Maryam Tilton

Sacral tumors often necessitate surgical resection via En Bloc Sacrectomy, a procedure requiring sub-millimeter precision to ensure complete tumor removal while preserving critical nerve roots. To address the inherent risks and limitations of conventional freehand techniques, this paper introduces a novel semi-autonomous robotic system for high-precision partial sacrectomy. The platform integrates a seven degree-of-freedom robotic manipulator with a commercial ultrasonic osteotome, an instrument specifically designed to cut hard bone while minimizing soft tissue damage. We conducted a comprehensive quantitative characterization of the system, demonstrating that sub-millimeter trajectory accuracy (Root Mean Squared Error ≤ 0.13 mm) is consistently maintained across a wide range of cutting speeds, from 0.5 mm/s to 3 mm/s. This robust performance enabled a significant reduction in total osteotomy procedure time (down to 28 s) with maximum interaction forces remaining low (1 N). Furthermore, the framework’s capability was validated in a clinically relevant case study on a custom CNC-machined human sacrum phantom, where the system executed complex, multi-pass cuts with an accuracy of 0.12 mm. These results confirm the system’s stability, speed, and precision, representing a significant technological step toward enabling safer and more efficient robotic assistance for complex orthopedic oncology procedures.

Conventional sacroiliac (SI) screw fixation relies on rigid drills and linear implant trajectories, which limits access to regions of high bone mineral density and contributes to screw loosening, misplacement, and neurovascular injury - particularly in osteoporotic patients. To address this issue, we present a Flexible Sacroiliac Screw (FSIS) and a complementary steerable drilling robot that enable SI joint fixation along curvilinear trajectories. We evaluate both systems by drilling J-shape trajectories and fixating our flexible implant into Sawbone phantoms with different densities simulating cortical and cancellous regions inside the bone. Moreover, we perform a case study on a CNC-machined phantom further demonstrating the system’s ability to accurately traverse the ilium, follow a curvilinear sacral corridor, and maintain safe boundaries under fluoroscopic visualization.

To address the screw loosening and pullout limitations of rigid pedicle screws in spinal fixation procedures, and to leverage our recently developed Concentric Tube Steerable Drilling Robot (CT-SDR) and Flexible Pedicle Screw (FPS), in this paper, we introduce the concept of Augmented Bridge Spinal Fixation (AB-SF). In this concept, two connecting J-shape tunnels are first drilled through pedicles of vertebra using the CT-SDR. Next, two FPSs are passed through this tunnel and bone cement is then injected through the cannulated region of the FPS to form an augmented bridge between two pedicles and reinforce strength of the fixated spine. To experimentally analyze and study the feasibility of AB-SF technique, we first used our robotic system (i.e., a CT-SDR integrated with a robotic arm) to create two different fixation scenarios in which two J-shape tunnels, forming a bridge, were drilled at different depth of a vertebral phantom. Next, we implanted two FPSs within the drilled tunnels and then successfully simulated the bone cement augmentation process.

In this study, we introduce a new concept for reconstruction of Volumetric Muscle Loss (VML) injuries and propose the spatial robotic embedded bioprinting technique. As opposed to the traditional layer-by-layer printing, we leverage the support-free nature of embedded bioprinting to print spatial and complex structures of fascicles in a fusiform muscle. To demonstrate feasibility of this concept, we first propose our robotic bioprinting framework including a robotic arm integrated with a custom-designed bioprinting injector. Complementary motion planning algorithms uniquely designed for this printing task are further proposed. Moreover, the effect of embedded bioprinting parameters, as well as the supporting bath and injecting materials compatibility on the uniformity and quality of the printed constructs has been analyzed. Finally, we perform a case study by printing a fusiform muscle-shape construct using the proposed concept and algorithms, and evaluate the quality of the printed structure.

