Surface Of PLGA Research Articles

PLGA Nanoparticle-Based Formulations to Cross the Blood-Brain Barrier for Drug Delivery: From R&D to cGMP.

The blood–brain barrier (BBB) is a natural obstacle for drug delivery into the human brain, hindering treatment of central nervous system (CNS) disorders such as acute ischemic stroke, brain tumors, and human immunodeficiency virus (HIV)-1-associated neurocognitive disorders. Poly(lactic-co-glycolic acid) (PLGA) is a biocompatible polymer that is used in Food and Drug Administration (FDA)-approved pharmaceutical products and medical devices. PLGA nanoparticles (NPs) have been reported to improve drug penetration across the BBB both in vitro and in vivo. Poly(ethylene glycol) (PEG), poly(vinyl alcohol) (PVA), and poloxamer (Pluronic) are widely used as excipients to further improve the stability and effectiveness of PLGA formulations. Peptides and other linkers can be attached on the surface of PLGA to provide targeting delivery. With the newly published guidance from the FDA and the progress of current Good Manufacturing Practice (cGMP) technologies, manufacturing PLGA NP-based drug products can be achieved with higher efficiency, larger quantity, and better quality. The translation from bench to bed is feasible with proper research, concurrent development, quality control, and regulatory assurance.

Biomineralization improves mechanical and osteogenic properties of multilayer‐modified PLGA porous scaffolds

Poly-(lactide-co-glycolide acid) (PLGA) has been widely investigated as scaffold material for bone tissue engineering owing to its biosafety, biodegradability, and biocompatibility. However, the bioinert surface of PLGA may fail in regulating cellular behavior and directing osteointegration between the scaffold and the host tissue. In this article, oxidized chondroitin sulfate (oCS) and type I collagen (Col I) were assembled onto PLGA surface via layer by layer technique (LbL) as an adhesive coating for the attachment of inorganic minerals. The multilayer-modified PLGA scaffold was mineralized in vitro to ensure the deposition of nanohydroxyapatite (nHAP). It was found that nHAP crystals were more uniformly and firmly attached on the multilayer-modified PLGA as compared with the pure PLGA scaffold, which remarkably improved PLGA surface and mechanical properties. Additionally, in vitro biocompatibility of PLGA scaffold, in terms of bone mesenchymal stem cells (BMSCs) attachment, spreading and proliferation was greatly enhanced by nHAP coating and multilayer deposition. Furthermore, the fabricated composite scaffold also shows the ability to promote the osteogenic differentiation of BMSCs through the up-regulation of osteogenic marker genes. Thus, this novel biomimetic composite scaffold might achieve a desirable therapeutic result for bone tissue regeneration. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2714-2725, 2018.

Characterization of rabies pDNA nanoparticulate vaccine in poloxamer 407 gel

Plasmid DNA (pDNA) vaccines have the potential for protection against a wide range of diseases including rabies but are rapid in degradation and poor in uptake by antigen-presenting cells. To overcome the limitations, we fabricated a pDNA nanoparticulate vaccine. The negatively charged pDNA was adsorbed onto the surface of cationic PLGA (poly (d, l-lactide-co-glycolide))-chitosan nanoparticles and were used as a delivery vehicle. To create a hydrogel for sustainable vaccine release, we dispersed the pDNA nanoparticles in poloxamer 407 gel which is liquid at 4 °C and turns into soft gels at 37 °C, providing ease of administration and preventing burst release of pDNA. Complete immobilization of pDNA to cationic nanoparticles was achieved at a pDNA to nanoparticles ratio (P/N) of 1/50. Cellular uptake of nanoparticles was both time and concentration dependent and followed a saturation kinetics with Vmax of 11.389 µg/mL h and Km of 139.48 µg/mL. The in vitro release studies showed the nanoparticulate vaccine has a sustained release for up to 24 days. In summary, pDNA PLGA-chitosan nanoparticles were non-cytotoxic, their buffering capacity and cell uptake were enhanced, and sustained the release of pDNA. We expect our pDNA vaccine’s potency will be greatly improved in the animal studies.

