Enhanced Osteogenesis in 2D and 3D Culture Systems Using RGD Peptide and α-TCP Phase Transition within Alginate-Based Hydrogel.

Cell-laden hydrogels have been extensively investigated in various tissue engineering fields by their potential capacity to deposit numerous types of cells in a specific area. They are largely used in soft-tissue engineering applications because of their low mechanical strength. In addition, sodium alginate is well-known for its encapsulation, loading capacity and for being easily controllable; however, it lacks cell-binding ligands and hence the ability to adhere cells. In this study, it is aimed to enhance osteogenesis in cells encapsulated in alginate and improve its mechanical properties by introducing a synthetic peptide and calcium phosphate phase transition. To increase cell-hydrogel interactions and increasing cell viability, an RGD peptide is added to a photocrosslinkable methacrylate-modified alginate, and alpha-tricalcium phosphate (α-TCP) is added to the hydrogel to increase its mechanical strength via phase transition. Cell proliferation, growth, and differentiation are assessed in both 2D and 3D cell cultures. The addition of α-TCP significantly improved the mechanical properties of the hydrogel. Moreover, the RGD peptide and α-TCP showed a synergistic effect with significantly improved cell adhesion and osteogenesis in both 2D and 3D cell cultures. Therefore, the functional hydrogel developed in this study can potentially be used for bone tissue regeneration.

Coaxial bioprinting of a stentable and endothelialized human coronary artery-sized in vitro model.

Atherosclerosis accounts for two-thirds of deaths attributed to cardiovascular diseases, which continue to be the leading cause of mortality. Current clinical management strategies for atherosclerosis, such as angioplasty with stenting, face numerous challenges, including restenosis and late thrombosis. Smart stents, integrated with sensors that can monitor cardiovascular health in real-time, are being developed to overcome these limitations. This development necessitates rigorous preclinical trials on either animal models or in vitro models. Despite efforts being made, a suitable human-scale in vitro model compatible with a cardiovascular stent has remained elusive. To address this need, this study utilizes an in-bath bioprinting method to create a human-scale, freestanding in vitro model compatible with cardiovascular stents. Using a coaxial nozzle, a tubular structure of human coronary artery (HCA) size is bioprinted with a collagen-based bioink, ensuring good biocompatibility and suitable rheological properties for printing. We precisely replicated the dimensions of the HCA, including its internal diameter and wall thickness, and simulated the vascular barrier functionality. To simplify post-processing, a pumpless perfusion bioreactor is fabricated to culture a HCA-sized model, eliminating the need for a peristaltic pump and enabling scalability for high-throughput production. This model is expected to accelerate stent development in the future.

Composition control of additively manufactured color-graded temporary veneer.

OBJECTIVE: The aim of this study was to design and assess composite resin composition for patient-specific esthetic color-graded temporary veneer. METHODS: Various compositions of composite structures (assorted by Ba2SiO4 filler, TiO2 pigment, and photoinitiator) were prepared via additive manufacturing with 3 s UV exposure (405 nm, 10 W/cm2) per 50 µm thick layer followed by 20 min post-curing treatment after fabrication. The effect of each component on the generated color shades was observed and compared to the commonly used VITA shade guide. The coloration was explored by staining aging treatment under dry, wet, artificial saliva environments, coffee, and cola. The mechanical properties were also evaluated. Color measurement and comparison were done using a colorimeter (lightness (L*), green-red color (a*), and blue-yellow color (b*)), and the changes were calculated by CIEDE2000 (ΔE00), translucency parameter (TP) and whiteness index (WID). The composition color analysis results were then applied to produce a color-graded temporary veneer for mimicking a natural look. RESULT: Mechanically, all composition result in adequate bending strength with maximum achievable strength of 111.64 MPa. At the same time, the composite color was affected by each constituent differently. The L* value, which indicates the color lightness of the composite, was considerably tuned by the TiO2 pigment, whereas Ba2SiO4 filler only triggered minor changes. Photoinitiator concentration significantly affected the yellowness, indicated by the increased b* value. Similar tendency also observed toward the calculated TP and WID as well. Based on these evaluations, color-graded temporary veneer successfully generated, matching the VITA A3, A2, and B1 shades gradation. However, the stability of the composite color decreased at high amounts of Ba2SiO4 and photoinitiator. SIGNIFICANCE: The study presents a composition guide for fabricating temporary patient-specific color-graded veneer. It provides insights on the effect of the constituent material on dental esthetics.