Abstract Oxygen deprivation within large or poorly vascularized bone defects remains a key barrier to successful regeneration, especially during the early postimplantation period before vascular ingrowth. Here, the development of COSnPPOD (CaO 2 –Silica NP Platform for Osteogenic Development) is reported, a visible light digital light processing‐printed hydrogel scaffold that integrates oxygen‐releasing nanoparticles (NPs) within a Primitive‐type triply periodic minimal surface architecture. The scaffold combines a gelatin methacrylate‐poly(ethylene glycol) diacrylate matrix with calcium peroxide (CaO 2 )‐loaded hollow silica NPs, enabling localized, short‐term oxygen release while preserving structural fidelity. COSnPPOD scaffolds demonstrate favorable degradation kinetics, tunable stiffness, and increased protein adsorption in vitro. In a preosteoblast model, COSnPPOD maintains cell viability and supports osteogenic gene expression without cytotoxic effects. While overall gene expression is comparable to controls, a 16‐fold increased expression of phosphoprotein 1 (Spp1) suggests scaffold‐driven activation of matrix remodeling pathways. In vivo, COSnPPOD scaffolds enhance bone regeneration in a murine calvarial defect model, with significantly greater bone formation and collagen deposition than untreated defects and hydrogel controls. Additionally, vascular endothelial growth factor immunostaining is increased within the defect, consistent with a proangiogenic response, and no systemic toxicity is observed. These findings establish COSnPPOD as a promising scaffold system that combines sustained oxygenation with biomimetic geometry to support localized bone regeneration.

Aging is increasingly recognized as a systemic process, yet the mechanisms by which senescent cells' signal from peripheral tissues accelerate brain aging remain poorly defined. Here, we used chronic exposure of human cerebral organoids to the secretome of senescent osteocytes to investigate how peripheral aging signals reshape brain tissue architecture. We combined spatially resolved optical fiberbased interferometry nanoindentation with transcriptomic and immunofluorescence profiling, demonstrating that bone-derived senescence-associated secretory phenotype (SASP) factors induce a biphasic mechanical response, early global tissue softening, followed by the emergence of discrete hyper-stiff microdomains. This spatially heterogeneous biomechanical remodeling was accompanied by upregulation of extracellular matrix (ECM), inflammatory, and senescence pathways, and suppression of neurodevelopmental and synaptic gene networks. Our results reveal that chronic paracrine SASP exposure from senescent osteocytes drives localized ECM reorganization and mechanical vulnerability in human brain tissue, providing mechanistic insight into how peripheral cellular senescence may contribute to regional brain fragility during aging.

To address the screw loosening and pullout limitations of rigid pedicle screws in spinal fixation procedures, and to leverage our recently developed Concentric Tube Steerable Drilling Robot (CT-SDR) and Flexible Pedicle Screw (FPS), in this paper, we introduce the concept of Augmented Bridge Spinal Fixation (AB-SF). In this concept, two connecting J-shape tunnels are first drilled through pedicles of vertebra using the CT-SDR. Next, two FPSs are passed through this tunnel and bone cement is then injected through the cannulated region of the FPS to form an augmented bridge between two pedicles and reinforce strength of the fixated spine. To experimentally analyze and study the feasibility of AB-SF technique, we first used our robotic system (i.e., a CT-SDR integrated with a robotic arm) to create two different fixation scenarios in which two J-shape tunnels, forming a bridge, were drilled at different depth of a vertebral phantom. Next, we implanted two FPSs within the drilled tunnels and then successfully simulated the bone cement augmentation process.

