Poly(lactide-co-glycolide) Acid Research Articles

Chitosan-alginate/R8 ternary polyelectrolyte complex as an oral protein-based vaccine candidate induce effective mucosal immune responses

Vaccination is the most effective method for preventing infectious diseases. Oral vaccinations have attracted much attention due to the ability to boost intestinal and systemic immunity. The focus of this study was to develop a poly (lactide-co-glycolide) acid (PLGA)-based ternary polyelectrolyte complex (PEC) with chitosan, sodium alginate, and transmembrane peptides R8 for the delivery of antigen proteins. In this study, the antigen protein (HBf), consisting of the Mycobacterium avium subspecies paratuberculosis (MAP) antigens HBHA, Ag85B, and Bfra, was combined with R8 to generate self-assembled conjugates. The results showed that PEC presented a cross-linked reticular structure to protect the encapsulated proteins in the simulated gastric fluid. Then, the nanocomposite separated into individual nanoparticles after entering the simulated intestinal fluid. The ternary PEC with R8 promoted the in vivo uptake of antigens by intestinal lymphoid tissue. Moreover, the ternary PEC administered orally to mice promoted the secretion of specific antibodies and intestinal mucosal IgA. In addition, in the mouse models of MAP infection, the ternary PEC enhanced splenic T cell responses, thus reducing bacterial load and liver pathology score. These results suggested that this ternary electrolyte complex could be a promising delivery platform for oral subunit vaccine candidates, not limited to MAP infection.

Fabrication of co-polymeric nanoformulation loaded with antibiotic ofloxacin and immunosuppressant tacrolimus for pediatric sepsis-induced acute lung injury

Sepsis is a complicated, life-threatening illness caused by pathogen invasion triggered by a dysfunctional immune system in the host. Even though several antimicrobial and immunosuppressive therapies have been suggested, no clinically viable treatments for sepsis exist. This research aims to find a way to cure sepsis and reduce its symptoms by creating a nanoparticle system that specifically targets inflammation and drug delivery sites. To fabricate the PLGA-NPs loaded with the wide-range antibiotic Ofloxacin (OFX) and the immunosuppressant anti-inflammatory Tacrolimus (TCR), we opted for the biocompatible and biodegradable PLGA (poly(lactide-co-glycolide acid)). Grafting the γ3 peptides that can selectively conjugate to the intercellular adhesion molecule-1 (ICAM-1), which is abundantly articulated on the inflammatory endothelium cells surface, confers the targeting capacity of the nanoparticles toward inflammatory areas. Good biocompatibility, lower hemolytic rate, and minimal toxicity are hallmarks of dual drug-loaded γ3@PLGA nanoparticles. This work presents a straightforward and sturdy nanoplatform for treating pediatric sepsis-induced acute lung injury as a first step toward developing multifunctional nanomedicine for clinical translation.

A "trained immunity" inducer-adjuvanted nanovaccine reverses the growth of established tumors in mice

Innate immune cells are critical in antitumor immune surveillance and the development of antitumor adaptive cellular immunity. Trained innate immune cells demonstrate immune memory-like characteristics, producing more vigorous immune responses to secondary homologous or heterologous stimuli. This study aimed to investigate whether inducing trained immunity is beneficial when using a tumor vaccine to promote antitumor adaptive immune responses. A biphasic delivery system was developed with the trained immunity inducer Muramyl Dipeptide (MDP) and specific tumor antigen human papillomavirus (HPV) E7 peptide encapsulated by poly(lactide-co-glycolide)-acid(PLGA) nanoparticles (NPs), and the NPs along with another trained immunity agonist, β-glucan, were further embedded in a sodium alginate hydrogel. The nanovaccine formulation demonstrated a depot effect for E7 at the injection site and targeted delivery to the lymph nodes and dendritic cells (DCs). The antigen uptake and maturation of DCs were significantly promoted. A trained immunity phenotype, characterized by increased production of IL-1β, IL-6, and TNF-α, was induced in vitro and in vivo in response to secondary homologous or heterologous stimulation. Furthermore, prior innate immune training enhanced the antigen-specific INF-γ-expressing immune cell response elicited by subsequent stimulation with the nanovaccine. Immunization with the nanovaccine completely inhibited the growth of TC-1 tumors and even abolished established tumors in mice. Mechanistically, the inclusion of β-glucan and MDP significantly enhanced the responses of tumor-specific effector adaptive immune cells. The results strongly suggest that the controlled release and targeted delivery of an antigen and trained immunity inducers with an NP/hydrogel biphasic system can elicit robust adaptive immunity, which provides a promising tumor vaccination strategy.Graphical