Bioinks for bioprinting using plant-derived biomaterials.

Three-dimensional (3D) bioprinting has revolutionized tissue engineering by enabling the fabrication of complex and functional human tissues and organs. An essential component of successful 3D bioprinting is the selection of an appropriate bioink capable of supporting cell proliferation and viability. Plant-derived biomaterials, because of their abundance, biocompatibility, and tunable properties, hold promise as bioink sources, thus offering advantages over animal-derived biomaterials, which carry immunogenic concerns. This comprehensive review explores and analyzes the potential of plant-derived biomaterials as bioinks for 3D bioprinting of human tissues. Modification and optimization of these materials to enhance printability and biological functionality are discussed. Furthermore, cancer research and drug testing applications of the use of plant-based biomaterials in bioprinting various human tissues such as bone, cartilage, skin, and vascular tissues are described. Challenges and limitations, including mechanical integrity, cell viability, resolution, and regulatory concerns, along with potential strategies to overcome them, are discussed. Additionally, this review provides insights into the potential use of plant-based decellularized ECM (dECM) as bioinks, future prospects, and emerging trends in the use of plant-derived biomaterials for 3D bioprinting applications. The potential of plant-derived biomaterials as bioinks for 3D bioprinting of human tissues is highlighted herein. However, further research is necessary to optimize their processing, standardize their properties, and evaluate their long-termin vivoperformance. Continued advancements in plant-derived biomaterials have the potential to revolutionize tissue engineering and facilitate the development of functional and regenerative therapies for diverse clinical applications.

Plant-Based Decellularization: A Novel Approach for Perfusion-Compatible Tissue Engineering Structures.

This study explores the potential of plant-based decellularization in regenerative medicine, a pivotal development in tissue engineering focusing on scaffold development, modification, and vascularization. Plant decellularization involves removing cellular components from plant structures, offering an eco-friendly and cost-effective alternative to traditional scaffold materials. The use of plant-derived polymers is critical, presenting both benefits and challenges, notably in mechanical properties. Integration of plant vascular networks represents a significant bioengineering breakthrough, aligning with natural design principles. The paper provides an in-depth analysis of development protocols, scaffold fabrication considerations, and illustrative case studies showcasing plant-based decellularization applications. This technique is transformative, offering sustainable scaffold design solutions with readily available plant materials capable of forming perfusable structures. Ongoing research aims to refine protocols, assess long-term implications, and adapt the process for clinical use, indicating a path toward widespread adoption. Plant-based decellularization holds promise for regenerative medicine, bridging biological sciences with engineering through eco-friendly approaches. Future perspectives include protocol optimization, understanding long-term impacts, clinical scalability, addressing mechanical limitations, fostering collaboration, exploring new research areas, and enhancing education. Collectively, these efforts envision a regenerative future where nature and scientific innovation converge to create sustainable solutions, offering hope for generations to come.

Atypical post-infectious glomerulonephritis with c-ANCA positivity followed by endocarditis.

Post-infectious glomerulonephritis (PIGN), an uncommon variety of glomerulonephritis (GN), is characterized by emergence of nephritic syndrome within a few weeks following an infectious event. PIGN typically presents as a mild condition and tends to resolve by the time of diagnosis for GN. Aggregatibacter actinomycetemcomitans belongs to the HACEK group of bacteria, which constitutes less than 3% of bacteria responsible for community-acquired infective endocarditis. We present a case of 29-year-old man suspected of lymphoma with B-symptoms along with severe splenomegaly and nephromegaly. Shortly after, he developed an episode of nephritic syndrome accompanied by acute kidney injury (AKI) and high titers of cytoplasmic ANCA (c-ANCA)-positivity. Kidney biopsy revealed PIGN with tubulointerstitial nephritis. Despite treatment with antibiotics and corticosteroid, he visited the emergency room due to worsening dyspnea and multi-organ failure. An echocardiogram showed a bicuspid aortic valve with vegetation unseen on previous echocardiogram. He underwent aortic valve replacement immediately without adverse events. Four months after valve replacement, his renal function and cardiac performance have remained stable. We report a case of PIGN with AKI and high titers of c-ANCA appearing later as an infective endocarditis due to Aggregatibacter actinomycetemcomitans. With careful clinical observation and appropriate and timely management, satisfactory outcomes for patient health are possible.

Enhancement of properties of a cell-laden GelMA hydrogel-based bioink via calcium phosphate phase transition.

