Development of Tri-Layered Biomimetic Atelocollagen Scaffolds with Interfaces for Osteochondral Tissue Engineering.

The development of biomimetic scaffolds containing cartilage, calcified cartilage, and bone regeneration for precise osteochondral repair remains a challenge. Herein, a novel tri-layered scaffold-with a top layer containing type II atelocollagen and chondroitin sulphate for cartilage regeneration, an intermediate layer with type II atelocollagen and hydroxyapatite for calcified cartilage formation, and a bottom layer with type I atelocollagen and hydroxyapatite for bone growth-that can be built using liquid-phase cosynthesis, is described. The tri-layered scaffolds are mechanically demonstrably superior and have a lower risk of delamination than monolayer scaffolds. This is due to higher cohesion arising from the interfaces between each layer. In vitro results show that although monolayer scaffolds can stimulate bone marrow stem cells to differentiate and form cartilage, calcified cartilage, and bone separately (detected using quantitative polymerase chain reaction analysis and staining with safranin-O and Alizarin Red S), the tri-layered scaffolds support the regeneration of cartilage, calcified cartilage, and bone simultaneously after 2 and 4 months of implantation (detected using gross and micro-computed tomography images, histological staining, and Avizo, a software used to detect microlevel defects in metals). This work presents data on a promising approach in devising strategies for the precise repair of osteochondral defects.

Biomimetic Trachea Engineering via a Modular Ring Strategy Based on Bone-Marrow Stem Cells and Atelocollagen for Use in Extensive Tracheal Reconstruction.

The fabrication of biomimetic tracheas with a architecture of cartilaginous rings alternately interspersed between vascularized fibrous tissue (CRVFT) has the potential to perfectly recapitulate the normal tracheal structure and function. Herein, the development of a customized chondroitin-sulfate-incorporating type-II atelocollagen (COL II/CS) scaffold with excellent chondrogenic capacity and a type-I atelocollagen (COL I) scaffold to facilitate the formation of vascularized fibrous tissue is described. An efficient modular ring strategy is then adopted to develop a CRVFT-based biomimetic trachea. The in vitro engineering of cartilaginous rings is achieved via the recellularization of ring-shaped COL II/CS scaffolds using bone marrow stem cells as a mimetic for native cartilaginous ring tissue. A CRVFT-based trachea with biomimetic mechanical properties, composed of bionic biochemical components, is additionally successfully generated in vivo via the alternating stacking of cartilaginous rings and ring-shaped COL I scaffolds on a silicone pipe. The resultant biomimetic trachea with pedicled muscular flaps is used for extensive tracheal reconstruction and exhibits satisfactory therapeutic outcomes with structural and functional properties similar to those of native trachea. This is the first study to utilize stem cells for long-segmental tracheal cartilaginous regeneration and this represents a promising method for extensive tracheal reconstruction.

Metallic Ions Encapsulated in Electrospun Nanofiber for Antibacterial and Angiogenesis Function to Promote Wound Repair.

Electrospun nanofiber is an attractive biomaterial for skin tissue engineering because it mimics the natural fibrous extracellular matrix structure and creates a physical structure suitable for skin tissue regeneration. However, endowing the nanofibrous membranes with antibacterial and angiogenesis functions needs to be explored. In the current study, we aimed to fabricate gelatin/polycaprolactone (GT/PCL) (GT/PCL-Ag-Mg) nanofibers loaded with silver (Ag) and magnesium (Mg) ions for antibacterial activity and pro-angiogenesis function for wound repair. The fabricated GT/PCL membranes had a nanofibrous structure with random arrangement and achieved sustained release of Ag and Mg ions. In vitro results indicated that the GT/PCL-Ag-Mg membranes presented satisfactory cytocompatibility with cell survival and proliferation. In addition, the membranes with Ag demonstrated good antibacterial capacity to both gram-positive and gram-negative bacteria, and the Mg released from the membranes promoted the tube formation of vascular endothelial cells. Furthermore, in vivo results demonstrated that the GT/PCL-Ag-Mg membrane presented an accelerated wound healing process compared with GT/PCL membranes incorporated with either Ag or Mg ions and pure GT/PCL alone. Superior epidermis formation, vascularization, and collagen deposition were also observed in GT/PCL-Ag-Mg membrane compared with the other membranes. In conclusion, a multifunctional GT/PCL-Ag-Mg membrane was fabricated with anti-infection and pro-angiogenesis functions, serving as a potential metallic ion-based therapeutic platform for applications in wound repair.

