Enhancing immunogenicity of antigens through sustained intradermal delivery using chitosan microneedles with a patch-dissolvable design.

Reducing the dosage required for vaccination is highly desirable, particularly in cases of epidemic emergencies. This study evaluated the potential of a chitosan microneedle (MN) system with a patch-dissolvable design for low-dose immunization. This system comprises antigen-loaded chitosan MNs and a hydrophilic polyvinyl alcohol/polyvinyl pyrrolidone supporting array patch, which provides extra strength to achieve complete MN insertion and then quickly dissolves in the skin to reduce patch-induced skin irritation. After insertion, MNs could be directly implanted in the dermal layer as an intradermal (ID) depot to allow a sustained release of the model antigen ovalbumin (OVA) for up to 28 days. We found that rats immunized with MNs containing low-dose OVA (approximately 200 µg) had persistently high antibody levels for 18 weeks, which were significantly higher than those observed after an intramuscular injection of full-dose OVA (approximately 500 µg), demonstrating at least 2.5-fold dose sparing. Moreover, OVA-encapsulated chitosan MNs had superior immunogenicity to OVA plus chitosan solution, indicating that MN-based delivery and prolonged skin exposure can further enhance chitosan's adjuvanticity. Therefore, this patch-dissolvable MN system offers a needle-free, accurate, and reliable ID delivery of antigens and has potential as a sustained ID delivery device to improve vaccine efficacy and facilitate dose sparing with existing vaccines. STATEMENT OF SIGNIFICANCE: This study developed implantable chitosan microneedles (MNs) with a patch-dissolvable design for the sustained intradermal (ID) delivery of antigens and demonstrated their antigen dose-sparing potential. We found that rats immunized with chitosan MNs containing low-dose OVA had persistently high antibody levels for 18 weeks, which were significantly higher than those observed after an intramuscular injection of full-dose OVA, demonstrating at least 2.5-fold dose sparing. Our results indicate that chitosan MNs can not only serve as an efficient vaccine delivery system but also exert their promising adjuvant activity by forming an ID depot for prolonged antigen exposure and activating dendritic cells for promoting immune responses.

Implantable polymeric microneedles with phototriggerable properties as a patient-controlled transdermal analgesia system.

Adequate pain control can be achieved using a patient-controlled drug delivery system that can provide analgesia to patients as needed. To achieve this objective, we developed a phototriggered microneedle (MN) system that enables the on-demand delivery of pain medications to the skin under external near-infrared (NIR) light stimulation. In this system, polymeric MNs, containing NIR absorbers and analgesics, are combined with a poly(l-lactide-co-d,l-lactide) supporting array. A "removable design" of the supporting array enables the quick implantation of the MNs into the skin to act as a drug depot, thus shortening the patch application time. Upon irradiation with NIR light, the NIR absorbers in the implanted MNs can absorb light energy and induce a phase transition in the MNs to activate drug release. We demonstrated that lidocaine release can be modulated or repeatedly triggered by varying the duration of irradiation and controlling the on and off status of the laser. Lidocaine delivered by the implanted MNs can be rapidly absorbed into the blood circulation within 10 min and has a bioavailability of at least 95% relative to the subcutaneous injection, showing that the proposed system has the potential to provide a rapid onset of pain relief. Such an implantable device may allow pain sufferers receiving the painkiller without the need for multiple needle injections, and may enable controlling pain more conveniently and comfortably.

Near-Infrared Light-Activatable Microneedle System for Treating Superficial Tumors by Combination of Chemotherapy and Photothermal Therapy.