OBJECTIVE: Current spinal fixation (SF) techniques face screw loosening and pullout challenges in osteoporotic patients. This can be attributed to conventional rigid pedicle screws (RPS) being forced to fixate along a constrained linear trajectory into low bone mineral density (BMD) areas of the vertebral body. This study proposes a synergistic patient-specific approach that integrates a steerable drilling robotic system with a novel Flexible Pedicle Screw (FPS) to enhance SF procedures by enabling curved screw fixation. METHODS: A patient-specific framework and synergistic design flowchart were developed to guide the synergistic design of the previously proposed Concentric Tube-Steerable Drilling Robot (CT-SDR) and the FPS. After, the novel FPS is designed based on critical design features and its design is validated using Finite Element Analysis (FEA). The FPS is then fabricated via Direct Metal Laser Sintering (DMLS). The FPS's morphability and self-tapping capability were experimentally assessed in Sawbones phantoms drilled by the CT-SDR system. RESULTS: The FPS successfully morphed to fixate in curvilinear paths, demonstrating effective morphability and self-tapping in simulated bone. CONCLUSION: By enabling a flexible, patient-specific approach to pedicle screw fixation, the FPS and CT-SDR system address key limitations of current SF procedures. This method enhances screw anchorage and fixation strength in osteoporotic vertebrae. SIGNIFICANCE: This work presents a transformative approach to SF, with potential clinical applications in improving surgical outcomes for osteoporotic patients. The integration of robotic-assisted drilling and flexible implants could significantly reduce fixation failure rates, advancing orthopedic and spinal surgical practices.

Aging-related bone loss significantly impacts the growing elderly population globally, leading to debilitating conditions such as osteoporosis. Senescent osteocytes play a crucial role in the aging process of bone. This longitudinal study examines the impact of continuous local and paracrine exposure to senescence-associated secretory phenotype (SASP) factors on biophysical and biomolecular markers in osteocytes. Significant cytoskeletal stiffening in irradiated (IR) osteocytes are found, accompanied by expansion of F-actin areas and a decline in dendritic integrity. These changes, correlating with alterations in pro-inflammatory cytokine levels and osteocyte-specific gene expression, support the reliability of biophysical markers for identifying senescent osteocytes. Notably, local accumulation of SASP factors have a more pronounced impact on osteocyte biophysical properties than paracrine effects, suggesting that the interplay between local and paracrine exposure can substantially influence cellular aging. This study underscores the importance of osteocyte mechanical and morphological properties as biophysical markers of senescence, highlighting their time dependence and differential effects of local and paracrine SASP exposure. Collectively, the investigation into biophysical senescence markers offers unique and reliable functional hallmarks for the non-invasive identification of senescent osteocytes, providing insights that can inform therapeutic strategies to mitigate aging-related bone loss.

Abstract Senescent osteocytes are key contributors to age‐related bone loss and fragility; however, the impact of mechanobiological changes in these cells remains poorly understood. This study provides a novel analysis of these changes in primary osteocytes following irradiation‐induced senescence. By integrating subcellular mechanical measurements with gene expression analyses, we identified significant, time‐dependent alterations in the mechanical properties of senescent bone cells. Increases in classical markers such as SA‐β‐Gal activity and p16 Ink4a expression levels confirmed the senescence status post‐irradiation. Our key findings include a time‐dependent increase in cytoskeletal Young's modulus and altered viscoelastic properties of the plasma membrane, affecting the contractility of primary osteocytes. Additionally, we observed a significant increase in Sclerostin ( Sost ) expression 21 days post‐irradiation. These biophysical changes may impair osteocyte mechanosensation and mechanotransduction, contributing to bone fragility. This is the first study to time‐map senescence‐associated mechanical changes in the osteocyte cytoskeleton. Our findings highlight the potential of biophysical markers as indicators of cellular senescence, providing more specificity than traditional, variable biomolecular markers. We believe these results may support biomechanical stimulation as a potential therapeutic strategy to rejuvenate aging osteocytes and enhance bone health.