Polymeric nanoparticles for the delivery of Sonoran desert propolis: Synthesis, characterization and antiproliferative activity on cancer cells

Sonoran propolis (SP) exerts remarkable biological activities attributed to its polyphenolic composition, mostly described as poplar-type flavonoids. It is known that polyphenols present low bioavailability derived of their molecular weight, glycosylation level, metabolic conversion, as well as interaction with the intestinal microbiota, affording limitations for possible in vivo applications. The aim of this work was to synthesize Poly-(lactide-co-glycolide) acid (PLGA) nanoparticles for encapsulation of SP, as a matrix to increase solubility of their polyphenolic compounds and improve delivery, for the evaluation of its antiproliferative activity on cancer cells. The Sonoran propolis-loaded PLGA nanoparticles (SP-PLGA NPs) were synthesized (by nanoprecipitation), and their physicochemical parameters were determined (size, morphology, zeta potential, stability, and drug release). Additionally, the antiproliferative activity of SP-PLGA nanoparticles was evaluated in vitro against murine (M12.C3.F6) and human (HeLa) cancer cell lines, including a non-cancer human cell line (ARPE-19) as control. SP-PLGA NPs presented a mean size of 152.6 ± 7.1 nm with an average negative charge of − 21.2 ± 0.7 mV. The encapsulation yield of SP into PLGA system was approximately 68.2 ± 6.0% with an in vitro release of 45% of propolis content at 48 h. SP-PLGA NPs presented antiproliferative activity against both cancer cell lines tested, with lower IC50 values in M12.C3.F6 and HeLa cell lines (7.8 ± 0.4 and 3.8 ± 0.4 μg/mL, respectively) compared to SP (24.0 ± 4.3 and 7.4 ± 0.4 μg/mL, respectively). In contrast, the IC50 of SP-PLGA NPs and SP against ARPE-19 was higher than 50 µg/mL. Cancer cells treated with SP and SP-PLGA NPs presented morphological features characteristic of apoptosis (cellular shrinkage and membrane blebs), as well as elongated cells, effect previously reported for SP, meanwhile, no morphological changes were observed with ARPE-19 cells. The obtained delivery system demonstrates appropriate encapsulation characteristics and antiproliferative activity to be used in the field of nanomedicine, therefore SP could be potentially used in antitumoral in vivo assays upon its encapsulation into PLGA nanoparticles.

ISMN-loaded PLGA-PEG nanoparticles conjugated with anti-Staphylococcus aureus α-toxin inhibit Staphylococcus aureus biofilms in chronic rhinosinusitis.

Background:Staphylococcus aureus biofilms were linked to negative postsurgical outcomes of chronic rhinosinusitis (CRS). This study aims to develop a targeted nanoparticle and characterize its bactericidal effects. Methods: The authors prepared ISMN-loaded poly-lactide-co-glycolide acid (PLGA) and polyethylene glycol (PEG) nanoparticles conjugated with anti-S. aureus α-toxin (AA;ISMN-PLGA-PEG-AA), and determined its bactericidal and toxic effects. The antibiofilm propriety of ISMN-PLGA-PEG-AA was further investigated in a sheep CRS model. Results: ISMN-PLGA-PEG-AA had no toxic effect, while ISMN, ISMN-PLGA-PEG and ISMN-PLGA-PEG-AA had significantly anti-S. aureus effects. The blood concentrations and mRNA levels in sinus tissues of IL-4, IL-8and IFN-γ in the sheep CRS model were significantly low. Conclusion: ISMN-PLGA-PEG-AA can effectively inhibit S. aureus biofilm, and is a promising drug for CRS treatment.

