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Bitter orange is a citrus fruit rich in bioactive compounds, but its waste is currently underutilized. One potential solution is to encapsulate these bioactive compounds. This research aims to synthesize gelatin nanoparticles loaded with an ethanolic extract of bitter orange peel and to evaluate their in vitro antioxidant and antibacterial activities. Coaxial electrospray was used to encapsulate the ethanolic extract of bitter orange with bovine gelatin as wall material, considering a voltage of 15 kV, a wall solution flow rate of 0.1 mL/h, and a core solution flow rate of 0.08 mL/h. Characterization of the nanoparticles was performed using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR). Antioxidant activity was evaluated by the total phenolic content, flavonoids, and antioxidant capacity by the DPPH•, ABTS•+, and FRAP assays. Antibacterial activity was assessed by the well diffusion technique on Mueller–Hinton agar against Listeria monocytogenes and Escherichia coli O157:H7 bacteria. SEM images confirmed that the nanoparticles were spherical in shape, while FT-IR analysis indicated that the incorporation of the extract did not alter the amide bonds of the gelatin protein. The nanoparticles containing the extract exhibited higher antioxidant activity and heightened inhibition against E. coli O157:H7, indicating their potential food applications. [ABSTRACT FROM AUTHOR]

This study reports the successful synthesis of core-shell microparticles utilizing coaxial electrospray techniques, with zeolitic imidazolate framework-8 (ZIF-8) encapsulating rhodamine B (RhB) in the core and a phase change material (PCM) shell composed of a eutectic mixture of lauric acid (LA) and stearic acid (SA). ZIF-8 is well-recognized for its pH-responsive degradation and biocompatibility, making it an ideal candidate for targeted drug delivery. The LA-SA PCM mixture, with a melting point near physiological temperature (39 °C), enables temperature-triggered drug release, enhancing therapeutic precision. The structural properties of the microparticles were extensively characterized through scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). Drug release studies revealed a dual-stimuli response, where the release of RhB was significantly influenced by both temperature and pH. Under mildly acidic conditions (pH 4.0) at 40 °C, a rapid and complete release of RhB was observed within 120 h, while at 37 °C, the release rate was notably slower. Specifically, the release at 40 °C was 79% higher than at 37 °C, confirming the temperature sensitivity of the system. Moreover, at physiological pH (7.4), minimal drug release occurred, demonstrating the system's potential for minimizing premature drug release under neutral conditions. This dual-stimuli approach holds promise for improving therapeutic outcomes in cancer treatment by enabling precise control over drug release in response to both pH and localized hyperthermia, reducing off-target effects and improving patient compliance. [ABSTRACT FROM AUTHOR]

Cancer is one of the major threats to human health and lives. However, effective cancer treatments remain a great challenge in clinical medicine. As a common approach for cancer treatment, chemotherapy has saved the life of millions of people; however, patients who have gone through chemotherapy often suffer from severe side effects owing to the inherent cytotoxicity of anti-cancer drugs. Stabilizing the blood concentration of an anti-cancer drug will reduce the occurrence or severity of side effects, and relies on using an appropriate drug delivery system (DDS) for achieving sustained or even on-demand drug delivery. However, this is still an unmet clinical challenge since the mainstay of anti-cancer drugs is small molecules, which tend to be diffused rapidly in the body, and conventional DDSs exhibit the burst release phenomenon. Here, we establish a class of DDSs based on biodegradable core–shell microspheres with encapsulated doxorubicin hydrochloride-loaded gold nanoparticles (DOX@Au@MSs), with the core–shell microspheres being made of poly(lactic-co-glycolic acid) in the current study. By harnessing the physical barrier of the biodegradable shell of core–shell microspheres, DOX@Au@MSs can provide a sustained release of the anti-cancer drug in the test duration (which is 21 days in the current study). Thanks to the photothermal properties of the encapsulated gold nanoparticle carriers, the core–shell biodegradable microspheres can be ruptured through remotely controlled near-infrared (NIR) light, thereby achieving an NIR-controlled triggered release of the anti-cancer drug. Furthermore, the route of the DOX-Au@MS-enabled controlled release of the anti-cancer drug can provide durable cancer cell ablation for the long period of 72 h. [ABSTRACT FROM AUTHOR]

Vancomycin in sustained release still needs to be investigated. Polylactic acid (PLA) was widely used in the biomedical field for its good biocompatibility, especially in the field of controlled drug release. In this study, polyethylene glycol (PEG) was used to modify PLA to improve the hydrophilicity of the material. The synthesis of the block copolymers was proven by infrared and nuclear magnetic characterization and the hydrophilicity was tested. Vancomycin was immobilized by coaxial electrospray, and PLA-PEG6000-PLA was used as the shell layer. The parameters of coaxial electrospray under this material were explored, and the effects of concentration, voltage and temperature on the formation of microspheres were systematically studied. The optimum parameters were determined as follows: concentration 20 wt%, temperature 35 °C and voltage 14 kV. The maximum encapsulation rate and drug loading were calculated to be 89.54 ± 1.22 % and 15.33 ± 0.97 %, respectively, and the cumulative release of drug-loaded microspheres was less than 45 % in 24 h with a slow releasing time of more than one month. The drug loaded microspheres showed good sustained release and good control of burst release. [ABSTRACT FROM AUTHOR]

Berberine (BBR) has many pharmacological activities including anti-inflammatory, antibacterial, and against many diseases, especially cancer. However, berberine is a large molecular compound with rapid metabolism by enzymes in the human body and limited bioavailability. To improve, berberine needs to be coated and combined with other active ingredients. In this study, berberine is successfully encapsulated by coaxial electrosprayed chitosan (CS) without cross-linked agents, and piperine (PPR) is also added via two different approaches to improve absorption and bioavailability. The obtained nanoparticles have a spherical shape with smooth surface and relatively wide particle size distribution observed by SEM. The uniform core/shell structure with clear boundary, non-agglomerated encapsulated spheres, and average diameter of 143.67 ± 56.16 nm (for BBR-PPR@CS) and 152.91 ± 48.50 nm (for BBR@PPR-CS) were analyzed by TEM images. The appearance of some weak peaks and slight shifting of major peaks in FTIR spectra of BBR-PPR@CS and BBR@PPR-CS samples and their high encapsulation efficiencies reflected their successful encapsulation. In drug release experiments, as expected, with the combination of PPR and CS in the outer shell (BBR@PPR-CS), the release of PPR was faster than that of BBR. The amount of drug in all samples was released more strongly at pH 5.0 than at pH 7.4, and the release time extended up to 36 h. Therefore, coaxial electrosprayed chitosan nanoparticle is a promising functional material for the simultaneous encapsulation of BBR and PPR, as well as increasing bioavailability of BBR, contributing to the development of controlled drugs toward effective anticancer. [ABSTRACT FROM AUTHOR]