Degenerative spine pathologies, including intervertebral disc (IVD) degeneration, present a significant healthcare challenge due to their association with chronic pain and disability. This study explores an innovative approach to IVD regeneration utilizing 3D bioprinting technology, specifically visible light-based digital light processing (VL-DLP), to fabricate tissue scaffolds that closely mimic the native architecture of the IVD. Utilizing a hybrid bioink composed of gelatin methacrylate (GelMA) and poly (ethylene glycol) diacrylate (PEGDA) at a 10% concentration, we achieved enhanced printing fidelity and mechanical properties suitable for load-bearing applications such as the IVD. Preconditioning rat bone marrow-derived mesenchymal stem cell (rBMSC) spheroids with chondrogenic media before incorporating them into the GelMA-PEGDA scaffold further promoted the regenerative capabilities of this system. Our findings demonstrate that this bioprinted scaffold not only supports cell viability and integration but also contributes to the restoration of disc height in a rat caudal disc model without inducing adverse inflammatory responses. The study underscores the potential of combining advanced bioprinting techniques and cell preconditioning strategies to develop effective treatments for IVD degeneration and other musculoskeletal disorders, highlighting the need for further research into the dynamic interplay between cellular migration and the hydrogel matrix.

Spinal fixation procedures are currently limited by the rigidity of the existing instruments and pedicle screws leading to fixation failures and rigid pedicle screw pull out. Leveraging our recently developed Concentric Tube Steerable Drilling Robot (CT-SDR) in integration with a robotic manipulator, to address the aforementioned issue, here we introduce the transformative concept of Spatial Spinal Fixation (SSF) using a unique Flexible Pedicle Screw (FPS). The proposed SSF procedure enables planar and out-of-plane placement of the FPS throughout the full volume of the vertebral body. In other words, not only does our fixation system provide the option of drilling in-plane and out-of-plane trajectories, it also enables implanting the FPS inside linear (represented by an I-shape) and/or non-linear (represented by J-shape) trajectories. To thoroughly evaluate the functionality of our proposed robotic system and the SSF procedure, we have performed various experiments by drilling different I-J and J-J drilling trajectory pairs into our custom-designed L3 vertebral phantoms and analyzed the accuracy of the procedure using various metrics.

Aging-related bone loss significantly impacts the growing elderly population globally, leading to debilitating conditions such as osteoporosis. Senescent osteocytes play a crucial role in the aging process of bone. This longitudinal study examines the impact of continuous local and paracrine exposure to senescence-associated secretory phenotype (SASP) factors on senescence-associated biophysical and biomolecular markers in osteocytes. We found significant cytoskeletal stiffening in irradiated osteocytes, accompanied by expansion of F-actin areas and a decline in dendritic integrity. These changes, correlating with alterations in pro-inflammatory cytokine levels and osteocyte-specific gene expression, support the reliability of biophysical markers for identifying senescent osteocytes. Notably, local accumulation of SASP factors had a more pronounced impact on osteocyte properties than paracrine effects, suggesting that the interplay between local and paracrine exposure could substantially influence cellular aging. This study underscores the importance of osteocyte mechanical and morphological properties as biophysical markers of senescence, highlighting their time-dependence and differential effects of local and paracrine SASP exposure. Collectively, our investigation into biophysical senescence markers offer unique and reliable functional hallmarks for non-invasive identification of senescent osteocytes, providing insights that could inform therapeutic strategies to mitigate aging-related bone loss.

Abstract Senescent osteocytes are key contributors to age-related bone loss and fragility; however, the impact of mechanobiological changes in these cells remains poorly understood. This study provides a novel analysis of these changes in primary osteocytes following irradiation-induced senescence. By integrating sub-cellular mechanical measurements with gene expression analyses, we identified significant, time-dependent alterations in the mechanical properties of these cells. Increases in SA-β-Gal activity and p16Ink4a expression levels confirmed the senescence status post-irradiation. Key findings include a time-dependent increase in cytoskeletal Young’s modulus and altered viscoelastic properties of the plasma membrane, affecting the contractility of primary osteocytes. Additionally, we observed a significant increase in Sclerostin (Sost) expression 21 days post-irradiation. These mechanobiological changes may impair osteocyte mechanosensation and mechanotransduction, contributing to bone fragility. This is the first study to time-map senescence-associated mechanical changes in the osteocyte cytoskeleton. Our findings highlight the potential of biophysical markers as indicators of cellular senescence, providing more specificity than traditional, variable biomolecular markers. We believe these results support biomechanical stimulation as a potential therapeutic strategy to rejuvenate aging osteocytes and enhance bone health.

