Identification and classification of reverse transcriptases in bacterial genomes and metagenomes.

Reverse transcriptases (RTs) are found in different systems including group II introns, Diversity Generating Retroelements (DGRs), retrons, CRISPR-Cas systems, and Abortive Infection (Abi) systems in prokaryotes. Different classes of RTs can play different roles, such as template switching and mobility in group II introns, spacer acquisition in CRISPR-Cas systems, mutagenic retrohoming in DGRs, programmed cell suicide in Abi systems, and recently discovered phage defense in retrons. While some classes of RTs have been studied extensively, others remain to be characterized. There is a lack of computational tools for identifying and characterizing various classes of RTs. In this study, we built a tool (called myRT) for identification and classification of prokaryotic RTs. In addition, our tool provides information about the genomic neighborhood of each RT, providing potential functional clues. We applied our tool to predict RTs in all complete and draft bacterial genomes, and created a collection that can be used for exploration of putative RTs and their associated protein domains. Application of myRT to metagenomes showed that gut metagenomes encode proportionally more RTs related to DGRs, outnumbering retron-related RTs, as compared to the collection of reference genomes. MyRT is both available as a standalone software (https://github.com/mgtools/myRT) and also through a website (https://omics.informatics.indiana.edu/myRT/).

Information optimization of laser scanning microscopes for real-time feedback-driven single particle tracking.

Real-time feedback-driven single particle tracking (RT-FD-SPT) is a class of microscopy techniques that uses measurements of finite excitation/detection volume in a feedback control loop to actuate that volume and track with high spatio-temporal resolution a single particle moving in three dimensions. A variety of methods have been developed, each defined by a set of user-defined choices. Selection of those values is typically done through ad hoc, off-line tuning for the best perceived performance. Here we present a mathematical framework, based on optimization of the Fisher information, to select those parameters such that the best information is acquired for estimating parameters of interest, such as the location of the particle, specifics of the excitation beam such as its dimensions or peak intensity, or the background noise. For concreteness, we focus on tracking of a fluorescently-labeled particle and apply this framework to determine the optimal parameters for three existing fluorescence-based RT-FD-SPT techniques with respect to particle localization.

Anti-leishmanial, immunomodulatory and additive potential effect of Piperine on Leishmania major: The in silico and in vitro study of Piperine and its combination.

BACKGROUND: Piperine (Pn), an indole alkaloid compound found in pepper, is an effective compound with anti-leishmanial medications that administered alone or in combination. This study aimed to use Pn for possible biochemical targets and to assess mechanisms of anti-leishmanial action and immunomodulatory roles. METHODS: The ability of Pn to bind to interleukin-12P40 (IL-12P40) and interferon-γ (IFN-γ) was investigated using molecular docking. The leishmanicidal effect of Pn, meglumine antimoniate (Glucantime®; MA), and Pn plus MA was assessed on Leishmania major promastigotes and amastigotes. A real-time PCR was applied to quantify cytokines gene expression in drug-treated murine macrophages. RESULTS: The molecular docking findings indicated that Pn could bind to IL-12P40/IFN-γ. In silico analyses showed an affinity of Pn to IL-12P40/IFN-γ, with the MolDock score of -236.91 and -64.87 kcal/mol, respectively. Pn plus MA reduced the proliferation rate of promastigote and amastigote forms of L. major compared to each drug alone (IC50 = 43.22 and 19.41 µg/mL, respectively). Moreover, the combination drug demonstrated no cytotoxicity as the selectivity index (SI) was 14.81. Also, Th1-related cytokines were upregulated, while Th2-related cytokines were downregulated in Pn combination-treated murine macrophages. CONCLUSIONS: The superior effectiveness of combination therapy on L. major warrants further investigations on the clinical potential of this combination in the treatment of leishmaniasis.

A mechanistic overview of sulforaphane and its derivatives application in diabetes and its complications.

Sulforaphane (SFN) is a type of phytochemical found in many cruciferous vegetables that has been shown to positively benefit the control of Type 2 Diabetes Mellitus (T2DM). The search was done from 2000 until December 2022 using PubMed, Scopus, Web of Sciences, and Google Scholar databases. We included all in vitro, in vivo, and clinical trials. Sulforaphane has been demonstrated to activate the PI3K/AKT and AMP-activated protein kinase pathways and the glucose transporter type 4 to increase insulin production and reduce insulin resistance. Interestingly, SFN possesses protective effects against diabetes complications, such as diabetic-induced hepatic damage, vascular inflammation and endothelial dysfunction, nephropathy, and neuropathy via nuclear factor erythroid 2-related factor 2 activation that leads to the translation of several anti-oxidant enzymes and regulation glycolysis, pentose phosphate pathway, fatty acid metabolism, glutamine metabolism, and glutathione metabolism. Furthermore, multiple clinical trial studies emphasized the ameliorating effects of SFN on T2DM patients. This review provides sufficient evidence for further research and development of sulforaphane as a hypoglycemic drug.