To improve the properties of the hydrogel-based bioinks, a calcium phosphate phase transition was applied, and the products were examined. We successfully enhanced the mechanical properties of the hydrogels by adding small amounts (< 0.5 wt%) of alpha-tricalcium phosphate (α-TCP) to photo-crosslinkable gelatin methacrylate (GelMA). As a result of the hydrolyzing calcium phosphate phase transition involvingα-TCP, which proceeded for 36 h in the cell culture medium, calcium-deficient hydroxyapatite was produced. Approximately 18 times the compressive modulus was achieved for GelMA with 0.5 wt%α-TCP (20.96 kPa) compared with pure GelMA (1.18 kPa). Although cell proliferation decreased during the early stages of cultivation, both osteogenic differentiation and mineralization activities increased dramatically when the calcium phosphate phase transition was performed with 0.25 wt%α-TCP. The addition ofα-TCP improved the printability and fidelity of GelMA, as well as the structural stability and compressive modulus (approximately six times higher) after three weeks of culturing. Therefore, we anticipate that the application of calcium phosphate phase transition to hydrogels may have the potential for hard tissue regeneration.

Signal-Decay Based Approach for Visualization of Buried Defects in 3-D Printed Ceramic Components Imaged with Help of Optical Coherence Tomography.

We propose the use of Optical Coherence Tomography (OCT) as a tool for the quality control of 3-D-printed ceramics. Test samples with premeditated defects, namely single- and two-component samples of zirconia, titania, and titanium suboxides, were printed by stereolithography-based DLP (Digital Light Processing) processes. The OCT tomograms obtained on the green samples showed the capability of the method to visualize variations in the layered structure of the samples as well as the presence of cracks and inclusions at depths up to 130 µm, as validated by SEM images. The structural information was visible in cross-sectional images as well as in plan-view images. The optical signal measured from the printed zirconia oxide and titanium oxide samples showed strong attenuation with depth and could be fit with an exponential decay curve. The variations of the decay parameter correlated very well with the presence of defects and material variation. When used as an imaging quantity, the decay parameter projects the position of the defects into 2-D (X,Y) coordinates. This procedure can be used in real time, it reduces the data volume up to 1000 times, and allows for faster subsequent data analysis and transfer. Tomograms were also obtained on sintered samples. The results showed that the method can detect changes in the optical properties of the green ceramics caused by sintering. Specifically, the zirconium oxide samples became more transparent to the light used, whereas the titanium suboxide samples became entirely opaque. In addition, the optical response of the sintered zirconium oxide showed variations within the imaged volume, indicating material density variations. The results presented in this study show that OCT provides sufficient structural information on 3-D-printed ceramics and can be used as an in-line tool for quality control.

142Development of an alginate-gelatin bioink enhancing osteogenic differentiation by gelatin release.

Hydrogels are natural bioink options for cellular printing due to their high-water content and permeable three-dimensional (3D) polymeric structure, which are favorable for cellular anchoring and metabolic activities. To increase the functionality of hydrogels as bioinks, biomimetic components are often incorporated, such as proteins, peptides, and growth factors. In this study, we aimed to enhance the osteogenic activity of a hydrogel formulation by integrating both the release and retention of gelatin so that gelatin serves as both an indirect support for released ink component on cells nearby and a direct support for encapsulated cells inside a printed hydrogel, thereby fulfills two functions. Methacrylate-modified alginate (MA-alginate) was chosen as the matrix because it has a low cell adhesion effect due to the absence of ligands. The gelatin-containing MA-alginate hydrogel was fabricated, and gelatin was found to remain in the hydrogel for up to 21 days. The gelatin remaining in the hydrogel had positive effects on encapsulated cells, especially on cell proliferation and osteogenic differentiation. The gelatin released from the hydrogel affected the external cells, showing more favorable osteogenic behavior than the control sample. It was also found that the MA-alginate/gelatin hydrogel could be used as a bioink for printing with high cell viability. Therefore, we anticipate that the alginate-based bioink developed in this study could potentially be used to induce osteogenesis in bone tissue regeneration.

Fabrication of color-graded feldspathic dental prosthetics for aesthetic and restorative dentistry.