FABRICATION OF POLY(LACTIC-CO-GLYCOLIC ACID)/CALCIUM PHOSPHATE BONE CEMENT COMPOSITE: SYNTHESIZATION OF CALCIUM PHOSPHATE FROM CRAB SHELLS

Crab shells are waste product, rich with calcium compound. Calcium element is often used as a material for bone scaffold due to its bioactive and biodegradation properties. In this study, calcium phosphate (CaP) nanoparticles were synthesized from crab shells through a wet chemical route. The CaP nanoparticles were then sintered and mixed with poly(lactic-co-glycolic acid) (PLGA) to form a bone cement composite. The mixture was casted in a cylinder shape and it was characterized through ATR-FTIR, XRD, FESEM, contact angle and DSC analyses. The CaP pellet and the CaP/PLGA bone cement composite were then subjected to in vitro simulated body fluid (SBF) bioactivity test. The CaP/PLGA bone cement composite was found to have a composition of crystal CaP and PLGA with a tolerable glass transition state, suitable to be used in a physiological environment. The CaP nanoparticles were agglomerated on the 3D interconnected surface of PLGA. The hydrophobicity of the CaP was increased (66.94%) with the addition of PLGA as a binder matrix where this composite has induced the formation of apatite layer. This bioactive property is crucial in fabricating a bone substitute material as it can promotes cell penetration, attachment and proliferation..

Comparison Study on Four Biodegradable Polymer Coatings for Controlling Magnesium Degradation and Human Endothelial Cell Adhesion and Spreading.

Magnesium (Mg)-based bioresorbable cardiovascular scaffold (BCS) is a promising alternative to conventional permanent cardiovascular stents, but it faces the challenges of rapid degradation and poor endothelium recovery after device degradation. To address these challenges, we investigated poly(l-lactic acid) (PLLA), poly(lactic-co-glycolic acid) (PLGA) (90:10), PLGA (50:50), and polycaprolactone (PCL) coatings on Mg, respectively, and evaluated their surface and biological properties. Intact polymer coatings with complete coverage on Mg substrate were achieved. The biological performance of the materials was evaluated by culturing with human umbilical vein endothelial cells (HUVECs) in vitro using the direct culture method. The pH of the culture media and Mg2+ and Ca2+ ion concentrations in the media were measured after culture to characterize the degradation rate of the materials in vitro. The results showed that the PLGA (50:50) coating improved the adhesion and spreading of HUVECs the most among the four polymer coatings. Moreover, we found three possible factors that promoted HUVECs directly attached on the surface of PLGA (50:50)-coated Mg: (1) the higher concentration of Mg2+ ions released into culture media with a concentration range of 9-15 mM; (2) the lower Ca2+ ion concentration in culture media at 1.3-1.6 mM; and (3) the favorable surface conditions of PLGA (50:50), when compared with the other sample groups. This in vitro study provided the first evidence that the PLGA (50:50) is a promising coating material for Mg-based biodegradable metals toward potential cardiovascular or neurovascular applications.

Synthesis and characterization of magnesium gluconate contained poly(lactic-co-glycolic acid)/chitosan microspheres

Abstract The goal of this study was to fabricate and investigate the chitosan coated poly(lactic- co -glycolic acid) (PLGA) microspheres for the development of controlled release magnesium delivery system. PLGA based microspheres are ideal vehicles for many controlled release drug delivery applications. Chitosan is a naturally occurring biodegradable and biocompatible polysaccharide, which can coat the surface of PLGA to alter the release of drugs. Magnesium gluconate (MgG) was encapsulated in the PLGA and PLGA/chitosan microspheres by utilizing the double emulsion solvent evaporation technique for controlled release study. The microspheres were tested with respect to several physicochemical and biological properties, including morphology, chemical structure, chitosan adsorption efficiency, magnesium encapsulation efficiency, in vitro release of magnesium ions, and cellular compatibility using both human adipose-derived stem cells (ASCs) and PC12 cells. Chitosan coated PLGA microspheres can significantly control the release of magnesium ions compared to uncoated PLGA microspheres. Both coated and uncoated microspheres showed good cellular compatibility.

