Enhancing Craniofacial Bone Reconstruction with Clinically Applicable 3D Bioprinted Constructs.

Medical imaging and 3D bioprinting can be used to create patient-specific bone scaffolds with complex shapes and controlled inner architectures. This study investigated the effectiveness of a biomimetic approach to scaffold design by employing geometric control. The biomimetic scaffold with a dense external layer showed improved bone regeneration compared to the control scaffold. New bone filled the defected region in the biomimetic scaffolds, while the control scaffolds only presented new bone at the boundary. Histological examination also shows effective bone regeneration in the biomimetic scaffolds, while fibrotic tissue ingrowth is observed in the control scaffolds. These findings suggest that the biomimetic bone scaffold, designed to minimize competition for fibrotic tissue formation in the bony defect, can enhance bone regeneration. This study underscores the notion that patient-specific anatomy can be accurately translated into a 3D bioprinting strategy through medical imaging, leading to the fabrication of constructs with significant clinical relevance.

Rapid mineralization of graphene-based 3D porous scaffolds by semi-dry electrodeposition for photothermal treatment of tumor-induced bone defects.

Graphene-based three-dimensional (3D) porous scaffolds have been extensively investigated in the photothermal treatment of tumor-induced bone defects due to their photothermal and osteogenic capacity. However, scaffold processing destroys conjugated graphene structure and reduces its photothermal conversion efficiency. In this study, a graphene-based 3D scaffold (GS) with intact conjugated structure was prepared by chemical vapor deposition (CVD). GS was rapidly mineralized biomimetically by a newly developed semi-dry electrochemical deposition method to form a hydroxyapatite (HA) incorporated graphene scaffold (HA-GS). The simulation of the charged particle dynamics provides a better understanding of the mechanism of semi-dry electrodeposition. This scaffold exhibits high photothermal sensitivity that generates sufficient thermal energy for photothermal therapy even under near-infrared irradiation (980 nm) with extremely low power density (0.2 W/cm2). Moreover, osteogenic activity was improved by HA-GS compared with GS. Compared with the blank GS, the HA-GS scaffold deposited with HA also showed regulation of macrophage-derived chemokine (MDC) and remodeled the immune microenvironment of the wound after photothermal therapy. In vivo experiments further verified that HA-GS can ablate osteosarcoma through a photothermal effect. These results suggest that the as-prepared HA-GS may be adopted as a promising multifunctional bone scaffold against tumor-induced bone defect. STATEMENT OF SIGNIFICANCE: The hydroxyapatite (HA) incorporated graphene scaffold (HA-GS) scaffold was prepared by semi-dry electrodeposition first time. The prepared HA-GS has a high photothermal conversion efficiency (it can rise to 48 °C under the 5 min irradiation of 980 nm near-infrared laser at 0.2 W/cm2). The mineralized layer prepared by semi-dry electrodeposition is not only osteoinductive, but also reduces the inflammatory response after photothermal therapy. This modulates the immune microenvironment at the bone tumor invasion site, thereby promoting defect repair.

Release Kinetics and In Vitro Characterization of Sodium Percarbonate and Calcium Peroxide to Oxygenate Bioprinted Tissue Models.

Oxygen-generating materials have been used in several tissue engineering applications; however, their application as in situ oxygen supply within bioprinted constructs has not been deeply studied. In this study, two oxygen-generating materials, sodium percarbonate (SPO) and calcium peroxide (CPO), were studied for their oxygen release kinetics under a 0.1% O2 condition. In addition, a novel cell-culture-insert setup was used to evaluate the effects of SPO and CPO on the viability of skeletal muscle cells under the same hypoxic condition. Results showed that SPO had a burst oxygen release, while CPO had a more stable oxygen release than SPO. Both SPO and CPO reduced cell viability when used alone. The addition of catalase in SPO and CPO increased the oxygen release rate, as well as improving the viability of skeletal muscle cells; however, CPO still showed cytotoxicity with catalase. Additionally, the utilization of 1 mg/mL SPO and 20 U catalase in a hydrogel for bioprinting significantly enhanced the cell viability under the hypoxic condition. Moreover, bioprinted muscle constructs could further differentiate into elongated myotubes when transferring back to the normoxic condition. This work provides an excellent in vitro model to test oxygen-generating materials and further discover their applications in bioprinting, where they represent promising avenues to overcome the challenge of oxygen shortage in bioprinted constructs before their complete vascularization.

Extrusion-Based Bioprinting: Current Standards and Relevancy for Human-Sized Tissue Fabrication.

The field of bioengineering has long pursued the goal of fabricating large-scale tissue constructs for use both in vitro and in vivo. Recent technological advances have indicated that bioprinting will be a key technique in manufacturing these specimens. This chapter aims to provide an overview of what has been achieved to date through the use of microextrusion bioprinters and what major challenges still impede progress. Microextrusion printer configurations will be addressed along with critical design characteristics including nozzle specifications and bioink modifications. Significant challenges within the field with regard to achieving long-term cell viability and vascularization, and current research that shows promise in mitigating these challenges in the near future are discussed. While microextrusion is a broad field with many applications, this chapter aims to provide an overview of the field with a focus on its applications toward human-sized tissue constructs.

Bioprinted trachea constructs with patient-matched design, mechanical and biological properties.

Tracheal stenosis is a rare but life-threatening disease. Primary clinical procedures for treating this disease are limited if the region requiring repair is long or complex. This study is the first of its kind to fabricate bioprinted tracheal constructs with separate cartilage and smooth muscle regions using polycaprolactone (PCL) and human mesenchymal stem cell (hMSC)-laden hydrogels. Our final bioprinted trachea showed comparable elastic modulus and yield stress compared to native tracheal tissue. In addition, both cartilage and smooth muscle formation were observed in the desired regions of our bioprinted trachea through immunohistochemistry and western blot after two weeks of in vitro culture. This study demonstrates a novel approach to manufacture tissue engineered trachea with mechanical and biological properties similar to native trachea, which represents a step closer to overcoming the clinical challenges of treating tracheal stenosis.

A 3D bioprinting system to produce human-scale tissue constructs with structural integrity.

A challenge for tissue engineering is producing three-dimensional (3D), vascularized cellular constructs of clinically relevant size, shape and structural integrity. We present an integrated tissue-organ printer (ITOP) that can fabricate stable, human-scale tissue constructs of any shape. Mechanical stability is achieved by printing cell-laden hydrogels together with biodegradable polymers in integrated patterns and anchored on sacrificial hydrogels. The correct shape of the tissue construct is achieved by representing clinical imaging data as a computer model of the anatomical defect and translating the model into a program that controls the motions of the printer nozzles, which dispense cells to discrete locations. The incorporation of microchannels into the tissue constructs facilitates diffusion of nutrients to printed cells, thereby overcoming the diffusion limit of 100-200 µm for cell survival in engineered tissues. We demonstrate capabilities of the ITOP by fabricating mandible and calvarial bone, cartilage and skeletal muscle. Future development of the ITOP is being directed to the production of tissues for human applications and to the building of more complex tissues and solid organs.