The encapsulation and the oxidative stability of cod liver fish oil (CLO) within coaxial electrosprayed (ethyl cellulose/CLO) core–(octenyl succinic anhydride, OSA-modified starch) shell, and monoaxial electrosprayed ethyl cellulose/CLO microcapsules were investigated. Core–shell (H-ECLO) and monoaxial (ECLO) electrosprayed microcapsules with an average diameter of 2.8 ± 1.8 µm, and 2.2 ± 1.4 µm, respectively, were produced. Confocal microscopy confirmed not only the core–shell structure of the H-ECLO microcapsules, but also the location of the CLO in the core. However, for the ECLO microcapsules, the CLO was distributed on the microcapsules' surface, as also confirmed by Raman spectroscopy. Atomic force microscopy showed that the average surface adhesion of the H-ECLO microcapsules was significantly lower (5.41 ± 0.31 nN) than ECLO microcapsules (18.18 ± 1.07 nN), while the H-ECLO microcapsules showed a remarkably higher Young's modulus (33.84 ± 4.36 MPa) than the ECLO microcapsules (6.64 ± 0.84 MPa). Differential scanning calorimetry results confirmed that the H-ECLO microcapsules enhanced the oxidative stability of encapsulated CLO by about 15 times, in comparison to non-encapsulated oil, mainly by preventing the presence of the fish oil at the surface of the microcapsules, while ECLO microcapsules enhanced the oxidative stability of CLO about 2.9 times due to the hydrophobic interactions of the oil and ethyl cellulose. Furthermore, the finite element method was also used to evaluate the electric field strength distribution, which was substantially higher in the vicinity of the collector and lower in the proximity of the nozzle when the coaxial electrospray process was employed in comparison to the monoaxial process. [ABSTRACT FROM AUTHOR]

Bitter orange is a citrus fruit rich in bioactive compounds, but its waste is currently underutilized. One potential solution is to encapsulate these bioactive compounds. This research aims to synthesize gelatin nanoparticles loaded with an ethanolic extract of bitter orange peel and to evaluate their in vitro antioxidant and antibacterial activities. Coaxial electrospray was used to encapsulate the ethanolic extract of bitter orange with bovine gelatin as wall material, considering a voltage of 15 kV, a wall solution flow rate of 0.1 mL/h, and a core solution flow rate of 0.08 mL/h. Characterization of the nanoparticles was performed using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FT-IR). Antioxidant activity was evaluated by the total phenolic content, flavonoids, and antioxidant capacity by the DPPH•, ABTS•+, and FRAP assays. Antibacterial activity was assessed by the well diffusion technique on Mueller–Hinton agar against Listeria monocytogenes and Escherichia coli O157:H7 bacteria. SEM images confirmed that the nanoparticles were spherical in shape, while FT-IR analysis indicated that the incorporation of the extract did not alter the amide bonds of the gelatin protein. The nanoparticles containing the extract exhibited higher antioxidant activity and heightened inhibition against E. coli O157:H7, indicating their potential food applications.

Cancer is one of the major threats to human health and lives. However, effective cancer treatments remain a great challenge in clinical medicine. As a common approach for cancer treatment, chemotherapy has saved the life of millions of people; however, patients who have gone through chemotherapy often suffer from severe side effects owing to the inherent cytotoxicity of anti-cancer drugs. Stabilizing the blood concentration of an anti-cancer drug will reduce the occurrence or severity of side effects, and relies on using an appropriate drug delivery system (DDS) for achieving sustained or even on-demand drug delivery. However, this is still an unmet clinical challenge since the mainstay of anti-cancer drugs is small molecules, which tend to be diffused rapidly in the body, and conventional DDSs exhibit the burst release phenomenon. Here, we establish a class of DDSs based on biodegradable core–shell microspheres with encapsulated doxorubicin hydrochloride-loaded gold nanoparticles (DOX@Au@MSs), with the core–shell microspheres being made of poly(lactic-co-glycolic acid) in the current study. By harnessing the physical barrier of the biodegradable shell of core–shell microspheres, DOX@Au@MSs can provide a sustained release of the anti-cancer drug in the test duration (which is 21 days in the current study). Thanks to the photothermal properties of the encapsulated gold nanoparticle carriers, the core–shell biodegradable microspheres can be ruptured through remotely controlled near-infrared (NIR) light, thereby achieving an NIR-controlled triggered release of the anti-cancer drug. Furthermore, the route of the DOX-Au@MS-enabled controlled release of the anti-cancer drug can provide durable cancer cell ablation for the long period of 72 h.

Nanoparticles (NPs) are referred to as tiny materials in size ranging from 1 to 100 nm. Unique characteristics of the NPs, including small sizes and high surface area, appropriate reactivity, proper stability, great strength, and many more, have resulted in their wide use in numerous fields. Among different techniques reported for synthesizing the nanoparticles, electro-hydrodynamic atomization or electrospray has been identified as a well-practiced and high efficient technique for the formation of fine and homogenous NPs from a liquid under the influence of electrical forces. This process allows feasible encapsulation of different drugs, vitamins, and proteins applicable in the targeted drug delivery systems. Since the release rate of the loaded pharmaceutical materials could be easily tuned via varying the properties of core and shell components. Herein, we summarized the importance of the electrospray technique for the production of drug-loaded nanoparticles applicable in controlled drug delivery systems.

