Coating Chitosan Thin Shells: A Facile Technique to Improve Dispersion Stability of Magnetoliposomes.

Magnetoliposomes (ML) have been emerging as a novel multifunctional nanoparticle with a wide range of biomedical and therapeutic applications over the past decade. Although the ML system has shown excellent performances, the stability and lipid peroxidation of liposomal components are still remaining as key issues and need to be solved for intensive applications. Changing zeta potential of nanoparticles' surface can be seen as a potential way to achieve the stable dispersion. In this work, we have employed the positive charged, abundant and cheap chitosan to coat ML in order to change the zeta potential of the ML system and examined the stability of chitosan@magnetoliposomes (CML) in long-term storage. The combining of pH-sensitive chitosan with temperature-sensitive phospholipid formed a novel pH- and temperature-sensitive nanoparticles which can be promisingly used as controllable drug release applications. These novel CML with chitosan thin shells showed excellent stability in long-term storage; meanwhile, the bare ML sample showed aggregations and forming micrometer-size particles. The CML system can achieve a drug encapsulation efficiency of nearly 50% and an enhanced drug release behavior under pH 5 at 45 °C.

In vitro study on apoptotic cell death by effective magnetic hyperthermia with chitosan-coated MnFe2O4.

Magnetic nanoparticles (MNPs) have been widely investigated as a hyperthermic agent for cancer treatment. In this study, thermally responsive Chitosan-coated MnFe2O4 (Chitosan-MnFe2O4) nanoparticles were developed to conduct localized magnetic hyperthermia for cancer treatment. Hydrophobic MnFe2O4 nanoparticles were synthesized via thermal decomposition and modified with 2,3-dimercaptosuccinic acid (DMSA) for further conjugation of chitosan. Chitosan-MnFe2O4 nanoparticles exhibited high magnetization and excellent biocompatibility along with low cell cytotoxicity. During magnetic hyperthermia treatment (MHT) with Chitosan-MnFe2O4 on MDA-MB 231 cancer cells, the targeted therapeutic temperature was achieved by directly controlling the strength of the external AC magnetic fields. In vitro Chitosan-MnFe2O4-assisted MHT at 42 °C led to drastic and irreversible changes in cell morphology and eventual cellular death in association with the induction of apoptosis through heat dissipation from the excited magnetic nanoparticles. Therefore, the Chitosan-MnFe2O4 nanoparticles with high biocompatibility and thermal capability can be an effective nano-mediated agent for MHT on cancer.

A literature review of bioactive substances for the treatment of periodontitis: In vitro, in vivo and clinical studies.

Periodontitis is a common chronic inflammatory disease of the supporting tissues of the tooth that involves a complex interaction of microorganisms and various cell lines around the infected site. To prevent and treat this disease, several options are available, such as scaling, root planning, antibiotic treatment, and dental surgeries, depending on the stage of the disease. However, these treatments can have various side effects, including additional inflammatory responses, chronic wounds, and the need for secondary surgery. Consequently, numerous studies have focused on developing new therapeutic agents for more effective periodontitis treatment. This review explores the latest trends in bioactive substances with therapeutic effects for periodontitis using various search engines. Therefore, this study aimed to suggest effective directions for therapeutic approaches. Additionally, we provide a summary of the current applications and underlying mechanisms of bioactive substances, which can serve as a reference for the development of periodontitis treatments.

Cellulose nanofibrils reinforced chitosan-gelatin based hydrogel loaded with nanoemulsion of oregano essential oil for diabetic wound healing assisted by low level laser therapy.