A Biomimetic Biphasic Scaffold Consisting of Decellularized Cartilage and Decalcified Bone Matrixes for Osteochondral Defect Repair.

It is challenging to develop a biphasic scaffold with biomimetic compositional, structural, and functional properties to achieve concomitant repair of both superficial cartilage and subchondral bone in osteochondral defects (OCDs). This study developed a biomimsubchondraletic biphasic scaffold for OCD repair via an iterative layered lyophilization technique that controlled the composition, substrate stiffness, and pore size in each phase of the scaffold. The biphasic scaffold consisted of a superficial decellularized cartilage matrix (DCM) and underlying decalcified bone matrix (DBM) with distinct but seamlessly integrated phases that mimicked the composition and structure of osteochondral tissue, in which the DCM phase had relative low stiffness and small pores (approximately 134 µm) and the DBM phase had relative higher stiffness and larger pores (approximately 336 µm). In vitro results indicated that the biphasic scaffold was biocompatible for bone morrow stem cells (BMSCs) adhesion and proliferation, and the superficial DCM phase promoted chondrogenic differentiation of BMSCs, as indicated by the up-regulation of cartilage-specific gene expression (ACAN, Collagen II, and SOX9) and sGAG secretion; whereas the DBM phase was inducive for osteogenic differentiation of BMSCs, as indicated by the up-regulation of bone-specific gene expression (Collagen I, OCN, and RUNX2) and ALP deposition. Furthermore, compared with the untreated control group, the biphasic scaffold significantly enhanced concomitant repair of superficial cartilage and underlying subchondral bone in a rabbit OCD model, as evidenced by the ICRS macroscopic and O'Driscoll histological assessments. Our results demonstrate that the biomimetic biphasic scaffold has a good osteochondral repair effect.

Electrospun nanofibrous membrane of fish collagen/polycaprolactone for cartilage regeneration.

Tissue engineering technology provides a promising approach for cartilage repair, and in this strategy, scaffolds play a pivotal role in directing cartilage regeneration. Fish collagen (FC) is currently considered an alternative source of mammalian collagen (MC) for tissue engineering due to its excellent biocompatibility, suitable biodegradability, inert immunogenicity, rich sources, low price and lack of risk for the transmission of zoonosis. Here, we fabricated three types of electrospun nanofibrous membranes composed of FC and polycaprolactone (PCL) with three different FC/PCL ratios (9/1, 7/3, 5/5) and investigated the feasibility of using the membranes with chondrocytes in cartilage regeneration. Our results demonstrated that increases in the FC content were associated with improvements in biodegradability, absorption, and cell adhesion capacity, but weaker mechanical properties. In addition, all three nanofibrous membranes showed satisfactory biocompatibility as evidenced by supporting chondrocyte proliferation and cartilage formation in vitro. Furthermore, all three membranes seeded with chondrocytes formed mature cartilage-like tissue after 8 weeks of in vivo culture, but satisfactory homogeneous cartilage regeneration was only achieved with the F9P1 group. The current results demonstrated that the electrospun FC/PCL membrane is a promising scaffold for cartilage regeneration and that the F9P1 group might represent a relatively suitable ratio. The research models established in the current study provide detailed information for the regeneration of cartilage and other tissue based on electrospun FC/PCL membranes.