[Display omitted] Enhancing the efficacy and reducing the toxicity of chemotherapeutic agents like doxorubicin (DOX) is crucial in cancer treatment. Core-shell nanoparticles (NPs) fabricated by coaxial electrospraying offer controlled release of anticancer agents with the polymer shell protecting drug molecules from rapid degradation, prolonging therapeutic effect. This study developed DOX-loaded poly(lactic-co-glycolic acid) (PLGA) NPs. NPs were fabricated with matrix or core–shell structure via single needle or coaxial electrospraying, respectively. Core-shell NPs exhibited high encapsulation efficiency (>80 %) with controlled DOX distribution. Compared to matrix NPs, core–shell NPs demonstrated slower sustained release (69 % in 144 h) after reduced initial burst (22 % in 8 h). Release kinetics followed a diffusion mechanism when compared to free drug and matrix DOX-loaded NPs. In vitro assays showed core–shell NPs' enhanced cytotoxicity against breast cancer cells MCF-7, with higher uptake observed by fluorescence microscopy and flow cytometry. The IC 50 for core-shell NPs displayed a significant drop (0.115 μg/mL) compared to matrix NPs (0.235 μg/mL) and free DOX (1.482 μg/mL) after 72 h. Coaxial electrospraying enables the production of therapeutically advantageous core–shell NPs, offering controlled drug release with high encapsulation efficiency, potentially improving clinical anticancer chemotherapy. [ABSTRACT FROM AUTHOR]

To improve the antitumor effect of combined capecitabine (CAP) and osimertinib (OSI) therapy and quickly and efficiently reduce tumor volumes for preoperative chemotherapy, we designed a compound CAP colon-targeted microparticle (COPMP) prepared by coaxial electrospray. COPMP is a core–shell microparticle composed of a Eudragit S100 outer layer and a CAP/OSI-loaded PLGA core. In this study, we characterized its size distribution, drug loading (DL), encapsulation efficiency (EE), differential scanning calorimetry (DSC), Fourier transform infrared spectra (FTIR), in vitro release, formula ratio, cellular growth inhibition, and in vivo antitumor efficacy. COPMP is of spherical appearance with a size of 1.87 ± 0.23 μm. The DLs of CAP and OSI are 4.93% and 4.95%, respectively. The DSC showed that the phase state of CAP and OSI changed after encapsulation. The FTIR results indicated good compatibility between the drug and excipients. The release curve showed that CAP and OSI were released in a certain ratio. They were barely released prior to 2 h (pH 1.0), less than 50% was released between 3 and 5 h (pH 6.8), and sustained release of up to 80% occurred between 6 and 48 h (pH 7.4). CAP and OSI demonstrated a synergistic effect on HCT-116 cells. In a colon tumor model, the tumor inhibition rate after oral administration of COPMP reached 94% within one week. All the data suggested that COPMP promotes the sustained release of CAP and OSI in the colon, which provides a preoperative chemotherapy scheme for the treatment of colon cancer. [ABSTRACT FROM AUTHOR]

Abstract Background The oral colon-targeting drug delivery vehicle is vital for the efficient application of curcumin (Cur) in ulcerative colitis (UC) treatment because of its lipophilicity and instability in the gastrointestinal tract. Methods The core–shell microparticle (MP) system composed of eco-friendly materials, zein and shellac, was fabricated using a coaxial electrospray technique. In this manner, Cur was loaded in the zein core, with shellac shell coating on it. The colon-targeting efficiency and accumulation capacity of shellac@Cur/zein MPs were evaluated using a fluorescence imaging test. The treatment effects of free Cur, Cur/zein MPs, and shellac@Cur/zein MPs in acute experimental colitis were compared. Results With the process parameters optimized, shellac@Cur/zein MPs were facilely fabricated with a stable cone-jet mode, exhibiting standard spherical shape, uniform size distribution (2.84 ± 0.15 µm), and high encapsulation efficiency (95.97% ± 3.51%). Particularly, with the protection of shellac@zein MPs, Cur exhibited sustained drug release in the simulated gastrointestinal tract. Additionally, the in vivo fluorescence imaging test indicated that the cargo loaded in shellac@zein MPs improves the colon-targeting efficiency and accumulation capacity at the colonitis site. More importantly, compared with either free Cur or Cur/zein MPs, the continuous oral administration of shellac@Cur/zein MPs for a week could efficiently inhibit inflammation in acute experimental colitis. Conclusion The shellac@Cur/zein MPs would act as an effective oral drug delivery system for UC management.

Polyelectrolyte-doped microcapsules (PDM) was fabricated by coaxial electrospray of a mixture of glycerol and water containing 10 mg/mL cationic polyelectrolyte poly(allylamine hydrochloride) (PAH) fed as the core phase solution, and a N,N-dimethylacetylamide solution of 10 wt% polyurethane fed as the shell phase solution. Multi-enzyme system involving Candida Antarctica lipase B (CALB), glucose oxidase (GOD), and horseradish peroxidase (HRP) for cascade reaction was assembled in the PDM at three different places, namely, surface, shell, and lumen. Placing of enzyme inside aqueous lumen of the PDM was realized by in situ encapsulation through adding the enzyme in the core-phase solution for coaxial electrospray. By ion-pairing of enzyme with cationic surfactant CTAB, an organic soluble enzyme-CTAB complex was prepared, so that in situ embedding of enzyme in the shell of the PDM was realized by adding it into the shell phase solution. Surface attachment of enzymes was achieved by layer-by-layer (LbL) technology, which is based on the ion-exchange interactions between oppositely charged enzymes and PAH that was doped in PDM. The enzyme-decorated microcapsule was then studied as a micro-bioreactor, in which 1-Oxododecyla-α-glucopyranoside was converted by CALB to glucose, which was oxidised by GOD to gluconolactone in a second step. The hydrogen peroxide produced was then used by HRP to oxidize ABTS to form coloured radical cation ABTS•+ for activity analysis. The successful fabrication of the PDM and precise localization of enzymes in the PDM by different strategies were fully characterized. By varying the immobilization strategy, totally six PDM bioreactors with three enzymes precisely positional assembled in different strategies were constructed and their activities for the cascade reaction were investigated and compared. The PDM micro-bioreactor prepared by novel electrospray technologies provide a smart platform for positional assembly of multi-enzyme cascade reaction in a precise and well-controlled manner.