Cytotoxicity properties of plant-mediated synthesized K-doped ZnO nanostructures.

In this study, potassium-doped zinc oxide nanoparticles (K-doped ZnO NPs) were green-synthesized using pine pollen extracts based on bioethics principles. The synthesized NPs were analyzed using X-ray diffraction (XRD), inductively coupled plasma atomic emission spectroscopy (ICP-AES), scanning electron microscopy (SEM), energy-dispersive X-ray analysis (EDXA), and transmission electron microscopy (TEM). The cytotoxicity of these nanoparticles (NPs) on normal macrophage cells and cancer cell lines was evaluated. In the same concentrations of K-doped ZnO and pure ZnO NPs, K-doped ZnO NPs demonstrated higher toxicity. The results confirmed that the doped potassium could increase cytotoxicity. The IC50 of K-doped ZnO NPs, pure ZnO NPs, and the examined control drug were 497 ± 15, 769 ± 12, and 606 ± 19 µg/mL, respectively. Considering the obtained IC50 of K-doped ZnO NPs, they were more toxic to the cancer cell lines and had less cytotoxicity on normal macrophage cells.

MyDGR: a server for identification and characterization of diversity-generating retroelements.

MyDGR is a web server providing integrated prediction and visualization of Diversity-Generating Retroelements (DGR) systems in query nucleotide sequences. It is built upon an enhanced version of DGRscan, a tool we previously developed for identification of DGR systems. DGR systems are remarkable genetic elements that use error-prone reverse transcriptases to generate vast sequence variants in specific target genes, which have been shown to benefit their hosts (bacteria, archaea or phages). As the first web server for annotation of DGR systems, myDGR is freely available on the web at http://omics.informatics.indiana.edu/myDGR with all major browsers supported. MyDGR accepts query nucleotide sequences in FASTA format, and outputs all the important features of a predicted DGR system, including a reverse transcriptase, a template repeat and one (or more) variable repeats and their alignment featuring A-to-N (N can be C, T or G) substitutions, and VR-containing target gene(s). In addition to providing the results as text files for download, myDGR generates a visual summary of the results for users to explore the predicted DGR systems. Users can also directly access pre-calculated, putative DGR systems identified in currently available reference bacterial genomes and a few other collections of sequences (including human microbiomes).

Leishmanicidal activities of biosynthesized BaCO3 (witherite) nanoparticles and their biocompatibility with macrophages.

The aim of this study was cost-effective and greener synthesis of barium carbonate (BaCO3 or witherite) nanoparticles with economic importance, and to evaluate their therapeutic potentials and biocompatibility with immune cells. Barium carbonate nanoparticles were biosynthesized using black elderberry extract in one step with non-toxic precursors and simple laboratory conditions; their morphologies and specific structures were analyzed using field emission scanning electron microscopy with energy dispersive X-ray spectroscopy (FESEM-EDX). The therapeutic capabilities of these nanoparticles on the immune cells of murine macrophages J774 and promastigotes Leishmania tropica were evaluated. BaCO3 nanoparticles with IC50 = 46.6 µg/mL were more effective than negative control and glucantium (positive control) in reducing promastigotes (P < 0.01). Additionally, these nanoparticles with a high value of cytotoxicity concentration 50% (CC50) were less toxic to macrophage cells than glucantime; however, they were significantly different at high concentrations compared to the negative control.

Synaptic plasticity in hippocampal CA1 neurons and learning behavior in acute kidney injury, and estradiol replacement in ovariectomized rats.