Assessment of the Effect of PLGA Co-polymers and PEG on the Formation and Characteristics of PLGA-PEG-PLGA Co-block Polymer Using Statistical Approach.

Purpose: To assess the effect of the lactic acid (LA)-to-glycolic acid (GA) molar ratio and polyethylene glycol (PEG) concentration on the formation of poly-lactide co-glycolide acid (PLGA)-PEG-PLGA co-block polymers simultaneously using statistical approach. Methods: A 22 full factorial design with the addition of a point in the center of the design, namely curvature, was applied. Fourier transform infrared (FTIR), differential scanning calorimetry (DSC), and nuclear magnetic resonance (NMR) were performed to confirm the formation of the co-block polymer. Simvastatin (SMV), a drug model was incorporated into the nano-polymeric micellar (NpM) of PLGA-PEG-PLGA followed by solubility phase, particle size, zeta potential, and entrapment efficiency characterizations. Results: FTIR, DSC, and NMR successfully confirmed the formation of co-block polymers. Solubility of SMV increased from 2 to 44-folds depending on co-block concentration with entrapment efficiency of 59%-80%. The NpM had size in the range of 206 to 402 nm with negative zeta potential. LA to GA ratio had greater effect on particle size reduction and increasing of co-polymer length. In addition, it had higher contributions on increasing of solubility and entrapment efficiency of SMV than PEG. Conclusion: According to these findings, the LA to GA ratio and PEG concentration gained a great consideration in order to prepare the PLGA-PEG-PLGA co-block which fulfilled the quality target product profile of NpM delivery system.

Novel and safe single-dose treatment of cutaneous leishmaniasis with implantable amphotericin B-loaded microparticles

The development of an effective amphotericin B (AmB) topical formulation to replace the systemically toxic injections currently used in cutaneous leishmaniasis (CL) treatment is challenging due to poor absorption through the skin. Aiming at an effective local chemotherapy, we designed PLGA (poly(lactide-co-glycolide acid) microparticles loaded with deoxycholate amphotericin B (d-AmB) for both macrophage intracellular targeting and sustained extracellular release. For that, d-AmB/PLGA microparticles with sizes ranging from 0.5 μm to 20 μm were synthesized and tested both in vitro and in vivo. In vitro, d-AmB/PLGA was more selectively active against intracellular amastigotes of Leishmania amazonensis than free d-AmB (selectivity index = 50 and 25, respectively). In vivo, the efficacy of a single intralesional (i.l) injection with d-AmB/PLGA was determined in early and established BALB/c mouse ear lesions. In early lesions, a single injection given on day 10 of infection was more effective in controlling parasite growth than eight i.l. injections with free d-AmB, as measured on day 120. Such d-AmB/PLGA injection was also effective in established lesions (day 30), leading to 97% parasite burden reduction, as compared with d-AmB or liposomal AmB (Ambisome®) i.l. injection containing the same AmB dose. Pharmacokinetic studies showed that following d-AmB/PLGA injection, AmB leaked slower from non-infected than infected ears, yet remaining in the ear tissue for as long as 30 days. Of interest, AmB was not detectable in the circulating plasma for at least two weeks of d-AmB/PLGA injection, contrasting with the rapid and durable (2 days) detection after free d-AmB injection. Despite the transient ear swelling and local cell infiltration, no alterations in AST, ALT and creatinine serum levels was induced by d-AmB/PLGA. For its approved components, local efficacy, and single-dose applicability, this novel and safe AmB microparticle depot formulation has strong potential as a new therapy for human CL.