Diabetic foot ulcers (DFUs) are a significant challenge in the clinical care of diabetic patients, often necessitating limb amputation and compromising the quality of life and life expectancy of this cohort. Minimally invasive therapies, such as modular scaffolds, are at the forefront of current DFU treatment, offering an efficient approach for administering therapeutics that accelerate tissue repair and regeneration. In this study, we report a facile method for fabricating granular nanofibrous microspheres (NMs) with predesigned structures and porosities. The proposed technology combines electrospinning and electrospraying to develop a therapeutic option for DFUs. Specifically, porous NMs were constructed using electrospun poly(lactic- co -glycolic acid) (PLGA):gelatin short nanofibers, followed by gelatin cross-linking. These NMs demonstrated enhanced cell adhesion to human dermal fibroblasts (HDF) during an in vitro cytocompatibility assessment. Notably, porous NMs displayed superior performance owing to their interconnected pores compared to nonporous NMs. Cell-laden NMs demonstrated higher Young's modulus values than NMs without loaded cells, suggesting improved material resiliency attributed to the reinforcement of cells and their secreted extracellular matrix. Dynamic injection studies on cell-laden NMs further elucidated their capacity to safeguard loaded cells under pressure. In addition, porous NMs promoted host cell infiltration, neovascularization, and re-epithelialization in a diabetic mouse wound model, signifying their effectiveness in healing diabetic wounds. Taken together, porous NMs hold significant potential as minimally invasive, injectable treatments that effectively promote tissue integration and regeneration.

Despite its high efficacy and good patient compliance, the only long-acting injectable (LAI) contraceptive currently available in the US, depot medroxyprogesterone acetate (DMPA), is limited by significant side effects and a delayed return to fertility for up to 10 months after its intended duration of action. To overcome these limitations, we sought to develop an injectable poly(D, l -lactide) (PLA) microparticle for sustained release of contraceptive hormone, etonogestrel (ENG). A one-step technique, coaxial electrospray method was applied to prepare uniform ENG loaded core-shell structured and slow-degrading PLA microparticles (ENG-cs-MPs) to provide release control while minimizing polymer content. By adjusting voltage, polymer concentration and flow rate of the coaxial jetting solution, the prepared ENG-cs-MPs exhibited uniformly small particle size with volume mean diameter of 14.7 ± 0.5 μm and a shell thickness of 2.5 ± 0.1 μm, high drug loading of ~54%, high encapsulation efficiency of ~99%, and initial 1-day burst release of just ~10%. Long-term in vitro release of ENG was continuous for more than 3 months without change of the shell structure in 6 months. In PK studies, ENG-cs-MPs achieved a steady and continuous drug release for approximately 3 months and then quickly tapered off within 3 weeks. Hence, ENG-cs-MPs prepared by the coaxial electrospray method may be useful as a LAI contraceptive with an improved PK profile relative to DMPA. [Display omitted] • LAI contraceptives hold great promise to reduce worldwide unintended pregnancy. • An unmet need exists for LAI contraceptive products with fewer side effects and rapid return to fertility. • Coaxial electrospraying can produce uniform microparticles with a well-defined core-shell structure. • Reservoir design with slow-degrading PLA shell enables high drug loading and steady release via diffusion through the polymer. • PLA core-shell microparticles releases steroids for ~3 months followed by short PK tail. [ABSTRACT FROM AUTHOR]

In this study, a coaxial electrospray process was used to produce nanoparticles, in which thymol was encapsulated in a core–shell of zein and shellac. We then investigated fluid velocity and solution concentrations on the electrospray. Scanning electron microscopy revealed that the thymol/zein/shellac particles prepared at a flow rate ratio of 0.6:0.3 mL/h were rounder with smaller diameters. The highest encapsulation efficiency of thymol was 81.34%, the average particle size was 606.53 nm, and the retention rate of thymol was 68.74% after 30 days. Moreover, we investigated the thermal stability, antimicrobial activity, and simulated gastrointestinal release curves of the nanoparticles. Finally, the nanoparticles were used as a coating for fresh-cut cantaloupe. Compared with the blank nanoparticle control group, the coating significantly extended the shelf life of cantaloupe to 16 days. Our results indicate that these nanoparticles have great potential for application in food preservation.

Background: The oral colon-targeting drug delivery vehicle is vital for the efficient application of curcumin (Cur) in ulcerative colitis (UC) treatment because of its lipophilicity and instability in the gastrointestinal tract. Methods: The core–shell microparticle (MP) system composed of eco-friendly materials, zein and shellac, was fabricated using a coaxial electrospray technique. In this manner, Cur was loaded in the zein core, with shellac shell coating on it. The colon-targeting efficiency and accumulation capacity of shellac@Cur/zein MPs were evaluated using a fluorescence imaging test. The treatment effects of free Cur, Cur/zein MPs, and shellac@Cur/zein MPs in acute experimental colitis were compared. Results: With the process parameters optimized, shellac@Cur/zein MPs were facilely fabricated with a stable cone-jet mode, exhibiting standard spherical shape, uniform size distribution (2.84 ± 0.15 µm), and high encapsulation efficiency (95.97% ± 3.51%). Particularly, with the protection of shellac@zein MPs, Cur exhibited sustained drug release in the simulated gastrointestinal tract. Additionally, the in vivo fluorescence imaging test indicated that the cargo loaded in shellac@zein MPs improves the colon-targeting efficiency and accumulation capacity at the colonitis site. More importantly, compared with either free Cur or Cur/zein MPs, the continuous oral administration of shellac@Cur/zein MPs for a week could efficiently inhibit inflammation in acute experimental colitis. Conclusion: The shellac@Cur/zein MPs would act as an effective oral drug delivery system for UC management. [ABSTRACT FROM AUTHOR]

Natamycin (NAT) is the drug of choice for the treatment of fungal keratitis (FK). However, its inherent shortcomings, such as poor solubility, high dosing frequency, and long treatment cycle, need to be urgently addressed by designing a new delivery to widen its clinical utility. Growing research has confirmed that clotrimazole (CLZ) plays a significant role in fungal growth inhibition. Hence, coaxial electrospray (CO-ES) technology is used herein to prepare nano-systems with an average hydrodynamic particle size of 309-406 nm for the co-delivery of NAT and CLZ in chitosan (CTS) and poly(lactic-co-glycolic acid) (PLGA). The resulting NAT/CLZ@CTS/PLGA formulations were characterized by a transmission electron microscope (TEM) and in vitro release test. The results show that the formulations had obvious core-shell structures, uniform particle distribution, and also can sustain the release of drugs over 36 h. Furthermore, in vitro hemolysis, in vivo corneal irritation test, local allergenic test, and antifungal activity analyses are performed to evaluate the safety and efficiency of the formulations. Thus, good biosafety along with a significant anti-candidiasis effect are found in the NAT/CLZ@CTS/PLGA nanoparticles (NPs). Taken together, the results suggest that this design may provide a promising drug delivery system and a new option for the treatment of FK.