BACKGROUND: Neurological complications may occur in patients with acute or chronic renal failure; however, in cases of acute renal failure, the signs and symptoms are usually more pronounced, and progressed rapidly. Oxidative stress and nitric oxide in the hippocampus, following kidney injury may be involved in cognitive impairment in patients with uremia. Although many women continue taking hormone therapy for menopausal symptom relief, but there are also some controversies about the efficacy of exogenous sex hormones, especially estrogen therapy alone, in postmenopausal women with kidney injury. Herein, to the best of our knowledge for the first time, spatial memory and synaptic plasticity at the CA1 synapse of a uremic ovariectomized rat model of menopause was characterized by estradiol replacement alone. RESULTS: While estradiol replacement in ovariectomized rats without uremia, promotes synaptic plasticity, it has an impairing effect on spatial memory through hippocampal oxidative stress under uremic conditions, with no change on synaptic plasticity. It seems that exogenous estradiol potentiated the deleterious effect of acute kidney injury (AKI) with increasing hippocampal oxidative stress. CONCLUSIONS: Although, estrogen may have some positive effects on cognitive function in healthy subjects, but its efficacy in menopause subjects under uremic states such as renal transplantation, needs to be further investigated in terms of dosage and duration.

Differential expression of TLRs 2, 4, 9, iNOS and TNF-α and arginase activity in peripheral blood monocytes from glucantime unresponsive and responsive patients with anthroponotic cutaneous leishmaniasis caused by Leishmania tropica.

BACKGROUND: Detection of the mechanism of host/parasite interactions in unresponsive forms of anthroponotic cutaneous leishmaniasis (ACL) caused by Leishmania tropica is helpful for immunotherapy and vaccine development. In the present study, the gene expression of toll-like receptors (TLRs), TNF-α, iNOS and also arginase (ARG) activity in monocytes from Glucantime unresponsive in comparison to responsive patients infected with L. tropica was investigated. METHODS: In this case-control study, patients with unresponsive (n = 10) and responsive (n = 10) ACL were recruited. Gene expression of TLR2, TLR4, TLR9, TNF-α and iNOS was analyzed in L. tropica-exposed monocytes. The level of ARG activity in both isolated promastigotes and the lysates of monocytes was also determined. RESULTS: L. tropica-exposed monocytes represented higher expression of all three TLRs and TNF-α and lower expression of iNOS compared to unexposed ones in both groups of patients. Results revealed a significant down-regulation of TLR2 and TNF-α and up-regulation of TLR9 expression in unresponsive isolates in comparison to responsive ones. Besides, ARG level showed a significant increase in L. tropica-stimulated monocytes and cultured promastigotes from unresponsive isolates versus responsive ones. CONCLUSIONS: The decreased TLR2, TLR4, TNF-α and iNOS and the increased level of TLR9 expression in L. tropica-exposed monocytes from unresponsive isolates and also the increment in ARG activity in their promastigotes and monocytes, might possibly be involved in the severity of the disease and leading to Glucantime unresponsiveness.

Leishmanicidal, cytotoxic and apoptotic effects of Gossypium hirsutum bulb extract and its separated fractions on Leishmania major.

BACKGROUND & OBJECTIVES: Leishmaniasis is a major global health problem with no safe and effective therapeutic drugs. This study evaluated the cytotoxic and apoptotic effects of crude extract and fractions of Gossypium hirsutum bulb on Leishmania major stages using advanced experimental models. METHODS: Bulbs of G. hirsutum were collected from the Kerman province of Iran. The bulb was extracted using Soxhlet apparatus and different fractions were obtained by column chromatography (CC). Different concentrations of the extract and the fractions were evaluated against L. major and compared with Glucantime®. The cytotoxicity and apoptotic values were analysed by flow cytometry. The fractions obtained in CC were monitored by thin layer chromatography, and fractions with similar chromatographic patterns were mixed. RESULTS: The extract and two fractions, F4 and F5 inhibited the proliferation of L. major promastigotes and amastigotes in a dose-dependent manner at 72 h post-treatment. No significant cytotoxic effects were observed for extract and fractions, as the selectivity index was over 1000, far beyond >10. The mean apoptotic values for L. major were superior to those of Glucantime®. INTERPRETATION & CONCLUSION: Both the crude extract and fractions (F4 and F5) had significant antileishmanial effects on L. major stages, and were were superior relative to Glucantime®. No cytotoxic effects were associated with the extract or fractions and they showed excellent apoptotic index, a possible mechanism behind inducing parasite death. Further investigations are essential to study the effect of G. hirsutum bulb fractions in animal model and clinical settings for planning strategies for the prevention and control of leishmaniasis.

Vascular apoptosis associated with meglumine antimoniate: In vivo investigation of a chick embryo model.