Diabetic foot ulcers are imperfections in the process of wound healing due to hyperglycemic conditions. Here, a nanoemulgel fabricated with oregano essential oil nanoemulsion, assisted by low-level laser therapy, was investigated for its efficacy in diabetic wound healing. A hydrogel- based healing patch, fabricated using biological polymers namely chitosan and gelatin and, polyvinyl pyrollidone. The hydrogel was reinforced with cellulose nanofibrils for enhanced stability and barrier properties. Nanoemulsion of oregano essential oil, with an average particle size of 293.7 ± 8.3 nm, was prepared via homogenization with chitosan as the coating agent. Nanoemulsion impregnated hydrogel, termed as the nanoemulgel, was assessed for its physio-mechanical properties and healing efficiency. The strong linkages in nanoemulgel demonstrated its large swelling capacity, high mechanical strength, and maximum thermal stability. The optimized conditions for low-level laser therapy using 808 nm were 1 W. cm-2 and 5 min. The optimized drug concentration of 128 µg. mL-1 exhibited viability of NIH/3 T3 fibroblasts as 75.5 ± 1.2 % after 24 h. Cell migration assay demonstrated that dual therapy facilitated wound healing, with a maximum closure rate of 100 % at 48 h. In vivo results revealed the rapid healing effects of the dual therapy in diabetic rat models with foot ulcers: a maximum healing rate of 97.5 %, minimum scar formation, increased granulation, enhanced reepithelialization, and a drastic decrease in inflammation and neutrophil infiltration within the treatment period compared to monotherapy and control. In summary, the combinatorial therapy of nanoemulgel and low-level laser therapy is a promising regimen for managing diabetic foot ulcers with a rapid healing effect.

Facile synthesis of black phosphorus-zinc oxide nanohybrids for antibacterial coating of titanium surface.

Bacterial infection is a major complication associated with bioimplant materials, including titanium (Ti) based orthopedic joints and dental implants. Thus, the fabrication of Ti surfaces with antibacterial activity is highly important. Black phosphorus (BP) is a recently discovered promising two-dimensional semiconductor for various biomedical applications due to its tunable bandgap and physicochemical properties. The present study aimed to synthesize zinc oxide (ZnO) laden BP nanohybrids (NH) and their coatings on a Ti bioimplant surface for improving the antibacterial activities against pathogenic bacteria with and without near-infrared (NIR) light irradiation. Nanohybrids were produced with the slightly oxidized BP NF and electrostatically laden ZnO NP. The produced BP-ZnO NH was a NIR active nanomaterial (up to ∼1000 nm), demonstrating a photothermal effect against bacterial infection and showing improved activity by damaging the cell membrane towards S. aureus in comparison to E. coli. Ti surface coated with BP-ZnO NH embedded chitosan (CS) demonstrated better antibacterial activity than BP NF, especially with NIR light treatment. Additionally, the produced BP nanoflakes and BP-ZnO NH, and their coatings over the Ti surface were found to be toxic at a negligible level. Electrochemical studies revealed the high corrosion resistance of the Ti surface coated with the synthesized antibacterial agents without altering its characteristic passive behavior. Owing to the interactions between the charged groups between chitosan and cell surfaces, a slight increase in antibacterial activities was noticed. Chitosan-based coating matrix embedded with nanoagents has adhered well over the Ti surface due to its inherent film-forming and high adhesion properties.

Demineralized Dentin Matrix Particle-Based Bio-Ink for Patient-Specific Shaped 3D Dental Tissue Regeneration.

Demineralized dentin matrix (DDM)-based materials have been actively developed and are well-known for their excellent performance in dental tissue regeneration. However, DDM-based bio-ink suitable for fabrication of engineered dental tissues that are patient-specific in terms of shape and size, has not yet been developed. In this study, we developed a DDM particle-based bio-ink (DDMp bio-ink) with enhanced three-dimensional (3D) printability. The bio-ink was prepared by mixing DDM particles and a fibrinogen-gelatin mixture homogeneously. The effects of DDMp concentration on the 3D printability of the bio-ink and dental cell compatibility were investigated. As the DDMp concentration increased, the viscosity and shear thinning behavior of the bio-ink improved gradually, which led to the improvement of the ink's 3D printability. The higher the DDMp content, the better were the printing resolution and stacking ability of the 3D printing. The printable minimum line width of 10% w/v DDMp bio-ink was approximately 252 µm, whereas the fibrinogen-gelatin mixture was approximately 363 µm. The ink's cytocompatibility test with dental pulp stem cells (DPSCs) exhibited greater than 95% cell viability. In addition, as the DDMp concentration increased, odontogenic differentiation of DPSCs was significantly enhanced. Finally, we demonstrated that cellular constructs with 3D patient-specific shapes and clinically relevant sizes could be fabricated through co-printing of polycaprolactone and DPSC-laden DDMp bio-ink.

Cellulose nanocrystals/nanofibrils loaded astaxanthin nanoemulsion for the induction of apoptosis via ROS-dependent mitochondrial dysfunction in cancer cells under photobiomodulation.

