Biofabrication and biomanufacturing are rapidly transforming how materials, therapeutics, and functional biological constructs are produced. These fields integrate developments in sustainable biomaterials, precision fabrication, biological systems, and data-driven engineering to produce scalable, efficient, and environmentally aligned production pathways. This review high-lights recent scientific advances led by researchers in Singapore, focusing on three interconnected pillars: sustainable bio-derived materials, enabling fabrication and manufacturing technologies, and emerging applications. We first examine the expanding use of biomass-derived feedstocks, including human hair keratin, aquaculture side-streams, and plant-derived polysaccharides, which support circular and resource-conscious material development. We then present advances in biofabrication technologies, including electrospinning, three-dimensional bioprinting, and metal additive manufacturing, that enable improved control over the structure, function, and manufacturability of biomedical and functional constructs. Emerging applications, such as machine learning-assisted additive manufacturing, food biomanufacturing, regenerative cell therapy, microneedles, and bioelectronics, exemplify how biofabrication and biomanufacturing are increasingly interrelated across the health, materials, and technological domains. These research contributions from Singapore exemplify how sustainable feedstocks, digital and automated fabrication platforms, and biologically driven applications are shaping the evolving landscape of biofabrication and biomanufacturing. The convergence of materials science, biological engineering, and advanced manufacturing continues to enable new opportunities for innovation in biomedical, industrial, and societal contexts.

Microneedles have gained increasing attention as an ideal device for transdermal drug delivery; however, their application to proteins has been a challenging task because of their inadequate tip-loading and unsatisfactory stability. Herein, we report the development of a microgel-integrated microneedle array patch (MI-MAP) capable of not only localizing protein cargoes in the tip portion but also stabilizing them via protein-phenolic interactions. Incorporation of protein-capturing microgels allows enrichment of protein cargoes within the tip region of microneedles, resulting in a significant enhancement in their transdermal delivery performance. MI-MAP enables more effective stabilization of labile recombinant proteins, such as botulinum neurotoxin, human interferon alpha-2a, and SARS-CoV-2 spike receptor-binding domain (RBD), during storage at 25 °C for 28 days compared to the counterpart lacking the microgels (HA-MAP). Transdermal delivery of CpG-adjuvanted RBD using MI-MAP elicits a more rapid neutralizing antibody response and germinal center B-cell proliferation in mice than HA-MAP and subcutaneous injection, without causing any severe inflammation. This study presents a potential strategy for improving tip-loading and storage stability of recombinant proteins within microneedle structures.

Superficial fungal skin infections are common but often misdiagnosed, which may result in inappropriate treatment and the worsening of symptoms. An accurate and timely diagnosis is essential to differentiate these infections from similar conditions such as secondary syphilis, annular psoriasis, and pityriasis rosea. This single-centre prospective cohort study at the National Skin Centre, Singapore, evaluated the diagnostic agreement between direct microscopy and fungal culture. Between August and December 2022, 268 skin scrape samples were collected from 149 patients with suspected fungal infections. Microscopy identified 67 (25.0%) positives, while fungal culture detected 42 (16.7%) positives. Among the 252 samples tested with both methods, 213 (84.5%) showed concordant results (κ = 0.487, p < 0.0001), a finding that indicates moderate agreement. The most commonly cultured organisms were Trichophyton rubrum and T. mentagrophytes. Our findings suggest that both microscopy and fungal culture may be performed to prevent true-positive cases from being missed. However, in cases where cost is a concern, microscopy can be selected as an initial diagnostic tool. Should microscopy be negative in cases with high clinical suspicion for fungal infection or when empirical treatment fails, culture remains a valuable follow-up test. These findings support a stepwise diagnostic approach—using microscopy first, then followed by culture when necessary—to improve diagnostic accuracy while enabling timely treatment.