The vasculo-toxic effect of meglumine antimoniate (MA) was confirmed in our previous investigation. The current study investigates the association of this effect with altered VEGF-A and VEGF-R2 expression. Additional mechanisms by which MA causes vascular toxicity are not clearly understood. We hypothesized that MA may alter normal expression of apoptotic genes and cause vascular toxicity. The current investigation was designed to address this issue using a chick embryo model. Fertile chicken eggs were treated with MA and the extra-embryonic membrane (EEM) vasculature was evaluated by morphometric, molecular and immunohistochemistry assays. The results showed that MA not only altered apoptotic gene expression, but that this alteration may disturb the normal development of the vascular network and cause embryo malformation. The relative expression level of the CASP3, CASP7, CASP9, APAF1, AIF1 and TP53 genes increased in drug-exposed EEMs. In addition, IHC assay confirmed the low expression BCL2 and increased expression of Bax, which are associated with a high rate of apoptosis. We suggest that induction of an apoptotic signaling pathway can lead to vascular defects during embryo development and the consecutive cascade of events can lead to the embryo malformation.

Comparison of autologous fibrin glue versus nylon sutures for securing conjunctival autografting in pterygium surgery.

PURPOSE: To compare the safety, operating time, postoperative ocular signs, symptoms, overall patient satisfaction, complications rate and recurrence rate of autologous fibrin glue (AFG) and nylon suturing (NS) for attaching conjunctival autografts in pterygium surgery. METHODS: A prospective, randomized, interventional study was performed among 120 patients (120 eyes) with primary pterygium. Superior conjunctival autograft was harvested and transferred on to bare sclera after pterygium excision. For attaching the autograft, AFG (n = 60 eyes) and NS (n = 60 eyes) were used. The patients were followed up for 12 months. The groups were compared for the safety, operative time, postoperative ocular signs, symptoms, overall patient satisfaction, recurrence and complications rate. RESULTS: All conjunctival autografts in both groups were successfully attached. The average operating time for the AFG group was significantly shorter (P < 0.001). Postoperative symptoms were fewer for the AFG group than the suture group. After 12 months of follow-up, no recurrence was reported for the AFG group, but 8.3% of patients experienced recurrence in the NS group. CONCLUSIONS: Our study demonstrated the superiority of AFG to NS in saving operating time and elimination of recurrence without any complications in pterygium surgery. AFG can obviate the need for suturing and increases the viability of tissue flaps. Additional studies are necessary to determine the long-term effects considering factors such as primary or recurrent status of pterygium, age and sex of patient, dose and duration of treatment for mitomycin C.

Protein cold adaptation: Role of physico-chemical parameters in adaptation of proteins to low temperatures.

During years 2007 and 2008, we published three papers (Jahandideh, 2007a, JTB, 246, 159-166; Jahandideh, 2007b, JTB, 248, 721-726; Jahandideh, 2008, JTB, 255, 113-118) investigating sequence and structural parameters in adaptation of proteins to low temperatures. Our studies revealed important features in cold-adaptation of proteins. Here, we calculate values of a new set of physico-chemical parameters and perform a comparative systematic analysis on a more comprehensive database of psychrophilic-mesophilic homologous protein pairs. Our obtained results confirm that psychrophilicity rules are not merely the inverse rules of thermostability; for instance, although contact order is reported as a key feature in thermostability, our results have shown no significant difference between contact orders of psychrophilic proteins compared to mesophilic proteins. We are optimistic that these findings would help future efforts to propose a strategy for designing cold-adapted proteins.

Pneumatic retinopexy for primary repair of rhegmatogenous retinal detachments.

PURPOSE: To evaluate anatomical and functional outcomes of pneumatic retinopexy (PR) for primary repair of rhegmatogenous retinal detachments and to determine demographic and ocular risk factors for failure of the procedure. DESIGN: Retrospective interventional case series. METHODS: A chart review on 97 eyes of 97 consecutive patients who had undergone PR as the initial procedure for unilateral recent primary retinal detachments with causative break(s) in the superior two-third of the retina. After injection of 0.35 mL to 0.60 mL of sulfur hexafluoride, 6 hours face-down positioning, and the steamroller maneuver, the patient's head position was changed so that the gas could efficiently tamponade the retinal break(s). Pre- and post-gas injection laser photocoagulation of break(s) and 360° peripheral retina was applied. RESULTS: Seventy-two male and 25 female patients were included in this study. The single-operation success rate was 82.5%. The pars plana vitrectomy procedure as the second intervention was not adversely affected by the preceding PR. Presence of proliferative vitreoretinopathy (odds ratio: 58.7, 95% confidence interval: 7.8-443.5) and delay between initial symptoms and PR (odds ratio: 1.21 per each additional day, 95% confidence interval: 1.07-1.37) were the only independent predictors for the failure. CONCLUSION: With proper selection of cases, PR is a good surgical option for primary rhegmatogenous retinal detachments with acceptable success rate, minimal discomfort to the patient, and minimal surgical risks.

