Guided tissue regeneration (GTR) is a promising treatment for periodontal tissue defects, which generally uses a membrane to build a mechanical barrier from the gingival epithelium and hold space for the periodontal regeneration especially the tooth-supporting bone. However, existing membranes possess insufficient mechanical properties and limited bioactivity for periodontal bone regenerate. Herein, fish collagen and polyvinyl alcohol (Col/PVA) dual-layer membrane were developed via a combined freezing/thawing and layer coating method. This dual-layer membrane had a clear but contact boundary line between collagen and PVA layers, which were both hydrophilic. The dual membrane had an elongation at break of 193 ± 27% and would undergo an in vitro degradation duration of more than 17 days. Further cell experiments showed that compared with the PVA layer, the collagen layer not only presented good cytocompatibility with rat bone marrow-derived mesenchymal stem cells (BMSCs), but also promoted the osteogenic genes (RUNX2, ALP, OCN, and COL1) and protein (ALP) expression of BMSCs. Hence, the currently developed dual-layer membranes could be used as a stable barrier with a stable degradation rate and selectively favor the bone tissue to repopulate the periodontal defect. The membranes could meet the challenges encountered by GTR for superior defect repair, demonstrating great potential in clinical applications.

Hydrogel microspheres have been widely used as cell carriers and 3D cell culture matrices. However, these microspheres are associated with several unfavorable properties for bone tissue engineering applications, e.g. their surface is too smooth to attach cells and they do not contain inorganic materials. This manuscript presents a new method to overcome these disadvantages by depositing CaCO3 crystals on the hydrogel microsphere surface. Specifically, we used a non-planar flow-focusing microfluidic device to produce Gelatin Methacrylate (GelMA)/Na2CO3 based microspheres. We subsequently obtained CaCO3 crystals via a chemical reaction between Na2CO3 and CaCl2. The efficacy of this method was demonstrated by in vitro experiments with human umbilical vein endothelial cells (HUVECs) and immortalized mouse embryonic fibroblast (iMEF). Cell culture on GelMA/CaCO3 microspheres showed that cells can easily attach and adhere to GelMA/CaCO3 microspheres and maintain high viability. Alkaline phosphatase (ALP) expression was increased as well. These results suggest that this novel microsphere has a high potential for bone tissue engineering applications.

The current study, inspired by the immunosuppressive property of rapamycin (Rapa) and the benefit of microspheres both as drug delivery system and cell carriers, was designed to develop an efficient Rapa delivery system with tunable controllability to facilitate its local administration. A capillary-based two-phase microfluidic device was designed to prepare monodisperse poly(lactide-co-glycolide) (PLGA) microspheres to load Rapa (PLGA-Rapa-M). The physical and chemical properties of PLGA-Rapa-M were characterized, and the Rapa loading capacity and release profile were explored. Chondrocytes were chosen as a cell model to evaluate the adhesion and proliferation on these microspheres. Controllability over the microsphere properties was illustrated. The PLGA-Rapa-M is averagely 63.91 μm in size with a narrow size distribution and a CV of 2.44%. The encapsulation efficiency of Rapa within microspheres via the current microfluidics was around 98%, and Rapa loading could be easily varied with a maximum value of ∼20%. The PLGA-Rapa-M has a sustained Rapa release duration of ∼3 months. These microspheres could not only successfully be used for Rapa sustained release but also as cell carriers for cell therapy since they can support the attachment/proliferation of chondrocytes. Hence, improved therapeutic index could be expected by using the current developed Rapa-release system.