There is a significant clinical need to develop effective treatments for bone defects in patients with diabetes mellitus (DM), as they are at higher risk of fractures and impaired healing. Guided bone tissue engineering using biocompatible and biodegradable polymers is a promising approach. However, current diabetic bone regenerative therapies often fail due to the accumulation of advanced glycation products, which can affect the integration of traditional tissue engineering scaffolds with native bone. Therefore, novel approaches are needed to improve the efficacy of diabetic bone regeneration. This study presents a proof-of-concept development of a multifunctional polymer composite coating tailored towards restoring diabetes-related damage in osteoblast differentiation. Our composite system involves 3D-printed poly(caprolactone fumarate) (PCLF) and poly(caprolactone) (PCL) blend scaffolds coated with multifunctional chitosan methacrylate (chiMA). The chiMA coating is embedded with a sustained-release formulation of glucose oxidase (GOx) from MIL-127 metal-organic frameworks making the coating a stimuli-responsive biomolecule delivery system. The multifunctional coating is designed for the sustained release of GOx and sodium pyruvate for in vitro glucose modulation and oxidative stress reduction, respectively. We propose that sustained release of GOx from MIL-127 embedded chiMA coatings can modulate the high glucose (HG) cellular milieu towards normal glucose (NG), enhancing osteoblast (OB) differentiation via downstream effects. Our results show successful synthesis of MIL-127, encapsulation of GOx, and fabrication of composite coating on the PCLF/PCL scaffolds with effective enzyme activity measured as a function of lowering glucose concentration in HG media for 144 h to normal levels. In vitro evaluation of OB viability, attachment, proliferation, and differentiation showed an overall decrease in cellular activity in HG conditions, which was restored through the glucose-modulating functionality of the GOx-releasing MIL-127 coatings. Our results also presented preliminary evidence of a statistical correlation between DM-related gene markers and osteogenic markers in vitro that requires further exploration. Although this proof-of-concept study holds promise for advancing precision biomaterials development for diabetic tissue engineering and meeting the unmet clinical need for effective treatments and warrants future in vivo evaluation of the composite coating and molecular biology understanding of correlations between DM and osteogenic markers.

In this paper, we explore the feasibility of developing a novel flexible pedicle screw (FPS) for enhanced spinal fixation of osteoporotic vertebrae. Vital for spinal fracture treatment, pedicle screws have been around since the early 20th century and have undergone multiple iterations to enhance internal spinal fixation. However, spinal fixation treatments tend to be problematic for osteoporotic patients due to multiple inopportune variables. The inherent rigid nature of the pedicle screw, along with the forced linear trajectory of the screw path, frequently leads to the placement of these screws in highly osteoporotic regions of the bone. This results in eventual screw slippage and causing neurological and respiratory problems for the patient. To address this problem, we focus on developing a novel FPS that is structurally capable of safely bending to fit curved trajectories drilled by a steerable drilling robot and bypass highly osteoporotic regions of the vertebral body. Afterwards, we simulate its morphability capabilities using finite element analysis (FEA). We then additively manufacture the FPS using stainless steel (SS) 316L alloy through direct metal laser sintering (DMLS). Finally, the fabricated FPS is experimentally evaluated for its bending performance and compared with the FEA results for verification. Results demonstrate the feasibility of additive manufacturing of FPS using DMLS approach and agreement of the developed FEA with the experiments.