Facile fabrication of next-generation sustainable brick and mortar through geopolymerization of construction debris.

Waste from construction and demolition (also known as CDW) is one of the most harmful environmental issues. This study's primary goal is to produce new mortar and brick materials from recycled concrete powder (RCP) and recycled brick powder (RBP), two of the most popular CDW. Geopolymeric mortar and brick samples were produced by passing RCP and RBP through sieve No. 50 (with sand filler if necessary) and combining them with an alkaline solution made of water glass (WG) and NaOH. In this study, the mixture was then cured for three days at 80 °C in an oven. The effects of filler, RBP amount, WG amount, and the concentration of NaOH alkaline solution on the samples' strength were examined. Additionally, XRF and SEM/XRD tests were performed to verify the materials' composition and microstructure. The mechanical strength of the samples showed an increase with the increase of RCP values, so the brick sample with filler showed the highest compressive strength, measuring 59.53 MPa. The study's samples exhibited strong mechanical properties. Additionally, all of the bricks' water absorption fell within the standard range. In summary, according to different standards, both waste concrete and waste brick can be used to produce geopolymer materials especially bricks for construction and paving purposes.

Gallocin-derived Engineered Peptides Targeting EGFR and VEGFR in Colorectal Cancer: A Bioinformatic Approach.

BACKGROUND: Colorectal cancer (CRC) treatment using time-saving and cost-effective targeted therapies with high selectivity and low toxicity drugs, is a great challenge. In primary investigations on Gallocin, as a most proposed factor in CRC pathogenesis caused by Streptococcus gallolyticus, it was surprisingly found that this bacteriocin has four α-helix structures and some anti- cancer sequences. OBJECTIVE: The aim of this study was to determine the ability of Gallocin-based anticancer peptides (ACPs) against epidermal growth factor receptor (EGFR) and vascular epidermal growth factor receptor (VEGFR) and the evaluation of their pharmacokinetics properties using bioinformatics approaches. METHODS: Support vector machine algorithm web-based tools were used for predicting ACPs. The physicochemical characteristics and the potential of anti-cancer activity of Gallocin-derived ACPs were determined by in silico tools. The 3D structure of predicted ACPs was modeled using modeling tools. The interactions between predicted ACPs and targets were investigated by molecular docking exercises. Then, the stability of ligand-receptor interactions was determined by molecular dynamic simulation. Finally, ADMET analysis was carried out to check the pharmacokinetic properties and toxicity of ACP. RESULTS: Four amino acid sequences with anti-cancer potential were selected. Through molecular docking, Pep2, and Pep3 gained the best scores, more binding affinity, and strong attachments by the formation of reasonable H-bonds with both EGFR and VEGFR. Molecular simulation confirmed the stability of Pep3- EGFR. According to pharmacokinetic analysis, the ACPs were safe and truthful. CONCLUSION: Designed peptides can be nominated as drugs for CRC treatment. However, different in-vitro and in-vivo assessments are required to approve this claim.

Synergistic activity of crocin and crocin loaded in niosomes alone and in combination with fluconazole against Candida albicans isolates: In vitro and in silico study.

INTRODUCTION: Since the drug resistance in Candida species is becoming a serious clinical challenge, novel alternative therapeutic options, particularly herbal medicine, have attracted increasing interest. This study aimed to pinpoint the potential antifungal activity of crocin (Cro), the efficacy of the niosomal formulation of Cro (NCro), and the synergistic activity of both formulations in combination with fluconazole (FLC) against susceptible and resistant C. albicans isolates. MATERIAL AND METHODS: NCro was formulated using the heating method. The in vitro antimycotic activity of Cro, NCro, and FLC was evaluated. Checkerboard and isobologram assays evaluated the interaction between both formulations of Cro and FLC. Necrotic and apoptotic effects of different agents were analyzed using the flow cytometry method. In silico study was performed to examine the interactions between Lanosterol 14 alpha-demethylase and Cro as a part of our screening compounds with antifungal properties. RESULTS: NCro exhibited high entrapment efficiency up to 99.73 ± 0.54, and the mean size at 5.224 ± 0.618 µm (mean ± SD, n = 3). Both formulations of Cro were shown to display good anticandidal activity against isolates. The synergistic effect of the NCro in combination with FLC is comparable to Cro (P-value =0.03). Apoptotic indicators confirmed that tested compounds caused cell death in isolates. The docking study indicated that Cro has interactivity with the protein residue of 14α-demethylase. CONCLUSION: The results showed a remarkable antifungal effect by NCro combined with FLC. Natural compounds, particularly nano-sized carrier systems, can act as an effective therapeutic option for further optimizing fungal infection treatment.