Because of the aggressive and recurrent nature of cancers, repeated and multimodal treatments are often necessary. Traditional cancer therapies have a risk of serious toxicity and side effects. Hence, it is crucial to develop an alternative treatment modality that is minimally invasive, effectively treats cancers with low toxicity, and can be repeated as required. We developed a light-activatable microneedle (MN) system that can repeatedly and simultaneously provide photothermal therapy and chemotherapy to superficial tumors and exert synergistic anticancer effects. This system consists of embeddable polycaprolactone MNs containing a photosensitive nanomaterial (lanthanum hexaboride) and an anticancer drug (doxorubicin; DOX), and a dissolvable poly(vinyl alcohol)/polyvinylpyrrolidone supporting array patch. Because of this supporting array, the MNs can be completely inserted into the skin and embedded within the target tissue for locoregional cancer treatment. When exposed to near-infrared light, the embedded MN array uniformly heats the target tissue to induce a large thermal ablation area and then melts at 50 °C to release DOX in a broad area, thus destroying tumors. This light-activated heating and releasing behavior can be precisely controlled and switched on and off on demand for several cycles. We demonstrated that the MN-mediated synergistic therapy completely eradicated 4T1 tumors within 1 week after a single application of the MN and three cycles of laser treatment. No tumor recurrence and no significant body weight loss of mice were observed. Thus, the developed light-activatable MN with a unique embeddable feature offers an effective, user-friendly, and low-toxicity option for patients requiring long-term and multiple cancer treatments.

Poly-γ-glutamic acid microneedles with a supporting structure design as a potential tool for transdermal delivery of insulin.

Incomplete insertion is a common problem associated with polymer microneedles (MNs) that results in a limited drug delivery efficiency and wastage of valuable medication. This paper presents a fully insertable MN system that is composed of poly-γ-glutamic acid (γ-PGA) MNs and polyvinyl alcohol (PVA)/polyvinyl pyrrolidone (PVP) supporting structures. The PVA/PVP supporting structures were designed to provide an extended length for counteracting skin deformation during insertion and mechanical strength for fully inserting the MNs into the skin. When inserted into the skin, both the supporting structures and MNs can be dissolved in the skin within 4min, thus quickly releasing the entire drug load from the MNs. To evaluate the feasibility and reproducibility of using the proposed system for treating diabetes, we administered insulin-loaded MNs to diabetic rats once daily for 2days. The results indicated that the hypoglycemic effect in the rats receiving insulin-loaded MNs was comparable to that observed in rats receiving subcutaneous insulin injections. The relative pharmacological availability and relative bioavailability of the insulin were in the range of 90-97%, indicating that the released insulin retained its pharmacological activity. We observed no significant differences in the plasma insulin concentration profiles between the first and second administrations, confirming the stability and accuracy of using the proposed MN system for insulin delivery. These results indicated that the γ-PGA MNs containing the supporting structure design enable complete and efficient delivery of encapsulated bioactive molecules and have great potential for the relatively rapid and convenient transdermal delivery of protein drugs. STATEMENT OF SIGNIFICANCE: Incomplete insertion of microneedles largely limits drug delivery efficiency and wastage of valuable medication. To address this problem, we developed a fully insertable poly-glutamic acid microneedles with a supporting structure design to ensure complete and efficient delivery of encapsulated drugs. The supporting structures were designed to provide an extended length for counteracting skin compressive deformation during puncture and mechanical strength for fully inserting the microneedles into the skin. When inserted into the skin, both the supporting structures and microneedles can be dissolved in the skin within 4min, thus quickly releasing the entire drug load. This study demonstrated that the proposed microneedle system featuring this unique design allows more convenient and efficient self-administration of drugs into the skin.

Near-infrared light-responsive composite microneedles for on-demand transdermal drug delivery.

This study presents near-infrared (NIR) light-responsive polymer-nanostructure composite microneedles used for on-demand transdermal drug delivery. Silica-coated lanthanum hexaboride (LaB6@SiO2) nanostructures were incorporated into polycaprolactone microneedles, serving as an NIR absorber. When the microneedles were irradiated with NIR light, light-to-heat transduction mediated by the LaB6@SiO2 nanostructures caused the microneedle melting at 50 °C. This increased the mobility of the polymer chains, enabling drug release from the matrix. Drug release from the microneedles was evaluated for four laser on/off cycles. In each cycle, the samples were irradiated until the temperature reached 50 °C for 3 min (laser on); the laser was then turned off for 30 min (laser off). The results showed that light-induced phase transition in the polymer triggered drug release from the melted microneedles. A stepwise drug-release behavior was observed after multiple cycles of NIR light exposure. No notable drug leakage was found in the off state. This NIR-light-triggerable device exhibits excellent reproducibility, low off-state leakage, and noninvasive triggerability and, thus, represents an advance in transdermal delivery technology.