Pharmaceutical co-crystals (CCs) are multicomponent materials that enable the development of novel therapeutic products by enhancing the properties of active pharmaceutical ingredients, such as solubility, permeability and bioavailability. Currently, CCs are a commercial reality; nonetheless, their industrial production remains a challenge due to problems related to scale up, control and mode of preparation, which usually relies on batch production rather than continuous. This paper describes the implementation of a concurrent coaxial antisolvent electrospray (Co-E), as a new manufacturing technique, for the synthesis of CCs in a rapid, continuous and controlled manner. The features of Co-E were sized against other co-crystallization methods such as antisolvent crystallization, neat and liquid assisted grinding. Three pairs of amino acids were used as model compounds to demonstrate the features of this new system. The Co-E displayed exclusive product characteristics, including spherical particle morphology and enhanced CC formation. This technique exhibited robustness against process disturbances, displaying consistent product characteristics. Co-E represents a new alternative for the reliable production of CCs and other pharmaceutical products. [ABSTRACT FROM AUTHOR]

Aim: To reduce the contamination arising from abuse of commercial pesticide formulations, the coaxial electrospray (CES) method was used for one-step microencapsulation and spraying of pesticides. Methods: After optimisation of process parameters, polymeric microcapsules with different structures were fabricated as the carriers of azoxystrobin (AZS). For the resultant microcapsules, the sustained pesticide release was verified in vitro and the adhesion properties were investigated through a normalised rinsing test. Results: The maximum encapsulation efficiency of the fabricated AZS-loaded microcapsules was 99.14%. Compared to commercial AZS aqueous suspension, the microcapsules fabricated by the CES method exhibited improved sustained release performance of AZS, which could be readily controlled by adjusting the shell thicknesses. Moreover, highly enhanced adhesion performance was observed for the AZS-loaded microcapsules directly sprayed in CES process. Conclusions: The CES process is promising to be applied as a one-step microencapsulation and spraying technology for improving pesticide utilisation and reducing environmental pollution. [ABSTRACT FROM AUTHOR]

Abstract Appropriate administration of growth factors is of great importance for directing cell behavior in regenerative medicine, which usually needs suitable carriers to protect the growth factors and to control their releases in specific spatiotemporal manners. Electrospray techniques, particularly emulsion electrospray and coaxial electrospray, have been proven effective to generate capsular/core-shell structured microspheres for growth factor delivery with superior convenience and high bioactivity retention. However, the difference in the release behavior of growth factors from emulsion electrosprayed microspheres and coaxial electrosprayed microspheres remains ambiguous, which causes the difficulty in selecting appropriate approach for certain growth factor-based therapy. In our investigation, vascular endothelial growth factor (VEGF) was used as a model molecule to be encapsulated in the microspheres prepared respectively by emulsion electrospray and coaxial electrospray using the same biodegradable poly(lactic- co -glycolic acid) (PLGA) polymer, for which size distribution, structure, morphology and in vitro degradation properties were studied and found to be tunable by different electrospray techniques. PLGA microspheres fabricated by emulsion electrospray presented nanoporous surface morphology and interior multi-compartment cores, resulting in sustained VEGF release. In comparison, the microspherical vehicles made by coaxial electrospray showing golf-ball-like textured surface morphology and interior monolithic cores led to initial fast release of VEGF. Vascular endothelial cells responded differently under the stimuli of locally released VEGF from different types of electrosprayed PLGA microspheres. The underlying mechanisms for different release behaviors of the encapsulated growth factors that were affected by microspherical vehicles formed by different electrospray techniques were presented, which would offer the design rationale for growth factor delivery vehicles with specific release kinetics, suiting for different therapeutic purposes. Highlights • Comparisons in the structure and properties of microspheres made by emulsion electrospray or coaxial electrospray are made. • Microspheres via different techniques have different morphologies and structures, hence different biodegradation property. • Growth factor release can be modulated by using microspheres made by different electrospray techniques. • Growth factor-encapsulated microspheres via two different electrospray techniques show different therapeutic potentials. • A design rationale for microspheres with specific release kinetics suiting for different therapeutic purposes is offered. [ABSTRACT FROM AUTHOR]

Coaxial electrospray has been used effectively for several dual-emitter applications, but has not been utilized for the study of rapid in-source chemistry. In this paper, we report the fabrication of a coaxial, micro-volume dual-emitter through the modification of a manufacturer’s standard electrospray probe. This modification creates rapid mixing inside the Taylor cone and the ability to manipulate fast reactions using a variety of solvents and analytes. We demonstrate its potential as a low-cost, dual-emitter assembly for diverse applications through three examples: relative ionization in a biphasic electrospray, hydrogen-deuterium exchange, and protein supercharging.Graphical Abstract [ABSTRACT FROM AUTHOR]

Abstract: Polymeric core–shell microspheres have been widely applied in dual‐drug release system for their convenience to treat diseases through multiple therapies. In this contribution, acrylamide‐modified chitin (AMC)/chitosan (CS) core–shell microspheres were prepared by a coaxial electrospray method under mild conditions. The dimension of the microspheres and thickness of the layer can be delicately controlled by changing the feed rate of spray solutions and the applied voltage. The core–shell structure of the microspheres was characterized by Fourier transform infrared spectra (FT‐IR), scanning electron microscope (SEM), and stereo fluorescence microscope. To investigate the dual‐drug release behavior of the core–shell microspheres, small molecular FITC‐dextran and macromolecular dextran blue 2000 were chosen as model drugs. The release rate of small molecular FITC‐dextran was higher than macromolecular dextran blue 2000 regardless of the location of FITC‐dextran in the microspheres. The biocompatibility, biodegradability, and dual‐drug release behavior of the core–shell microspheres endow it with potential applications in combination therapies. [ABSTRACT FROM AUTHOR]