Isolation and characterization of two Staphylococcus aureus lytic bacteriophages "Huma" and "Simurgh".

Nowadays finding the new antimicrobials is necessary due to the emerging of multidrug resistant strains. The present study aimed to isolate and characterize bacteriophages against S. aureus. Strains Huma and Simurgh were the two podovirus morphology phages which isolated and then characterized. Huma and Simurgh had a genome size of 16,853 and 17,245 bp, respectively and both were Rosenblumvirus with G + C content of 29%. No lysogeny-related genes, nor virulence genes were identified in their genomes. They were lytic only against two out of four S. aureus strains. They also were able to inhibit S. aureus for 8 h in-vitro. Both showed a rapid adsorption. Huma and Simurgh had the latent period of 80 and 60 m and the burst sizes of 45 and 40 PFU/ml and also, they showed very low cell toxicity of 1.23%-1.79% on HT-29 cells, respectively. Thus, they can be considered potential candidates for biocontrol applications.

Global Dilemma and Needs Assessment Toward Achieving Sustainable Development Goals in Controlling Leishmaniasis.

Leishmaniasis is a disease of poverty that imposes a devastating medical, social, and economic burden on over 1 billion people nationwide. To date, no in-depth study to analyze the major global challenges and needs assessment has been carried out. This investigation aimed to explore a comprehensive narrative review of leishmaniasis's main challenges and initially highlight obstacles that might impede the implementation of control measures. Also, we propose a specific list of priorities for needs assessment. The presence of socioeconomic factors, multiple clinical and epidemiological forms, various Leishmania species, the complexity of the life cycle, the absence of effective drugs and vaccines, and the lack of efficient vector and reservoir control make this organism unique and sophisticated in playing a tangled role to react tricky with its surrounding environments, despite extensive efforts and implementation of all-inclusive former control measures. These facts indicate that the previous strategic plans, financial support, and basic infrastructures connected to leishmaniasis surveillance are still insufficient. Strengthening the leishmaniasis framework in a context of accelerated programmatic action and intensification of cross-cutting activities along with other neglected tropical diseases (NTDs) is confidently expected to result in greater effectiveness, cost-benefit, and fruitful management. Sensitive diagnostics, effective therapeutics, and efficacious vaccines are vital to accelerating advancement toward elimination, and reducing morbidity/mortality and program costs. Collective actions devoted by all sectors and policy-makers can hopefully overcome technical and operational barriers to guarantee that effective and coordinated implementation plans are sustained to meet the road map for NTDs 2021- 2030 goals.

Precise Engineering of Growth Factor Presentation Using Extracellular Microenvironment-Mimicking Microfluidic Microparticles.

One of the main challenges in tissue engineering is finding a way to deliver specific growth factors (GFs) with precise spatiotemporal control over their presentation. Here, we report a novel strategy for generating microscale carriers with enhanced affinity for high content loading suitable for the sustained and localized delivery of GFs. Our developed microparticles can be injected locally and sustainably release encapsulated growth factors for up to 28 days. Fine-tuning of particles' size, affinity, microstructures, and release kinetics is achieved using a microfluidic system along with bioconjugation techniques. We also describe an innovative 3D micromixer platform to control the formation of core-shell particles based on superaffinity using a polymer-peptide conjugate for further tuning of release kinetics and delayed degradation. Chitosan shells block the burst release of encapsulated GFs and enable their sustained delivery for up to 10 days. The matched release profiles and degradation provide the local tissues with biomimetic, developmental-biologic-compatible signals to maximize regenerative effects. The versatility of this approach is verified using three different therapeutic proteins, including human bone morphogenetic protein-2 (rhBMP-2), vascular endothelial growth factor (VEGF), and stromal cell-derived factor 1 (SDF-1α). As in vivo morphogenesis is typically driven by the combined action of several growth factors, the proposed technique can be developed to generate a library of GF-loaded particles with designated release profiles.