Current wound therapies struggle to dynamically regulate immune responses and angiogenesis, often resulting in impaired healing, scarring, and poor tissue regeneration. The development of smart hydrogel scaffolds offers an opportunity to precisely modulate the wound healing process. Here, we present a pioneering wireless immunomodulatory strategy by integrating amino-modified barium titanate (BTN) nanoparticles with a natural collagen matrix, using oxidized gellan gum (OG) as a crosslinker, to fabricate a tilapia collagen (Col)-based biomimetic piezoelectric hydrogel scaffold (Col/OG/BTN). The hydrogel scaffold exhibits skin-like mechanical properties, controlled biodegradability, and ultrasound (US)-activated piezoelectricity, while providing a three-dimensional porous microenvironment for cell migration and signaling. Under US, the hydrogel scaffold reprograms pro-inflammatory M1 macrophages toward pro-healing M2 macrophages by modulating the phosphoinositide 3-kinase (PI3K) /protein kinase B (Akt) and tumor necrosis factor (TNF) signaling pathways, as revealed by transcriptomics. This immunoregulation synergizes with endothelial cell crosstalk to amplify pro-angiogenic factor secretion. Importantly, in vivo application of the Col/OG/BTN hydrogel scaffold significantly reduces inflammation, enhances angiogenesis, promotes collagen deposition, and stimulates hair follicle regeneration, ultimately achieving high-quality wound healing with functional restoration. In conclusion, this study demonstrates a spatiotemporally controllable approach to modulate the immune microenvironment of inflammatory wounds while promoting vascular regeneration, offering a clinically translatable strategy for regenerative medicine. STATEMENT OF SIGNIFICANCE: Current wound therapies face challenges in dynamically regulating immune responses and angiogenesis. We developed a tilapia collagen-based piezoelectric hydrogel scaffold integrated with oxidized gellan gum and amino-modified barium titanate nanoparticles (Col/OG/BTN hydrogel scaffold). This ultrasound-activated system uniquely reprograms pro-inflammatory macrophages to pro-healing phenotypes via PI3K/Akt and TNF pathways, synergistically enhancing angiogenesis and hair follicle regeneration. The scaffold eliminates implanted electrodes, offering wireless immunomodulation and vascular restoration, enabling high-quality wound healing with functional skin appendage recovery. This work provides a clinically translatable strategy for inflammatory wound repair through bioelectrical signaling.

Collagen is a highly favorable macromolecule for biomedical applications and is often made into functional forms via gelation. Conventional gelation approach relies on the self-assembly of collagen molecules via pH and temperature adjustment, which is slow and tedious, and often associated with poor physical strength. To overcome these challenges, we present a facile and rapid collagen gelation strategy using epigallocatechin gallate (EGCG), a natural polyphenol from green tea. Upon simple mixing with solubilized collagen, EGCG induced the spontaneous formation of stable and injectable hydrogels via hydrophobic and π-π interactions. Capitalizing on the hydrogel's thixotropic behavior and the established skin health benefits of collagen and EGCG, we investigated its translational potential. The hydrogel exhibited excellent skin compatibility, prolonged surface retention with moisturizing effects, and erosion-controlled codelivery of proteins and EGCG. Collectively, these findings highlight the potential of the collagen-EGCG hydrogel as a promising platform for topical applications.

Tissue engineered skin equivalents are increasingly recognized as potential alternatives to traditional skin models such as human ex vivo skin or animal skin models. However, most of the currently investigated human skin equivalents (HSEs) are constructed using mammalian collagen which can be expensive and difficult to extract. Fish skin is a waste product produced by fish processing industries and identified as a cost-efficient and sustainable source of type I collagen. In this work, we describe a method for generating highly stable HSEs based on fibrin fortified tilapia fish collagen. The fortified fish collagen (FFC) formulation is optimized to enable reproducible fabrication of full-thickness HSEs that undergo limited contraction, facilitating the incorporation of human donor-derived skin cells and formation of biomimetic dermal and epidermal layers. The morphology and barrier function of the FFC HSEs are compared with a commercial skin model and validated with immunohistochemical staining and transepithelial electrical resistance testing. Finally, the potential of a high throughput screening platform with FFC HSE is explored by scaling down its fabrication to 96-well format.

In recent years, we have witnessed remarkable progress in the field of regenerative medicine, in large part fuelled by developments in advanced biofabrication technologies such as three-dimensional (3D) bioprinting [...].