It is of great significance to efficiently prepare core-shell nanoparticles. In this study, we directly and efficiently prepared core-shell structured PCL/PEG nanoparticles based on shell insertion strategy by using coaxial electrospray. The key points to ensure the success of this new strategy include: the preparation of PCL particles with size less than 100 nm which worked as core structure; the insertion of PEG into PCL surface which worked as shell structure. We directly fabricated PCL particles with size less than 100 nm by using shell evaporation coaxial electrospray method. It means that PCL nanoparticles could be prepared and used as core structure through coaxial electrospray method. PEG solution worked as shell layer, PCL solution worked as core layer, PCL/PEG core shell structured microparticles were obtained through coaxial electrospray technique. Because of the solvent diffusion during the coaxial electrospray process, PEG could insert into PCL when TFE was used as the solvent of both core and shell layer. Then, after removing uninserted PEG, PCL/PEG nanoparticles with core-shell structure were fabricated successfully. The core-shell structured nanoparticles were stored at 4 °C and were stable without changing size for at least 7 days. This method can easily prepare core-shell structured nanoparticle with different size by changing the injection speed of PCL solution. [ABSTRACT FROM AUTHOR]

Abstract: A temperature‐regulated system for the controlled release of nerve growth factor (NGF) to promote neurite outgrowth is reported. The system is based upon microparticles fabricated using coaxial electrospray, with the outer solution containing a phase‐change material (PCM) and the inner solution encompassing payload(s). When the temperature is kept below the melting point of the PCM, there is no release due to the extremely slow diffusion through a solid matrix. Upon increasing the temperature to slightly pass the melting point, the encapsulated payload(s) can be readily released from the melted PCM. By leveraging the reversibility of the phase transition, the payload(s) can be released in a pulsatile mode through on/off heating cycles. The controlled release system is evaluated for potential use in neural tissue engineering by sandwiching the microparticles, coloaded with NGF and a near‐infrared dye, between two layers of electrospun fibers to form a trilayer construct. Upon photothermal heating with a near‐infrared laser, the NGF is released with well‐preserved bioactivity to promote neurite outgrowth. By choosing different combinations of PCM, biological effector, and scaffolding material, this controlled release system can be applied to a wide variety of biomedical applications. [ABSTRACT FROM AUTHOR]

Improvement of thrombo-resistant properties is vital for vascular substitutes. Heparin, an effective natural anticoagulant, attracted concerns about its incorporate way, releasing profile, and the biological function when applied onto vascular substitutes. Coaxial electrospraying technology was used to develop and form PCL-PEG encapsulated heparin microspheres in this study. Heparin can maintain its activity and sustained release in 15 days with the package and protection of shell layer. When the microspheres were sprayed onto the PCL porous membrane, the materials showed sufficient property in avoiding the platelet adhesion and blood coagulation. [ABSTRACT FROM AUTHOR]

Alginate-based microencapsulation of cells has made a significant impact on the fields of regenerative medicine and tissue engineering mainly because of its ability to provide immunoisolation for the encapsulated material. This characteristic has allowed for the successful transplantation of non-autologous cells in several clinical trials for life threatening conditions, such as diabetes, myocardial infarction, and neurodegenerative disorders. Methods for alginate hydrogel microencapsulation have been well developed for various types of cells and can generate microcapsules of different diameters, degradation time, and composition. It appears the most prominent and successful method in clinical applications is the coaxial electrospray method, which can be used to generate both homogenous and non-homogeneous microcapsules with uniform size on the order of 100 μm. The present review aims to discuss why alginate hydrogel is an ideal biomaterial for the encapsulation of cells, how alginate-based microcapsules are generated, and methods of modifying the microcapsules for specific clinical treatments. This review will also discuss clinical applications that have utilized alginate-based microencapsulation in the treatment of diabetes, ischemic heart disease, and neurodegenerative diseases. [ABSTRACT FROM PUBLISHER]

The objective of this study was to investigate the combined influence of independent variables in the preparation of folic acid-chitosan-methotrexate nanoparticles (FA-Chi-MTX NPs). These NPs were designed and prepared for targeted drug delivery in tumor. The NPs of each batch were prepared by coaxial electrospray atomization method and evaluated for particle size (PS) and particle size distribution (PSD). The independent variables were selected to be concentration of FA-chitosan, ratio of shell solution flow rate to core solution flow rate, and applied voltage. The process design of experiments (DOE) was obtained with three factors in three levels by Design expert software. Box-Behnken design was used to select 15 batches of experiments randomly. The chemical structure of FA-chitosan was examined by FTIR. The NPs of each batch were collected separately, and morphologies of NPs were investigated by field emission scanning electron microscope (FE-SEM). The captured pictures of all batches were analyzed by ImageJ software. Mean PS and PSD were calculated for each batch. Polynomial equation was produced for each response. The FE-SEM results showed the mean diameter of the core-shell NPs was around 304 nm, and nearly 30% of the produced NPs are in the desirable range. Optimum formulations were selected. The validation of DOE optimization results showed errors around 2.5 and 2.3% for PS and PSD, respectively. Moreover, the feasibility of using prepared NPs to target tumor extracellular pH was shown, as drug release was greater in the pH of endosome (acidic medium). Finally, our results proved that FA-Chi-MTX NPs were active against the human epithelial cervical cancer (HeLa) cells. [ABSTRACT FROM AUTHOR]

BACKGROUND Coaxial electrospray is an emerging technology to produce multilayer micro- and nano-particles ( NPs) with well controlled shape and size. ALA-PDT is a fascinating technology used in the treatment for skin and other cancers. In this work, ALA-loaded poly(lactide-co-glycolide) ( PLGA) ( PLGA-ALA) NPs with high drug loading efficiency were produced via coaxial electrospray and used for the PDT treatment in HSC-3 cells. RESULTS Results showed that 200-1000 nm PLGA-ALA NPs with 60-75% ALA loading efficiency were successfully produced. ALA and PLGA were well integrated and no extra chemical reaction occurred during the electrospray. PLGA-ALA NPs displayed a delayed release of ALA in PBS solution up to 7 days. A massive accumulation of PpIX in HSC-3 cells happened after the incubation of PLGA-ALA NPs. With light irradiation, these PLGA-ALA NPs showed a remarkable cytotoxicity against HSC-3 cells, leading to 40-70% cell death depending on the incubation dose. CONCLUSIONS Coaxial electrospraying is a facile method to prepare PLGA-ALA NPs with high drug loading content and efficiency, and PLGA-ALA NPs turned out to be an effective vehicle for delivering ALA to tumor cells. © 2015 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Coaxial electrospray is a well established procedure to generate coaxial micro- and nano-structures (microcapsules and coaxial fibers). In this work we show the capabilities of a high-precision numerical code using dynamic mapping, analytic Jacobian formulation and the Taylor–Melcher leaky dielectric model (LDM) to deal with real coaxial electrospray configurations. Since the number of parameters involved in the problem is large, we focus on an experimentally tested liquid pair configuration where the thickness, relative position, and end sharpness of the coaxial feeding tubes are geometrical parameters of interest. The effects of those parameters and the voltage applied at the tubes on the coaxial liquid menisci, liquid streamlines, size of jet and core, and electric current are discussed. • The influence of the outlet tube geometry on coaxial electrosprays is studied. • Accurate numerical solutions are given using Taylor–Melcher Leaky Dielectric model • Protruding, flush and recessed tube outlet setups, and edge sharpening are explored • Physical details (streamline patterns, menisci geometries, stresses) are provided [ABSTRACT FROM AUTHOR]