The growing aging population, with its associated chronic diseases, underscores the urgency for effective tissue regeneration strategies. Biomaterials play a pivotal role in the realm of tissue reconstruction and regeneration, with a distinct shift toward minimally invasive (MI) treatments. This transition, fueled by engineered biomaterials, steers away from invasive surgical procedures to embrace approaches offering reduced trauma, accelerated recovery, and cost-effectiveness. In the realm of MI tissue repair and cargo delivery, various techniques are explored. While in situ polymerization is prominent, it is not without its challenges. This narrative review explores diverse biomaterials, fabrication methods, and biofunctionalization for injectable pre-formed scaffolds, focusing on their unique advantages. The injectable pre-formed scaffolds, exhibiting compressibility, controlled injection, and maintained mechanical integrity, emerge as promising alternative solutions to in situ polymerization challenges. The conclusion of this review emphasizes the importance of interdisciplinary design facilitated by synergizing fields of materials science, advanced 3D biomanufacturing, mechanobiological studies, and innovative approaches for effective MI tissue regeneration.

Spinal fixation procedures are currently limited by the rigidity of the existing instruments and pedicle screws leading to fixation failures and rigid pedicle screw pull out. Leveraging our recently developed Concentric Tube Steerable Drilling Robot (CT-SDR) in integration with a robotic manipulator, to address the aforementioned issue, here we introduce the transformative concept of Spatial Spinal Fixation (SSF) using a unique Flexible Pedicle Screw (FPS). The proposed SSF procedure enables planar and out-of-plane placement of the FPS throughout the full volume of the vertebral body. In other words, not only does our fixation system provide the option of drilling in-plane and out-of-plane trajectories, it also enables implanting the FPS inside linear (represented by an I-shape) and/or non-linear (represented by J-shape) trajectories. To thoroughly evaluate the functionality of our proposed robotic system and the SSF procedure, we have performed various experiments by drilling different I-J and J-J drilling trajectory pairs into our custom-designed L3 vertebral phantoms and analyzed the accuracy of the procedure using various metrics.

3D stem cell spheroids have immense potential for various tissue engineering applications. However, current spheroid fabrication techniques encounter cell viability issues due to limited oxygen access for cells trapped within the core, as well as nonspecific differentiation issues due to the complicated environment following transplantation. In this study, functional 3D spheroids are developed using mesenchymal stem cells with 2D hetero-nanostructures (HNSs) composed of single-stranded DNA (ssDNA) binding carbon nanotubes (sdCNTs) and gelatin-bind black phosphorus nanosheets (gBPNSs). An osteogenic molecule, dexamethasone (DEX), is further loaded to fabricate an sdCNTgBP-DEX HNS. This approach aims to establish a multifunctional cell-inductive 3D spheroid with improved oxygen transportation through hollow nanotubes, stimulated stem cell growth by phosphate ions supplied from BP oxidation, in situ immunoregulation, and osteogenesis induction by DEX molecules after implantation. Initial transplantation of the 3D spheroids in rat calvarial bone defect shows in vivo macrophage shifts to an M2 phenotype, leading to a pro-healing microenvironment for regeneration. Prolonged implantation demonstrates outstanding in vivo neovascularization, osteointegration, and new bone regeneration. Therefore, these engineered 3D spheroids hold great promise for bone repair as they allow for stem cell delivery and provide immunoregulative and osteogenic signals within an all-in-one construct.

LAPONITE®-based drug delivery systems offer many advantages due to the unique ionic and physical properties of LAPONITE®. The high ionicity and large surface area of LAPONITE® nanoparticles enable the intercalation and dissolution of biomolecules. In this study, we explored the potential of LAPONITE® as a carrier for FG-4592 to support angiogenesis and as a carrier for bone morphogenic protein-2 (BMP-2) to support osteogenesis. Interestingly, we found that LAPONITE® promoted the FG-4592 induced upregulation of vascular endothelial growth factor (VEGF) gene expression of human umbilical cord endothelial cells (HUVECs). Additionally, we observed that LAPONITE® could provide a sustained release of BMP-2 and significantly potentiate the osteogenic effects of BMP-2 on adipose derived mesenchymal stem cells (AMSCs). Overall, current findings on the LAPONITE®-drug/protein model system provide a unique way to potentiate the angiogenic activities of FG-4592 on HUVECs and osteogenic effects of BMP-2 on AMSCs for tissue engineering application. Future studies will be directed towards gaining a deeper understanding of these effects on a co-culture system of HUVECs and AMSCs.