Bone induction by biomimetic PLGA copolymer loaded with a novel synthetic RADA16-P24 peptide in vivo

Bone morphogenetic protein-2 (BMP-2) is a key bone morphogenetic protein, and poly(lactic-co-glycolic acid) (PLGA) has been widely used as scaffold for clinical use to carry treatment protein. In the previous studies, we have synthesized BMP-2-related peptide (P24) and found its capacity of inducing bone regeneration. In this research, we have synthesized a new amphiphilic peptide Ac-RADA RADA RADA RADA S[PO4]KIPKASSVPTELSAISTLYLDDD-CONH2 (RADA16-P24) with an assembly peptide RADA16-Ion the P24 item of BMP2 to form divalent ion-induced gelatin. Two methods of physisorption and chemical cross-linking were used to bind RADA16-P24 onto the surface of the copolymer PLGA to synthesize RADA16-P24–PLGA, and its capacity of attaching bone marrow stromal cells (BMSCs) was evaluated in vitro and inducing ectopic bone formation was examined in vivo. In vitro our results demonstrated that RADA16-P24–PLGA copolymer prepared by physisorbing or prepared by chemical cross-linking had a peptide binding rate of (2.0180±0.5296)% or (10.0820±0.8405)% respectively (P<0.05). In addition the BMSCs proliferated vigorously in the RADA16-P24–PLGA biomaterials. Significantly the percentage of BMSCs attached to RADA16-P24–PLGA composite prepared by chemical cross-linking and physisorbing were (71.4±7.5) % or (46.7±5.8) % (P<0.05). The in vivo study showed that RADA16-P24–PLGA chemical cross-linking could better induce ectopic bone formation compared with RADA16-P24–PLGA physisorbing and PLGA. It is concluded that the PLGA copolymer is a good RADA16-P24 carrier. This novel RADA16-P24–PLGA composite has strong osteogenic capability.

Studies on stabilization mechanism and stealth effect of poloxamer 188 onto PLGA nanoparticles

In nanoparticulate engineering for drug delivery systems, poloxamers tri block copolymers are employed as adsorbing molecules to modify the aggregation state and impart stability to products. The aim was to prepare nanoparticles using poloxamer188 as stabiliser and investigate the mechanism of stabilisation of the prepared particles. Nanoparticles were prepared by solvent diffusion method with poloxamer 188 as stabiliser. Hydrodynamic thickness and zeta potential of the prepared nanoparticles were determined by photon correlation spectroscopy. To study the extent of adsorption of poloxamer onto the prepared nanoparticles, adsorption isotherms were constructed. The adsorbed amount of poloxamer 188 onto the particles was determined by depletion method. Macrophageal uptake study was performed to assess the uptake of the prepared nanoparticles using RAW 264.7 cell lines. Nanoparticles were prepared with slight increase in particle size and in absolute value of zeta potential compared to uncoated particles suggesting that this effect was due to adsorption of poloxamer 188. TEM studies and surface area analysis supported the results obtained from particle size analysis indicating preparation of particles with a thin layer of adsorbed poloxamer 188. Adsorption kinetics modeling suggested that at low concentrations (0.001-0.010g/L), Langmuir monolayer equation fits quite well and at higher concentrations (above 0.010g/L) multilayer adsorption of poloxamer 188 onto the prepared particles occurred. Thus the nanoparticles had multilayer of poloxamer 188 adsorbed onto the non uniform surface of PLGA. Results of macrophageal uptake and liver cell study exhibits adsorbed concentration dependent bypass of RES uptake of nanoparticles. Hence, results substantiate the application of adsorption isotherms for designing nanoparticles possessing potential to exhibit prolonged circulation when administered in vivo.

Bone marrow stromal cells cultured on poly (lactide‐co‐glycolide)/nano‐hydroxyapatite composites with chemical immobilization of Arg‐Gly‐Asp peptide and preliminary bone regeneration of mandibular defect thereof

Polyethyleneimine (PEI) was used to create active groups on the poly (lactide-co-glycolide)/nano-hydroxyapatite (PLGA/NHA) surface and Arg-Gly-Asp (RGD) was grafted on the active groups and novel PLGA/NHA 2-D membranes and 3D scaffolds modified with RGD were obtained. X-ray photoelectron spectrum (XPS) results show that sulfur displays only on the modified surface. The RGD-modified PLGA/NHA materials also have much lower static water contact angle and much higher water-absorption ability, which shows that after chemical treatment, the modified materials show better hydrophilic properties. Atomic force microscope (AFM) shows that after surface modification, the surface morphology of PLGA is greatly changed. All these results indicate that RGD peptide has successfully grafted on the surface of PLGA. Rabbit bone marrow stromal cells (MSCs) were seeded in the 2D membranes and 3D scaffolds materials. The influences of the RGD on the cell attachment, growth and differentiation, and proliferation on the different materials were studied. The modified scaffolds were implanted into rabbits to observe preliminary application in regeneration of mandibular defect. The PLGA/NHA-RGD presents better results in bone regeneration in rabbit mandibular defect.