The present study investigated effects of low-level laser therapy with cellulose nanocrystals/cellulose nanofibrils loaded in nanoemulsion (NE) against skin cancer cells on apoptosis. The nanoemulsion was fabricated and characterized by the standard methods. The toxicity level by cytotoxicity assays, generation of reactive singlet oxygen (ROS) and antioxidant potential, cell proliferation and migration were confirmed by using standard assays. The cellular uptake efficacy was evaluated by differential staining. The protein levels of EGFR, PI3K, AKT, ERK, GAPDH, and ß-actin were detected by western blot. The samples showed a spherical shaped structure with the average size confirmed strong and stable hydrogen bonding forces with high degradation temperature and endothermic transition peaks. The fabricated samples showed no toxicity and high cell proliferation by generating more singlet oxygen levels and antioxidants. The intracellular signaling pathways was regulated with high protein expression levels, which was stimulated by specific molecules for cell proliferation, migration, and differentiation in cancer cells. The results proved that combined treatment regulated the intracellular signaling pathways in cancer cells. The current study showed a novel strategy for improving therapeutic efficacy of nanoemulsion by using low-level laser therapy. Further, the current favorable outcomes will be evaluated in in vivo animal models.

In Vitro Mechanical and Biological Properties of 3D Printed Polymer Composite and ß-Tricalcium Phosphate Scaffold on Human Dental Pulp Stem Cells.

3D printed biomaterials have been extensively investigated and developed in the field of bone regeneration related to clinical issues. However, specific applications of 3D printed biomaterials in different dental areas have seldom been reported. In this study, we aimed to and successfully fabricated 3D poly (lactic-co-glycolic acid)/ß-tricalcium phosphate (3D-PLGA/TCP) and 3D ß-tricalcium phosphate (3D-TCP) scaffolds using two relatively distinct 3D printing (3DP) technologies. Conjunctively, we compared and investigated mechanical and biological responses on human dental pulp stem cells (hDPSCs). Physicochemical properties of the scaffolds, including pore structure, chemical elements, and compression modulus, were characterized. hDPSCs were cultured on scaffolds for subsequent investigations of biocompatibility and osteoconductivity. Our findings indicate that 3D printed PLGA/TCP and ß-tricalcium phosphate (ß-TCP) scaffolds possessed a highly interconnected and porous structure. 3D-TCP scaffolds exhibited better compressive strength than 3D-PLGA/TCP scaffolds, while the 3D-PLGA/TCP scaffolds revealed a flexible mechanical performance. The introduction of 3D structure and ß-TCP components increased the adhesion and proliferation of hDPSCs and promoted osteogenic differentiation. In conclusion, 3D-PLGA/TCP and 3D-TCP scaffolds, with the incorporation of hDPSCs as a personalized restoration approach, has a prospective potential to repair minor and critical bone defects in oral and maxillofacial surgery, respectively.

Fabrication and biological activity of polycaprolactone/phlorotannin endotracheal tube to prevent tracheal stenosis: An in vitro and in vivo study.

Prolonged endotracheal intubation is the most common cause of tracheal stenosis, which may lead to serious airway obstruction. Development of an endotracheal tube coated with biomaterials that exhibit anti-inflammatory or anti-fibrogenic effects may prevent tracheal stenosis. This study demonstrates that an endotracheal tube coated with phlorotannin, which is present in extracts of the brown alga Ecklonia cava, can prevent tracheal stenosis in a rabbit model. An in vitro study shows that phlorotannin inhibits proliferation of human tracheal fibroblasts treated with transforming growth factor ß1. Phlorotannin-coated endotracheal tubes show steady release of phlorotannin for up to 7 days, and removal of the tube 1 week after insertion reveals a reduction in both fibrogenesis and thickening of tracheal submucosa. Western blot analysis of tracheal tissues after removal of the phlorotannin-coated tube shows decreased protein expression levels of phenotypic markers of fibrosis such as collagen type I and α-smooth muscle actin. The ability of phlorotannin-coated endotracheal tube to prevent tracheal stenosis caused by endotracheal intubation indicates that phlorotannin may be considered as a candidate biomaterial for coating the cuff of endotracheal tubes to prevent tracheal stenosis.

Bioprinting of three-dimensional dentin-pulp complex with local differentiation of human dental pulp stem cells.