OBJECTIVE: Feasibility investigation of natural teeth shades replication on dental prosthetics fabricated via functionally graded additive manufacturing (FGAM) using combination of feldspathic porcelain (FP) and yttrium aluminum garnet cerium (Y3Al5O12:Ce, YAG:Ce) as a promising esthetic restoration option. METHODS: Color-graded feldspathic crown fabrication parameter through FGAM method was comprehensively examined from the slurry rheology, cure depth, debinding to sintering temperature. Effect of light absorbent also checked towards overcuring reaction during UV exposure by the shape comparison. Lastly, the flexural bending strength measured following ISO 6872:2015 to assure the applicability. Applying the studied parameter, natural teeth shades then imitated and investigated by alteration of FP and FP + 0.1 wt% YAG:Ce (Y-FP). Generated color across the structure captured through mobile camera, interpreted through the CIELAB coordinate and the gradation confirmed by the color differences (ΔE00) calculated using CIEDE2000 formula. RESULT: Parameter study indicated that 70 wt% of FP slurry with 3 wt% dispersant and 0.2 wt% light absorbent is favored. It produces excellent flowability in our FGAM system with less overcuring justified by edge margin reduction from 95.65° to 90.00° after UV exposure on rectangle shapes masking. The obtain structure also offers adequate flexural bending strength of 106.26 MPa (FP) and 101.36 MPa (Y-FP) after sintering at 780 °C. This validated the materials as class 2 dental prosthetics citing ISO 6872:2015. Color gradation was verified by the yellow b* value reduction (14.8 to -3.33) as it shifted from cervical to incisal area while ΔE00 further affirmed the differences from each segment in comparison with the FP and Y-FP. SIGNIFICANCE: Color gradation was successfully replicated by FP and YAG:Ce composition shift via FGAM technique. This result highlights the potential of FGAM as an alternative for fabricating dental prosthetics with high efficiency and improved esthetic appeal.

Support-less ceramic 3D printing of bioceramic structures using a hydrogel bath.

Volumetric bone tissue defects are beyond the intrinsic regenerative capacity of bone tissue. With the recent development of ceramic 3D printing, various bioceramic scaffolds that can induce bone regeneration are being actively developed. However, hierarchical bone is complex, with overhanging structures that require additional sacrificial support during ceramic 3D printing. Not only can this increase the overall process time and material consumption, but breaks and cracks may occur when sacrificial supports are removed from fabricated ceramic structures. In this study, a support-less ceramic printing (SLCP) process using a hydrogel bath was developed to facilitate the manufacture of complex bone substitutes. A hydrogel bath, consisting of pluronic P123 with temperature-sensitive properties, mechanically supported the fabricated structure when the bioceramic ink was extruded into the bath and promoted the cement reaction to cure the bioceramic. SLCP enables the fabrication of complex bone constructs with overhanging structures, such as the mandible and maxillofacial bones, with reduced overall processing time and material consumption. Scaffolds fabricated by SLCP showed more cell adhesion, higher cell growth rate, and osteogenic protein expression due to their rougher surface than conventionally printed scaffolds. Hybrid scaffolds were fabricated by SLCP to co-print cells and bioceramics, and SLCP provided a cell-friendly environment, exhibiting high cell viability. SLCP enables control of the shape of various cells, bioactive substances, and bioceramics and thus can be used as an innovative 3D bioprinting technique to manufacture complex hierarchical bone structures.

Fabrication of multifunctional alginate microspheres containing hydroxyapatite powder for simultaneous cell and drug delivery.

Novel alginate-hydroxyapatite hybrid microspheres were developed for simultaneous delivery of drugs and cells as a multifunctional bone substitute for osteoporotic bone tissue regeneration. The microspheres were used to enhance osteogenesis and to carry and deliver quercetin, a representative phytoestrogen that controls bone tissue regeneration metabolism in osteoporosis patients, through sustained release over a long period. To overcome quercetin's hydrophobicity and low solubility in aqueous environments, we added it to the surface of hydroxyapatite (HAp) nanoparticles before mixing them with an alginate solution. The homogeneous distribution of the HAp nanoparticles in the alginate solution was essential for preventing nozzle clogging and achieving successfully fabricated hybrid microspheres. To this end, a 3D ultrasonic treatment was applied. Electrostatic microencapsulation was then used to fabricate hybrid alginate-HAp microspheres containing quercetin and cells. The microspheres were approximately 290.7 ± 42.5 µm (aspect ratio of 1). The sustained release of quercetin was confirmed during a test period of 20 weeks. The cells in the hybrid microspheres maintained good cell viability during the entire testing period, and their osteogenic differentiation behavior was boosted by the presence of HAp. Thus, osteogenic differentiation could be greatly improved by adding quercetin. These novel multi-biofunctional hybrid microspheres have great potential for the regeneration of osteoporotic bone tissue at indeterminate defect sites.