[Display omitted] • A coaxial electrospray system was developed for diabetes drug co-encapsulation. • Encapsulation and delivery of multiple drugs from a single-dosage form carrier. • Near 100% co-encapsulation in different layers of PLGA nanoparticles was achieved. • A SGLT2 inhibitor and a thiazide-like diuretic were released in a controlled manner. • Improved sustained and controlled release of multidrugs with distinctive profile. Coaxial electrospray is advantageous for the production of multidrug-releasing nanocarriers because it permits precise control over particle size, inhibits initial burst release, and offers moderate preparation conditions. In this study, a single-step coaxial electrospray technique is presented that achieves over 90 % co-encapsulation of the saxagliptin and dapagliflozin, two drugs treating type 2 diabetes, into biodegradable poly (d , l -lactide- co -glicolide) (PLGA) nanoparticles. Scanning electron microscopy reveals spherical and smooth shapes with diameters ranging from 534.8 to 708.6 nm. Transmission electron microscopy revealed clear core–shell and trilayer nanostructures. Fourier transform infrared spectroscopy confirmed the presence of PLGA, saxagliptin, and dapagliflozin in all the evaluated formulations. The results of the drug release investigation indicated the prolonged and regulated release of saxagliptin and dapagliflozin from bi- and trilayer structures, as compared to monolayer particles. Computational modelling showed good agreement with the experimental drug release profile in vitro. Further, cytotoxicity assay demonstrates that the formulated nanoparticles display good cytocompatibility. This study indicates that with the controllable and distinctive sustained release profiles, the hybrid nanoparticle-based drug delivery system can effectively co-encapsulate multiple drugs treating type 2 diabetes in a protectively shell of PLGA for therapeutically-benefit controlled release. [ABSTRACT FROM AUTHOR]

Enteric-coated formulations can delay the release of drugs until they have passed through the stomach. However, high concentration of drugs caused by rapidly released in the small intestine leads to the intestinal damage, and frequent administration would increase the probability of missing medication and reduce the patient compliance. To solve the above-mentioned problems, aspirin-loaded enteric-coated sustained-release nanoparticles with core-shell structure were prepared via one-step method using coaxial electrospray in this study. Eudragit L100-55 as pH-sensitive polymer and Eudragit RS as sustained-release polymer were used for the outer coating and inner core of the nanoparticles, respectively. The maximum loading capacity of nanoparticles was 23.66 % by changing the flow rate ratio of outer/inner solutions, and the entrapment efficiency was nearly 100 %. Nanoparticles with core-shell structure were observed via fluorescence microscope and transmission electron microscope. And pH-sensitive and sustained drug release profiles were observed in the media with different pH values (1.2 and 6.8). In addition, mild cytotoxicity in vitro was detected, and the nanoparticles could be taken up by Caco-2 cells within 1.0 h in cellular uptake study. These results indicate that prepared enteric-coated sustained-release nanoparticles would be a more safety and effective carrier for oral drug delivery. [ABSTRACT FROM AUTHOR]

Recent developments in multimodal imaging and image-guided therapy requires multilayered microparticles that encapsulate several imaging and therapeutic agents in the same carrier. However, commonly used microencapsulation processes have multiple limitations such as low encapsulation efficiency and loss of bioactivity for the encapsulated biological cargos. To overcome these limitations, we have carried out both experimental and theoretical studies on coaxial electrospray of multilayered microparticles. On the experimental side, an improved coaxial electrospray setup has been developed. A customized coaxial needle assembly combined with two ring electrodes has been used to enhance the stability of the cone and widen the process parameter range of the stable conejet mode. With this assembly, we have obtained poly(lactide-co-glycolide) microparticles with fine morphology and uniform size distribution. On the theoretical side, an instability analysis of the coaxial electrified jet has been performed based on the experimental parameters. The effects of process parameters on the formation of different unstable modes have been studied. The reported experimental and theoretical research represents a significant step toward quantitative control and optimization of the coaxial electrospray process for microencapsulation of multiple drugs and imaging agents in multimodal imaging and image-guided therapy. [ABSTRACT FROM AUTHOR]

Coaxial electrospray is an electrohydrodynamic process that produces multilayer microparticles and nanoparticles by introducing coaxial electrified jets. In comparison with other microencapsulation/nanoencapsulation processes, coaxial electrospray has several potential advantages such as high encapsulation efficiency, effective protection of bioactivity and uniform size distribution. However, process control in coaxial electrospray is challenged by the multiphysical nature of the process and the complex interplay of multiple design, process and material parameters. This paper reviews the previous works and the recent advances in design, modeling and control of a coaxial electrospray process. The review intends to provide general guidance for coaxial electrospray and stimulate further research and development interests in this promising microencapsulation/nanoencapsulation process. [ABSTRACT FROM AUTHOR]

Cell-based therapies require a large number of cells, as well as appropriate methods to deliver the cells to damaged tissue. Microcarriers provide an optimal platform for large-scale cell culture while also improving cell retention during cell delivery. However, this technology still presents significant challenges due to low-throughput fabrication methods and an inability of the microcarriers to recreate the properties of human tissue. This work proposes, for the first time, the use of methacryloyl platelet lysates (PLMA), a photocrosslinkable material derived from human platelet lysates, to produce porous microcarriers. Initially, high quantities of PLMA/alginate core-shell microcapsules are produced using coaxial electrospray. Subsequently, the microcapsules are collected, irradiated with ultraviolet light, washed, and freeze dried yielding PLMA microsponges. These microsponges are able to support the adhesion and proliferation of human adipose-derived stem cells, while also displaying potential in the assembly of autologous microtissues. Cell-laden microsponges were shown to self-organize into aggregates, suggesting possible applications in bottom-up tissue engineering applications. Impact Statement Microcarriers have increasingly been used as delivery platforms in cell therapy. Herein, the encapsulation of human-derived proteins in alginate microcapsules is proposed as a method to produce microcarriers from photopolymerizable materials. The capsules function as a template structure, which is then processed into spherical microparticles, which can be used in cell culture, cell delivery, and bottom-up assembly. As a proof of concept, this method was combined with lyophilization to process methacryloyl platelet lysates into injectable microsponges for cell delivery.