Development of PLGA microparticles with high immunoglobulin G-loaded levels and sustained-release properties obtained by spray-drying a water-in-oil emulsion

In this study, the possibility of producing highly antibody-loaded microparticles with sustained-release properties was evaluated. Polyclonal immunoglobulin G (IgG) was used as a model of antibody and its encapsulation into poly(lactide-co-glycolide) acid (PLGA) microparticles was performed by spray-drying a water-in-oil (w/o) emulsion. It was demonstrated that the use of the Resomer® RG505 PLGA allowed an IgG loading of 20% w/w with an encapsulation efficiency higher than 85%. The produced microparticles were characterized by a mean diameter lower than 10 µm. The burst effect was shown to reach a maximal value of 40%. IgG stability after encapsulation was also assessed. The use of this single PLGA provided a lag time of 3 months which dramatically slowed down the release rate after the initial release of the encapsulated IgG. Using blends of PLGA characterized by different inherent viscosities allowed decreasing the lag time and modulating the dissolution profile of the IgG from the spray-dried microparticles. Therefore, spray-drying a water-in-oil emulsion appeared to be a promising strategy to produce highly antibody-loaded microparticles characterized by sustained-release properties.

Modified PLGA nanofibers as a nerve regenerator with Schwann cells

Polylactide-co-glycolide acid (PLGA) is known as a biodegradable and biocompatible polymer. This polymer has been highly used in tissue engineering. In this study, the biological behavior of Schwann cells (Rat) was investigated in co-culture with L lysine/gelatine coated PLGA nano-fiber. In this study, PLGA was dissolved in a hexafluoro propanol based solvent and nanofiber prepared by an electronic method. They were coated with gelatin and poly-L-lysine individually. These polymer properties were investigated by Scanning Electron Microscopy (SEM) analysis and contact angle measurement. After extraction of rat Schwann cells, the cells were cultured in three groups of nano-fiber; nano-fiber PLGA, nano-fiber gelatine coated PLGA and nano-fiber poly-L-lysine coated PLGA. Cell death and Cell proliferation were evaluated by Acridine orange staining (living cell with a green nucleus and dead cell with an orange nucleus) and morphology was investigated by SEM in 2, 4 and 6 days. The diameter of electronic nanofiber PLGA was between 270 to 700 nm. Average contact angles of PLGA, PLGA coated with gelatine, coated with poly-L-lysine and PLGA were 40.12, 64.58 and 107.66degrees, respectively. The findings showed a significant reduction of cell proliferation in PLGA nanofiber ( it was important than PLGA without nano-fiber (P <0.05)). But, this amount was increased in nanofiber which coated with poly-L-lysine and gelatine. PLGA nanofiber-poly-L-lysine was more biocompatible than PLGA nanofiber-gelatine and this comparison was done with rat Schwann cells.

Nanovaccine Incorporated with Hydroxychloroquine Enhances Antigen Cross-Presentation and Promotes Antitumor Immune Responses.