Polycaprolactone fumarate (PCLF) is a cross-linkable PCL derivative extensively considered for tissue engineering applications. Although injection molding has been widely used to develop PCLF scaffolds, platforms developed using such technique lack precise control on architecture, design, and porosity required to ensure adequate cellular and tissue responses. In particular, the scaffolds should provide a suitable surface for cell attachment and proliferation, and facilitate cell-cell communication and nutrient flow. 3D printing technologies have led to new architype for biomaterial development with micro-architecture mimicking native tissue. Here, we developed a method for 3D printing of PCLF structures using the extrusion printing technique. The crosslinking property of PCLF enabled the unique post-processing of 3D printed scaffolds resulting in highly porous and flexible PCLF scaffolds with compressive properties imitating natural features of cancellous bone. Generated scaffolds supported excellent attachment and proliferation of mesenchymal stem cells (MSC). The high porosity of PCLF scaffolds facilitated vascularized membrane formation demonstrable with the stringency of the ex ovo chicken chorioallantoic membrane (CAM) implantation. Furthermore, upon implantation to rat calvarium defects, PCLF scaffolds enabled an exceptional new bone formation with a bone mineral density of newly formed bone mirroring native bone tissue. These studies suggest that the 3D-printed highly porous PCLF scaffolds may serve as a suitable biomaterial platform to significantly expand the utility of the PCLF biomaterial for bone tissue engineering applications.

Pre-formed hydrogel scaffolds have emerged as favorable vehicles for tissue regeneration, promoting minimally invasive treatment of native tissue. However, due to the high degree of swelling and inherently poor mechanical properties, development of complex structural hydrogel scaffolds at different dimensional scales has been a continuous challenge. Herein, we take a novel approach at the intersections of engineering design and bio-ink chemistry to develop injectable pre-formed structural hydrogel scaffolds fabricated via visible light (VL) induced digital light processing (DLP). In this study, we first determined the minimum concentration of poly(ethylene glycol) diacrylate (PEGDA) to be added to the gelatin methacrylate (GelMA) bio-ink in order to achieve scalable and high printing-fidelity with desired cell adhesion, viability, spreading, and osteogenic differentiation characteristics. Despite the advantages of hybrid GelMA-PEGDA bio-ink in improving scalability and printing-fidelity, compressibility, shape-recovery, and injectability of the 3D bioprinted scaffolds were compromised. To restore these needed characteristics for minimally invasive tissue regeneration applications, we performed topological optimization to design highly compressible and injectable pre-formed (i.e., 3D bioprinted) microarchitectural scaffolds. The designed injectable pre-formed microarchitectural scaffolds showed a great capacity to retain the viability of the encapsulated cells (>72% after 10 cycles of injection). Lastly, ex ovo chicken chorioallantoic membrane (CAM) studies revealed that the optimized injectable pre-formed hybrid hydrogel scaffold is biocompatible and supports angiogenic growth.

Current bone cement systems often demand free radical or metal-related initiators and/or catalysts for the crosslinking process, which may cause serious toxicity to the human body. In addition, the resultant dense scaffolds may have a prolonged degradation time and are difficult for cells to infiltrate and form new tissue. In this study, we developed a porous “click” organic–inorganic nanohybrid (PO-click-ON) cement that crosslinks via metal-free biorthogonal click chemistry and forms porous structures mimicking the native bone tissue via particulate leaching. Strain-promoted click reaction enables fast and efficient crosslinking of polymer chains with the exclusion of any toxic initiator or catalyst. The resulting PO-click-ON implants supported exceptional in vitro stem cell adhesion and osteogenic differentiation with a large portion of stem cells infiltrated deep into the scaffolds. In vivo study using a rat cranial defect model demonstrated that the PO-click-ON system achieved outstanding cell adsorption, neovascularization, and bone formation. The porous click cement developed in this study serves as a promising platform with multifunctionality for bone and other tissue engineering applications.