Remotely triggered release of small molecules from LaB6@SiO2-loaded polycaprolactone microneedles.

We established near-infrared (NIR)-light-triggered transdermal delivery systems by encapsulating NIR absorbers, silica-coated lanthanum hexaboride (LaB6@SiO2) nanostructures and the cargo molecule to be released in biodegradable polycaprolactone (PCL) microneedles. Acting as a local heat source when exposed to an NIR laser, these nanostructures cause a phase transition of the microneedles, thereby increasing the mobility of the polymer chains and triggering drug release from the microneedles. On IR thermal images, the light-triggered melting behavior of the LaB6@SiO2-loaded microneedles was observed. By adjusting the irradiation time and the laser on/off cycles, the amount of molecules released was controlled accurately. Drug release was switched on and off for at least three cycles, and a consistent dose was delivered in each cycle with high reproducibility. The designed microneedles were remotely triggered by laser irradiation for the controlled release of a chemotherapeutic drug, doxorubicin hydrochloride, in vivo. This system would enable dosages to be adjusted accurately to achieve a desired effect, feature a low off-state drug leakage to minimize basal effects and can increase the flexibility of pharmacotherapy performed to treat various medical conditions.

Dissolving polymer microneedle patches for rapid and efficient transdermal delivery of insulin to diabetic rats.

This study presents a dissolving microneedle patch, composed of starch and gelatin, for the rapid and efficient transdermal delivery of insulin. The microneedles completely dissolve after insertion into the skin for 5 min, quickly releasing their encapsulated payload into the skin. A histological examination shows that the microneedles have sufficient mechanical strength to be inserted in vitro into porcine skin to a depth of approximately 200 µm and in vivo into rat skin to 200-250 µm depth. This penetration depth does not induce notable skin irritation or pain sensation. To evaluate the feasibility of using these dissolving microneedles for diabetes treatment insulin-loaded microneedles were administered to diabetic rats using a homemade applicator. Pharmacodynamic and pharmacokinetic results show a similar hypoglycemic effect in rats receiving insulin-loaded microneedles and a subcutaneous injection of insulin. The relative pharmacological availability and relative bioavailability of insulin were both approximately 92%, demonstrating that insulin retains its pharmacological activity after encapsulation and release from the microneedles. Storage stability analysis confirms that more than 90% of the insulin remained in the microneedles even after storage at 25 or 37°C for 1 month. These results confirm that the proposed starch/gelatin microneedles enable stable encapsulation of bioactive molecules and have great potential for transdermal delivery of protein drugs in a relatively painless, rapid, and convenient manner.

Fully embeddable chitosan microneedles as a sustained release depot for intradermal vaccination.

This study introduces a microneedle transdermal delivery system, composed of embeddable chitosan microneedles and a poly(L-lactide-co-D,L-lactide) (PLA) supporting array, for complete and sustained delivery of encapsulated antigens to the skin. Chitosan microneedles were mounted to the top of a strong PLA supporting array, providing mechanical strength to fully insert the microneedles into the skin. When inserted into rat skin in vivo, chitosan microneedles successfully separated from the supporting array and were left within the skin for sustained drug delivery without requiring a transdermal patch. The microneedle penetration depth was approximately 600 µm (i.e. the total length of the microneedle), which is beneficial for targeted delivery of antigens to antigen-presenting cells in the epidermis and dermis. To evaluate the utility of chitosan microneedles for intradermal vaccination, ovalbumin (OVA; MW = 44.3 kDa) was used as a model antigen. When the OVA-loaded microneedles were embedded in rat skin in vivo, histological examination showed that the microneedles gradually degraded and prolonged OVA exposure at the insertion sites for up to 14 days. Compared to traditional intramuscular immunization, rats immunized by a single microneedle dose of OVA showed a significantly higher OVA-specific antibody response which lasted for at least 6 weeks. These results suggest that embeddable chitosan microneedles are a promising depot for extended delivery of encapsulated antigens to provide sustained immune stimulation and improve immunogenicity.

Chitosan microneedle patches for sustained transdermal delivery of macromolecules.