Induction of effective antigen-specific CD8+ T-cell responses is critical for cancer immunotherapy success. Hydroxychloroquine (HCQ) is a widely used classical antimalarial and antirheumatic drug. HCQ is also an endosomal membrane disrupting agent that can lead to vesicular swelling and membrane permeabilization, which likely facilitates the release of therapeutic agents from lysosomes into the cytoplasm. Here, we develop a minimalistic nanovaccine, which is composed of poly(lactide- co-glycolide)acid (PLGA) nanoparticles (NPs) encapsulating a physical mixture of ovalbumin (OVA, a model antigen) and HCQ (HCQ-OVA-PLGA NPs). We tested whether HCQ could spatiotemporally control the cytosolic delivery of antigens, enhance antigen processing and presentation via the major histocompatibility complex (MHC)-I pathway, and thus generate a sufficient antitumor cytotoxic T-cell response. The results of in vitro experiments showed that HCQ-OVA-PLGA NPs significantly enhanced OVA escape from lysosomes into the cytoplasm within bone-marrow-derived dendritic cells. We also observed that HCQ-OVA-PLGA NPs enhanced the expression level of MHC-I on dendritic cells and improved cross-presentation of antigen, compared to free OVA or OVA-PLGA NPs. Results of in vivo experiments confirmed that HCQ initiated Th1-type responses and strong CD8+ T-cell responses that induced tumor cell apoptosis. Moreover, vaccination of mice with HCQ-OVA-PLGA NPs effectively generated memory immune responses in vivo and prevented tumor progression. We conclude that co-encapsulation of HCQ with antigens in nanovaccines can boost antigen-specific antitumor immune responses, particularly through CD8+ T-cells, serving as a simple and effective platform for the treatment of tumors and infectious diseases.

Microneedles as Enhancer of Drug Absorption Through the Skin and Applications in Medicine and Cosmetology.

The microneedles technology has found applications in many health-related fields. For example, their application in drugs and vaccines delivery as well, as the determination of biomarkers, has been reported. They also have a place in the dermatology and cosmetic areas such as the treatment of wounds from burns, scars, acne, depigmentation, and alopecia will be shown. Microneedles are used in therapeutic applications and are manufactured using materials such as metal (steel, titanium, nickel), polymer (oly-glycolic acid (PGA), poly-lactide-co-glycolide acid (PLGA), poly-L-lactic acid (PLA), chitosan), glass, silicon, ceramic, carbohydrates (trehalose, sucrose, mannitol). Examples of application of microneedles and their advantages and disadvantages are discussed.This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.

Pore shape and size dependence on cell growth into electrospun fiber scaffolds for tissue engineering: 2D and 3D analyses using SEM and FIB-SEM tomography

Electrospun nanofibers have ability to boost cell proliferation in tissue engineered scaffolds as their structure remind cells extra cellular matrix of the native tissue. The complex architecture and network of nanofibrous scaffolds requires advanced characterization methods to understand interrelationship between cells and nanofibers. In our study, we used complementary 2D and 3D analyses of electrospun polylactide-co-glycolide acid (PLGA) scaffolds in two configurations: aligned and randomly oriented nanofibers. Sizes of pores and fibers, pores shapes and porosity, before and after cell culture, were verified by imaging with scanning electron microscopy (SEM) and combination of focus ion beam (FIB) and SEM to obtain 3D reconstructions of samples. Using FIB-SEM tomography for 3D reconstructions and 2D analyses, a unique set of data allowing understanding cell proliferation mechanism into the electrospun scaffolds, was delivered. Critically, the proliferation of cells into nanofibers network depends mainly on the pore shape and pores interconnections, which allow deep integration between cells and nanofibers. The proliferation of cells inside the network of fibers is much limited for aligned fibers comparing to randomly oriented fibers. For random fibers cells have easier way to integrate inside the scaffold as the circularity of pores and their sizes are larger than for aligned scaffolds. The complex architecture of electrospun scaffolds requires appropriate, for tissue engineering needs, cell seeding and culture methods, to maximize tissue growth in vitro environment.

Evaluation of the effectiveness of a bMSC and BMP-2 polymeric trilayer system in cartilage repair