To optimize the anti-tumor efficacy of combination therapy with paclitaxel (PTX) and imatinib (IMN), we used coaxial electrospray to prepare sequential-release core–shell microparticles composed of a PTX-loaded sodium hyaluronate outer layer and an IMN-loaded PLGA core. The morphology, size distribution, drug loading, differential scanning calorimetry (DSC), Fourier transform infrared spectra (FTIR), in vitro release, PLGA degradation, cellular growth inhibition, in vivo vaginal retention, anti-tumor efficacy, and local irritation in a murine orthotopic cervicovaginal tumor model after vaginal administration were characterized. The results show that such core–shell microparticles were of spherical appearance, with an average size of 14.65 μm and a significant drug-loading ratio (2.36% for PTX, 19.5% for IMN, w/w), which might benefit cytotoxicity against cervical-cancer-related TC-1 cells. The DSC curves indicate changes in the phase state of PTX and IMN after encapsulation in microparticles. The FTIR spectra show that drug and excipients are compatible with each other. The release profiles show sequential characteristics in that PTX was almost completely released in 1 h and IMN was continuously released for 7 days. These core–shell microparticles showed synergistic inhibition in the growth of TC-1 cells. Such microparticles exhibited prolonged intravaginal residence, a >90% tumor inhibitory rate, and minimal mucosal irritation after intravaginal administration. All results suggest that such microparticles potentially provide a non-invasive local chemotherapeutic delivery system for the treatment of cervical cancer by the sequential release of PTX and IMN. [ABSTRACT FROM AUTHOR]

Streptokinase, a clot-dissolving agent, is widely used in treatment of cardiovascular diseases such as blood clots and deep thrombosis. Streptokinase is a cost-effective drug with a short biological half-life (i.e. 15 to 30 min). In addition, due to its prokaryotic source, the immune response quickly reacts to the drug. Despite these limitations, streptokinase is still the first choice for diseases associated with thrombosis. In this work, streptokinase was encapsulated in mPEG-PLGA nanoparticles to improve its pharmacokinetic properties. The nanoparticles containing the enzyme were prepared by coaxial electrospray and their physicochemical properties, blood compatibility, circulation time and cell toxicity were evaluated. The results showed that the use of mPEG-PLGA nanoparticles to encapsulate the enzyme resulted in prolonged circulation time (up to 120 min) with a slight decrease in its activity. In vivo studies also showed that the nanoparticles containing streptokinase did not have adverse effect on blood biochemistry parameters as well as liver and kidney tissues. As a result, the mPEG-PLGA nanoparticles showed the potential for increasing the biological activity of streptokinase with no important adverse effect. Unlabelled Image • Coaxial electrospray was used to prepare nanoparticles of mPEG-PLGA/SK. • These NPs had an acceptable degree of blood compatibility and were not cytotoxic. • The NPs showed to be effective in increasing biological life time of the enzyme. [ABSTRACT FROM AUTHOR]

Hypothesis: Developing oral formulations to enable effective release of poorly water-soluble drugs like progesterone is a major challenge in pharmaceutics. Coaxial electrospray can generate drug-loaded nanoparticles of strategic compositions and configurations to enhance physiological dissolution and bioavailability of poorly water-soluble drug progesterone., Experiments: Six formulations comprising nanoparticles encapsulating progesterone in different poly(lactide-co-glycolide) (PLGA) matrix configurations and compositions were fabricated and characterized in terms of morphology, molecular crystallinity, drug encapsulation efficiency and release behavior., Findings: A protocol of fabrication conditions to achieve 100% drug encapsulation efficiency in nanoparticles was developed. Scanning electron microscopy shows smooth and spherical morphology of 472.1±54.8 to 588.0±92.1 nm in diameter. Multiphoton Airyscan super-resolution confocal microscopy revealed core-shell nanoparticle configuration. Fourier transform infrared spectroscopy confirmed presence of PLGA and progesterone in all formulations. Diffractometry indicated amorphous state of the encapsulated drug. UV-vis spectroscopy showed drug release increased with hydrophilic copolymer glycolide ratio while core-shell formulations with progesterone co-dissolved in PLGA core exhibited enhanced release over five hours at 79.9±1.4% and 70.7±3.5% for LA:GA 50:50 and 75:25 in comparison with pure progesterone without polymer matrix in the core at 67.0±1.7% and 57.5±2.8%, respectively. Computational modeling showed good agreement with the experimental drug release behavior in vitro., Competing Interests: The authors report no conflicts of interest in this work., (© 2020 Zhang et al.)

Minimally invasive therapies avoiding surgical complexities evoke great interest in developing injectable biomedical devices. Herein, a versatile approach is reported for engineering injectable and biomimetic nanofiber microspheres (NMs) with tunable sizes, predesigned structures, and desired compositions via gas bubble-mediated coaxial electrospraying. The sizes and structures of NMs are controlled by adjusting processing parameters including air flow rate, applied voltage, distance, and spinneret configuration in the coaxial setup. Importantly, unlike the self-assembly method, this technique can be used to fabricate NMs from any material feasible for electrospinning or other nanofiber fabrication techniques. To demonstrate the versatility, open porous NMs are successfully fabricated that consist of various short nanofibers made of poly(ε-caprolactone), poly(lactic-co-glycolic acid), gelatin, methacrylated gelatin, bioglass, and magneto-responsive polymer composites. Open porous NMs support human neural progenitor cell growth in 3D with a larger number and more neurites than nonporous NMs. Additionally, highly open porous NMs show faster cell infiltration and host tissue integration than nonporous NMs after subcutaneous injection to rats. Such a novel class of NMs holds great potential for many biomedical applications such as tissue filling, cell and drug delivery, and minimally invasive tissue regeneration., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)