Numerous approaches have been introduced to regenerate artificial dental tissues. However, conventional approaches are limited when producing a construct with three-dimensional patient-specific shapes and compositions of heterogeneous dental tissue. In this research, bioprinting technology was applied to produce a three-dimensional dentin-pulp complex with patient-specific shapes by inducing localized differentiation of human dental pulp stem cells within a single structure. A fibrin-based bio-ink was designed for bioprinting with the human dental pulp stem cells. The effects of fibrinogen concentration within the bio-ink were investigated in terms of printability, human dental pulp stem cell compatibility, and differentiation. The results show that micro-patterns with human dental pulp stem cells could be achieved with more than 88% viability. Its odontogenic differentiation was also regulated according to the fibrinogen concentration. Based on these results, a dentin-pulp complex having patient-specific shape was produced by co-printing the human dental pulp stem cell-laden bio-inks with polycaprolactone, which is a bio-thermoplastic used for producing the overall shape. After culturing with differentiation medium for 15 days, localized differentiation of human dental pulp stem cells in the outer region of the three-dimensional cellular construct was successfully achieved with localized mineralization. This result demonstrates the possibility to produce patient-specific composite tissues for tooth tissue engineering using three-dimensional bioprinting technology.

Investigation of thermal degradation with extrusion-based dispensing modules for 3D bioprinting technology.

Recently, numerous three-dimensional (3D) bioprinting systems have been introduced for the artificial regeneration of tissues. Among them, the extrusion-based dispensing module is the most widely used because of the processability it gives various biomaterials. The module uses high forces and temperature to dispense materials through a micro-nozzle. Generally, the harsh conditions induce thermal degradation of the material in the dispensing procedure. The thermal degradation affects the properties of the materials, and the change of the properties should be carefully controlled, because it severely affects the regeneration of tissues. Therefore, in this research, the relationship between the dispensing module and the thermal degradation of material was investigated. Extrusion-based dispensing modules can be divided into the syringe type (ST) and filament type (FT) based on working principles. We prepared a poly lactic-co-glycolic acid (PLGA) scaffold with the two methods at various time points. Then, the characteristics of the printed scaffolds were assessed by measuring molecular weight (M w), glass transition temperature (T g), in vitro degradation, compressive modulus, and cytocompatibility. The results showed that the PLGA scaffold with the FT dispensing module maintained its properties regardless of printing time points. In contrast, severe thermal degradation was observed in the scaffold group prepared by the ST dispensing module. Consequentially, it was obvious that the FT dispensing module was more suitable for producing scaffolds without severe thermal degradation.

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.

Bioprinting Cellularized Constructs Using a Tissue-specific Hydrogel Bioink.

Bioprinting has emerged as a versatile biofabrication approach for creating tissue engineered organ constructs. These constructs have potential use as organ replacements for implantation in patients, and also, when created on a smaller size scale as model "organoids" that can be used in in vitro systems for drug and toxicology screening. Despite development of a wide variety of bioprinting devices, application of bioprinting technology can be limited by the availability of materials that both expedite bioprinting procedures and support cell viability and function by providing tissue-specific cues. Here we describe a versatile hyaluronic acid (HA) and gelatin-based hydrogel system comprised of a multi-crosslinker, 2-stage crosslinking protocol, which can provide tissue specific biochemical signals and mimic the mechanical properties of in vivo tissues. Biochemical factors are provided by incorporating tissue-derived extracellular matrix materials, which include potent growth factors. Tissue mechanical properties are controlled combinations of PEG-based crosslinkers with varying molecular weights, geometries (linear or multi-arm), and functional groups to yield extrudable bioinks and final construct shear stiffness values over a wide range (100 Pa to 20 kPa). Using these parameters, hydrogel bioinks were used to bioprint primary liver spheroids in a liver-specific bioink to create in vitro liver constructs with high cell viability and measurable functional albumin and urea output. This methodology provides a general framework that can be adapted for future customization of hydrogels for biofabrication of a wide range of tissue construct types.

Bioprinted Scaffolds for Cartilage Tissue Engineering.