Identification of Survival-Specific Genes in Clear Cell Renal Cell Carcinoma Using a Customized Next-Generation Sequencing Gene Panel.

PURPOSE: Although mutations are associated with carcinogenesis, little is known about survival-specific genes in clear cell renal cell carcinoma (ccRCC). We developed a customized next-generation sequencing (NGS) gene panel with 156 genes. The purpose of this study was to investigate whether the survival-specific genes we found were present in Korean ccRCC patients, and their association with clinicopathological findings. MATERIALS AND METHODS: DNA was extracted from the formalin-fixed, paraffin-embedded tissue of 22 ccRCC patients. NGS was performed using our survival-specific gene panel with an Illumina MiSeq. We analyzed NGS data and the correlations between mutations and clinicopathological findings and also compared them with data from the Cancer Genome Atlas-Kidney Renal Clear Cell Carcinoma (TCGA-KIRC) and Renal Cell Cancer-European Union (RECA-EU). RESULTS: We found a total of 100 mutations in 37 of the 156 genes (23.7%) in 22 ccRCC patients. Of the 37 mutated genes, 11 were identified as clinicopathologically significant. Six were novel survival-specific genes (ADAMTS10, CARD6, NLRP2, OBSCN, SECISBP2L, and USP40), and five were top-ranked mutated genes (AKAP9, ARID1A, BAP1, KDM5C, and SETD2). Only CARD6 was validated as an overall survival-specific gene in this Korean study (p = 0.04, r = -0.441), TCGA-KIRC cohort (p = 0.0003), RECA-EU (p = 0.0005). The 10 remaining gene mutations were associated with clinicopathological findings; disease-free survival, mortality, nuclear grade, sarcomatoid component, N-stage, sex, and tumor size. CONCLUSIONS: We discovered 11 survival-specific genes in ccRCC using data from TCGA-KIRC, RECA-EU, and Korean patients. We are the first to find a correlation between CARD6 and overall survival in ccRCC. The 11 genes, including CARD6, NLRP2, OBSCN, and USP40, could be useful diagnostic, prognostic, and therapeutic markers in ccRCC.

Mechanically and biologically promoted cell-laden constructs generated using tissue-specific bioinks for tendon/ligament tissue engineering applications.

Tendon and ligament tissues provide stability and mobility crucial for musculoskeletal function, but are particularly prone to injury. Owing to poor innate healing capacity, the regeneration of mature and functional tendon/ligament (T/L) poses a formidable clinical challenge. Advanced bioengineering strategies to develop biomimetic tissue implants are highly desired for the treatment of T/L injuries. Here, we presented a cell-based tissue engineering strategy to generate cell-laden tissue constructs comprising stem cells and tissue-specific bioinks using 3D cell-printing technology. We implemented anin vitropreconditioning approach to guide semi-organized T/L-like formation before thein vivoapplication of cell-printed implants. Duringin vitromaturation, tissue-specific decellularized extracellular matrix-based cellular constructs facilitated long-termin vitroculture with high cell viability and promoted tenogenesis with enhanced cellular/structural anisotropy. Moreover, we demonstrated improved cell survival/retention uponin vivoimplantation of pre-matured constructs in nude mice with de novo tendon formation and improved mechanical strength. Althoughin vivomechanical properties of the cell-printed implants were lower than those of human T/L tissues, the results of this study may have significant implications for future cell-based therapies in tendon and ligament regeneration and translational medicine.

Selective Detection of an Infection Biomarker by an Osteo-Friend Scaffold: Development of a Multifunctional Artificial Bone Substitute.

Developments in three-dimensional (3D) printing technologies have led to many potential applications in various biomedical fields, especially artificial bone substitutes (ABSs). However, due to the characteristics of artificial materials, biocompatibility and infection remain issues. Here, multifunctional ABSs have been designed to overcome these issues by the inclusion of a biochemical modality that allows simultaneous detection of an infection biomarker by osteo-friend 3D scaffolds. The developed multifunctional scaffolds consist of calcium-deficient hydroxyapatite (CDHA), which has a similar geometric structure and chemical composition to human bone, and gold nanoparticles (Au NPs), which assists osteogenesis and modulates the fluorescence of labels in their microenvironment. The Au NPs were subsequently conjugated with fluorescent dye-labeled probe DNA, which allowed selective interaction with a specific target biomarker, and the fluorescent signal of the dye was temporally quenched by the Au NP-derived Förster resonance energy transfer (FRET). When the probe DNA unfolded to bind to the target biomarker, the fluorescence signal was recovered due to the increased distance between the dye and Au NPs. To demonstrate this sensing mechanism, a microbial oligonucleotide was selected as a target biomarker. Consequently, the multifunctional scaffold simultaneously facilitated osteogenic proliferation and the detection of the infection biomarker.