Biomineralization of the surface of PLGA-(ASP-PEG) modified with the K16 and RGD-containing peptide

K16 and RGD-containing peptide was used to modify the surface of three-dimensional PLGA-(ASP-PEG) matrix, then the modified PLGA-(ASP-PEG) was incubated in modified simulated body fluid (SBF). The biomineralization of the modified PLGA-(ASP-PEG) was explored, and the peptide was synthesized with solid phase synthesis technology and linked covalently to PLGA-(ASP-PEG) through cross-linker (Sulfo-LC-SPDP), which was characterized with XPS. The modified PLGA-(ASP-PEG) (Experiment group, EG) and PLGA-(ASP-PEG) (Control group, CG) were all incubated into SBF for 10 d, and the growth of hydroxyapatite (HA) nanocrystals was confirmed with XRD, EDS and SEM. HPLC shows that peptide purity is 94.13%, while MS analysis shows that molecular value of peptide is 2741.26. Binding energy of the sulphur in EG was 164 eV is detected by XPS, and the ratio of carbon and sulphur is 99.746:0.1014. SEM analysis demonstrates the better growth of bonelike HA nanocrystals in EG than that in CG. The component of mineral in EG consisted mainly of hydroxyapatite containing low crystalline nanocrystals, and the Ca/P ratio is about 1.60, which is similar to that of natural bone, while the Ca/P ratio in CG is 1.52. PLGA-(ASP-PEG) modified with peptide provided enough functional groups for biomineralization, and possessed the bonelike structure.

Electrochemical characterization of deoxyribonucleic acid on aluminum(III)/poly( l-glutamic acid) film and its application for the detection of phosphinothricin acetyltransferase gene-specific sequence

A simple strategy of transgenic sequence-specific detection without a special amplification procedure was developed on the basis of aluminum(III)/poly( l-glutamic acid) (PLGA) film. An aluminum ion (Al(III)) thin film was assembled on the surface of PLGA via the electrostatic binding of Al(III) with carboxyl, namely Al(III)/PLGA. The immobilization of deoxyribonucleic acid (DNA) was carried out on this Al(III)/PLGA film by Al(III)-single strand DNA (ssDNA) interaction. Surface hybridization between the immobilized ssDNA and its complementary ssDNA was monitored by electrochemical impedance spectroscopy (EIS) using [Fe(CN) 6] 3−/4− as a redox probe. Under the optimal conditions, this DNA electrochemical sensor was applied to determine the specific gene sequence related to phosphinothricin acetyltransferase transgene (PAT) in the transgenic plants by label-free EIS.

Accelerating calcium phosphate growth on NaOH-treated poly-(lactic- co-glycolic acid) by evaporation-induced surface crystallization

Poly(lactic- co-glycolic acid) (PLGA) is a promising material for the regeneration of bone tissue, but its surface properties are not optimal for the application. Coating the surface of PLGA with a continuous layer of calcium phosphate is an effective approach to address the limitation. Current coating techniques for PLGA require immersion in supersaturated calcium phosphate solutions for days to weeks. In this study, we report a simple technique to accelerate the coating process to only 2 h immersion in supersaturated solutions. PLGA pellets were first treated with NaOH to increase their hydrophilicity. The NaOH-treated PLGA pellets were repeatedly dipped in a supersaturated calcium phosphate solution and dried in air. After 10 times of the dip-and-dry treatment, a layer of calcium phosphate crystallites uniformly covered the surfaces of the pellets. After the crystallite-covered pellets were immersed in the supersaturated solution for 2 h, about 5-μm thick continuous calcium phosphate coatings formed on the surfaces. The dip-and-dry technique was also applied on a variety of metals and porous structures. An evaporation-induced surface crystallization process was suggested as the mechanism for the dip-and-dry treatment.