In this study a poly(lactide-co-glycolide) acid (PLGA) tri-layer scaffold is proposed for cartilage repair. The trilayer system consists of a base layer formed by a tablet of PLGA microspheres, a second layer composed of a microsphere suspension placed on top of the tablet, and the third layer, which constitutes an external electrospun PLGA thin polymeric membrane. Combinations of bone morphogenetic protein-2 (BMP-2) encapsulated in the microspheres of the suspension layer, and bone marrow mesenchymal stem cells (bMSC) seeded on the electrospun membrane, are evaluated by histologic analyses and immunohistochemistry in a critical size osteochondral defect in rabbits. Five experimental groups, including a control group (empty defect), a blank group (blank scaffold), a bMSC treated group, two groups treated with 2.5 μg or 8.5 μg of BMP-2 and another two groups implanted with bMSC-BMP-2 combination are evaluated. The repair area increases throughout the experimental time (24 weeks). The repair observed in the treated groups is statistically higher than in control and blank groups. However, the bMSC-BMP-2 combination does not enhance the BMP-2 response. In conclusion, BMP-2 and bMSC repaired effectively the osteochondral defect in the rabbits. The bMSC-BMP-2 combination did not produce synergism.

Effects of different fluid shear stress patterns on the in vitro degradation of poly(lactide-co-glycolide) acid membranes.

The applications of poly (lactide-co-glycolide) acid (PLGA) for coating or fabricating polymeric biodegradable stents (BDSs) have drawn more attention. The fluid shear stress has been proved to affect the in vitro degradation process of PLGA membranes. During the maintenance, BDSs could be suffered different patterns of fluid shear stress, but the effect of these different patterns on the whole degradation process is unclear. In this study, in vitro degradation of PLGA membranes was examined with steady, sinusoid, and squarewave fluid shear stress patterns in 150 mL deionized water at 37°C for 20 days, emphasizing on the changes in the viscosity of the degradation solution, mechanical, and morphological properties of the samples. The unsteady fluid shear stress with the same average magnitude as the steady one accelerate the in vitro degradation process of PLGA membranes in terms of maximum fluid shear stress and "window" of effectiveness. Maximum fluid shear stress accelerates the in vitro degradation of molecular fragments that diffused out in the solution while the "window" of effectiveness affects too in the early stage. Besides, maximum fluid shear stress and "window" of effectiveness accelerates the in vitro loss of tensile modulus and ultimate strength of the PLGA membranes while the maximum fluid shear stress plays the leading role in the decrease of tensile modulus at the early degradation stage. This study could help advance the degradation design of PLGA membranes under different fluid shear stress patterns for biomedical applications like stents and drug release systems. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 23-30, 2017.

The next-generation nicotine vaccine: a novel and potent hybrid nanoparticle-based nicotine vaccine.

Owing to the urgent need for more effective treatment against nicotine addiction, a hybrid nanoparticle-based nicotine vaccine (NanoNiccine) was developed in this study. NanoNiccine was composed of a poly(lactide-co-glycolide) acid (PLGA) core, keyhole limpet hemocyanin (KLH) as an adjuvant protein enclosed within the PLGA core, a lipid layer, and nicotine haptens conjugated to the outer surface of the lipid layer. In contrast to the traditional nicotine vaccine, NanoNiccine is not a nicotine-protein conjugate vaccine. Instead, the nicotine hapten and protein are separately located in the nanostructure to minimize antibody production towards KLH. The cellular uptake study demonstrated that NanoNiccine was ideal for internalization and processing by dendritic cells (DCs). Mice immunized with NanoNiccine produced much lower IgG level against KLH as compared to that immunized with the traditional nicotine-KLH (Nic-KLH) vaccine. In addition, NanoNiccine achieved up to a 400% higher titer of anti-nicotine IgG than the positive control, Nic-KLH. Additionally, the Th1/Th2 index of NanoNiccine suggested that the immune response induced by NanoNiccine was antibody response dominant. Furthermore, NanoNiccine was found to be safe in mice.

The effect of fluid shear stress on the in vitro degradation of poly(lactide-co-glycolide) acid membranes.