Promoting Long-Term Cultivation of Motor Neurons for 3D Neuromuscular Junction Formation of 3D In Vitro Using Central-Nervous-Tissue-Derived Bioink.

3D cell printing technology is in the spotlight for producing 3D tissue or organ constructs useful for various medical applications. In printing of neuromuscular tissue, a bioink satisfying all the requirements is a challenging issue. Gel integrity and motor neuron activity are two major characters because a harmonious combination of extracellular materials essential to motor neuron activity consists of disadvantages in mechanical properties. Here, a method for fabrication of 3D neuromuscular tissue is presented using a porcine central nervous system tissue decellularized extracellular matrix (CNSdECM) bioink. CNSdECM retains CNS tissue-specific extracellular molecules, provides rheological properties crucial for extrusion-based 3D cell printing, and reveals positive effects on the growth and maturity of axons of motor neurons compared with Matrigel. It also allows long-term cultivation of human-induced-pluripotent-stem-cell-derived lower motor neurons and sufficiently supports their cellular behavior to carry motor signals to muscle fibers. CNSdECM bioink holds great promise for producing a tissue-engineered motor system using 3D cell printing.

Application of 3D bioprinting in the prevention and the therapy for human diseases.

Rapid development of vaccines and therapeutics is necessary to tackle the emergence of new pathogens and infectious diseases. To speed up the drug discovery process, the conventional development pipeline can be retooled by introducing advanced in vitro models as alternatives to conventional infectious disease models and by employing advanced technology for the production of medicine and cell/drug delivery systems. In this regard, layer-by-layer construction with a 3D bioprinting system or other technologies provides a beneficial method for developing highly biomimetic and reliable in vitro models for infectious disease research. In addition, the high flexibility and versatility of 3D bioprinting offer advantages in the effective production of vaccines, therapeutics, and relevant delivery systems. Herein, we discuss the potential of 3D bioprinting technologies for the control of infectious diseases. We also suggest that 3D bioprinting in infectious disease research and drug development could be a significant platform technology for the rapid and automated production of tissue/organ models and medicines in the near future.

Multifunctional Calcium-Deficient Hydroxyl Apatite-Alginate Core-Shell-Structured Bone Substitutes as Cell and Drug Delivery Vehicles for Bone Tissue Regeneration.

In this work, we fabricated unique coiled-structured bioceramics contained in hydrogel beads for simultaneous drug and cell delivery using a combination of bone cement chemistry and bioprinting and characterized them. The core of the calcium-deficient hydroxyl apatite (CDHA) contains quercetin, which is a representative phytoestrogen isolated from onions and apples, to control the metabolism of bone tissue regeneration through sustained release over a long period of time. The shell consists of an alginate hydrogel that includes preosteoblast MC3T3-E1 cells. Ceramic paste and hydrogel were simultaneously extruded to fabricate core-shell beads through the inner and outer nozzles, respectively, of a concentric nozzle system based on a material-extruding-based three-dimensional (3D) printing system. The formation of beads and the coiled ceramic core is related to both alginate concentration and printing conditions. The size of the microbeads and the thickness of the coiled structure could be controlled by adjusting the nozzle conditions. The whole process was carried out at physiological conditions (37 °C) to be gentle on the cells. The alginate shell undergoes solidification by cross-linking in CaCl2 or monocalcium phosphate monohydrate (MCPM) solution, while the hardening and cementation of the α-tricalcium phosphate (α-TCP) core to CDHA are subsequently initiated by immersion in phosphate-buffered saline solution. This process replaces the typical sintering of ceramic processing to prevent damage to the hydrogel, cells, and drugs in the beads. The cell-loaded beads were then cultured in cell culture media where the cells could maintain good viability during the entire testing period, which was over 50 days. Cell growth and elongation were observed even in the alginate along the CDHA coiled structure over time. Sustained release of quercetin without any initial burst was also confirmed during a test period of 120 days. These novel structured microbeads with multibiofunctionality can be used as new bone substitutes for hard tissue regeneration in indeterminate defect sites.