Microneedles (MNs) is an emerging technology that employs needles ranging from 10 to 1000 μm in height, as a minimally invasive technique for various procedures such as therapeutics, disease monitoring and diagnostics. The commonly used method of fabrication, micromolding, has the advantage of scalability, however, micromolding is unable to achieve rapid customizability in dimensions, geometries and architectures, which are the pivotal factors determining the functionality and efficacy of the MNs. 3D printing offers a promising alternative by enabling MN fabrication with high dimensional accuracy required for precise applications, leading to improved performance. Furthermore, enabled by its customizability and one-step process, there is propitious potential for growth for 3D-printed MNs especially in the field of personalized and on-demand medical devices. This review provides an overview of considerations for the key parameters in designing MNs, an introduction on the various 3D-printing techniques for fabricating this new generation of MNs, as well as highlighting the advancements in biomedical applications facilitated by 3D-printed MNs. Lastly, we offer some insights into the future prospects of 3D-printed MNs, specifically its progress towards translation and entry into market.

Collagen is the primary component of the extracellular matrix in humans. Traditionally commercial collagen is confined to bovine and porcine sources which have concerns of pathogenic transfer. Marine wastage accounts up to 85% by weight in the fishing industry. Extraction of collagen from these wastes for economic value and environmental sustainability is clear. Marine collagens have several advantages such as excellent biocompatibility, lower zoonotic risks, less immunological risk for patients allergic to mammalian products, and less religious restrictions. However, the properties of marine collagen-based constructs are highly dependent on the methods of fabrication. This article reviews advances in the design and fabrication of marine collagen-based constructs for medical applications. The potential applications of marine collagen in the regeneration of skin, bone and cartilage were also highlighted.

In vitro three-dimensional (3D) skin tissue models are critical tools in advancing our understanding of basic skin physiology and function as well as in specific applications such as toxicity testing of dermatological compounds. However, the utilization of such skin models is often limited by the structural instability of the construct, lack of physiologically relevant features and weak barrier function. In this review, we highlight the current research efforts in hydrogel biomaterial selection and scaffold design that allow for maturation of engineered skin in vitro, with special emphasis on matured full-thickness (including epidermal and dermal compartments) skin. The different types of scaffold biomaterials, broadly categorized as natural, synthetic, or composite will also be discussed. At the same time, we will outline strategies for next-generation biomimetic skin templates incorporating skin appendages or perfusion systems that can more closely reflect the native skin environment. STATEMENT OF SIGNIFICANCE: In vitro 3D human skin models are critical tools in advancing our understanding of skin physiology and function. Many of the existing reconstructed models are limited in terms of structure and complexity, thus failing to recapitulate native human skin. In order to address this, hydrogels have been identified as useful scaffold materials for fabricating the dermal equivalent of 3D skin models, allowing for greater flexibility and control in scaffold properties and cellular incorporation. This review aims to provide a critical discussion of the biomaterial selection and design strategies in the construction of hydrogel-based full-thickness skin equivalents. At the same time, we will offer insights into the future developments and technological advances which can accelerate the progress in this field.

Skin microbiome sampling is currently performed with tools such as swabs and tape strips to collect microbes from the skin surface. However, these conventional approaches may be unable to detect microbes deeper in the epidermis or in epidermal invaginations. We describe a sampling tool with a depth component, a transepidermal microprojection array (MPA), which captures microbial biomass from both the epidermal surface and deeper skin layers. We leveraged the rapid customizability of 3D printing to enable systematic optimization of MPA for human skin sampling. Evaluation of sampling efficacy on human scalp revealed the optimized MPA was comparable in sensitivity to swab and superior to tape strip, especially for nonstandard skin surfaces. We observed differences in species diversity, with the MPA detecting clinically relevant fungi more often than other approaches. This work delivers a tool in the complex field of skin microbiome sampling to potentially address gaps in our understanding of its role in health and disease.