Apatite Formation on α-Tricalcium Phosphate/Poly(D,L-Lactide-Co-Glycolide) Nanocomposite

In this study, a biocomposite comprising nanostructured α-tricalcium phosphate (α-TCP) in a poly(D,L-lactic-co-glycolic acid) (PLGA) matrix was fabricated by a modified solution evaporation method. As a potential temporary bone fixation and substitution material, its bioactivity was evaluated by its ability to form bone-like apatite layer in simulated body fluid (SBF). Owing to the increased surface area covered by the osteoconductive bioceramic of α-TCP, rapid apatite formation was observed. After 7 days of immersion, enhanced nucleation of apatite was observed on the nanocomposite. At day 14, dense lamellar-like apatite was formed on the nanocomposite whilst apatite nucleation had only just started to develop on the surface of pure PLGA. At the same time, a preliminary in-vitro cell culture study was conducted using human osteoblast-like (HOB) cells. A significant increase in cell number with culturing time was observed for the nanocomposite. After 9 days incubation, a confluent lamellar-like apatite layer was formed on the composite surface. This apatite layer was also shown beneath the proliferating HOB cells at Day 16.

Surface modification and characterization of chitosan or PLGA membrane with laminin by chemical and oxygen plasma treatment for neural regeneration

Attachment to and proliferation on the substrate are deemed important considerations when Schwann cells (SCs) are to be seeded in synthetic nerve grafts. Good attachment is a prerequisite for the SCs to survive. Fast proliferation will yield large numbers of SCs in a short time, which appears to be promising for stimulating peripheral nerve regeneration. However, surface properties are the dominating factor in influencing the interactions between cells and synthetic nerve grafts. The aim of this study was to investigate the surface effects of laminin modified PLGA and chitosan membranes after chemical method and plasma treatment. Laminin, the extracellular matrix protein, is a permissive protein for SCs adhesion used in neural regeneration. The surface properties of laminin modified membranes were assayed by BCA, FTIR and XPS analysis. Results showed that laminin was covalently bonded onto the surface of both PLGA and chitosan membranes either by chemical method or by oxygen plasma treatment. The cell affinity of the laminin modified membranes was verified by Schwann cells culturing. Our results also indicate that oxygen plasma is indeed a better method to incorporate laminin onto the surface of membrane. Laminin-modified chitosan membrane significantly increases SCs attachment and affinity for directing peripheral nerve regeneration.

The degradation of the three layered nano-carbonated hydroxyapatite/collagen/PLGA composite membrane in vitro

Objective The purpose of this paper was to investigate the in vitro biodegradation of a guided tissue regeneration composite membrane, nano-carbonated hydroxyapatite/collagen/poly(lactic-co-glycolic acid) (nCHAC/PLGA). Especially for periodontal therapy, the functional graded material (FGM) nCHAC/PLGA membrane was prepared that consisted of three layers with 8 wt% nCHAC + PLGA/4 wt% nCHAC + PLGA/PLGA, where one face of the membrane is porous, thereby allowing cell growth thereon and the opposite face of the membrane smooth, thereby inhibiting cell adhesion. Methods For evaluation, in vitro degradation specimens of nCHAC/PLGA were immersed into artificial saliva solution at 37 °C for 1, 2, 4, 8 and 12 weeks to detect the weight loss over the period, and set pure PLGA membrane as control to compare the degraded behaviors. pH value and calcium concentration of the residual solution were measured, and morphology change was investigated by scanning electron microscopy (SEM). Results During the experimental period in vitro, the whole shape of the membrane could be kept for 4 weeks, after that it became powder at between 8 and 12 weeks. The results demonstrated that weight loss increased continuously with a reduction in mass of 23.1% after 4 weeks and 88% after 12 week for the nCHAC/PLGA three FGM layers composite membrane. The calcium concentration in the residual solution showed a significant increase after 4 weeks, which referred to the nano-carbonated hydroxyapatite degradation. Moreover, the pH value in the solution of the nCHAC/PLGA membrane was a little higher than that of the pure PLGA membrane, which demonstrated the possible neutralization effect from nCHAC composite for the acid outcome of PLGA in the solution. The pore structure of 8 wt% nCHAC + PLGA was enlarged on the porous surface, while the nonporous surface of pure PLGA also showed a small porous structure after increased time. Significance Degradation of the composite membrane is appropriate for practical periodontal repair. Moreover, the new mineral formation on the surface of the composite membrane referred to the possible positive effect in vivo for new bone tissue regeneration.