Poly(lactide-co-glycolide) acid (PLGA) has been widely used as a biodegradable polymer material for coating stents or fabricating biodegradable stents. Its mechanism of degradation has been extensively investigated, especially with regard to how tensile and compressive loadings may affect the in vitro degradation of PLGA. Fluid shear stress is also one of the most important factors in the development of atherosclerosis and restenosis. But the effect of fluid shear stress on the degradation process is still unclear. The purpose of this study was to characterize the in vitro degradation of PLGA membranes that experienced different fluid shear stresses in 150 mL of deionized water at 37°C for 20 days. Particular emphasis was given to changes in the viscosity of the degradation solution, as well as the mechanical and morphological properties of the samples. The viscosity of the degradation solution with the mechanical loaded specimens was more severely affected than that of the control group. Increasing the fluid shear stress could accelerate the loss of the ultimate strength of PLGA membranes while it slowed down the change of the tensile elastic modulus in the early period. With regard to morphology, the surface roughness was more obviously reduced in the loaded groups. This indicated that the fluid shear stress could affect the in vitro degradation of PLGA membranes. Therefore, this study could help improve the design of PLGA membranes for biomedical applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2016.

PH-sensitive nanoparticles for colonic delivery of curcumin in inflammatory bowel disease

Nano-scaled particles have been found to preferentially accumulate in inflamed regions. Local delivery of anti-inflammatory drugs loaded in nanoparticles to the inflamed colonic site is of great interest for inflammatory bowel disease (IBD) treatment. Curcumin (CC) is an anti-inflammatory local agent, which presents poor ADME properties. Hence, we evaluated, both in vitro and in vivo, the local delivery of CC using pH-sensitive polymeric nanoparticles (NPs) combining both poly(lactide-co-glycolide) acid (PLGA) and a polymethacrylate polymer (Eudragit® S100). CC-NPs significantly enhanced CC permeation across Caco-2 cell monolayers when compared to CC in suspension. CC-NPs significantly reduced TNF-α secretion by LPS-activated macrophages (J774 cells). In vivo, CC-NPs significantly decreased neutrophil infiltration and TNF-α secretion while maintaining the colonic structure similar to the control group in a murine DSS-induced colitis model. Our results support the use of nanoparticles made of PLGA and Eudragit® S100 combination for CC delivery in IBD treatment.

Local Foreign-Body Reaction to Commercial Biodegradable Implants: An in Vivo Animal Study

Biodegradable plates have been used extensively in fracture fixation since the 1960s. They rarely cause stress-protection atrophy or problems requiring secondary plate removal, common complications seen with metallic plates. However, aseptic foreign-body reactions have been reported, sometimes years after the original implantation. Both inadequate polymer degradation and debris accumulation have been implicated as causes. The current generation of commercial biodegradable plates is formulated to minimize this complication by altering the ratio of polylactic and polyglycolic acids. This in vivo study compares the degree of local foreign-body reaction of two commercially available resorbable plates in rabbits. Two types of biodegradable plates were examined: poly(D/L)lactide acid (PDLLA) and polylactide-co-glycolide acid (PLGA). Each plate was placed into a periosteal pericalvarial pocket created beneath the anterior or posterior scalp of a rabbit. Humane killing occurred at 3, 6, and 12 months postoperatively. Foreign-body reaction was evaluated histologically. The PDLLA plates demonstrated marked local foreign-body reactions within the implant capsule as early as 3 months after implantation, with presence of inflammatory cells and granulomatous giant cells in close association with the implant material. All local foreign-body reactions were subclinical with no corresponding tissue swelling requiring drainage. PLGA plates did not demonstrate any signs of inflammatory reactions. In addition, the PLGA plates did not appear to resorb or integrate at 12 months. Neither PDLLA nor PLGA plates demonstrated inflammation of the soft tissue or adjacent bone outside the implant capsule. In our study, the PDLLA plates demonstrated histological evidence of foreign-body reaction that is confined within the implant capsule, which was not seen with the PLGA plates. This finding may be attributable to the lack of significant resorption seen in the PLGA plates. Both PDLLA and PLGA plates were biocompatible with the rabbit tissue environment and should be considered for continued use in craniofacial, maxillofacial, and orthopedic reconstruction.