Three-dimensional (3D) printed scaffolds have recently emerged as an innovative treatment option for patients with critical-sized skin wounds. Current approaches to managing life-threatening wounds include skin grafting and application of commercially sourced skin substitutes. However, these approaches are not without several challenges. Limited donor tissue and donor site morbidity remain a concern for tissue grafting, while engineered skin substitutes fail to fully recapitulate the complex native environment required for wound healing. The implementation of 3D printed dermal scaffolds offers a potential solution for these shortcomings. Spatial control over scaffold structure, the ability to incorporate multiple materials and bioactive ingredients, enables the creation of conditions specifically optimized for wound healing. Three-dimensional bioprinting, a subset of 3D printing, allows for the replacement of lost cell populations and secreted active compounds that contribute to tissue repair and recovery. The replacement of damaged and lost cells delivers beneficial effects directly, or synergistically, supporting injured tissue to recover its native state. Despite encouraging results, the promise of 3D printed scaffolds has yet to be realized. Further improvements to current material formulations and scaffold designs are required to achieve the goal of clinical adoption. Herein, we provide an overview of 3D printing techniques and discuss several strategies for healing of full-thickness wounds by using 3D printed acellular scaffolds or bioprinted cellular scaffolds, aimed at translating this technology to the clinical management of skin lesions. We identify the challenges associated with designing and optimizing printed tissue replacements, and discuss the future perspectives of this emerging option for managing patients who present with critical-sized life-threatening cutaneous wounds. Impact statement Chronic wounds and burn injuries often present with the full-thickness loss of skin, threatening the life of the patient and generating significant socioeconomic burden for these patients, their treating clinicians, and the wider community in which these patients live. Effective clinical management that permits damaged skin tissue to repair and restore its native functional state reduces the strain on health care systems. Three-dimensional (3D) printed scaffolds have been proposed as a potential solution and could be instrumental in facilitating the recovery and healing process. In this review, we will summarize the current research approaches, technologies, and limitations of 3D printed scaffolds as an efficient and effective approach to managing cutaneous wound healing.

3D printed scaffolds have recently emerged as an innovative treatment option for patients with critical-sized skin wounds. Current approaches to managing life-threatening wounds include skin grafting and application of commercially sourced skin substitutes. However, these approaches are not without several challenges. Limited donor tissue and donor site morbidity remain a concern for tissue grafting, while engineered skin substitutes fail to fully recapitulate the complex native environment required for wound healing. The implementation of 3D printed dermal scaffolds offers a potential solution for these shortcomings. Spatial control over scaffold structure, the ability to incorporate multiple materials and bioactive ingredients, enable the creation of conditions specifically optimized for wound healing. 3D bioprinting, a subset of 3D printing, allows for the replacement of lost cell populations and secreted active compounds that contribute to tissue repair and recovery. The replacement of damaged and lost cells delivers beneficial effects directly, or synergistically, supporting injured tissue to recover its native state. Despite encouraging results, the promise of 3D printed scaffolds has yet to be realized. Further improvements to current material formulations and scaffold designs are required to achieve the goal of clinical adoption. Herein, we provide an overview of 3D printing techniques and discuss several strategies for healing of full-thickness wounds by using 3D printed acellular scaffolds or bioprinted cellular scaffolds, aimed at translating this technology to the clinical management of skin lesions. We identify the challenges associated with designing and optimizing printed tissue replacements, and discuss the future perspectives of this emerging option for managing patients who present with critical-sized life-threatening cutaneous wounds.

Diabetes mellitus (DM) is a chronic metabolic disease with increasing prevalence worldwide. Diabetic foot ulcers (DFUs) are a serious complication of DM. It is estimated that 15–25% of DM patients develop DFU at least once in their lifetime. The lack of effective wound dressings and targeted therapy for DFUs often results in prolonged hospitalization and amputations. As the incidence of DM is projected to rise, the demand for specialized DFU wound management will continue to increase. Hence, it is of great interest to improve and develop effective DFU-specific wound dressings and therapies. In the last decade, 3D bioprinting technology has made a great contribution to the healthcare sector, with the development of personalized prosthetics, implants, and bioengineered tissues. In this review, we discuss the challenges faced in DFU wound management and how 3D bioprinting technology can be applied to advance current treatment methods, such as biomanufacturing of composite 3D human skin substitutes for skin grafting and the development of DFU-appropriate wound dressings. Future co-development of 3D bioprinting technologies with novel treatment approaches to mitigate DFU-specific pathophysiological challenges will be key to limiting the healthcare burden associated with the increasing prevalence of DM.

