerative profile. These results indicate that the biomimetic electrospun membranes functioned as architectural templates that provided favorable microenvironments for cell colonization, angiogenesis, and early bone regeneration in a preclinical in vivo model. Zn-M membranes appear suitable for early osteogenic stimulation, while Dox-M membranes may be advantageous in clinical contexts requiring modulation of inflammation and osteoclastic activity.