Surface modification of biomimetic PLGA-(ASP-PEG) matrix with RGD-containing peptide: a new non-viral vector for gene transfer and tissue engineering

RGD-containing peptide (K16-GRGDSPC), characterized as non-viral gene vectors, was fabricated to modify the surface of PLGA-[ASP-PEG] matrix, which offered the foundation for gene transfer with porous matrix of gene activated later. Peptide was synthesized and matrix was executed into chips A, B and chip C. Chip C was regarded as control. Chips A and B were reacted with cross-linker. Then chip A was reacted with peptide. MS and HPLC were used to detect the MW and purity of peptide. Sulphur, existing on the surface of biomaterials, was detected by XPS. The purity of un-reacted peptide in residual solution was detected by a spectrophotometer. HPLC shows that the peptide purity was 94%–95%, and MS shows that the MW was 2 741.3307. XPS reveals that the binding energy of sulphur was 164 eV and the ratio of carbon to sulphur (C/S) was 99.746:0.1014 in reacted chip A. The binding energy of sulphur in reacted chip B was 164 eV and 162 eV, C/S was 99.574:0.4255, and there was no sulphur in chip C. Peptide was manufactured and linked to the surface of biomimetic and 3-D matrix, which offered the possibilities for gene transfer and tissue engineering with this new kind of non-viral gene vector.

Characterization of surface property of poly(lactide- co-glycolide) after oxygen plasma treatment

In this study, poly (lactide- co-glycolide) (PLGA) films were treated by oxygen plasma. The surface structure, topography and surface chemistry of treated PLGA films were characterized by contact angle measurement, scanning electron microscope observation, atomic force microscopy, and X-ray photoelectron spectrum analysis. The cell affinity of the oxygen plasma treated films was evaluated under dynamic conditions by Parallel Plate Flow Chamber (PPFC). The results showed that the hydrophilicity increased greatly after oxygen plasma treatment. High quantities of –C–O groups, such as hydroxyl and peroxyl groups could be incorporated into the surface of PLGA (70/30) by controlling appropriate plasma treatment conditions. Moreover, the oxygen plasma treatment resulted in formation of peaks and valleys on the sample surfaces, and the roughness increased with treatment time. Cells stretched very well and the ability to endure the shear stress was improved greatly after the PLGA (70/30) was modified by appropriate plasma treatment, i.e. under 50 W for 2 or 10 min. However, when the treatment time was increased to 20 min, the percentage of adherent cells on the roughest surface decreased because the content of polar groups incorporated onto the surface decreased. The results showed that improved cell adhesion was attributed to the combination of surface chemistry and surface morphology of PLGA during plasma etching.

Controllable surface modification of poly(lactic-co-glycolic acid) (PLGA) by hydrolysis or aminolysis I: physical, chemical, and theoretical aspects.

While biodegradable, biocompatible polyesters such as poly (lactic-co-glycolic acid) (PLGA) are popular materials for the manufacture of tissue engineering scaffolds, their surface properties are not particularly suitable for directed tissue growth. Although a number of approaches to chemically modify the PLGA surface have been reported, their applicability to soft tissue scaffolds, which combine large volumes, complex shapes, and extremely fine structures, is questionable. In this paper, we describe two wet-chemical methods, base hydrolysis and aminolysis, to introduce useful levels of carboxylic acid or primary and secondary amine groups, respectively, onto the surface of PLGA with minimal degradation. The effects of temperature, concentration, pH, and solvent type on the kinetics of these reactions are studied by following changes in the wettability of the PLGA using contact angle measurements. In addition, the treated surfaces are studied using X-ray photoelectron spectroscopy (XPS) to determine the effect on the surface chemical structure. Furthermore, we show using XPS analysis that these carboxyl and amine groups are readily activated to allow the covalent attachment of biological macromolecules.