It has been shown that in vivo tissues are highly crowded by proteins, nucleic acids, ribonucleoproteins, polysaccharides, etc. The following protocol applies a macromolecular crowding (MMC) technique to mimic this physiological crowding through the addition of neutral polymers (crowders) to cell cultures in vitro. Previous studies using Ficoll or dextran as crowders demonstrate that the expression of collagen I and fibronectin in WI38 and WS-1 cell lines are significantly enhanced using the MMC technique. However, this technique has not been validated in primary hypertrophic scar-derived human skin fibroblasts (hHSFs). As hypertrophic scarring arises from the excessive deposition of collagen, this protocol aims to construct a collagen-rich in vitro hypertrophic scar model by applying the MMC technique with hHSFs. This optimized MMC model has been shown to possess more similarities with in vivo scar tissue compared to traditional 2-dimensional (2-D) cell culture systems. In addition, it is cost-effective, time-efficient, and ethically desirable compared to animal models. Therefore, the optimized model reported here offers an advanced "in vivo-like" model for hypertrophic scar-related studies.

Acute myeloid leukemia (AML) is an aggressive malignancy that leads to a poor prognosis even with intensive chemotherapy. As the key feature of AML is the blockade of hematopoietic cell maturation, considerable attention has been paid to 'differentiation therapy' aimed at transforming AML cells into more mature, benign phenotypes using pharmacological agents. Here we report a hyaluronic acid-(-)-epigallocatechin-3-O-gallate (HA-EGCG) conjugate as a unique anti-leukemic agent, capable of selectively killing AML cells as well as promoting their terminal differentiation into monocytes and granulocytes. This 'two-pronged' effect of the HA-EGCG conjugate was demonstrated in two different AML cell lines (NB4 and HL60), but absent in a physical mixture (HA + EGCG), highlighting the importance of HA conjugation for targeting of EGCG moieties to AML cells. Moreover, administration of the HA-EGCG conjugate not only suppressed AML progression, but also prolonged survival in the HL60 xenograft mouse model. Our study suggests new opportunities for designing two-pronged anti-leukemic agents for more effective AML treatment.

The recent advances in the design of injectable hydrogels for stem cell delivery, especially for in vivo applications, are overviewed in this review.

Low drug loading and instability in blood circulation are two key challenges that impede the successful clinical translation of nanomedicine, as they result in only marginal therapeutic efficacy and toxic side effects associated with premature drug leakage, respectively. Herein, highly stable and ultrahigh drug loading micellar nanocomplexes (MNCs) based on the self-assembly of the anticancer drug doxorubicin (DOX) and a poly(ethylene glycol)-epigallocatechin-3-O-gallate (EGCG) conjugate are developed. The formation of these MNCs is facilitated by strong favorable intermolecular interactions between the structurally similar aromatic EGCG and DOX molecules, which impart exceptionally high drug-loading capability of up to 88% and excellent thermodynamic and kinetic stability. Unlike two clinical formulations of DOX-free DOX and liposomal DOX, which are not effective below their lethal dosages, these DOX-loaded MNCs demonstrate significant tumor growth inhibition in vivo on a human liver cancer xenograft mouse model with minimal unwanted toxicity. Overall, these MNCs can represent a safe and effective strategy to deliver DOX for cancer therapy.

Additive manufacturing (AM) is a fast-growing manufacturing approach that comes with the promise of delivering personalized medicine to treat individual patients. However, large-scale commercial applications in the pharmaceutical industry have been limited. Here, some of the challenges are discussed along with some pharmaceutical products where AM has the potential to make a tangible impact and pave the way for more rapid adoption are highlighted.

Despite the burgeoning interest in three-dimensional (3D) printing for the manufacture of customizable oral dosage formulations, a U.S. Food and Drug Administration-approved tablet notwithstanding, the full potential of 3D printing in pharmaceutical sciences has not been realized. In particular, 3D-printed drug-eluting devices offer the possibility for personalization in terms of shape, size, and architecture, but their clinical applications have remained relatively unexplored. We used 3D printing to manufacture a tailored oral drug delivery device with customizable design and tunable release rates in the form of a mouthguard and, subsequently, evaluated the performance of this system in the native setting in a first-in-human study. Our proof-of-concept work demonstrates the immense potential of 3D printing as a platform for the development and translation of next-generation drug delivery devices for personalized therapy.