Researchers are focusing on bioprinting technology as a viable option to overcome current difficulties in cartilage tissue engineering. Bioprinting enables a three-dimensional (3-D), free-form, computer-designed structure using biomaterials, biomolecules, and/or cells. The inner and outer shape of a scaffold can be controlled by this technology with great precision. Here, we introduce a hybrid bioprinting technology that is a co-printing process of multiple materials including high-strength synthetic polymer and cell-laden hydrogel. The synthetic polymer provides mechanical support for shape maintenance and load bearing, while the hydrogel provides the biological environment for artificial cartilage regeneration. This chapter introduces the procedures for printing of a 3-D scaffold using our hybrid bioprinting technology and includes the source materials for preparation of 3-D printing.

A 3D bioprinted complex structure for engineering the muscle-tendon unit.

Three-dimensional integrated organ printing (IOP) technology seeks to fabricate tissue constructs that can mimic the structural and functional properties of native tissues. This technology is particularly useful for complex tissues such as those in the musculoskeletal system, which possess regional differences in cell types and mechanical properties. Here, we present the use of our IOP system for the processing and deposition of four different components for the fabrication of a single integrated muscle-tendon unit (MTU) construct. Thermoplastic polyurethane (PU) was co-printed with C2C12 cell-laden hydrogel-based bioink for elasticity and muscle development on one side, while poly(ϵ-caprolactone) (PCL) was co-printed with NIH/3T3 cell-laden hydrogel-based bioink for stiffness and tendon development on the other. The final construct was elastic on the PU-C2C12 muscle side (E = 0.39 ± 0.05 MPa), stiff on the PCL-NIH/3T3 tendon side (E = 46.67 ± 2.67 MPa) and intermediate in the interface region (E = 1.03 ± 0.14 MPa). These constructs exhibited >80% cell viability at 1 and 7 d after printing, as well as initial tissue development and differentiation. This study demonstrates the versatility of the IOP system to create integrated tissue constructs with region-specific biological and mechanical characteristics for MTU engineering.

A hydrogel bioink toolkit for mimicking native tissue biochemical and mechanical properties in bioprinted tissue constructs.

Advancement of bioprinting technology is limited by the availability of materials that both facilitate bioprinting logistics as well as support cell viability and function by providing tissue-specific cues. Herein we describe a modular hyaluronic acid (HA) and gelatin-based hydrogel toolbox comprised of a 2-crosslinker, 2-stage polymerization technique, and the capability to provide tissue specific biochemically and mechanically accurate signals to cells within biofabricated tissue constructs. First, we prepared and characterized several tissue-derived decellularized extracellular matrix-based solutions, which contain complex combinations of growth factors, collagens, glycosaminoglycans, and elastin. These solutions can be incorporated into bioinks to provide the important biochemical cues of different tissue types. Second, we employed combinations of PEG-based crosslinkers with varying molecular weights, geometries (linear, 4-arm, and 8-arm), and functional groups to yield hydrogel bioinks that supported extrusion bioprinting and the capability to achieve final construct shear stiffness values ranging from approximately 100 Pa to 20 kPa. Lastly, we integrated these hydrogel bioinks with a 3-D bioprinting platform, and validated their use by bioprinting primary liver spheroids in a liver-specific bioink to create in vitro liver constructs with high cell viability and measurable functional albumin and urea output. This hydrogel bioink system has the potential to be a versatile tool for biofabrication of a wide range of tissue construct types. STATEMENT OF SIGNIFICANCE: Biochemical and mechanical factors both have important implications in guiding the behavior of cells in vivo, yet both realms are rarely considered together in the context of biofabrication in vitro tissue construct models. We describe a modular hydrogel system that (1) facilitates extrusion bioprinting of cell-laden hydrogels, (2) incorporates tissue-specific factors derived from decellularized tissue extracellular matrix, thus mimicking biochemical tissue profile, and (3) allows control over mechanical properties to mimic the tissue stiffness. We believe that employing this technology to attend to both the biochemical and mechanical profiles of tissues, will allow us to more accurately recapitulate the in vivo environment of tissues while creating functional 3-D in vitro tissue constructs that can be used as disease models, personalized medicine, and in vitro drug and toxicology screening systems.

Indirect three-dimensional printing of synthetic polymer scaffold based on thermal molding process.