A novel strategy of spine defect repair with a degradable bioactive scaffold preloaded with adipose-derived stromal cells

Background contextAlthough the use of mesenchymal stem cells (MSC) with scaffolds for bone repair has been considered an effective method, the interactions between implanted materials and bone tissues have not been fully elucidated. At some specific sites, such as the vertebral body (VB) of the spine, the process of bone repair with implanted biomaterials is rarely reported. Recently, adipose tissue was found to be an alternative source of MSC besides bone marrow. However, the strategy of using adipose-derived stromal (ADS) cells with bioactive scaffold for the repair of spinal bone defects has seldom been studied. PurposeTo use a sintered poly(lactide-co-glycolide) acid (PLGA) microspheres scaffold seeded with induced rat ADS cells to repair a bone defect of the VB in a rat model. Study designBasic science and laboratory study. MethodsA sintered porous microspheres scaffold was manufactured by PLGA. ADS cells were isolated from Fischer 344 rats and then induced by osteogenic medium with growth and differentiation factor 5 (GDF5) in vitro. Before implantation, cells were cultured with inductive media for 2 weeks as a monolayer situation and 1 more week on a PLGA scaffold as a three-dimensional structure. These assembled bioactive scaffolds then were implanted in lumbar VB bone defects in Fischer 344 rats. The ex vivo differentiation of the cells was confirmed by von Kossa staining and real-time polymerase chain reaction. The performance of cells on the scaffold was detected by scanning electron microscopy and (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay. In vivo bone formation was quantitatively measured by computed tomography study. And the effect of tissue repair was also evaluated by histological studies. ResultsProliferation and differentiation of cells were confirmed before in vivo implantation. Quantification of bone formation in vivo through serial three-dimensional computed tomography images revealed that the VB implanted with GDF5-induced cells demonstrated more bone formation than the control groups. Besides the bone formation period that occurred between 2 and 4 weeks in all groups, a second bone formation period was found to occur only in the groups that received cells with previous induction in vitro. This second period of significant bone formation happened simultaneously with collapsing of the scaffolds. It was then demonstrated histologically that vascularization early in the process and cooperation between host bone and implanted cells accompanied by collapse of the scaffold may be the factors that influence bone formation. This study not only provides a therapeutic strategy of using biomaterial for bone repair in the spine, but also may lead to a technological method for studying the relationship between implanted stem cells and host tissue. ConclusionsAdipose-derived stromal cells maintained in culture on a scaffold and treated with osteogenic induction with growth factor ex vivo could be used to enhance bone repair in vivo.

Optimization of Formulation Variables to Increase Antigen Entrapment in PLGA Particles

Efficient antigen entrapment is a key factor in preparation of poly (lactide-co-glycolide) acid (PLGA) vaccine formulations when the antigen is of short supply. This study presents a systematic approach in the testing of formulation variables with the objective to increase antigen entrapment in particles when the antigen stock concentration was low. Some of the experimental variables tested were poly (vinyl) alcohol (PVA) concentration in the inner (W1) and outer (W2) aqueous phase, W1/oil (O) phase ratio and choice of organic solvent. The double emulsion solvent evaporation technique was applied to prepare PLGA particles with sonication as the emulsifying force. To measure antigen entrapment efficiency, the antigen (bovine serum albumin, BSA) was isotope labeled with 125iodine (125I). Our results demonstrated that a low PVA concentration in the inner aqueous (W1) phase was beneficial to achieve a high encapsulation efficiency of antigen. On the contrary, in the outer aqueous (W2) phase, a high PVA concentration favored antigen entrapment. We also demonstrated that decreasing the W1 to O/polymer ratio contributed to increased entrapment efficiency. Testing different organic solvents (ethyl acetate, dichloromethane and chloroform), either alone or in combination, revealed that using chloroform as solvent resulted in the highest encapsulation of antigen and the highest production yield. Some of the results presented in this work are in disagreement with well-established formulation variables from previous studies.