This paper introduces a chitosan microneedle patch for efficient and sustained transdermal delivery of hydrophilic macromolecules. Chitosan microneedles have sufficient mechanical strength to be inserted in vitro into porcine skin at approximately 250 µm in depth and in vivo into rat skin at approximately 200 µm in depth. Bovine serum albumin (BSA, MW=66.5 kDa) was used as a model protein to explore the potential use of chitosan microneedles as a transdermal delivery device for protein drugs. In vitro drug release showed that chitosan microneedles can provide a sustained release of BSA for at least 8 days (approximately 95% of drugs released in 8 days). When the Alexa Fluor 488-labeled BSA (Alexa 488-BSA)-loaded microneedles were applied to the rat skin in vivo, confocal microscopic images showed that BSA can gradually diffuse from the puncture sites to the dermal layer and the fluorescence of Alexa 488-BSA can be observed at the depth of 300 µm. In addition, encapsulation of BSA within the microneedle matrix did not alter the secondary structure of BSA, indicating that the gentle nature of the fabrication process allowed for encapsulation of fragile biomolecules. These results suggested that the developed chitosan microneedles may serve as a promising device for transdermal delivery of macromolecules in a sustained manner.

A novel in-situ-gelling liquid suppository for site-targeting delivery of anti-colorectal cancer drugs.

In order to avoid anti-cancer drugs undergoing a first-pass effect and reduce their toxicity, and to solve conventional suppositories defects, we developed an in-situ-gelling and injectable Pluronic-poly(acrylic acid) (Pluronic-PAA) liquid suppository, which could gel fast in the physiological state and had suitable gel strength and bioadhesive force. The liquid suppositories were inserted into the rectum of rabbits without difficulty and leakage, and retained in the rectum for at least 6 h and while releasing the drug. The toxicity and cytotoxic tests indicated that Pluronic and PAA were non-toxic materials and could inhibit colon cancer cells when oxaliplatin was incorporated. C max and AUC0→12h values of oxaliplatin after rectal administration of a oxaliplatin suppository were higher than those for an oxaliplatin solution administered orally. These results suggest that an in-situ-gelling and injectable liquid suppository for humans can be further developed as a more convenient and effective rectal dosage form.

PH-sensitive Dioctadecylamine-501 polymeric micelles for delivery of insulin.

In this work, fluorescently labeled smart micelle copolymers which consist of Dioctadecylamine-501 (DODA-501) as the hydrophobic segment, N-isopropylacrylamide (NIPAAm) as well as acrylic acid (AAc) as the hydrophilic segments were prepared. These micelles showed both thermo- and pH-sensitive properties due to the nature properties of NIPAAm and AAc, respectively. The particle size of the prepared micelles ranged from 94 approximately 200 nm and was found to increase with DODA-501 concentration. The size of particles varied in different pH mediums or different temperatures suggesting these micelles were pH- and thermo-sensitive. The image of confocal laser scanning microscopy (CLSM) illustrates these micelles had the ability to encapsulate rhodamine solution. From CLSM observation, fluorescein isothiocyanate (FITC) expression was found on the surface of micelles indicating the target detecting ability of these micelles. In drug loading and release studies, these micelles had the ability to encapsulate insulin and its release was pH sensitive, being more rapid under intestinal fluid environment, but resisting the drug release at gastric fluid environment. Stability test indicates these micelles had good stability during storage. These results suggest the pH-sensitivity of the DODA-501 polymeric micelles may be an interesting candidate for oral drug delivery system.

High strength and low friction of a PAA-alginate-silica hydrogel as potential material for artificial soft tissues.

In this study, a series of poly(acrylic) acid (PAA)-based hydrogels was prepared by UV polymerization. Hydrogels with an interpenetrating network structure were formed by combining PAA and alginate (Alg) solutions. The incorporation of nano-silica into these gel solutions significantly increased their compressive strength and fracture toughness but lowered their cross-linking density and friction coefficient. The prepared hydrogels were considerably hydrophilic for water content greater than 98%, which is in accordance with the nature of soft tissues such as cartilage. The preliminary cell culture of adipose stem cells (ADSCs) on PAA-Alg-Si hydrogels results in good biological safety. These features suggest that the PAA-Alg-Si hydrogels prepared in this study can be used as artificial soft tissues.