One of the major issues in tissue engineering has been the development of three-dimensional (3D) scaffolds, which serve as a structural template for cell growth and extracellular matrix formation. In scaffold-based tissue engineering, 3D printing (3DP) technology has been successfully applied for the fabrication of complex 3D scaffolds by using both direct and indirect techniques. In principle, direct 3DP techniques rely on the straightforward utilization of the final scaffold materials during the actual scaffold fabrication process. In contrast, indirect 3DP techniques use a negative mold based on a scaffold design, to which the desired biomaterial is cast and then sacrificed to obtain the final scaffold. Such indirect 3DP techniques generally impose a solvent-based process for scaffold fabrication, resulting in a considerable increase in the fabrication time and poor mechanical properties. In addition, the internal architecture of the resulting scaffold is affected by the properties of the biomaterial solution. In this study, we propose an advanced indirect 3DP technique using projection-based micro-stereolithography and an injection molding system (IMS) in order to address these challenges. The scaffold was fabricated by a thermal molding process using IMS to overcome the limitation of the solvent-based molding process in indirect 3DP techniques. The results indicate that the thermal molding process using an IMS has achieved a substantial reduction in scaffold fabrication time and has also provided the scaffold with higher mechanical modulus and strength. In addition, cell adhesion and proliferation studies have indicated no significant difference in cell activity between the scaffolds prepared by solvent-based and thermal molding processes.

Annealing-free, flexible silver nanowire-polymer composite electrodes via a continuous two-step spray-coating method.

For the realization of high-efficiency flexible optoelectronic devices, transparent electrodes should be fabricated through a low-temperature process and have the crucial feature of low surface roughness. In this paper, we demonstrated a two-step spray-coating method for producing large-scale, smooth and flexible silver nanowire (AgNW)-poly3,4-ethylenedioxythiophene:polystyrenesulfonate (PEDOT:PSS) composite electrodes. Without the high-temperature annealing process, the conductivity of the composite film was improved via the lamination of highly conductive PEDOT:PSS modified by dimethyl sulfoxide (DMSO). Under the room temperature process condition, we fabricated the AgNW-PEDOT:PSS composite film showing an 84.3% mean optical transmittance with a 10.76 Ω sq(-1) sheet resistance. The figure of merit Φ(TC) was higher than that obtained from the indium tin oxide (ITO) films. The sheet resistance of the composite film slightly increased less than 5.3% during 200 cycles of tensile and compression folding, displaying good electromechanical flexibility for use in flexible optoelectronic applications.

Microstereolithography-based computer-aided manufacturing for tissue engineering.

Various solid freeform fabrication technologies have been introduced for constructing three-dimensional (3-D) freeform structures. Of these, microstereolithography (MSTL) technology performs the best in 3-D space because it not only has high resolution, but also fast fabrication speed. Using this technology, 3-D structures with mesoscale size and microscale resolution are achievable. Many researchers have been trying to apply this technology to tissue engineering to construct medically applicable scaffolds, which require a 3-D shape that fits a defect with a mesoscale size and microscale inner architecture for efficient regeneration of artificial tissue. This chapter introduces the principles of MSTL technology and representative systems. It includes fabrication and computer-aided design/computer-aided manufacturing (CAD/CAM) processes to show the automation process by which measurements from medical images are used to fabricate the required 3-D shape. Then, various tissue engineering applications based on MSTL are summarized.

Development of an indirect stereolithography technology for scaffold fabrication with a wide range of biomaterial selectivity.

Tissue engineering, which is the study of generating biological substitutes to restore or replace tissues or organs, has the potential to meet current needs for organ transplantation and medical interventions. Various approaches have been attempted to apply three-dimensional (3D) solid freeform fabrication technologies to tissue engineering for scaffold fabrication. Among these, the stereolithography (SL) technology not only has the highest resolution, but also offers quick fabrication. However, a lack of suitable biomaterials is a barrier to applying the SL technology to tissue engineering. In this study, an indirect SL method that combines the SL technology and a sacrificial molding process was developed to address this challenge. A sacrificial mold with an inverse porous shape was fabricated from an alkali-soluble photopolymer by the SL technology. A sacrificial molding process was then developed for scaffold construction using a variety of biomaterials. The results indicated a wide range of biomaterial selectivity and a high resolution. Achievable minimum pore and strut sizes were as large as 50 and 65 µm, respectively. This technology can also be used to fabricate three-dimensional organ shapes, and combined with traditional fabrication methods to construct a new type of scaffold with a dual-pore size. Cytotoxicity tests, as well as nuclear magnetic resonance and gel permeation chromatography analyses, showed that this technology has great potential for tissue engineering applications.